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WO2024026076A2 - Substituted 3,4-dihydroquinolinone inhibitors of tshr - Google Patents

Substituted 3,4-dihydroquinolinone inhibitors of tshr Download PDF

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Publication number
WO2024026076A2
WO2024026076A2 PCT/US2023/028950 US2023028950W WO2024026076A2 WO 2024026076 A2 WO2024026076 A2 WO 2024026076A2 US 2023028950 W US2023028950 W US 2023028950W WO 2024026076 A2 WO2024026076 A2 WO 2024026076A2
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WO
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Prior art keywords
alkyl
alkoxy
compound
halo
optionally substituted
Prior art date
Application number
PCT/US2023/028950
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French (fr)
Other versions
WO2024026076A3 (en
Inventor
Steven J. MCKERRALL
Daniel D. Long
Xiaohui Du
Deniz AYDIN
Matthew Volgraf
Original Assignee
Septerna, Inc.
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Publication of WO2024026076A2 publication Critical patent/WO2024026076A2/en
Publication of WO2024026076A3 publication Critical patent/WO2024026076A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/20—Oxygen atoms
    • C07D215/22—Oxygen atoms attached in position 2 or 4
    • C07D215/227—Oxygen atoms attached in position 2 or 4 only one oxygen atom which is attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/78—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in position 2
    • C07D239/80—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C07D241/38—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems with only hydrogen or carbon atoms directly attached to the ring nitrogen atoms
    • C07D241/40—Benzopyrazines
    • C07D241/44—Benzopyrazines with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/52—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings condensed with carbocyclic rings or ring systems
    • C07D263/54—Benzoxazoles; Hydrogenated benzoxazoles
    • C07D263/58—Benzoxazoles; Hydrogenated benzoxazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
    • C07D265/34—1,4-Oxazines; Hydrogenated 1,4-oxazines condensed with carbocyclic rings
    • C07D265/36—1,4-Oxazines; Hydrogenated 1,4-oxazines condensed with carbocyclic rings condensed with one six-membered ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/10—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems

Definitions

  • TSH-Receptor This pathological activation of TSH-Receptor (TSHR) leads to uncontrolled production of thyroid hormones (e.g., T3 and T4) causing hyperthyroidism.
  • TSH and the TSHR are important proteins for controlling thyroid function.
  • TSHR is primarily expressed in follicular epithelial cells of the thyroid gland, but also is expressed in a variety of additional cell types, such as retro-orbital fibroblasts, kidney cells, adipocytes and bone cells.
  • TSH binds to its receptor and leads to the stimulation of second messenger pathways involving predominantly cAMP. Inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) pathways are also activated at higher TSH concentrations.
  • IP3 Inositol 1,4,5-triphosphate
  • DAG diacylglycerol
  • thyrostatic drugs that block the production of thyroid hormones. These medications play a role further downstream in the signal cascade of the thyroid upon activation of the TSHR. Since thyroid hormones T3 and T4 are secreted in the thyroid gland, thyrostatic drugs induce an inhibition of their synthesis. Thus, the current primary anti-thyroid treatment does not target the causative molecular activation of the TSHR by autoantibodies and patients are therefore burdened by a rate of at least 5% adverse effects. This demands frequent controls of the thyroid hormone levels and adjustments of thyrostatics dosage. In contrast to these drugs, which regulate the thyroid hormone level, another promising target is the TSHR itself.
  • TSHR pathological activation of TSHR leads to the production of the extracellular matrix by involvement of hyaluronanic acid, fibrosis and swelling of extraocular muscle as well as adipogenesis of orbital fibroblasts (orbital fat expansion).
  • the increase in tissue volume in the orbit often causes diplopia and compression of the optic nerve and exophthalmos.
  • Grave’s ophthalmology and thyroid eye disease Accordingly, there is a need in the art to provide additional means for the treatment of hyperthyroidism, in particular compounds that act as TSHR antagonists.
  • A is (C 6 -C 10 )aryl, 5- to 10-membered heteroaryl, (C 3- C 8 )cycloalkyl, or 4- to 7-membered heterocycloalkyl;
  • n is 0, 1, 2, 3, 4, or 5;
  • R 1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4-
  • a compound has the structure of Formula (II): or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R 1 is independently for each occurrence (C 1- C 6 )alkyl, (C 2- C 6 )alkynyl, (C 2- C 6 )alkenyl, (C 3- C 8 )cycloalkyl, (C 3- C 8 )cycloalkoxy, (C 6 -C 10 )aryl, (C 6 -C 10 )aryloxy, (C 6 -C 10 )aryl(C 1- C 6 )alkyl, 4- to 7-membered heterocycloalkyl, (C 1- C 6 )alkoxy, (C 1- C 6 )alkoxy(C 1- C 6 )alkyl, halo,
  • a compound has the structure of Formula (III): or a pharmaceutically acceptable salt thereof, wherein: A is (C 6 -C 10 )aryl, 5- to 10-membered heteroaryl, (C 3- C 8 )cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R 1 is independently for each occurrence (C 1- C 6 )alkyl, (C 2- C 6 )alkynyl, (C 2- C 6 )alkenyl, (C 3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy,
  • a reference to “A and/or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.
  • “or” should be understood to have the same meaning as “and/or” as defined above.
  • the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e., “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.
  • the phrase “at least one,” in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements.
  • At least one of A and B can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.
  • compositions of the present invention may exist in particular geometric or stereoisomeric forms.
  • polymers of the present invention may also be optically active.
  • the present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
  • “Geometric isomer” means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration.
  • R,” “S,” “S*,” “R*,” “E,” “Z,” “cis,” and “trans,” indicate configurations relative to the core molecule.
  • Certain of the disclosed compounds may exist in “atropisomeric” forms or as “atropisomers.”
  • Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers.
  • the compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from a mixture of isomers.
  • Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods.
  • a particular enantiomer of compound of the present invention may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
  • the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.
  • Percent purity by mole fraction is the ratio of the moles of the enantiomer (or diastereomer) or over the moles of the enantiomer (or diastereomer) plus the moles of its optical isomer.
  • the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure relative to the other stereoisomers.
  • the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure.
  • the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure.
  • a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound or mixtures enriched in one enantiomer relative to its corresponding optical isomer.
  • prodrug as used herein encompasses compounds that, under physiological conditions, are converted into therapeutically active agents.
  • a common method for making a prodrug is to include selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule.
  • the prodrug is converted by an enzymatic activity of the host animal.
  • phrases “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject chemical from one organ or portion of the body, to another organ or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, not injurious to the patient, and substantially non- pyrogenic.
  • materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum
  • compositions of the present invention are non-pyrogenic, i.e., do not induce significant temperature elevations when administered to a patient.
  • pharmaceutically acceptable salts refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting a purified compound(s) in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed.
  • Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts, and the like.
  • sulfate bisulfate
  • phosphate nitrate
  • acetate valerate
  • oleate palmitate
  • stearate laurate
  • benzoate lactate
  • phosphate tosylate
  • citrate maleate
  • fumarate succinate
  • tartrate naphthylate
  • mesylate glucoheptonate
  • lactobionate lactobionate
  • laurylsulphonate salts and the like.
  • the compounds useful in the methods of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases.
  • pharmaceutically acceptable salts refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine.
  • Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like.
  • Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).
  • pharmaceutically acceptable cocrystals refers to solid coformers that do not form formal ionic interactions with the small molecule.
  • a “therapeutically effective amount” (or “effective amount”) of a compound with respect to use in treatment refers to an amount of the compound in a preparation which, when administered as part of a desired dosage regimen (to a mammal, preferably a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit/risk ratio applicable to any medical treatment.
  • a desired dosage regimen to a mammal, preferably a human
  • the term “prophylactic or therapeutic” treatment is art-recognized and includes administration to the host of one or more of the subject compositions.
  • the treatment is prophylactic, (i.e., it protects the host against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic, (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof).
  • the term “patient” or “subject” refers to a mammal in need of a particular treatment.
  • a patient is a primate, canine, feline, or equine.
  • a patient is a human.
  • An aliphatic chain comprises the classes of alkyl, alkenyl and alkynyl defined below.
  • a straight aliphatic chain is limited to unbranched carbon chain moieties.
  • the term “aliphatic group” refers to a straight chain, branched-chain, or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups, such as an alkyl group, an alkenyl group, or an alkynyl group.
  • “Alkyl” refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having the number of carbon atoms specified, or up to 30 carbon atoms if no specification is made.
  • alkyl of 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, and those moieties which are positional isomers of these moieties.
  • Alkyl of 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl and tetracosyl.
  • a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chains, C3-C30 for branched chains), and more preferably 20 or fewer.
  • Alkyl goups may be substituted or unsubstituted.
  • heteroalkyl refers to an alkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms.
  • haloalkyl refers to an alkyl group as hereinbefore defined substituted with at least one halogen.
  • hydroxyalkyl refers to an alkyl group as hereinbefore defined substituted with at least one hydroxyl.
  • alkylene refers to an alkyl group having the specified number of carbons, for example from 2 to 12 carbon atoms, that contains two points of attachment to the rest of the compound on its longest carbon chain.
  • alkylene groups include methylene -(CH 2 )-, ethylene -(CH 2 CH 2 )-, n-propylene -(CH 2 CH 2 CH 2 )-, isopropylene - (CH 2 CH(CH 3 ))-, and the like.
  • Alkylene groups can be cyclic or acyclic, branched or unbranched carbon chain moiety, and may be optionally substituted with one or more substituents.
  • Cycloalkyl means mono- or bicyclic or bridged or spirocyclic, or polycyclic saturated carbocyclic rings, each having from 3 to 12 carbon atoms. Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3-6 carbons in the ring structure. Cycloalkyl groups may be substituted or unsubstituted. As used herein, the term “halocycloalkyl” refers to an cycloalkyl group as hereinbefore defined substituted with at least one halogen.
  • Cycloheteroalkyl refers to an cycloalkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms.
  • Preferred cycloheteroalkyls have from 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4-6 carbons and heteroatoms in the ring structure. Cycloheteroalkyl groups may be substituted or unsubstituted.
  • lower alkyl means an alkyl group, as defined above, but having from one to ten carbons, more preferably from one to six carbon atoms in its backbone structure such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.
  • lower alkenyl and “lower alkynyl” have similar chain lengths.
  • preferred alkyl groups are lower alkyls.
  • a substituent designated herein as alkyl is a lower alkyl.
  • Alkenyl refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain moiety having the number of carbon atoms specified, or up to 26 carbon atoms if no limitation on the number of carbon atoms is specified; and having one or more double bonds in the moiety.
  • Alkenyl of 6 to 26 carbon atoms is exemplified by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosoenyl, docosenyl, tricosenyl, and tetracosenyl, in their various isomeric forms, where the unsaturated bond(s) can be located anywhere in the moiety and can have either the (Z) or the (E) configuration about the double bond(s).
  • Alkynyl refers to hydrocarbyl moieties of the scope of alkenyl, but having one or more triple bonds in the moiety.
  • aryl as used herein includes 3- to 12-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon (i.e., carbocyclic aryl) or where one or more atoms are heteroatoms (i.e., heteroaryl).
  • aryl groups include 5- to 12-membered rings, more preferably 6- to 10-membered rings
  • aryl also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.
  • Carboycyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.
  • Heteroaryl groups include substituted or unsubstituted aromatic 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, whose ring structures include one to four heteroatoms.
  • Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
  • Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic.
  • halo means halogen and includes, for example, and without being limited thereto, fluoro, chloro, bromo, iodo and the like, in both radioactive and non-radioactive forms. In a preferred embodiment, halo is selected from the group consisting of fluoro, chloro and bromo.
  • heterocyclyl or “heterocyclic group” or “heterocycloalkyl” refer to 3- to 12- membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10- membered rings, whose ring structures include one to four heteroatoms.
  • Heterocycles can be monocyclic, bicyclic, spirocyclic, or polycyclic.
  • Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, chromane, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine,
  • the heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF 3 , -CN, and the like.
  • substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino
  • substituted refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds.
  • the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds.
  • the permissible substituents can be one or more and the same or different for appropriate organic compounds.
  • the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms.
  • Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety
  • the substituents on substituted alkyls are selected from C1-6 alkyl, C3-6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on substituted alkyls are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants.
  • small molecules refers to small organic or inorganic molecules of molecular weight below about 3,000 Daltons. In general, small molecules useful for the invention have a molecular weight of less than 3,000 Daltons (Da).
  • the small molecules can be, e.g., from at least about 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da).
  • a “small molecule” refers to an organic, inorganic, or organometallic compound typically having a molecular weight of less than about 1000.
  • a small molecule is an organic compound, with a size on the order of 1 nm.
  • small molecule drugs of the invention encompass oligopeptides and other biomolecules having a molecular weight of less than about 1000.
  • An “effective amount” is an amount sufficient to effect beneficial or desired results.
  • a therapeutic amount is one that achieves the desired therapeutic effect. This amount can be the same or different from a prophylactically effective amount, which is an amount necessary to prevent onset of disease or disease symptoms.
  • An effective amount can be administered in one or more administrations, applications or dosages.
  • a therapeutically effective amount of a composition depends on the composition selected. The compositions can be administered from one or more times per day to one or more times per week; including once every other day.
  • treatment of a subject with a therapeutically effective amount of the compositions described herein can include a single treatment or a series of treatments.
  • the terms “decrease,” “reduce,” “reduced”, “reduction”, “decrease,” and “inhibit” are all used herein generally to mean a decrease by a statistically significant amount relative to a reference.
  • “reduce,” “reduction” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level and can include, for example, a decrease by at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, up to and including, for example, the complete absence of the given entity or parameter ascompared to the reference level, or any decrease between 10-99% as compared to the absence of a given treatment.
  • the terms “increased”, “increase” or “enhance” or “activate” are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms “increased”, “increase” or “enhance” or “activate” means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10- fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level.
  • the term “modulate” includes up-regulation and down-regulation, e.g., enhancing or inhibiting a response.
  • a “radiopharmaceutical agent,” as defined herein, refers to a pharmaceutical agent which contains at least one radiation-emitting radioisotope. Radiopharmaceutical agents are routinely used in nuclear medicine for the diagnosis and/or therapy of various diseases.
  • the radiolabelled pharmaceutical agent for example, a radiolabelled antibody, contains a radioisotope (RI) which serves as the radiation source.
  • RI radioisotope
  • the term “radioisotope” includes metallic and non-metallic radioisotopes. The radioisotope is chosen based on the medical application of the radiolabeled pharmaceutical agents.
  • the radioisotope is a metallic radioisotope
  • a chelator is typically employed to bind the metallic radioisotope to the rest of the molecule.
  • the radioisotope is a non-metallic radioisotope
  • the non-metallic radioisotope is typically linked directly, or via a linker, to the rest of the molecule.
  • the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.
  • A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl;
  • n is 0, 1, 2, 3, 4, or 5;
  • R 1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C 8 )cycloalkyl, (C 3- C 8 )cycloalkoxy, (C 6 -C 10 )aryl, (C 6 -C 10 )aryloxy, (C 6 -C 10 )aryl(C 1- C 6 )alkyl, 4- to 7-membered heterocycloalkyl, (C 1- C 6 )alkoxy, (C 1- C 6 )alkoxy(C 1- C 6 )alkyl, 4- to 7-membered heterocycloalkyl, (C 1- C 6 )alkoxy, (C 1- C 6 )
  • A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4, or 5; R 1 is independently for each occurrence (C 1- C 6 )alkyl, (C 3- C 8 )cycloalkyl, 4- to 7- membered heterocycloalkyl, (C 1- C 6 )alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C 1- C 6 )alkyl, (C 3- C 8 )cycloalkyl, (C 1- C 6 )alkoxy, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy, or two vicinal occurrences
  • A is (C 6 -C 10 )aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R 1 is independently for each occurrence (C 1- C 6 )alkyl, (C 3- C 8 )cycloalkyl, , 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C1-C6)alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal
  • A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl;
  • n is 0, 1, 2, 3, 4, or 5;
  • R 1 is independently for each occurrence (C 1- C 6 )alkyl, (C 2- C 6 )alkynyl, (C 2- C 6 )alkenyl, (C 3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbon
  • A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R 1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7- membered heterocycloalkyl, (C 1- C 6 )alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C 1- C 6 )alkyl, (C 3- C 8 )cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C 1- C 6 )alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal occurrences
  • A is phenyl. In certain embodiments, A is a 5- to 7-membered heteroaryl comprising 1 or 2 nitrogen atoms. In certain embodiments, A is pyridyl or diazolyl. thiazolyl, pyrazinyl, pyrimidinyl, while in other embodiments, A is pyridyl or diazolyl. In some embodiments, A is wherein A is isoquinolinyl or indazolyl. In other embodiments, A is cyclopentyl, cyclohexyl, or tetrahydropyranyl.
  • R 1 is independently for each occurrence (C1-C6)alkyl, (C1- C 6 )alkoxy, fluoro, chloro, or cyano, wherein (C 1 -C 6 )alkyl and (C 1 -C 6 )alkoxy are optionally substituted with one or more substituents independently selected from fluoro and hydroxyl.
  • at least one R 1 is fluoro, chloro, or cyano.
  • at least one R 1 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from hydroxyl and fluoro.
  • R 1 is 2,2,2-trifluoroethyl-1-hydroxyethyl.
  • At least one R 1 is difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluropropyl, or 4,4,4- trifluorobutyl.
  • at least one R 1 is (C 1 -C 6 )alkoxy optionally substituted with at least one fluoro, preferably (C 1 -C 3 )alkoxy optionally substituted with at least one fluoro, more preferably difluoromethoxy or trifluoromethoxy.
  • R 1 is methoxy or isopropoxy.
  • R 1 is methyl optionally substituted with hydroxy or one or more fluoro.
  • R 1 is (C1-C3)alkoxy optionally substituted with at least one fluoro.
  • at least one R 1 is (C 3- C 8 )cycloalkyl, preferably cyclopropyl or cyclobutyl.
  • at least R 1 is (C 3- C 8 )cycloalkoxy, preferably cyclopentyloxy, cyclohexyloxy.
  • R 1 is (C2-C6)alkynyl optionally substituted with hydroxy.
  • two vicinal occurrences of R 1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C 1 -C 6 )alkyl, (C 3 - C 8 )cycloalkyl, and (C 1- C 6 )alkoxy.
  • A is phenyl comprising two vicinal occurrences of R 1 , wherein the two vicinal occurrences of R 1 , taken together with the atoms to which they are attached, form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring.
  • n is 2. In certain embodiments, n is 1. In certain embodiments, n is 0.
  • Q is CR 2c . In certain embodiments, Q is N.
  • R 2a , R 2b , and R 2c are each hydrogen.
  • R 2a , R 2b , and R 2c is fluoro, chloro, or cyano.
  • R 3 is (C1-C6)alkyl optionally substituted with hydroxy.
  • R 3 is t-butyl.
  • R 3 is (C 3 -C 8 )cycloalkyl optionally substituted with one or more (C 1- C 6 )alkyl, (C 1- C 6 )fluoroalkyl, fluoro, or (C 1- C 6 )alkoxy(C 1- C 6 )alkyl.
  • the (C 3 -C 8 )cycloalkyl is cyclobutyl, cyclopentyl, cylohexyl, bicyclo[1.1.1]pentanyl, bicyclo[2.2.1]heptanyl,bicycle[2.1.1]hexanyl, or bicyclo[3.1.0]hexanyl.
  • the (C3-C8)cycloalkyl is optionally substituted with one or more substituents selected from fluoro, trifluoromethyl, methoxymethyl, methyl, and hydroxyl.
  • R 3 is 4- to 7-membered heterocycloalkyl optionally substituted with one or more substituents selected from (C 1- C 6 )alkyl, (C 1- C 6 )fluoroalkyl, or fluoro.
  • the 4- to 7-membered heterocycloalkyl is etrahydropyranyl or oxetanyl.
  • Z is NR 4 , R 3 and R 4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C 1- C6)alkyl or halo.
  • the 4- to 7 membered heterocycloalkyl is azetidine or pyrrolidine,
  • X is NR 4 or O.
  • X is NH.
  • Z is NR 4 , or O.
  • Z is NH.
  • one and only one of X and Z is CH 2.
  • X and Z are each NH.
  • L is unsubstituted (C1-C6)alkylene.
  • L is (C1-C6)alkylene substituted with hydroxy.
  • L is (C1-C6)alkylene substituted with 1 to 3 halo atoms.
  • L is –CH2–, –-CH(CH3)–, –CH(CH2CH3)–, or –CH(CH2OH)–.
  • W a –W b represents: –C(R 6 ) 2 –C(R 7 ) 2 –.
  • R 6 is (C 1- C 6 )alkyl, wherein (C 1- C 6 )alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy.
  • At least one R 7 is (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, (C1- C6)alkoxy, and cyano.
  • each R 6 and each R 7 is hydrogen.
  • W a –W b represents: –O–C(R 8 ) 2 –.
  • R 8 is hydrogen.
  • W a –W b represents: –C(R 8 )2–O–, where each R8 is hydrogen.
  • W a –W b represents: –C(R 8 )2–NR 9 –.
  • each R 8 is hydrogen.
  • R 9 is (C1-C6)alkyl, preferably methyl.
  • R 10 is hydrogen.
  • R 10 is (C 1- C 6 )alkyl.
  • R 10 is methyl.
  • a compound is selected from Table 1: Table 1.
  • the compound is selected from Table 2. Table 2.
  • the compounds are atropisomers.
  • structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a 13 C- or 14 C-enriched carbon are within the scope of this invention.
  • Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention.
  • the (C1-C4)alkyl or the -O-(C1- C4)alkyl can be suitably deuterated (e.g., -CD3, -OCD3).
  • Any compound of the invention can also be radiolabed for the preparation of a radiopharmaceutical agent.
  • Methods of Treatment One aspect of the invention provides methods of inhibiting a thyroid stimulating hormone receptor, comnprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein.
  • Another aspect of the invention comprises treating or preventing a thyroid disease, comnprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein.
  • the subject has Graves' disease.
  • the subject has Graves' ophthalmopathy. In certain embodiments, the subject has Graves' dermopathy. In certain embodiments, the subject has thyroid cancer. While not being bound by theory, TSHR in thyroid cells, and likely in fibroblasts and adipocytes in the supporting tissue behind the eye (in the retro-orbital space), also are stimulated by TSHR-stimulating antibodies (TSAbs), resulting in Graves' disease.
  • TSHR-stimulating antibodies TSAbs
  • Graves' disease which is an autoimmune disease that occurs in 1% of the US population, has two important clinical components: 1) hyperthyroidism from stimulation of TSHR on thyroid cells and 2) Graves' orbitopathy (or Graves' ophthalmopathy or thyroid eye disease), which appears to result from stimulation of TSHR on retro-orbital fibroblasts and/or adipocytes.
  • Hyperthyroidism in particular Graves' hyperthyroidism, is a hypermetabolic state that affects virtually every tissue/cell in the body and can lead to, in particular, cardiovascular dysfunction and death.
  • Graves' ophthalmopathy also known as Graves' orbitopathy, occurs in 80% of Graves' hyperthyroid patients as diagnosed by computerized tomographic scan.
  • Graves dermopathy also known as Pretibial myxedema, thyroid dermopathy, Jadassohn- Dössekker disease or Myxoedema tuberosum, is an infiltrative dermopathy, resulting as a complication of Graves' disease, with an incidence rate of about 1-5% in patients.
  • the disease usually presents itself as a waxy, discolored induration of the skin on the anterior aspect of the lower legs.
  • a disease that can be treated o prevented by TSHR antagonists is thyroid cancer.
  • TSHR is expressed in thyroid cancer cells and regulates the growth, proliferation and metastatic potential of thyroid cancer cells.
  • the thyroid gland is, as is well known, one site of metabolic control within the body. Cancer of the thyroid gland is not particularly common, but the high rate of disease re- occurrence necessitates long-term surveillance. Usually, during treatment for cancer of the thyroid, the majority of the thyroid tumor is removed, but a small amount often remains that must be treated by radioactive iodide therapy. Indeed, thyroid cancer is characterized by a high likelihood of relapses in up to 30% of patients, even after successful therapy.
  • TSHR contains a hereditary mutation that makes it more active than the normal TSHR, resulting in hereditary non-immune hyperthyroidism.
  • TSHR antagonists could be effective treatment for these patients also.
  • a “TSHR antagonist” as described herein blocks or inhibits the action of the agonists (TSH or thyroid-stimulating antibodies for TSHR.
  • Small-molecule ligands for the TSHR (antagonists) typically bind to an intra-membrane domain of the receptor, and act by inducing a conformational change rather than simply competing for TSH binding to its extracellular site on the receptor.
  • the antagonists may be selective antagonists for TSHR (i.e, the compounds do not activate or modulate other hormone receptors, particularly luteinizing hormone/chorionic gonadotropin receptor (LHCGR) and follicle-stimulating hormone receptor (FSHR)).
  • the antagonists disclosed herein may be used for treating hyperthyroidism in a subject.
  • the antagonists may inhibit mutant TSHRs with higher than normal basal signaling activities (CAMs) that cause an unusual form of hyperthyroidism.
  • the antagonists may inhibit stimulation by antibodies found in Graves' disease, which is the most common form of hyperthyroidism.
  • the TSHR antagonists are useful for treating TSHR-mediated thyroid cancer or hyperthyroidism by blocking TSHR-stimulating antibodies (TSAbs) in Graves' hyperthyroidism.
  • the compound is administered orally to the subject.
  • the compound is administered parenterally to the subject.
  • the disease is prevented.
  • the disease is treated.
  • Pharmaceutical Compositions, Routes of Administration, and Dosing In certain embodiments, the invention is directed to a pharmaceutical composition, comprising a compound of the invention, e.g. a compound of Formula (1), and a pharmaceutically acceptable carrier.
  • the invention is directed to a pharmaceutical composition, comprising a compound of any of the disclosed embodiments, and a pharmaceutically acceptable carrier.
  • the invention is directed to a pharmaceutical composition, comprising a compound of Table 1 or 2, and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition comprises a plurality of compounds of the invention and a pharmaceutically acceptable carrier.
  • a pharmaceutical composition of the invention further comprises at least one additional pharmaceutically active agent other than a compound of the invention.
  • the at least one additional pharmaceutically active agent can be an agent useful in the treatment of ischemia-reperfusion injury.
  • compositions of the invention can be prepared by combining one or more compounds of the invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.
  • an “effective amount” refers to any amount that is sufficient to achieve a desired biological effect.
  • an effective prophylactic or therapeutic treatment regimen can be planned which does not cause substantial unwanted toxicity and yet is effective to treat the particular subject.
  • the effective amount for any particular application can vary depending on such factors as the disease or condition being treated, the particular compound of the invention being administered, the size of the subject, or the severity of the disease or condition.
  • intravenous administration of a compound may typically be from 0.1 mg/kg/day to 20 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 0.1 mg/kg/day to 2 mg/kg/day.
  • intravenous administration of a compound may typically be from 0.5 mg/kg/day to 5 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 1 mg/kg/day to 20 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 1 mg/kg/day to 10 mg/kg/day.
  • daily oral doses of a compound will be, for human subjects, from about 0.01 milligrams/kg per day to 1000 milligrams/kg per day. It is expected that oral doses in the range of 0.5 to 50 milligrams/kg, in one or more administrations per day, will yield therapeutic results.
  • Dosage may be adjusted appropriately to achieve desired drug levels, local or systemic, depending upon the mode of administration. For example, it is expected that intravenous administration would be from one order to several orders of magnitude lower dose per day. In the event that the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of the compound.
  • the therapeutically effective amount can be initially determined from animal models.
  • a therapeutically effective dose can also be determined from human data for compounds which have been tested in humans and for compounds which are known to exhibit similar pharmacological activities, such as other related active agents.
  • the applied dose can be adjusted based on the relative bioavailability and potency of the administered compound. Adjusting the dose to achieve maximal efficacy based on the methods described above and other methods as are well- known in the art is well within the capabilities of the ordinarily skilled artisan.
  • the formulations of the invention can be administered in pharmaceutically acceptable solutions, which may routinely contain pharmaceutically acceptable concentrations of salt, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.
  • an effective amount of the compound can be administered to a subject by any mode that delivers the compound to the desired surface.
  • Administering a pharmaceutical composition may be accomplished by any means known to the skilled artisan.
  • Routes of administration include but are not limited to intravenous, intramuscular, intraperitoneal, intravesical (urinary bladder), oral, subcutaneous, direct injection (for example, into a tumor or abscess), mucosal (e.g., topical to eye), inhalation, and topical.
  • a compound of the invention can be formulated as a lyophilized preparation, as a lyophilized preparation of liposome-intercalated or -encapsulated active compound, as a lipid complex in aqueous suspension, or as a salt complex.
  • Lyophilized formulations are generally reconstituted in suitable aqueous solution, e.g., in sterile water or saline, shortly prior to administration.
  • the compounds can be formulated readily by combining the active compound(s) with pharmaceutically acceptable carriers well known in the art.
  • Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated.
  • Pharmaceutical preparations for oral use can be obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP).
  • disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • the oral formulations may also be formulated in saline or buffers, e.g., EDTA for neutralizing internal acid conditions or may be administered without any carriers.
  • oral dosage forms of the above component or components may be chemically modified so that oral delivery of the derivative is efficacious.
  • the chemical modification contemplated is the attachment of at least one moiety to the component molecule itself, where said moiety permits (a) inhibition of acid hydrolysis; and (b) uptake into the blood stream from the stomach or intestine.
  • the increase in overall stability of the component or components and increase in circulation time in the body are also desired.
  • moieties include: polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline.
  • Abuchowski and Davis “Soluble Polymer- Enzyme Adducts”, In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, N.Y., pp.367-383 (1981); Newmark et al., J Appl Biochem 4:185-9 (1982).
  • Other polymers that could be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane.
  • polyethylene glycol moieties are suitable.
  • the location of release may be the stomach, the small intestine (the duodenum, the jejunum, or the ileum), or the large intestine.
  • the release will avoid the deleterious effects of the stomach environment, either by protection of the compound of the invention (or derivative) or by release of the biologically active material beyond the stomach environment, such as in the intestine.
  • a coating impermeable to at least pH 5.0 is essential.
  • cellulose acetate trimellitate hydroxypropylmethylcellulose phthalate
  • HPMCP 50 HPMCP 55
  • PVAP polyvinyl acetate phthalate
  • CAP cellulose acetate phthalate
  • shellac shellac
  • These coatings may be used as mixed films.
  • a coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings which make the tablet easier to swallow.
  • Capsules may consist of a hard shell (such as gelatin) for delivery of dry therapeutic (e.g., powder); for liquid forms, a soft gelatin shell may be used.
  • the shell material of cachets could be thick starch or other edible paper.
  • moist massing techniques can be used.
  • the therapeutic can be included in the formulation as fine multi-particulates in the form of granules or pellets of particle size about 1 mm.
  • the formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets.
  • the therapeutic could be prepared by compression. Colorants and flavoring agents may all be included.
  • the compound of the invention may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents.
  • an edible product such as a refrigerated beverage containing colorants and flavoring agents.
  • One may dilute or increase the volume of the therapeutic with an inert material.
  • These diluents could include carbohydrates, especially mannitol, ⁇ -lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch.
  • Certain inorganic salts may also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride.
  • Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.
  • Disintegrants may be included in the formulation of the therapeutic into a solid dosage form.
  • Materials used as disintegrates include but are not limited to starch, including the commercial disintegrant based on starch, Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used.
  • Another form of the disintegrants are the insoluble cationic exchange resins.
  • Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.
  • Binders may be used to hold the therapeutic agent together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic.
  • An anti-frictional agent may be included in the formulation of the therapeutic to prevent sticking during the formulation process.
  • Lubricants may be used as a layer between the therapeutic and the die wall, and these can include but are not limited to; stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000. Glidants that might improve the flow properties of the drug during formulation and to aid rearrangement during compression might be added. The glidants may include starch, talc, pyrogenic silica and hydrated silicoaluminate.
  • Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate.
  • anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate.
  • Cationic detergents which can be used and can include benzalkonium chloride and benzethonium chloride.
  • Non- ionic detergents that could be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the formulation of the compound of the invention or derivative either alone or as a mixture in different ratios.
  • Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added.
  • Microspheres formulated for oral administration may also be used. Such microspheres have been well defined in the art. All formulations for oral administration should be in dosages suitable for such administration.
  • the compositions may take the form of tablets or lozenges formulated in conventional manner.
  • the compound may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art.
  • Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral or pulmonary administration.
  • compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
  • the compound is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream.
  • Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl.5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) ( ⁇ 1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (a-1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins", Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado
  • Pat. No. 5,284,656 granulocyte colony stimulating factor; incorporated by reference.
  • a method and composition for pulmonary delivery of drugs for systemic effect is described in U.S. Pat. No.5,451,569 (incorporated by reference), issued Sep. 19, 1995 to Wong et al.
  • Contemplated for use in the practice of this invention are mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.
  • Some specific examples of commercially available devices suitable for the practice of this invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St.
  • each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material, in addition to the usual diluents, adjuvants and/or carriers useful in therapy. Also, the use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is contemplated.
  • Chemically modified compound of the invention may also be prepared in different formulations depending on the type of chemical modification or the type of device employed.
  • Formulations suitable for use with a nebulizer will typically comprise a compound of the invention (or derivative) dissolved in water at a concentration of about 0.1 to 25 mg of biologically active compound of the invention per mL of solution.
  • the formulation may also include a buffer and a simple sugar (e.g., for inhibitor stabilization and regulation of osmotic pressure).
  • the nebulizer formulation may also contain a surfactant, to reduce or prevent surface induced aggregation of the compound of the invention caused by atomization of the solution in forming the aerosol.
  • Formulations for use with a metered-dose inhaler device will generally comprise a finely divided powder containing the compound of the invention (or derivative) suspended in a propellant with the aid of a surfactant.
  • the propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof.
  • Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be useful as a surfactant.
  • Formulations for dispensing from a powder inhaler device will comprise a finely divided dry powder containing a compound of the invention (or derivative) and may also include a bulking agent, such as lactose, sorbitol, sucrose, or mannitol in amounts which facilitate dispersal of the powder from the device, e.g., 50 to 90% by weight of the formulation.
  • the compound of the invention (or derivative) should advantageously be prepared in particulate form with an average particle size of less than 10 micrometers ( ⁇ m), most preferably 0.5 to 5 ⁇ m, for most effective delivery to the deep lung. Nasal delivery of a pharmaceutical composition of the present invention is also contemplated.
  • Nasal delivery allows the passage of a pharmaceutical composition of the present invention to the blood stream directly after administering the therapeutic product to the nose, without the necessity for deposition of the product in the lung.
  • Formulations for nasal delivery include those with dextran or cyclodextran.
  • a useful device is a small, hard bottle to which a metered dose sprayer is attached.
  • the metered dose is delivered by drawing the pharmaceutical composition of the present invention solution into a chamber of defined volume, which chamber has an aperture dimensioned to aerosolize and aerosol formulation by forming a spray when a liquid in the chamber is compressed.
  • the chamber is compressed to administer the pharmaceutical composition of the present invention.
  • the chamber is a piston arrangement.
  • Such devices are commercially available.
  • a plastic squeeze bottle with an aperture or opening dimensioned to aerosolize an aerosol formulation by forming a spray when squeezed is used.
  • the opening is usually found in the top of the bottle, and the top is generally tapered to partially fit in the nasal passages for efficient administration of the aerosol formulation.
  • the nasal inhaler will provide a metered amount of the aerosol formulation, for administration of a measured dose of the drug.
  • the compounds, when it is desirable to deliver them systemically, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi- dose containers, with an added preservative.
  • compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.
  • Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran.
  • the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
  • the active compounds may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • the compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
  • a compound may also be formulated as a depot preparation.
  • Such long acting formulations may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
  • suitable polymeric or hydrophobic materials for example as an emulsion in an acceptable oil
  • ion exchange resins for example, as an emulsion in an acceptable oil
  • sparingly soluble derivatives for example, as a sparingly soluble salt.
  • the pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.
  • Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous or saline solutions for inhalation, microencapsulated, encochleated, coated onto microscopic gold particles, contained in liposomes, nebulized, aerosols, pellets for implantation into the skin, or dried onto a sharp object to be scratched into the skin.
  • the pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops or preparations with protracted release of active compounds, in whose preparation excipients and additives and/or auxiliaries such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners or solubilizers are customarily used as described above.
  • the pharmaceutical compositions are suitable for use in a variety of drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249:1527-33 (1990).
  • the compound of the invention and optionally other therapeutics may be administered per se (neat) or in the form of a pharmaceutically acceptable salt or cocrystal.
  • a pharmaceutically acceptable salt or cocrystal When used in medicine the salts or cocrystals should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts or cocrystals may conveniently be used to prepare pharmaceutically acceptable salts or cocrystals thereof.
  • Such salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulphuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluene sulphonic, tartaric, citric, methane sulphonic, formic, malonic, succinic, naphthalene-2-sulphonic, and benzene sulphonic.
  • such salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts of the carboxylic acid group.
  • Suitable buffering agents include: acetic acid and a salt (1-2% w/v); citric acid and a salt (1-3% w/v); boric acid and a salt (0.5-2.5% w/v); and phosphoric acid and a salt (0.8-2% w/v).
  • Suitable preservatives include benzalkonium chloride (0.003-0.03% w/v); chlorobutanol (0.3- 0.9% w/v); parabens (0.01-0.25% w/v) and thimerosal (0.004-0.02% w/v).
  • Pharmaceutical compositions of the invention contain an effective amount of a compound as described herein and optionally therapeutic agents included in a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier means one or more compatible solid or liquid filler, diluents or encapsulating substances which are suitable for administration to a human or other vertebrate animal.
  • carrier denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application.
  • the components of the pharmaceutical compositions also are capable of being commingled with the compounds of the present invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficiency.
  • the therapeutic agent(s) including specifically but not limited to a compound of the invention, may be provided in particles.
  • Particles as used herein means nanoparticles or microparticles (or in some instances larger particles) which can consist in whole or in part of the compound of the invention or the other therapeutic agent(s) as described herein.
  • the particles may contain the therapeutic agent(s) in a core surrounded by a coating, including, but not limited to, an enteric coating.
  • the therapeutic agent(s) also may be dispersed throughout the particles.
  • the therapeutic agent(s) also may be adsorbed into the particles.
  • the particles may be of any order release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof, etc.
  • the particle may include, in addition to the therapeutic agent(s), any of those materials routinely used in the art of pharmacy and medicine, including, but not limited to, erodible, nonerodible, biodegradable, or nonbiodegradable material or combinations thereof.
  • the particles may be microcapsules which contain the compound of the invention in a solution or in a semi-solid state.
  • the particles may be of virtually any shape.
  • Both non-biodegradable and biodegradable polymeric materials can be used in the manufacture of particles for delivering the therapeutic agent(s).
  • Such polymers may be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired.
  • Bioadhesive polymers of particular interest include bioerodible hydrogels described in Sawhney H S et al.
  • Macromolecules 26:581-7 the teachings of which are incorporated herein.
  • These include polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).
  • controlled release is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation are controlled. This refers to immediate as well as non-immediate release formulations, with non-immediate release formulations including but not limited to sustained release and delayed release formulations.
  • sustained release also referred to as “extended release” is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that preferably, although not necessarily, results in substantially constant blood levels of a drug over an extended time period.
  • delayed release is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug there from. “Delayed release” may or may not involve gradual release of drug over an extended period of time, and thus may or may not be “sustained release.” Use of a long-term sustained release implant may be particularly suitable for treatment of chronic conditions. “Long-term” release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30-60 days. Long-term sustained release implants are well-known to those of ordinary skill in the art and include some of the release systems described above.
  • Example 1 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme 1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (1 g, 5.204 mmol, 1 equiv) in DMF (20 mL) was added K2CO3 (2.154 g, 15.611 mmol, 3 equiv) and (bromomethyl)benzene (979 mg, 5.724 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt.
  • Example 2 1-(tert-butyl)-3-(2-oxo-1-(pyridin-2-ylmethyl)-1,2,3,4-tetrahydroquinolin-6- yl)urea 6-nitro-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2(1H)-one
  • K2CO3 1078.7 mg, 7.805 mmol, 3 equiv
  • 2- (bromomethyl)pyridine 492.3 mg, 2.862 mmol, 1 equiv).
  • 6-amino-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2-one 440 mg, 1.553 mmol, 1 equiv
  • EtOH 10 mL
  • H2O 2.5 mL
  • Fe 867.3 mg, 15.532 mmol, 10 equiv
  • NH 4 Cl 830.8 mg, 15.530 mmol, 10 equiv
  • Example 3 1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea 6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one
  • K2CO3 1078.7 mg, 7.805 mmol, 3 equiv
  • (1-bromoethyl)benzene 529.6 mg, 2.862 mmol, 1 equiv).
  • 6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one 400 mg, 1.350 mmol, 1 equiv
  • EtOH 10 mL
  • H2O 2.5 mL
  • Fe 753.8 mg, 13.499 mmol, 10 equiv
  • NH4Cl 722.0 mg, 13.499 mmol, 10 equiv
  • Example 4 1-(tert-butyl)-3-(1-((1-methyl-1H-pyrazol-3-yl)methyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea 1-[(1-methylpyrazol-3-yl)methyl]-6-nitro-3,4-dihydroquinolin-2-one
  • K2CO3 1086.6 mg, 7.805 mmol, 3 equiv
  • 3-(bromomethyl)-1- methylpyrazole 500.9 mg, 2.862 mmol, 1.1 equiv).
  • 6-amino-1-[(1-methylpyrazol-3-yl)methyl]-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(1H-pyrazol-3-ylmethyl)-3,4-dihydroquinolin-2-one (300 mg, 1.102 mmol, 1 equiv) in EtOH (10 mL) and H 2 O (2.5 mL) was added Fe (615.3 mg, 11.019 mmol, 10 equiv) and NH 4 Cl (589.3 mg, 11.019 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC.
  • reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-[(1-methylpyrazol-3-yl)methyl]-3,4- dihydroquinolin-2-one (101 mg, 36.03%) as a yellow solid.
  • Example 5 1-(1-benzyl-3-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme 1-benzyl-3-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (1 g, 3.54 mmol, 1 equiv) in THF (70 mL) was added 2M LiHMDS (in THF) (2.3 mL, 4.60 mmol, 1.3 equiv) at -20 °C under N2. The resulting mixture was stirred 30 min at -20 °C.
  • the reaction mixture was added MeI (653.6 mg, 4.60 mmol, 1.3 equiv) at -20 °C.
  • the resulting mixture was stirred 30 min at -20 °C and then stirred 16 h at rt.
  • the reaction mixture was quenched by water (50 mL) and extracted with EA (3 x 70 mL).
  • Example 6 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 1-(3-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • 1-(bromomethyl)-3-chlorobenzene 801.93 mg, 3.903 mmol, 1.5 equiv
  • Example 7 1-(4-benzyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea 4-benzyl-7-nitro-2H-1,4-benzoxazin-3-one
  • K 2 CO 3 860.4 mg, 6.181 mmol, 3 equiv
  • benzyl bromide 387.6 mg, 2.266 mmol, 1.1 equiv).
  • Example 8 1-(tert-butyl)-3-(1-(2-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 1-(2-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • K 2 CO 3 1086.6 mg, 7.805 mmol, 3 equiv
  • 1-(bromomethyl)-2- chlorobenzene 583.64 mg, 2.862 mmol, 1.1 equiv).
  • Example 9 tert-butyl (1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)carbamate Synthetic Scheme tert-butyl (1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)carbamate To a solution of 6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.396 mmol, 1 equiv) in DCM (6 mL) was added Boc2O (129.7 mg, 0.594 mmol, 1.5 equiv) and DMAP (72.6 mg, 0.594 mmol, 1.5 equiv).
  • Example 10 1-(1-benzyl-4,4-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme 4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one (0.4 g, 2.28 mmol, 1 equiv) in H2O (2 mL) and H2SO4 (9 mL) was added HNO3 (0.3 mL) at -10 °C. The reaction was stirred for 2 h at -10-0 °C.
  • Example 12 1-(1-benzyl-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert-butyl)ur ea Synthetic Scheme 2-(benzylamino)-5-nitrobenzonitrile To a solution of 2-fluoro-5-nitrobenzonitrile (5 g, 30.100 mmol, 1 equiv) in ACN (100 mL) was added benzylamine (4.84 g, 45.150 mmol, 1.5 equiv) and K 2 CO 3 (8.32 g, 60.200 mmol, 2 equiv). The reaction was stirred at rt for 1 hour.
  • Example 14 (R)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)u (R)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one
  • Example 15 1-(1-benzyl-4-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea 4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • H 2 O 2-mL
  • H 2 SO 4 9 mL
  • HNO 3 0.3 mL
  • the reaction was stirred for 2 h at -10-0 °C.
  • the reaction mixture was quenched by water.
  • Example 16 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(4-hydroxy-2- methylbutan-2-yl)urea Synthetic Scheme 6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one
  • a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2-one (400 mg, 1.417 mmol, 1 equiv) in EtOH (10 mL) and H 2 O (2 mL) was treated with Fe (791.29 mg, 14.170 mmol, 10 equiv) and NH 4 Cl (757.92 mg, 14.170 mmol, 10 equiv) for 2 h at 60 °C.The resulting mixture was concentrated under reduced pressure.
  • Desired product could be detected by LCMS.
  • the resulting mixture was concentrated under reduced pressure.
  • the crude product was purified by HPLC with the following conditions Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5 ⁇ m; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% B to 41% B in 10 min, 41% B; Wave Length: 254/220 nm; RT1(min): 10; Number Of Runs: 0; to afford 1-(1-benzyl-2-oxo-3,4- dihydroquinolin-6-yl)-3-(4-hydroxy-2-methylbutan-2-yl)urea (23.3 mg, 16.47%) as a white solid.
  • Example 17 1-(tert-butyl)-3-(1-(4-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 4-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile
  • 6-nitro-3,4-dihydroquinolin-2(1H)-one 500 mg, 2.602 mmol, 1 equiv
  • DMF 10 mL
  • K2CO3 1086.6 mg, 7.805 mmol, 3 equiv
  • 4- (bromomethyl)benzonitrile 558.09 mg, 2.862 mmol, 1.1 equiv).
  • Example 18 1-(tert-butyl)-3-(1-(3-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 3-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile
  • 6-nitro-3,4-dihydroquinolin-2(1H)-one 500 mg, 2.602 mmol, 1 equiv
  • DMF 10 mL
  • K 2 CO 3 1086.6 mg, 7.805 mmol, 3 equiv
  • 3- (bromomethyl)benzonitrile 558.09 mg, 2.862 mmol, 1.1 equiv).
  • Example 1 3-tert-butyl-1- ⁇ 1-[(2-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl ⁇ urea 1-[(2-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one
  • 6-nitro-3,4-dihydro-1H-quinolin-2-one (2 g, 10.407 mmol, 1 equiv)
  • 1-(bromomethyl)-2-fluorobenzene (3147.60 mg, 16.651 mmol, 1.6 equiv) in dimethylformamide (20 mL) was added K2CO3 (4346.47 mg, 31.221 mmol, 3 equiv).
  • Example 20 1-(tert-butyl)-3-(1-(2-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 2-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile
  • 2-(bromomethyl)benzonitrile 612.09 mg, 3.122 mmol, 1.5 equiv
  • potassium methaneperoxoate potassium (869.29 mg, 6.243 mmol, 3 equiv) at rt and stirred for overnight.
  • Example 21 1-(tert-butyl)-3-(1-(3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea 6-amino-1-[(3-methylphenyl)methyl]-3,4-dihydroquinolin-2-one
  • K2CO3 1086.6 mg, 7.862 mmol, 3 equiv
  • 1-(bromomethyl)-3- methylbenzene 529.6 mg, 2.862 mmol, 1.1 equiv).
  • Example 22 1-(tert-butyl)-3-(1-(3-methoxybenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- 1-[(3-methoxyphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one
  • K 2 CO 3 1078.7 mg, 7.862 mmol, 3 equiv
  • 1-(bromomethyl)-3- methoxybenzene 575.4 mg, 2.862 mmol, 1.1 equiv).
  • Example 23 1-(tert-butyl)-3-(2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4- tetrahydroquinolin-6-yl)urea 6-nitro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one
  • K2CO3 1086.6 mg, 7.862 mmol, 3 equiv
  • 1-(bromomethyl)-3- (trifluoromethyl)benzene 684.1 mg, 2.862 mmol, 1.1 equiv).
  • 6-amino-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-1- ⁇ [3-(trifluoromethyl)phenyl]methyl ⁇ -3,4-dihydroquinolin-2-one (500 mg, 1.427 mmol, 1 equiv) in EtOH (10 mL) and H 2 O (2.5 mL) was added Fe (797.1 mg, 14.274 mmol, 10 equiv) and NH 4 Cl (763.4 mg, 14.274 mmol, 10 equiv).
  • Example 24 3-tert-butyl-1- ⁇ 1-[(2,5-dichlorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6- yl ⁇ urea 1-[(2,5-dichlorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one
  • 2-(bromomethyl)-1,4-dichlorobenzene 936.34 mg, 3.903 mmol, 1.5 equiv
  • potassium carbonate (724.41 mg, 5.204 mmol, 2 equiv) at room temperature under nitrogen atmosphere.
  • Example 26 1-(tert-butyl)-3-(1-(3-fluorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea 1-(3-fluorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • K2CO3 1086.62 mg, 7.806 mmol, 3 equiv
  • bromomethyl)-3- fluorobenzene 737.72 mg, 3.903 mmol, 1.5 equiv
  • Example 28 1-(tert-butyl)-3-(2-oxo-1-(1-phenylpropyl)-1,2,3,4-tetrahydroquinolin-6- yl)urea 6-nitro-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one
  • K2CO3 863.0 mg, 6.244 mmol, 3 equiv
  • (1-bromopropyl)benzene 455.8 mg, 2.289 mmol, 1.1 equiv).
  • 6-amino-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one (130 mg, 0.419 mmol, 1 equiv) in EtOH (8 mL) and H2O (2 mL) was added Fe (233.9 mg, 4.189 mmol, 10 equiv) and NH4Cl (224.0 mg, 4.189 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C.
  • N-(2-bromophenyl)pivalamide To a stirred solution of O-bromoaniline (3 g, 17.439 mmol, 1 equiv) in anhydrous DCM (50 mL) was added triethylamine (2.29 g, 22.671 mmol, 1.3 equiv) followed by 2,2- dimethylpropanoyl chloride (2.10 g, 17.439 mmol, 1 equiv) at 0 °C. The reaction mixture was stirred for 16 h at rt. After completion of reaction, the reaction mixture was quenched by addition of water (50 mL). The aqueous layer was extracted with DCM (50 mL).
  • 6-amino-1-benzyl-3,3-dimethyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-3,3-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (130 mg, 0.419 mmol, 1 equiv) in 10 mL MeOH was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, then filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-benzyl-3,3-dimethyl-4H-quinolin-2-one (100 mg, 85.15%) as a white solid.
  • Example 30 1-(tert-butyl)-3-(1-(2-hydroxy-1-phenylethyl)-2-oxo-1,2,3,4-tetrahydroquinoli n-6-yl)urea Synthetic Scheme Into a 40 mL round-bottom flask was added 2-bromo-2-phenylethan-1-ol (3 g, 21.713 mmol, 1 equiv) and bromotrimethylsilane (3.66 g, 23.884 mmol, 1.1 equiv) at rt. The resulting mixture was stirred for additional overnight at rt. The resulting mixture was concentrated under reduced pressure.
  • Example 31 1-(1-benzyl-2-oxo-3H-indol-5-yl)-3-tert-butylurea Synthetic Scheme 1-benzyl-5-nitroindole-2,3-dione To a solution of 1H-indole-2,3-dione, 5-nitro (1 g, 5.205 mmol, 1 equiv) and benzyl bromide (979.24 mg, 5.726 mmol, 1.1 equiv) in dimethylformamide (10 mL) was added K2CO3 (1438.66 mg, 10.41 mmol, 2 equiv). The reaction was stirred at rt for 3h.
  • Example 33 3-tert-butyl-1- ⁇ 4-[(3-chlorophenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl ⁇ urea 4-[(3-chlorophenyl)methyl]-7-nitro-2H-1,4-benzoxazin-3-one
  • Example 34 and Example 35 (R)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro -2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme 7-nitro-4-(1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.575 mmol, 1 equiv) in DMF (20 mL) was added Cs 2 CO 3 (1683.41 mg, 5.150 mmol, 2 equiv) and
  • the reaction was stirred at rt for 1 days. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na 2 SO 4 , filtered, and concentrated.
  • Example 36 and Example 37 (R)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethyl)-3-oxo-3,4-di hydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethy l)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea 1-(1-bromoethyl)-3-chlorobenzene To a solution of 1-(3-chlorophenyl)ethanol (1.5 g, 9.578 mmol, 1 equiv) in Et2O (10 mL) was added PBr 3 (5.19 g, 19.156 mmol, 2 equiv) at 0 o C.
  • the reaction was stirred at rt for 1 day. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na 2 SO 4 , filtered, and concentrated.
  • Example 38 1-(3-benzyl-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)-3-(tert-butyl)urea 3-benzyl-6-nitrobenzo[d]oxazol-2(3H)-one
  • 6-nitrobenzo[d]oxazol-2(3H)-one 300 mg, 1.666 mmol, 1 equiv
  • CH3CN 6 mL
  • KI 76.49 mg, 1.666 mmol, 1 equiv
  • K2CO3 788.35 mg, 5.664 mmol, 3.4 equiv
  • benzyl bromide 112.4 mg, 0.657 mmol, 1.1 equiv.
  • Example 39 1-(tert-butyl)-3-(1-(2-fluoro-5-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahyd roquinolin-6-yl)urea 1-(2-fluoro-5-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • Cs 2 CO 3 1.695 g, 5.20 mmol, 2 equiv
  • 2-(bromomethyl)-1-fluoro- 4-(trifluoromethyl)benzene 735.1 mg, 2.860 mmol, 1.1 equiv).
  • Example 40 1-(tert-butyl)-3-(4-(3-methylbenzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazi n-7-yl)urea 4-(3-methylbenzyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one
  • 1-(bromomethyl)-3- methylbenzene 619.59 mg, 3.348 mmol, 1.3 equiv
  • Cs 2 CO 3 (2.52 g, 7.725 mmol, 3 equiv
  • Example 41 and Example 42 (R)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4] oxazin-7-yl)-3-(tert-butyl)urea and (S)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b ][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme 4-benzyl-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.441 mmol, 1 equiv) in DMF (10 mL) was added Cs2CO3 (941.99 mg, 2.882 mmol, 2 equiv) and benzyl bromide (369
  • Example 43 and Example 44 (R)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-methyl-3-oxo-3,4-di hydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-met hyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea 4-(3-chlorobenzyl)-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.402 mmol, 1 equiv) in DMF (10 mL) was added Cs 2 CO 3 (1569.98 mg, 4.804
  • Example 45 and Example 45 3-tert-butyl-1- ⁇ 3-methyl-2-oxo-1-[(1S)-1-phenylethyl]-4H- quinazolin-6-yl ⁇ urea and 3-tert-butyl-1- ⁇ 3-methyl-2-oxo-1-[(1R)-1-phenylethyl]-4H- quinazolin-6-yl ⁇ urea 3-methyl-6-nitro-1-(1-phenylethyl)-4H-quinazolin-2-one To a solution of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (600 mg, 2.896 mmol, 1 equiv) and (1-bromoethyl)benzene (800 mg, 4.323 mmol, 1.49 equiv) in dimethylformamide (30 mL) was added Cs2CO3 (2802.47 mg, 8.575 mmol, 2.96 equiv).
  • Example 46 and Example 52 (S)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3-methyl-2- oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea and (R)-1-(tert-butyl)-3-(1-(1-(4- chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea 1-(1-(3-chlorophenyl)ethyl)-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (500 mg, 2.413 mmol, 1 equiv) and Cs2CO3 (1.57 g, 4.826 mmol, 2 equiv
  • Example 47 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazo lin-6-yl)urea 3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one
  • KNO 3 0.50 g, 4.933 mmol, 0.8 equiv
  • Desired product could be detected by LCMS.
  • the mixture was diluted with water (100 mL).
  • the resulting mixture was extracted with EA (3 x 100 mL).
  • the combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure.
  • the residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 3-methyl-6- nitro-3,4-dihydroquinazolin-2(1H)-one (400 mg, 31.31%) as a white solid.
  • Desired product could be detected by LCMS.
  • the mixture was diluted with water (50 mL).
  • the resulting mixture was extracted with EA (3 x 50 mL).
  • the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated.
  • the residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-amino-1-(3-chlorobenzyl)-3- methyl-3,4-dihydroquinazolin-2(1H)-one (160 mg, 44.53%) as a yellow solid.
  • Example 48 1-(tert-butyl)-3-(1-(2-fluoro-3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin 1-(2-fluoro-3-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • 6-nitro-3,4-dihydro-1H-quinolin-2-one 300 mg, 1.561 mmol, 1 equiv
  • DMF 1-(bromomethyl)-2-fluoro-3-methylbenzene (412.08 mg, 2.029 mmol, 1.3 equiv) and Cs 2 CO 3 (1.53 g, 4.683 mmol, 3 equiv) at rt and stirred for overnight.
  • Example 49 1-(tert-butyl)-3-(1-(2-fluoro-5-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin 1-(2-fluoro-5-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • 2-(bromomethyl)-1-fluoro-4-methylbenzene 412.08 mg, 2.029 mmol, 1.3 equiv
  • Cs 2 CO 3 (1.53 g, 4.683 mmol, 3 equiv
  • Example 50 1-(tert-butyl)-3-(3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H-benzo[b] [1,4]oxazin-7-yl)urea 7-nitro-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one
  • Cs2CO3 1.678 g, 5.150 mmol, 2 equiv
  • 1- (bromomethyl)-3-(trifluoromethyl)benzene (676.8 mg, 2.832 mmol, 1.1 equiv).
  • Example 51 1-(1-benzyl-3-(hydroxymethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylic acid
  • 1-benzyl-6-nitro-3,4-dihydroquinolin-2- one 500 mg, 1.771 mmol, 1 equiv
  • tetrahydrofuran 5 mL, 0.069 mmol, 0.04 equiv
  • 2M LDA in THF
  • Example 53 3-tert-butyl-1- ⁇ 1-[(2-cyano-3-methylphenyl)methyl]-2-oxo-3,4-dihydroquinoli 2-methyl-6-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile
  • 2-(bromomethyl)-6-methylbenzonitrile 400 mg, 1.904 mmol, 1 equiv
  • 6-nitro-3,4-dihydro-1H-quinolin-2-one 548.88 mg, 2.856 mmol, 1.5 equiv
  • K 2 CO 3 (662.69 mg, 4.760 mmol, 2.5 equiv) in portions at room temperature under nitrogen atmosphere.
  • Example 54 1-(tert-butyl)-3-(1-(2-fluoro-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea 1-(2-fluoro-3-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • K2CO3 431.50 mg, 3.122 mmol, 2 equiv
  • 1- (bromomethyl)-2-fluoro-3-(trifluoromethyl)benzene (601.86 mg, 2.341 mmol, 1.5 equiv).
  • Example 55 3-tert-butyl-1-(1- ⁇ [2-cyano-5-(trifluoromethyl) phenyl] methyl ⁇ -2-oxo-3,4- dihydroquinolin-6-yl) urea Synthetic Scheme 1-bromo-2-(bromomethyl)-4-(trifluoromethyl)benzene To a stirred solution of [2-bromo-5-(trifluoromethyl) phenyl] methanol (2 g, 7.842 mmol, 1 equiv) in DCM (20 mL) was added phosphorus tribromide (2.12 g, 7.842 mmol, 1 equiv) in portions at 0°C under air atmosphere.
  • Example 58 3-tert-butyl-1- ⁇ 1-[(1S)-1-(2-fluorophenyl)ethyl]-2-oxo-3,4-dihydroquinolin-6-y l ⁇ ure 1-[(1S)-1-(2-fluorophenyl)ethyl]-6-nitro-3,4-dihydroquinolin-2-one
  • 6-amino-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one (285 mg, 0.782 mmol, 1 equiv) in methanol (3 mL) was added Pd/C (150 mg). The reaction was stirred at rt under H2 for 1h. The resulting mixture was stirred for 1 h at room temperature under hydrogen atmosphere. Desired product could be detected by LCMS.
  • Example 61 (S)-1-(tert-butyl)-3-(1-(1-(2-cyanophenyl)ethyl)-2-oxo-1,2,3,4-tetrahydroquino lin-6-yl)urea (S)-1-(1-(2-bromophenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • (1R)-1-(2-bromophenyl)ethanol (1 g, 4.974 mmol, 1 equiv) in THF (10 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (1.05 g, 5.471 mmol, 1.1 equiv), PPh 3 (1.30 g, 4.974 mmol, 1 equiv) at rt.
  • the crude product 50 mg was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5 ⁇ m; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 54% B in 7 min; Wave Length: 220 nm; RT1(min): 6.5; Number Of Runs: 0 ) to afford (S)-1-(tert-butyl)-3-(1-(1-(2-cyanophenyl)ethyl)- 2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (22.5 mg, 23.93%) as white solid.
  • Example 62 (S)-1-(tert-butyl)-3-(3-oxo-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)urea (S)-7-nitro-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one
  • (1R)-1-[3-(trifluoromethyl)phenyl]ethanol 500 mg, 2.629 mmol, 1 equiv
  • 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one 510.47 mg, 2.629 mmol, 1 equiv
  • PPh3 1034.46 mg, 3.944 mmol, 1.5 equiv
  • Example 64 1-(tert-butyl)-3-(1-(cyclohexylmethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)u rea 1-(cyclohexylmethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • DMF liquid-free fatty acid
  • Cs2CO3 1.53 g, 4.683 mmol, 3.0 equiv
  • (bromomethyl)cyclohexane (276.44 mg, 1.561 mmol, 1 equiv) at rt and stirred for overnight.
  • 6-amino-1-(cyclohexylmethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(cyclohexylmethyl)-6-nitro-3,4-dihydroquinolin-2-one (100 mg, 0.347 mmol, 1 equiv) in methanol (5 mL) was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. This resulted in 6-amino-1-(cyclohexylmethyl)-3,4-dihydroquinolin-2(1H)-one as a white solid.
  • the residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5 ⁇ m; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 50% B in 9 min, 50% B; Wave Length: 254/220 nm; RT1(min): 8.9;) to afford 1-(tert-butyl)-3-(1- (cyclohexylmethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (26.5 mg, 38.18%) as a white solid.
  • Example 65 1-(1-benzyl-7-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme 7-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • HNO 3 200.6 mg,3.185 mmol, 1.0 equiv)at 0 °C.
  • the resulting mixture was stirred at 0 °C until the starting material was totally consumed by LCMS, the reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash
  • reaction progress was monitored by LCMS.
  • the reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.
  • Example 66 1-(1-benzyl-7-chloro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea Synthesis of 7-chloro-6-nitro-3,4-dihydro-1H-quinolin-2-one To a stirred solution of 7-chloro-3,4-dihydro-1H-quinolin-2-one (2 g, 11.012 mmol, 1 equiv) in H2SO4 was added KNO3 (1.11 g, 11.012 mmol, 1 equiv) at 0°C. The final reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS.
  • 6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate (900 mg, 3.936 mmol, 1 equiv) in HCl (2M, in EA) (30 mL) at rt and stirred for 2 h. The precipitated solids were collected by filtration and washed with diethyl ether (3 x 50 mL). This resulted in 6-chloro-3,4- dihydro-1H-1,5-naphthyridin-2-one (600 mg, 83.49%) as a white solid.
  • 6-amino-1-benzyl-3,4-dihydro-1,5-naphthyridin-2(1H)-one Into a 20 mL bar were added tert-butyl N-(5-benzyl-6-oxo-7,8-dihydro-1,5-naphthyridin- 2-yl)carbamate (80 mg, 0.226 mmol, 1 equiv) and HCl (10 mL, 10.000 mmol, 44.18 equiv) at rt and strried for 3 h. The crude product/ resulting mixture was used in the next step directly without further purification.
  • Example 68 1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea Synthetic Scheme 1-benzyl-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (500 mg, 2.616 mmol, 1 equiv) in DMF (20 mL) was added benzyl bromide (671.10 mg, 3.924 mmol, 1.5 equiv) and K 2 CO 3 (728.31 mg, 5.232 mmol, 2 equiv). The reaction was stirred at rt for 1 hour.
  • Example 69 1-(1-benzyl-8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea 8-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one
  • HNO3 (190.7 mg, 3.027 mmol, 1.0 equiv)at -10 °C.
  • the resulting mixture was stirred at -10 °C until the starting material was totally consumed by LCMS,
  • the reaction mixture was quenched by water and adjusting PH with saturaed sodium bicarbonate aqueous solution and then extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na 2
  • Example 70 1-(tert-butyl)-3-(3-methyl-2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4-tetrahyd roquinazolin-6-yl)urea 3-methyl-6-nitro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinazolin-2(1H)-one
  • 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one 120 mg, 0.579 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3-(trifluoromethyl)benzene (152.29 mg, 0.637 mmol, 1.1 equiv) and Cs2CO3 (378.58 mg, 1.158 mmol, 2 equiv).
  • the residue was purified by silica gel column chromatography, eluted with EA (35%-46%) to afford crude product.
  • the residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30*150 mm, 5 ⁇ m; Mobile Phase A: Water (10 mmol/L NH 4 HCO 3 ), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 61% B in 9 min, 61% B; Wave Length: 220/254 nm) to afford1-(tert- butyl)-3-(4-(3-(difluoromethyl)benzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea (4 mg, 6.03%) as a white solid.
  • reaction was stirred at 90°C for 2 hours. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.
  • Example 157 3-tert-butyl-1-[(2R)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4- benzoxazin-7-yl]urea.
  • Synthetic Scheme (2R)-7-bromo-2-methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin-3-one To a stirred solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (400 mg, 1.652 mmol, 1 equiv) in DMF (4 mL) was added 1-(bromomethyl)-3-methylbenzene (458.70 mg, 2.478 mmol, 1.5 equiv) and Cs2CO3 (1080.10 mg, 3.304 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere.
  • the resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere.
  • the reaction was monitored by LCMS.
  • the mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.
  • Example 183 1-(tert-butyl)-3-((R)-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme (R)-7-bromo-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of(2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (270 mg, 1.115 mmol, 1 equiv) in DCM (10 mL) was added 1-[(1R)-1-bromoethyl]-3-chlorobenzene (9.07 mg, 0.041 mmol, 1 equiv) and PPh 3 (438.83 mg
  • reaction was stirred at 90 °C under N 2 for 3h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.
  • Example 191 1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(1-methylcyclobutyl)urea Synthetic Scheme benzyl-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (3 g, 15.695 mmol, 1 equiv) and benzyl bromide (5368.79 mg, 31.390 mmol, 2 equiv) in dimethylformamide (100 mL) was added Cs2CO3 (15388.46 mg, 47.085 mmol, 3 equiv). The reaction was stirred at rt for 3 h.
  • the reaction was added TEA (201.35 mg, 1.990 mmol, 5 equiv) and stirred at rt under N 2 for 1h.
  • the reaction was added 1-methylcyclobutan-1-amine (67.77 mg, 0.796 mmol, 2 equiv) and stirred at rt under N 2 for 1h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.
  • the reaction was stirred at 0 °C under N 2 for 1 h.
  • the reaction was added TEA (188.57 mg, 1.865 mmol, 5 equiv) and stirred at rt under N 2 for 1 h.
  • the reaction was added 3-amino-3- methylbutan-1-ol (153.80 mg, 1.492 mmol, 4 equiv) and stirred at rt under N 2 for 1 h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.
  • 6-nitro-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a mixture of 6-nitroquinoxalin-2(1H)-one (1.8 g, 9.417 mmol, 1.2 equiv) and 1-(bromomethyl)- 3-(trifluoromethoxy)benzene (2.00 g, 7.848 mmol, 1 equiv) in DMF (30 mL) was stirred and added K2CO3 (2.18 g, 15.695 mmol, 2 equiv). The reaction was stirred at rt for 1 h. The reaction mixture was quenched by ice water (30 mL) and extracted with EA (3 x 30 mL).
  • 6-amino-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a mixture of 6-nitro-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one (1.0 g, 2.738 mmol, 1 equiv) in EtOH (25 mL) and water (5 mL). Fe (1.53 g, 27.380 mmol, 10 equiv) and NH 4 Cl (1.46 g, 27.380 mmol, 10 equiv) was added into the mixture. The resulting solution was stirred at 60 °C for 1 h. The reaction was quenched with water (30 mL) at rt.
  • the reaction was stirred at rt for 1 h.
  • TEA 53 mg, 0.522 mmol, 2.5 equiv
  • the reaction was stirred at rt for 1 h.1-amino- 2-methylpropan-2-ol (23 mg, 0.251 mmol, 1.2 equiv) was added into the mixture at 0 °C.
  • the reaction was stirred at rt for 1 h.
  • the reaction mixture was quenched by 1N hydrochloric acid (5 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • Example 246 1-(tert-butyl)-3-(1-(1-(3-cyclobutylphenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme 1-(3-cyclobutylphenyl)ethan-1-one In a 100-mL round bottom flask, to a solution of 1-bromo-3-cyclobutylbenzene (500 mg, 2.369 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added dropwise n-BuLi (2.5 M, 1.1 mL, 2.843 mmol, 1.2 equiv) at -78 °C under N 2 atmosphere.
  • 1-bromo-3-cyclobutylbenzene 500 mg, 2.369 mmol, 1 equiv
  • tetrahydrofuran 5 mL
  • n-BuLi 2.5 M, 1.1 mL, 2.843
  • the resulting mixture was stirred for overnight at rt.
  • the reaction mixture was quenched by water (10 mL).
  • the resulting mixture was extracted with EA (5 mL x 3).
  • the combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure.
  • the crude product 30 mg was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5 ⁇ m; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to affored 3-tert-butyl-1- ⁇ 1-[1-(3- cyclobutylphenyl)ethyl]-2-oxoquinoxalin-6-yl ⁇ urea (7.8 mg, 14.70%) as a yellow solid.
  • the final reaction mixture was stirred overnight at 90 °C.
  • the reaction was monitored by LCMS.
  • the mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na 2 SO 4 , filtered and concentrated.
  • Example 268 and Example 269 1-(tert-butyl)-3-((S)-2-methyl-3-oxo-4-((S)-1-phenylethyl)- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4- ((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme 2-methyl-7-nitro-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (350 mg, 1.681 mmol, 1 equiv) in DCM (10 mL) was added (R)-1-
  • reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na 2 SO 4 and concentrated under vacuum.
  • the crude product was purified by silica gel column to afford 3-methyl-6-nitro-1-(1-(3- (trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)-one (100 mg, 27.31%) as a yellow solid.
  • Example 274 1-(tert-butyl)-3-(6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea Synthetic Scheme 6-chloro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of 6-chloro-3,4-dihydro-1H-1,5-naphthyridin-2-one (300 mg, 1.643 mmol, 1 equiv) in anhydrous dimethylformamide (5 mL) was added 1-(bromomethyl)-3- (trifluoromethyl)benzene (432 mg, 1.807 mmol, 1.1 equiv) and Cs 2 CO 3 (1.6 g, 4.929 mmol, 3 equiv) at rt and stirred for 2
  • reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum.
  • the crude product was purified by flash chromatography with PE/EA (3:1) to afford 6-chloro-1-(3- (trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (390 mg, 69.67%) as a light- yellow solid.
  • reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum.
  • the crude product was purified by flash chromatography with PE/EA (3:1) to afford tert-butyl (6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate (0.5 g, 96.01%) .
  • Example 276 (S)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3- (tert-butyl)urea (S)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea
  • 2-(6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)acetonitrile 70 mg, 0.240 mmol, 1 equiv
  • 2-isocyanato-2-methylpropane 72 mg, 0.720 mmol, 3.0 equiv
  • TEA 122 mg, 1.200 mmol, 5.0 equiv
  • reaction mixture was quenched by addition of water (15 mL).
  • the aqueous layer was extracted with ethyl acetate (3 x 30 mL).
  • the combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure.
  • the residue was purified by silica gel column, eluted with PE/EA (3:1) to give 2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (1.3 g, 90.17%) as a yellow solid.
  • reaction mixture was quenched by addition of water (15 mL).
  • the aqueous layer was extracted with DCM (3 x 20 mL).
  • the combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure.
  • the residue was purified by silica gel column, eluted with PE/EA (2:1) to give crude product.
  • Example 278 (S)-1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea (S)-1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea A solution of 7-amino-4-benzyl-2-ethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.531 mmol, 1 equiv), TEA (269 mg, 2.655 mmol, 5 equiv) and 2-isocyanato-2-methylpropane (263 mg, 2.655 mmol, 5 equiv) in DCM (10 mL) was stirred for overnight at r
  • reaction mixture was quenched by addition of water (15 mL).
  • the aqueous layer was extracted with DCM (3 x 20 mL).
  • the combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure.
  • the residue was purified by silica gel column, eluted with PE/EA (2:1) to give crude product.
  • Example 279 1-(1-benzyl-5-cyano-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-5-carbonitrile To a solution of 1-benzyl-5-chloro-6-nitro-3,4-dihydroquinolin-2-one (180 mg, 0.568 mmol, 1 equiv) in dimethylformamide (2 mL) was added zinc (5.57 mg, 0.085 mmol, 0.15 equiv), zinc cyanide (80.07 mg, 0.682 mmol, 1.2 equiv), Brettphos Pd G 3 (103.03 mg, 0.114 mmol, 0.2 equiv) and Brettphos (61.01 mg, 0.114 mmol, 0.2 equiv).
  • Example 280 (R)-1-(1-benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6- yl)-3-(tert-butyl)urea Synthetic Scheme ethyl (R)-2-((6-chloro-3-nitropyridin-2-yl)oxy)propanoate To a solution of 2,6-dichloro-3-nitropyridine (2.0 g, 10.364 mmol, 1 equiv) in tetrahydrofuran (20 mL) was added 60% NaH (0.5 g, 12.437 mmol, 1.2 equiv) at 0 °C.
  • 6-chloro-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one 300 mg, 1.643 mmol, 1 equiv
  • 6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one 300 mg, 1.643 mmol, 1 equiv
  • 1-(bromomethyl)-3-chlorobenzene (371 mg, 1.807 mmol, 1.1 equiv) and Cs2CO3 (1.6 g, 4.929 mmol, 3 equiv) at rt and stirred for 2 h.
  • Example 282 (S)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8- tetrahydro-1,5-naphthyridin-2-yl)urea Synthetic Scheme 6-chloro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a solution of 6-chloro-3,4-dihydro-1H-1,5-naphthyridin-2-one (400 mg, 2.190 mmol, 1 equiv) in DMF (10 mL) was added K 2 CO 3 (908 mg, 6.571 mmol, 3 equiv) and 1-(1-bromoethyl)-3- (trifluoromethyl)benzene (831 mg, 3.286 mmol
  • reaction progress was monitored by LCMS.
  • the reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.
  • Example 283 (R)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8- tetrahydro-1,5-naphthyridin-2-yl)urea (R)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea To a stirred solution of 6-amino-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (100 mg, 0.298 mmol, 1 equiv) in DCM (10.00 mL) was added 2- isocyanato-2-methylpropane
  • reaction progress was monitored by LCMS.
  • the reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.
  • Example 284 3-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-1-phenylurea 3-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-1-phenylurea
  • 6-amino-1-benzyl-3,4-dihydroquinolin-2-one 150 mg, 0.594 mmol, 1 equiv
  • triphosgene 59.98 mg, 0.202 mmol, 0.34 equiv
  • Example 285 N-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide N-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide
  • N-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide To a stirred solution of 3,3-dimethylbutanamide (35.15 mg, 0.305 mmol, 1.1 equiv) and HATU (137.13 mg, 0.360 mmol, 1.3 equiv) in DMF (1 mL) were added 6-amino-1-benzyl-3,4- dihydroquinolin-2-one (70 mg, 0.277 mmol, 1 equiv) and DIEA (107.57 mg, 0.831 mmol, 3 equiv) in portions
  • Example 286 (S)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-6-yl)urea Synthetic Scheme (S)-6-chloro-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of (S)-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (100 mg, 0.503 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-3-chlorobenzene (155 mg, 0.754 mmol, 1.5 equiv) and Cs 2 CO 3 (492 mg, 1.509 mmol, 3.0 equiv)
  • the resulting mixture was stirred for overnight at rt.
  • the reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • Example 287 (R)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-6-yl)urea (R)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)urea A solution of (R)-6-amino-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)- one (60 mg, 0.198 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (195 mg, 1.980 mmol, 10
  • the resulting mixture was stirred for overnight at rt.
  • the reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • the crude product 40 mg was purified by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IF, 2*25 cm, 5 ⁇ m; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 25% B to 25% B in 23 min; Wave Length: 220/254 nm; RT1(min): 13.384; RT2(min): 18.251; Sample Solvent: EtOH--HPLC; Injection Volume: 0.6 mL; Number Of Runs: 7) to afford (S)-1-(4-benzyl-2-(methoxymethyl)-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.0 mg, 21.32%) as a white solid.
  • the crude product 40 mg was purified by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IF, 2*25 cm, 5 ⁇ m; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 25% B to 25% B in 23 min; Wave Length: 220/254 nm; RT1(min): 13.384; RT2(min): 18.251; Sample Solvent: EtOH--HPLC; Injection Volume: 0.6 mL; Number Of Runs: 7) to afford (S)-1-(4-benzyl-2-(methoxymethyl)-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.0 mg, 21.32%) as a white solid.
  • tert-butyl (5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2- yl)carbamate
  • 6-chloro-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one 200 mg, 0.623 mmol, 1 equiv
  • Xantphos (36.0 mg, 0.062 mmol, 0.1 equiv)
  • Pd2(dba)3 (57.0 mg, 0.062 mmol, 0.1 equiv)
  • Cs2CO3 (608.6 mg, 1.868 mmol, 3 equiv).
  • the resulting mixture was placed at 90°C under a nitrogen atmosphere and stirred overnight until the starting material was totally consumed by LCMS. After cooled to room temperature the reaction mixture was quenched by ice water (10 mL) and extracted with EA (3 x 20 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
  • Example 292 1-(4-benzyl-2,2-dimethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea Synthetic Scheme 4-benzyl-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of 2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 1.350 mmol, 1 equiv) in anhydrous DMF (10 mL) was added K 2 CO (373 mg, 2.700 mmol, 2 equiv) and BnBr (346 mg, 2.025 mmol, 1.5 equiv) at rt.
  • K 2 CO 373 mg, 2.700 mmol, 2 equiv
  • BnBr 346 mg, 2.025 mmol, 1.5 equiv
  • reaction was stirred for overnight at rt.
  • the reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • Example 293 1-(tert-butyl)-3-(4-(1-(5-chloro-2-cyanophenyl)ethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme 1-bromo-2-(1-bromoethyl)-4-chlorobenzene To a mixture of 1-(2-bromo-5-chlorophenyl)ethan-1-ol (300 mg, 1.047 mmol, 1 equiv) in DCM (5 mL) was added PBr 3 (567 mg, 2.094 mmol, 2 equiv) at 0 °C.
  • the reaction was stirred for overnight at rt.
  • the reaction mixture was quenched by ice water (10 mL) and the pH value of the aqueous phase was adjusted to 8 with NaHCO3 (aq.).
  • the mixture was extracted with DCM (3 x 10 mL).
  • the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • the residue was purified by silica gel column, eluted with PE/EA (20:1) to give 1- bromo-2-(1-bromoethyl)-4-chlorobenzene (218 mg, 69.76%) as yellow oil.
  • reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford 4-(1-(2-bromo-5-chlorophenyl)ethyl)-7-nitro-2H-benzo[b][1,4]oxazin- 3(4H)-one (150 mg, 81.55%) as a yellow solid.

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Abstract

Disclosed are compounds that are thyroid stimulating hormone receptor antagonists, and methods useful for preventing or treating a thyroid disease.

Description

SUBSTITUTED 3,4-DIHYDROQUINOLINONE INHIBITORS OF TSHR CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63/393,411, filed July 29, 2022; the contents of which are hereby incorporated by reference in their entirety. BACKGROUND About 40% of hyperthyroidism patients suffer from Graves' disease (Morbus Basedow), an autoimmune disease in which autoantibodies activate the thyrotropin receptor, mimicking its natural hormone ligand, the thyroid-stimulating hormone (TSH). This pathological activation of TSH-Receptor (TSHR) leads to uncontrolled production of thyroid hormones (e.g., T3 and T4) causing hyperthyroidism. TSH and the TSHR are important proteins for controlling thyroid function. TSHR is primarily expressed in follicular epithelial cells of the thyroid gland, but also is expressed in a variety of additional cell types, such as retro-orbital fibroblasts, kidney cells, adipocytes and bone cells. TSH binds to its receptor and leads to the stimulation of second messenger pathways involving predominantly cAMP. Inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) pathways are also activated at higher TSH concentrations. The treatment of choice applied in the clinics for decades involves thyrostatic drugs that block the production of thyroid hormones. These medications play a role further downstream in the signal cascade of the thyroid upon activation of the TSHR. Since thyroid hormones T3 and T4 are secreted in the thyroid gland, thyrostatic drugs induce an inhibition of their synthesis. Thus, the current primary anti-thyroid treatment does not target the causative molecular activation of the TSHR by autoantibodies and patients are therefore burdened by a rate of at least 5% adverse effects. This demands frequent controls of the thyroid hormone levels and adjustments of thyrostatics dosage. In contrast to these drugs, which regulate the thyroid hormone level, another promising target is the TSHR itself. However, small allosteric antagonists acting directly at the TSHR are not available on the market yet. Additionally, about 25% of Graves' disease patients also develop an orbitopathy referred to as “Graves' ophthalmopathy”, a related organ-specific autoimmune disease affecting the appearance and functioning of the eyes. There is considerable evidence that expression of the TSHR in the orbital fibroblasts and orbital adipocytes behind the eye may contribute to this difficult-to-treat orbitopathy, and thyroid stimulating antibodies titer tend to correlate with severity of Grave’s ophthalmopathy. Orbital fibroblast has been recognized as primary target cells of autoimmune attack and TSHR acts as a primary autoantigen in Grave’s ophthalmopathy. The pathological activation of TSHR leads to the production of the extracellular matrix by involvement of hyaluronanic acid, fibrosis and swelling of extraocular muscle as well as adipogenesis of orbital fibroblasts (orbital fat expansion). The increase in tissue volume in the orbit often causes diplopia and compression of the optic nerve and exophthalmos. Thus, the TSHR is also potential target for pharmacological intervention of Grave’s ophthalmology and thyroid eye disease. Accordingly, there is a need in the art to provide additional means for the treatment of hyperthyroidism, in particular compounds that act as TSHR antagonists. SUMMARY One aspect of the invention provides compounds, compositions, and methods useful for preventing or treating a thyroid disease, such as hyperthyroidism, Grave’s disease, Grave’s opthalmopathy, or thyroid eye disease. Accordingly, provided herein in some embodiments is a compound having the structure of Formula (I):
Figure imgf000003_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. In some embodiments a compound has the structure of Formula (II):
Figure imgf000005_0001
or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. In some embodiments a compound has the structure of Formula (III):
Figure imgf000007_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1-C6)alkyl, (C1-C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1- C6)alkyl or halo, or when Z is C(R5)2, R3 and one R5 taken together with the carbon to which they are attached form a 4, (C3-C8)cycloalkyl optionally substituted with one or more (C1- C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; R10 is hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Other features, objects, and advantages of the invention will be apparent from the detailed description, and from the claims. DETAILED DESCRIPTION Definitions For convenience, before further description of the present invention, certain terms employed in the specification, examples and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of skill in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. In order for the present invention to be more readily understood, certain terms and phrases are defined below and throughout the specification. The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. The phrase “and/or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and/or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and/or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to “A and/or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc. As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and/or” as defined above. For example, when separating items in a list, “or” or “and/or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of” or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e., “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law. As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and/or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc. It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited. In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of” and “consisting essentially of” shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03. Certain compounds contained in compositions of the present invention may exist in particular geometric or stereoisomeric forms. In addition, polymers of the present invention may also be optically active. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention. “Geometric isomer" means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration. "R," "S," "S*," "R*," "E," "Z," "cis," and "trans," indicate configurations relative to the core molecule. Certain of the disclosed compounds may exist in “atropisomeric” forms or as “atropisomers.” Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from a mixture of isomers. Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods. If, for instance, a particular enantiomer of compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers. Percent purity by mole fraction is the ratio of the moles of the enantiomer (or diastereomer) or over the moles of the enantiomer (or diastereomer) plus the moles of its optical isomer. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least about 60%, about 70%, about 80%, about 90%, about 99% or about 99.9% by mole fraction pure. When a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound or mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry and has two or more chiral centers, it is to be understood that the name or structure encompasses a diastereomer free of other diastereomers, a number of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other diastereomer(s) or mixtures of diastereomers in which one or more diastereomer is enriched relative to the other diastereomers. The invention embraces all of these forms. Structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a 13C- or 14C-enriched carbon are within the scope of this invention. The term “prodrug” as used herein encompasses compounds that, under physiological conditions, are converted into therapeutically active agents. A common method for making a prodrug is to include selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the host animal. The phrase “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject chemical from one organ or portion of the body, to another organ or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, not injurious to the patient, and substantially non- pyrogenic. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer’s solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations. In certain embodiments, pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not induce significant temperature elevations when administered to a patient. The term “pharmaceutically acceptable salts” refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting a purified compound(s) in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts, and the like. (See, for example, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci.66:1-19.) In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term “pharmaceutically acceptable salts” in these instances refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra). The term “pharmaceutically acceptable cocrystals” refers to solid coformers that do not form formal ionic interactions with the small molecule. A “therapeutically effective amount” (or “effective amount”) of a compound with respect to use in treatment, refers to an amount of the compound in a preparation which, when administered as part of a desired dosage regimen (to a mammal, preferably a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit/risk ratio applicable to any medical treatment. The term “prophylactic or therapeutic” treatment is art-recognized and includes administration to the host of one or more of the subject compositions. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the host animal) then the treatment is prophylactic, (i.e., it protects the host against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic, (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof). The term “patient” or “subject” refers to a mammal in need of a particular treatment. In certain embodiments, a patient is a primate, canine, feline, or equine. In certain embodiments, a patient is a human. An aliphatic chain comprises the classes of alkyl, alkenyl and alkynyl defined below. A straight aliphatic chain is limited to unbranched carbon chain moieties. As used herein, the term “aliphatic group” refers to a straight chain, branched-chain, or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups, such as an alkyl group, an alkenyl group, or an alkynyl group. “Alkyl” refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having the number of carbon atoms specified, or up to 30 carbon atoms if no specification is made. For example, alkyl of 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, and those moieties which are positional isomers of these moieties. Alkyl of 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl and tetracosyl. In certain embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chains, C3-C30 for branched chains), and more preferably 20 or fewer. Alkyl goups may be substituted or unsubstituted. As used herein, the term “heteroalkyl” refers to an alkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. As used herein, the term “haloalkyl” refers to an alkyl group as hereinbefore defined substituted with at least one halogen. As used herein, the term “hydroxyalkyl” refers to an alkyl group as hereinbefore defined substituted with at least one hydroxyl. As used herein, the term “alkylene” refers to an alkyl group having the specified number of carbons, for example from 2 to 12 carbon atoms, that contains two points of attachment to the rest of the compound on its longest carbon chain. Non-limiting examples of alkylene groups include methylene -(CH2)-, ethylene -(CH2CH2)-, n-propylene -(CH2CH2CH2)-, isopropylene - (CH2CH(CH3))-, and the like. Alkylene groups can be cyclic or acyclic, branched or unbranched carbon chain moiety, and may be optionally substituted with one or more substituents. "Cycloalkyl" means mono- or bicyclic or bridged or spirocyclic, or polycyclic saturated carbocyclic rings, each having from 3 to 12 carbon atoms. Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3-6 carbons in the ring structure. Cycloalkyl groups may be substituted or unsubstituted. As used herein, the term “halocycloalkyl” refers to an cycloalkyl group as hereinbefore defined substituted with at least one halogen. "Cycloheteroalkyl" refers to an cycloalkyl moiety as hereinbefore defined which contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms in place of carbon atoms. Preferred cycloheteroalkyls have from 4-8 carbon atoms and heteroatoms in their ring structure, and more preferably have 4-6 carbons and heteroatoms in the ring structure. Cycloheteroalkyl groups may be substituted or unsubstituted. Unless the number of carbons is otherwise specified, “lower alkyl,” as used herein, means an alkyl group, as defined above, but having from one to ten carbons, more preferably from one to six carbon atoms in its backbone structure such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Likewise, “lower alkenyl” and “lower alkynyl” have similar chain lengths. Throughout the application, preferred alkyl groups are lower alkyls. In certain embodiments, a substituent designated herein as alkyl is a lower alkyl. “Alkenyl” refers to any cyclic or acyclic, branched or unbranched unsaturated carbon chain moiety having the number of carbon atoms specified, or up to 26 carbon atoms if no limitation on the number of carbon atoms is specified; and having one or more double bonds in the moiety. Alkenyl of 6 to 26 carbon atoms is exemplified by hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosoenyl, docosenyl, tricosenyl, and tetracosenyl, in their various isomeric forms, where the unsaturated bond(s) can be located anywhere in the moiety and can have either the (Z) or the (E) configuration about the double bond(s). “Alkynyl” refers to hydrocarbyl moieties of the scope of alkenyl, but having one or more triple bonds in the moiety. The term “aryl” as used herein includes 3- to 12-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon (i.e., carbocyclic aryl) or where one or more atoms are heteroatoms (i.e., heteroaryl). Preferably, aryl groups include 5- to 12-membered rings, more preferably 6- to 10-membered rings The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Carboycyclic aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. Heteroaryl groups include substituted or unsubstituted aromatic 3- to 12-membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10-membered rings, whose ring structures include one to four heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Aryl and heteroaryl can be monocyclic, bicyclic, or polycyclic. The term “halo”, “halide”, or “halogen” as used herein means halogen and includes, for example, and without being limited thereto, fluoro, chloro, bromo, iodo and the like, in both radioactive and non-radioactive forms. In a preferred embodiment, halo is selected from the group consisting of fluoro, chloro and bromo. The terms “heterocyclyl” or “heterocyclic group” or “heterocycloalkyl” refer to 3- to 12- membered ring structures, more preferably 5- to 12-membered rings, more preferably 5- to 10- membered rings, whose ring structures include one to four heteroatoms. Heterocycles can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, chromane, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. The heterocyclic ring can be substituted at one or more positions with such substituents as described above, as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF3, -CN, and the like. The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. In preferred embodiments, the substituents on substituted alkyls are selected from C1-6 alkyl, C3-6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on substituted alkyls are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants. As used herein, the definition of each expression, e.g., alkyl, m, n, etc., when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure. As used herein, “small molecules” refers to small organic or inorganic molecules of molecular weight below about 3,000 Daltons. In general, small molecules useful for the invention have a molecular weight of less than 3,000 Daltons (Da). The small molecules can be, e.g., from at least about 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da). In some embodiments, a “small molecule” refers to an organic, inorganic, or organometallic compound typically having a molecular weight of less than about 1000. In some embodiments, a small molecule is an organic compound, with a size on the order of 1 nm. In some embodiments, small molecule drugs of the invention encompass oligopeptides and other biomolecules having a molecular weight of less than about 1000. An “effective amount” is an amount sufficient to effect beneficial or desired results. For example, a therapeutic amount is one that achieves the desired therapeutic effect. This amount can be the same or different from a prophylactically effective amount, which is an amount necessary to prevent onset of disease or disease symptoms. An effective amount can be administered in one or more administrations, applications or dosages. A therapeutically effective amount of a composition depends on the composition selected. The compositions can be administered from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the compositions described herein can include a single treatment or a series of treatments. The terms “decrease,” “reduce,” “reduced”, “reduction”, “decrease,” and “inhibit” are all used herein generally to mean a decrease by a statistically significant amount relative to a reference. However, for avoidance of doubt, “reduce,” “reduction” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level and can include, for example, a decrease by at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, up to and including, for example, the complete absence of the given entity or parameter ascompared to the reference level, or any decrease between 10-99% as compared to the absence of a given treatment. The terms “increased”, “increase” or “enhance” or “activate” are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms “increased”, “increase” or “enhance” or “activate” means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10- fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. As used herein, the term “modulate” includes up-regulation and down-regulation, e.g., enhancing or inhibiting a response. A “radiopharmaceutical agent,” as defined herein, refers to a pharmaceutical agent which contains at least one radiation-emitting radioisotope. Radiopharmaceutical agents are routinely used in nuclear medicine for the diagnosis and/or therapy of various diseases. The radiolabelled pharmaceutical agent, for example, a radiolabelled antibody, contains a radioisotope (RI) which serves as the radiation source. As contemplated herein, the term “radioisotope” includes metallic and non-metallic radioisotopes. The radioisotope is chosen based on the medical application of the radiolabeled pharmaceutical agents. When the radioisotope is a metallic radioisotope, a chelator is typically employed to bind the metallic radioisotope to the rest of the molecule. When the radioisotope is a non-metallic radioisotope, the non-metallic radioisotope is typically linked directly, or via a linker, to the rest of the molecule. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Compounds of the Invention One aspect of the invention relates to a compound of Formula (I):
Figure imgf000020_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. In some embodiments of the compound of Formula (I), A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, (C1-C6)alkoxy, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy.: Another aspect of the invention relates to a compound Formula (II):
Figure imgf000023_0001
or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. In other embodiments or of the compound of Formula (II), A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, , 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C1-C6)alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; Wa–Wb represents: –C(R6)2–C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, or –N=C(R10) –; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy. Yet another aspect of the invention relates to a compound of Formula (III):
Figure imgf000026_0001
or a pharmaceutically acceptable salt thereof, A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1-C6)alkyl, (C1-C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1- C6)alkyl or halo, or when Z is C(R5)2, R3 and one R5 taken together with the carbon to which they are attached form a 4, (C3-C8)cycloalkyl optionally substituted with one or more (C1- C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; R10 is hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1- C6)alkyl, and (C1-C6)alkoxy. In some embodiments of the conmpound of Formula (III), A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C1-C6)alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; and R10 is hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy. In certain embodiments, A is phenyl. In certain embodiments, A is a 5- to 7-membered heteroaryl comprising 1 or 2 nitrogen atoms. In certain embodiments, A is pyridyl or diazolyl. thiazolyl, pyrazinyl, pyrimidinyl, while in other embodiments, A is pyridyl or diazolyl. In some embodiments, A is wherein A is isoquinolinyl or indazolyl. In other embodiments, A is cyclopentyl, cyclohexyl, or tetrahydropyranyl. In certain embodiments, R1 is independently for each occurrence (C1-C6)alkyl, (C1- C6)alkoxy, fluoro, chloro, or cyano, wherein (C1-C6)alkyl and (C1-C6)alkoxy are optionally substituted with one or more substituents independently selected from fluoro and hydroxyl. In certain embodiments, at least one R1 is fluoro, chloro, or cyano. In certain embodiments, at least one R1 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from hydroxyl and fluoro. In certain preferred embodiments, R1 is 2,2,2-trifluoroethyl-1-hydroxyethyl. In other preferred embodiments, at least one R1 is difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluropropyl, or 4,4,4- trifluorobutyl. In certain embodiments, at least one R1 is (C1-C6)alkoxy optionally substituted with at least one fluoro, preferably (C1-C3)alkoxy optionally substituted with at least one fluoro, more preferably difluoromethoxy or trifluoromethoxy. In other preferred embodiments, R1 is methoxy or isopropoxy. In certain embodiments, R1 is methyl optionally substituted with hydroxy or one or more fluoro. In certain embodiments, R1 is (C1-C3)alkoxy optionally substituted with at least one fluoro. In certain embodiments, at least one R1 is (C3-C8)cycloalkyl, preferably cyclopropyl or cyclobutyl. In certain embodiments, at least R1 is (C3-C8)cycloalkoxy, preferably cyclopentyloxy, cyclohexyloxy. In certain embodiments, R1 is (C2-C6)alkynyl optionally substituted with hydroxy. In certain embodiments, two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3- C8)cycloalkyl, and (C1-C6)alkoxy. In certain embodiments, A is phenyl comprising two vicinal occurrences of R1, wherein the two vicinal occurrences of R1, taken together with the atoms to which they are attached, form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring. In certain embodiments, n is 2. In certain embodiments, n is 1. In certain embodiments, n is 0. In certain embodiments, Q is CR2c. In certain embodiments, Q is N. In certain embodiments, R2a, R2b, and R2c are each hydrogen. In certain embodiments, at least one of R2a, R2b, and R2c is fluoro, chloro, or cyano. In certain embodiments, R3 is (C1-C6)alkyl optionally substituted with hydroxy. In certain preferred embodiments, R3 is t-butyl. In certain embodiments, R3 is (C3-C8)cycloalkyl optionally substituted with one or more (C1-C6)alkyl, (C1-C6)fluoroalkyl, fluoro, or (C1-C6)alkoxy(C1-C6)alkyl. In more particular embodiments, the (C3-C8)cycloalkyl is cyclobutyl, cyclopentyl, cylohexyl, bicyclo[1.1.1]pentanyl, bicyclo[2.2.1]heptanyl,bicycle[2.1.1]hexanyl, or bicyclo[3.1.0]hexanyl. Inb further particular embodiments, the (C3-C8)cycloalkyl is optionally substituted with one or more substituents selected from fluoro, trifluoromethyl, methoxymethyl, methyl, and hydroxyl. In certain embodiments, R3 is 4- to 7-membered heterocycloalkyl optionally substituted with one or more substituents selected from (C1-C6)alkyl, (C1-C6)fluoroalkyl, or fluoro. In more particular embodiments, the 4- to 7-membered heterocycloalkyl is etrahydropyranyl or oxetanyl. In some embodiments, Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1- C6)alkyl or halo. In some embodiments, the 4- to 7 membered heterocycloalkyl is azetidine or pyrrolidine, In certain embodiments, X is NR4 or O. In certain embodiments, X is NH. In certain embodiments, Z is NR4, or O. In certain embodiments, Z is NH. In certain embodiments, one and only one of X and Z is CH2. In certain embodiments, X and Z are each NH. In certain embodiments, L is unsubstituted (C1-C6)alkylene. In certain embodiments, L is (C1-C6)alkylene substituted with hydroxy. In certain embodiments, L is (C1-C6)alkylene substituted with 1 to 3 halo atoms. In certain embodiments, L is –CH2–, –-CH(CH3)–, –CH(CH2CH3)–, or –CH(CH2OH)–. In certain embodiments, Wa–Wb represents: –C(R6)2–C(R7)2–. In certain embodiments, R6 is (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy. In certain embodiments, at least one R7 is (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, (C1- C6)alkoxy, and cyano. In certain embodiments, each R6 and each R7 is hydrogen. In certain embodiments, Wa–Wb represents: –O–C(R8)2–. In certain embodiments, R8 is hydrogen. In certain embodiments, one R8 (C1-C6)alkyl, preferably methyl; and one R8 is hydrogen. In certain embodiments, Wa–Wb represents: –C(R8)2–O–, where each R8 is hydrogen. In certain embodiments, Wa–Wb represents: –C(R8)2–NR9–. In certain embodiments, each R8 is hydrogen. In certain embodiments, R9 is (C1-C6)alkyl, preferably methyl. In certain embodiments, Wa–Wb represents: –N=C(R10) –. In certain embodiments, R10 is hydrogen. In certain embodiments, R10 is (C1-C6)alkyl. In certain embodiments, R10 is methyl. In certain embodiments, a compound is selected from Table 1: Table 1.
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
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Figure imgf000060_0001
Figure imgf000061_0001
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Figure imgf000063_0001
Figure imgf000064_0001
Figure imgf000065_0001
Figure imgf000066_0001
Figure imgf000067_0001
258 259 260 261 262 263
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0001
Figure imgf000075_0001
Figure imgf000076_0001
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
Figure imgf000081_0001
Figure imgf000082_0001
Figure imgf000083_0001
Figure imgf000084_0001
Figure imgf000085_0001
Figure imgf000086_0001
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0002
In certain embodiments, the compound is selected from Table 2. Table 2.
Figure imgf000090_0001
Figure imgf000091_0001
Figure imgf000092_0001
Figure imgf000093_0001
Figure imgf000094_0001
Figure imgf000095_0001
or a pharmaceutically acceptable salt thereof. In certain embodiments, the compounds are atropisomers. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a 13C- or 14C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present invention. For example, in the case of variable R1, the (C1-C4)alkyl or the -O-(C1- C4)alkyl can be suitably deuterated (e.g., -CD3, -OCD3). Any compound of the invention can also be radiolabed for the preparation of a radiopharmaceutical agent. Methods of Treatment One aspect of the invention provides methods of inhibiting a thyroid stimulating hormone receptor, comnprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein. Another aspect of the invention comprises treating or preventing a thyroid disease, comnprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein. In certain embodiments, the subject has Graves' disease. In certain embodiments, the subject has Graves' ophthalmopathy. In certain embodiments, the subject has Graves' dermopathy. In certain embodiments, the subject has thyroid cancer. While not being bound by theory, TSHR in thyroid cells, and likely in fibroblasts and adipocytes in the supporting tissue behind the eye (in the retro-orbital space), also are stimulated by TSHR-stimulating antibodies (TSAbs), resulting in Graves' disease. Graves' disease, which is an autoimmune disease that occurs in 1% of the US population, has two important clinical components: 1) hyperthyroidism from stimulation of TSHR on thyroid cells and 2) Graves' orbitopathy (or Graves' ophthalmopathy or thyroid eye disease), which appears to result from stimulation of TSHR on retro-orbital fibroblasts and/or adipocytes. Hyperthyroidism, in particular Graves' hyperthyroidism, is a hypermetabolic state that affects virtually every tissue/cell in the body and can lead to, in particular, cardiovascular dysfunction and death. Graves' ophthalmopathy, also known as Graves' orbitopathy, occurs in 80% of Graves' hyperthyroid patients as diagnosed by computerized tomographic scan. Symptoms range from mild to moderate to severe to sight-threatening. Protrusion of the eyeball (proptosis) and varying degrees of extra-ocular muscle weakness or paralysis leading to double vision (diplopia) can be disfiguring and incapacitating. Graves dermopathy, also known as Pretibial myxedema, thyroid dermopathy, Jadassohn- Dössekker disease or Myxoedema tuberosum, is an infiltrative dermopathy, resulting as a complication of Graves' disease, with an incidence rate of about 1-5% in patients. The disease usually presents itself as a waxy, discolored induration of the skin on the anterior aspect of the lower legs. In certain embodiments, a disease that can be treated o prevented by TSHR antagonists is thyroid cancer.While not being bound by theory, TSHR is expressed in thyroid cancer cells and regulates the growth, proliferation and metastatic potential of thyroid cancer cells. The thyroid gland is, as is well known, one site of metabolic control within the body. Cancer of the thyroid gland is not particularly common, but the high rate of disease re- occurrence necessitates long-term surveillance. Usually, during treatment for cancer of the thyroid, the majority of the thyroid tumor is removed, but a small amount often remains that must be treated by radioactive iodide therapy. Indeed, thyroid cancer is characterized by a high likelihood of relapses in up to 30% of patients, even after successful therapy. In rare cases, the TSHR contains a hereditary mutation that makes it more active than the normal TSHR, resulting in hereditary non-immune hyperthyroidism. TSHR antagonists could be effective treatment for these patients also. A “TSHR antagonist” as described herein blocks or inhibits the action of the agonists (TSH or thyroid-stimulating antibodies for TSHR. Small-molecule ligands for the TSHR (antagonists) typically bind to an intra-membrane domain of the receptor, and act by inducing a conformational change rather than simply competing for TSH binding to its extracellular site on the receptor. In certain embodiments, the antagonists may be selective antagonists for TSHR (i.e, the compounds do not activate or modulate other hormone receptors, particularly luteinizing hormone/chorionic gonadotropin receptor (LHCGR) and follicle-stimulating hormone receptor (FSHR)). In certain embodiments, the antagonists disclosed herein may be used for treating hyperthyroidism in a subject. For example, the antagonists may inhibit mutant TSHRs with higher than normal basal signaling activities (CAMs) that cause an unusual form of hyperthyroidism. In another example, the antagonists may inhibit stimulation by antibodies found in Graves' disease, which is the most common form of hyperthyroidism. In certain embodiments, the TSHR antagonists are useful for treating TSHR-mediated thyroid cancer or hyperthyroidism by blocking TSHR-stimulating antibodies (TSAbs) in Graves' hyperthyroidism. In certain embodiments, the compound is administered orally to the subject. In certain embodiments, the compound is administered parenterally to the subject. In certain embodiments, the disease is prevented. In other embodiments, the disease is treated. Pharmaceutical Compositions, Routes of Administration, and Dosing In certain embodiments, the invention is directed to a pharmaceutical composition, comprising a compound of the invention, e.g. a compound of Formula (1), and a pharmaceutically acceptable carrier. In certain embodiments, the invention is directed to a pharmaceutical composition, comprising a compound of any of the disclosed embodiments, and a pharmaceutically acceptable carrier. In certain embodiments, the invention is directed to a pharmaceutical composition, comprising a compound of Table 1 or 2, and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds of the invention and a pharmaceutically acceptable carrier. In certain embodiments, a pharmaceutical composition of the invention further comprises at least one additional pharmaceutically active agent other than a compound of the invention. The at least one additional pharmaceutically active agent can be an agent useful in the treatment of ischemia-reperfusion injury. Pharmaceutical compositions of the invention can be prepared by combining one or more compounds of the invention with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents. As stated above, an “effective amount” refers to any amount that is sufficient to achieve a desired biological effect. Combined with the teachings provided herein, by choosing among the various active compounds and weighing factors such as potency, relative bioavailability, patient body weight, severity of adverse side-effects and mode of administration, an effective prophylactic or therapeutic treatment regimen can be planned which does not cause substantial unwanted toxicity and yet is effective to treat the particular subject. The effective amount for any particular application can vary depending on such factors as the disease or condition being treated, the particular compound of the invention being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound of the invention and/or other therapeutic agent without necessitating undue experimentation. A maximum dose may be used, that is, the highest safe dose according to some medical judgment. Multiple doses per day may be contemplated to achieve appropriate systemic levels of compounds. Appropriate systemic levels can be determined by, for example, measurement of the patient’s peak or sustained plasma level of the drug. “Dose” and “dosage” are used interchangeably herein. In certain embodiments, intravenous administration of a compound may typically be from 0.1 mg/kg/day to 20 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 0.1 mg/kg/day to 2 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 0.5 mg/kg/day to 5 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 1 mg/kg/day to 20 mg/kg/day. In one embodiment, intravenous administration of a compound may typically be from 1 mg/kg/day to 10 mg/kg/day. Generally, daily oral doses of a compound will be, for human subjects, from about 0.01 milligrams/kg per day to 1000 milligrams/kg per day. It is expected that oral doses in the range of 0.5 to 50 milligrams/kg, in one or more administrations per day, will yield therapeutic results. Dosage may be adjusted appropriately to achieve desired drug levels, local or systemic, depending upon the mode of administration. For example, it is expected that intravenous administration would be from one order to several orders of magnitude lower dose per day. In the event that the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of the compound. For any compound described herein the therapeutically effective amount can be initially determined from animal models. A therapeutically effective dose can also be determined from human data for compounds which have been tested in humans and for compounds which are known to exhibit similar pharmacological activities, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability and potency of the administered compound. Adjusting the dose to achieve maximal efficacy based on the methods described above and other methods as are well- known in the art is well within the capabilities of the ordinarily skilled artisan. The formulations of the invention can be administered in pharmaceutically acceptable solutions, which may routinely contain pharmaceutically acceptable concentrations of salt, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients. For use in therapy, an effective amount of the compound can be administered to a subject by any mode that delivers the compound to the desired surface. Administering a pharmaceutical composition may be accomplished by any means known to the skilled artisan. Routes of administration include but are not limited to intravenous, intramuscular, intraperitoneal, intravesical (urinary bladder), oral, subcutaneous, direct injection (for example, into a tumor or abscess), mucosal (e.g., topical to eye), inhalation, and topical. For intravenous and other parenteral routes of administration, a compound of the invention can be formulated as a lyophilized preparation, as a lyophilized preparation of liposome-intercalated or -encapsulated active compound, as a lipid complex in aqueous suspension, or as a salt complex. Lyophilized formulations are generally reconstituted in suitable aqueous solution, e.g., in sterile water or saline, shortly prior to administration. For oral administration, the compounds can be formulated readily by combining the active compound(s) with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Optionally the oral formulations may also be formulated in saline or buffers, e.g., EDTA for neutralizing internal acid conditions or may be administered without any carriers. Also specifically contemplated are oral dosage forms of the above component or components. The component or components may be chemically modified so that oral delivery of the derivative is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the component molecule itself, where said moiety permits (a) inhibition of acid hydrolysis; and (b) uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the component or components and increase in circulation time in the body. Examples of such moieties include: polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline. Abuchowski and Davis, “Soluble Polymer- Enzyme Adducts”, In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, N.Y., pp.367-383 (1981); Newmark et al., J Appl Biochem 4:185-9 (1982). Other polymers that could be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. For pharmaceutical usage, as indicated above, polyethylene glycol moieties are suitable. For the component (or derivative) the location of release may be the stomach, the small intestine (the duodenum, the jejunum, or the ileum), or the large intestine. One skilled in the art has available formulations which will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine. Preferably, the release will avoid the deleterious effects of the stomach environment, either by protection of the compound of the invention (or derivative) or by release of the biologically active material beyond the stomach environment, such as in the intestine. To ensure full gastric resistance a coating impermeable to at least pH 5.0 is essential. Examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings may be used as mixed films. A coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings which make the tablet easier to swallow. Capsules may consist of a hard shell (such as gelatin) for delivery of dry therapeutic (e.g., powder); for liquid forms, a soft gelatin shell may be used. The shell material of cachets could be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet triturates, moist massing techniques can be used. The therapeutic can be included in the formulation as fine multi-particulates in the form of granules or pellets of particle size about 1 mm. The formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets. The therapeutic could be prepared by compression. Colorants and flavoring agents may all be included. For example, the compound of the invention (or derivative) may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents. One may dilute or increase the volume of the therapeutic with an inert material. These diluents could include carbohydrates, especially mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Certain inorganic salts may also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell. Disintegrants may be included in the formulation of the therapeutic into a solid dosage form. Materials used as disintegrates include but are not limited to starch, including the commercial disintegrant based on starch, Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used. Another form of the disintegrants are the insoluble cationic exchange resins. Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants. Binders may be used to hold the therapeutic agent together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic. An anti-frictional agent may be included in the formulation of the therapeutic to prevent sticking during the formulation process. Lubricants may be used as a layer between the therapeutic and the die wall, and these can include but are not limited to; stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000. Glidants that might improve the flow properties of the drug during formulation and to aid rearrangement during compression might be added. The glidants may include starch, talc, pyrogenic silica and hydrated silicoaluminate. To aid dissolution of the therapeutic into the aqueous environment a surfactant might be added as a wetting agent. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents which can be used and can include benzalkonium chloride and benzethonium chloride. Potential non- ionic detergents that could be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the formulation of the compound of the invention or derivative either alone or as a mixture in different ratios. Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres have been well defined in the art. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner. For topical administration, the compound may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art. Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral or pulmonary administration. For administration by inhalation, compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch. Also contemplated herein is pulmonary delivery of the compounds disclosed herein (or salts thereof). The compound is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl.5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (a-1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins", Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor alpha) and Platz et al., U.S. Pat. No. 5,284,656 (granulocyte colony stimulating factor; incorporated by reference). A method and composition for pulmonary delivery of drugs for systemic effect is described in U.S. Pat. No.5,451,569 (incorporated by reference), issued Sep. 19, 1995 to Wong et al. Contemplated for use in the practice of this invention are mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art. Some specific examples of commercially available devices suitable for the practice of this invention are the Ultravent nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II nebulizer, manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Mass. All such devices require the use of formulations suitable for the dispensing of the compounds of the invention. Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material, in addition to the usual diluents, adjuvants and/or carriers useful in therapy. Also, the use of liposomes, microcapsules or microspheres, inclusion complexes, or other types of carriers is contemplated. Chemically modified compound of the invention may also be prepared in different formulations depending on the type of chemical modification or the type of device employed. Formulations suitable for use with a nebulizer, either jet or ultrasonic, will typically comprise a compound of the invention (or derivative) dissolved in water at a concentration of about 0.1 to 25 mg of biologically active compound of the invention per mL of solution. The formulation may also include a buffer and a simple sugar (e.g., for inhibitor stabilization and regulation of osmotic pressure). The nebulizer formulation may also contain a surfactant, to reduce or prevent surface induced aggregation of the compound of the invention caused by atomization of the solution in forming the aerosol. Formulations for use with a metered-dose inhaler device will generally comprise a finely divided powder containing the compound of the invention (or derivative) suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, and 1,1,1,2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be useful as a surfactant. Formulations for dispensing from a powder inhaler device will comprise a finely divided dry powder containing a compound of the invention (or derivative) and may also include a bulking agent, such as lactose, sorbitol, sucrose, or mannitol in amounts which facilitate dispersal of the powder from the device, e.g., 50 to 90% by weight of the formulation. The compound of the invention (or derivative) should advantageously be prepared in particulate form with an average particle size of less than 10 micrometers (μm), most preferably 0.5 to 5 μm, for most effective delivery to the deep lung. Nasal delivery of a pharmaceutical composition of the present invention is also contemplated. Nasal delivery allows the passage of a pharmaceutical composition of the present invention to the blood stream directly after administering the therapeutic product to the nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with dextran or cyclodextran. For nasal administration, a useful device is a small, hard bottle to which a metered dose sprayer is attached. In one embodiment, the metered dose is delivered by drawing the pharmaceutical composition of the present invention solution into a chamber of defined volume, which chamber has an aperture dimensioned to aerosolize and aerosol formulation by forming a spray when a liquid in the chamber is compressed. The chamber is compressed to administer the pharmaceutical composition of the present invention. In a specific embodiment, the chamber is a piston arrangement. Such devices are commercially available. Alternatively, a plastic squeeze bottle with an aperture or opening dimensioned to aerosolize an aerosol formulation by forming a spray when squeezed is used. The opening is usually found in the top of the bottle, and the top is generally tapered to partially fit in the nasal passages for efficient administration of the aerosol formulation. Preferably, the nasal inhaler will provide a metered amount of the aerosol formulation, for administration of a measured dose of the drug. The compounds, when it is desirable to deliver them systemically, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi- dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active compounds may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described above, a compound may also be formulated as a depot preparation. Such long acting formulations may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous or saline solutions for inhalation, microencapsulated, encochleated, coated onto microscopic gold particles, contained in liposomes, nebulized, aerosols, pellets for implantation into the skin, or dried onto a sharp object to be scratched into the skin. The pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops or preparations with protracted release of active compounds, in whose preparation excipients and additives and/or auxiliaries such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners or solubilizers are customarily used as described above. The pharmaceutical compositions are suitable for use in a variety of drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249:1527-33 (1990). The compound of the invention and optionally other therapeutics may be administered per se (neat) or in the form of a pharmaceutically acceptable salt or cocrystal. When used in medicine the salts or cocrystals should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts or cocrystals may conveniently be used to prepare pharmaceutically acceptable salts or cocrystals thereof. Such salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulphuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluene sulphonic, tartaric, citric, methane sulphonic, formic, malonic, succinic, naphthalene-2-sulphonic, and benzene sulphonic. Also, such salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts of the carboxylic acid group. Suitable buffering agents include: acetic acid and a salt (1-2% w/v); citric acid and a salt (1-3% w/v); boric acid and a salt (0.5-2.5% w/v); and phosphoric acid and a salt (0.8-2% w/v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w/v); chlorobutanol (0.3- 0.9% w/v); parabens (0.01-0.25% w/v) and thimerosal (0.004-0.02% w/v). Pharmaceutical compositions of the invention contain an effective amount of a compound as described herein and optionally therapeutic agents included in a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” means one or more compatible solid or liquid filler, diluents or encapsulating substances which are suitable for administration to a human or other vertebrate animal. The term “carrier” denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application. The components of the pharmaceutical compositions also are capable of being commingled with the compounds of the present invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficiency. The therapeutic agent(s), including specifically but not limited to a compound of the invention, may be provided in particles. Particles as used herein means nanoparticles or microparticles (or in some instances larger particles) which can consist in whole or in part of the compound of the invention or the other therapeutic agent(s) as described herein. The particles may contain the therapeutic agent(s) in a core surrounded by a coating, including, but not limited to, an enteric coating. The therapeutic agent(s) also may be dispersed throughout the particles. The therapeutic agent(s) also may be adsorbed into the particles. The particles may be of any order release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof, etc. The particle may include, in addition to the therapeutic agent(s), any of those materials routinely used in the art of pharmacy and medicine, including, but not limited to, erodible, nonerodible, biodegradable, or nonbiodegradable material or combinations thereof. The particles may be microcapsules which contain the compound of the invention in a solution or in a semi-solid state. The particles may be of virtually any shape. Both non-biodegradable and biodegradable polymeric materials can be used in the manufacture of particles for delivering the therapeutic agent(s). Such polymers may be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired. Bioadhesive polymers of particular interest include bioerodible hydrogels described in Sawhney H S et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein. These include polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate). The therapeutic agent(s) may be contained in controlled release systems. The term “controlled release” is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation are controlled. This refers to immediate as well as non-immediate release formulations, with non-immediate release formulations including but not limited to sustained release and delayed release formulations. The term “sustained release” (also referred to as “extended release”) is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that preferably, although not necessarily, results in substantially constant blood levels of a drug over an extended time period. The term “delayed release” is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug there from. “Delayed release” may or may not involve gradual release of drug over an extended period of time, and thus may or may not be “sustained release.” Use of a long-term sustained release implant may be particularly suitable for treatment of chronic conditions. “Long-term” release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30-60 days. Long-term sustained release implants are well-known to those of ordinary skill in the art and include some of the release systems described above. It will be understood by one of ordinary skill in the relevant arts that other suitable modifications and adaptations to the compositions and methods described herein are readily apparent from the description of the invention contained herein in view of information known to the ordinarily skilled artisan, and may be made without departing from the scope of the invention or any embodiment thereof. Having now described the present invention in detail, the same will be more clearly understood by reference to the following examples, which are included herewith for purposes of illustration only and are not intended to be limiting of the invention. EXAMPLES The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. Example 1: 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000110_0002
1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (1 g, 5.204 mmol, 1 equiv) in DMF (20 mL) was added K2CO3 (2.154 g, 15.611 mmol, 3 equiv) and (bromomethyl)benzene (979 mg, 5.724 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (500 mg, 34.04%) as a white solid.
Figure imgf000110_0001
6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (0.45 g, 1.590 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (888.0 mg, 15.90 mmol, 10 equiv) and NH4Cl (850.5 mg, 15.90 mmol, 10 equiv).The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6- amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one (350 mg, 87.28%) as a yellow solid. MS (ESI): mass calcd. for C16H16N2O: 252.13 m/z, found 253.10 [M+H]+.
Figure imgf000111_0001
1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one (150 mg, 0.594 mmol, 1 equiv) in DCM (8 mL) was added TEA (179.9 mg, 1.782 mmol, 3 equiv) and 2-isocyanato-2- methylpropane (176.65 mg, 1.782 mmol, 3 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 53% B in 9 min, 53% B; Wave Length: 254/220 nm; RT1(min): 8.52; Number Of Runs: 0) to afford 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)-3-(tert-butyl)urea (60.2 mg, 28.94%) as an off-white solid. MS (ESI): mass calcd. For C21H25N3O2, 351.19 m/z, found 352.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.08 (s, 1H), 7.26 – 7.34 (m, 3H), 7.16 – 7.25 (m, 3H), 6.95 (dd, J = 8.8, 2.5 Hz, 1H), 6.76 (d, J = 8.8 Hz, 1H), 5.90 (s, 1H), 5.09 (s, 2H), 2.87 (dd, J = 8.7, 5.9 Hz, 2H), 2.64 (dd, J = 8.7, 5.9 Hz, 2H), 1.26 (s, 9H). Example 2: 1-(tert-butyl)-3-(2-oxo-1-(pyridin-2-ylmethyl)-1,2,3,4-tetrahydroquinolin-6- yl)urea
Figure imgf000112_0001
6-nitro-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1078.7 mg, 7.805 mmol, 3 equiv) and 2- (bromomethyl)pyridine (492.3 mg, 2.862 mmol, 1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-nitro-1-(pyridin-2-ylmethyl)-3,4- dihydroquinolin-2(1H)-one (500 mg, 63.20%) as a white solid. MS (ESI): mass calcd. for C15H13N3O3283.10 m/z, found 284.05 [M+H]+.
Figure imgf000112_0002
6-amino-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2-one (440 mg, 1.553 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (867.3 mg, 15.532 mmol, 10 equiv) and NH4Cl (830.8 mg, 15.530 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-(pyridin-2-ylmethyl)-3,4- dihydroquinolin-2-one (200 mg, 46.52%) as a yellow solid. MS (ESI): mass calcd. for C15H15N3O, 253.12 m/z, found 254.15 [M+H]+.
Figure imgf000113_0001
1-(tert-butyl)-3-(2-oxo-1-(pyridin-2-ylmethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-(pyridin-2-ylmethyl)-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.793 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (234.8 mg, 2.369 mmol, 3 equiv) and TEA (79.9 mg, 0.790 mmol, 1 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 22% B to 34% B in 10.5 min; Wave Length: 254/220 nm; RT1(min): 11.3; Number Of Runs: 4) to afford 1-(tert-butyl)-3-(2-oxo-1-(pyridin-2- ylmethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea (83.1 mg, 29.82%) as a white solid. MS (ESI): mass calcd. for C20H24N4O2, 352.19 m/z, found 353.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.52 (d, J = 4.8 Hz, 1H), 8.11 (d, J = 3.5 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H), 7.34 (s, 1H), 7.25 (q, J = 5.1 Hz, 1H), 7.21 – 7.21 (m, 1H), 6.97 (d, J = 8.8 Hz, 1H), 6.69 – 6.78 (m, 1H), 5.92 (s, 1H), 5.13 (s, 2H), 2.89 (d, J = 8.0 Hz, 2H), 2.64 (d, J = 8.4 Hz, 2H), 1.26 (s, 9H). Example 3: 1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000113_0002
6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1078.7 mg, 7.805 mmol, 3 equiv) and (1-bromoethyl)benzene (529.6 mg, 2.862 mmol, 1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (400 mg, 45.17%) as a white solid.MS (ESI): mass calcd. for C17H16N2O3296.12 m/z, found 297.05 [M+H]+.
Figure imgf000114_0001
6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (400 mg, 1.350 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (753.8 mg, 13.499 mmol, 10 equiv) and NH4Cl (722.0 mg, 13.499 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2- one (200 mg, 55.63%) as a white solid.
Figure imgf000114_0002
1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (200 mg, 0.751 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (223.3 mg, 2.253 mmol, 3 equiv) and TEA (75.9 mg, 0.751 mmol, 1 equiv).The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 38% B to 50% B in 10.5 min; Wave Length: 254/220 nm; RT1(min): 10.7; Number Of Runs: 3) to afford 1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4- tetrahydroquinolin-6-yl)urea (57.7 mg, 21.02%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2, 365.21 m/z, found 366.15 [M+H]+ .1H NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.18 – 7.39 (m, 6H), 6.81 (d, J = 8.8 Hz, 1H), 6.42 – 6.52 (m, 1H), 6.15 (d, J = 8.0 Hz, 1H), 5.91 (s, 1H), 2.80 (d, J = 5.6 Hz, 2H), 2.54 – 2.70 (m, 2H), 1.67 (t, J = 5.7 Hz, 3H), 1.24 (dd, J = 4.1, 2.2 Hz, 9H). Example 4: 1-(tert-butyl)-3-(1-((1-methyl-1H-pyrazol-3-yl)methyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea
Figure imgf000115_0001
1-[(1-methylpyrazol-3-yl)methyl]-6-nitro-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1086.6 mg, 7.805 mmol, 3 equiv) and 3-(bromomethyl)-1- methylpyrazole (500.9 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-[(1-methylpyrazol-3-yl)methyl]-6-nitro- 3,4-dihydroquinolin-2-one (500 mg, 46.59%) as a white solid. MS (ESI): mass calcd. for C14H14N4O3: 286.11 m/z, found 287.00 [M+H]+.
Figure imgf000115_0002
6-amino-1-[(1-methylpyrazol-3-yl)methyl]-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(1H-pyrazol-3-ylmethyl)-3,4-dihydroquinolin-2-one (300 mg, 1.102 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (615.3 mg, 11.019 mmol, 10 equiv) and NH4Cl (589.3 mg, 11.019 mmol, 10 equiv).The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-[(1-methylpyrazol-3-yl)methyl]-3,4- dihydroquinolin-2-one (101 mg, 36.03%) as a yellow solid.
Figure imgf000116_0001
1-(tert-butyl)-3-(1-((1-methyl-1H-pyrazol-3-yl)methyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea To a solution of 6-amino-1-[(1-methylpyrazol-3-yl)methyl]-3,4-dihydroquinolin-2-one (50 mg, 0.195 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (58.0 mg, 0.585 mmol, 3 equiv) and TEA (19.7 mg, 0.195 mmol, 1 equiv).The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-((1-methyl-1H- pyrazol-3-yl)methyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (9.0 mg, 12.23%) as a white solid. MS (ESI): mass calcd. for C19H25N5O2, 355.20 m/z, found 356.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.10 (d, J = 5.2 Hz, 1H), 7.48 – 7.62 (m, 1H), 7.29 (s, 1H), 7.01 (s, 2H), 5.85 – 6.04 (m, 2H), 4.95 (s, 2H), 3.78 – 3.82 (m, 3H), 2.81 (s, 2H), 2.51 (s, 2H), 1.17 – 1.37 (m, 9H). Example 5: 1-(1-benzyl-3-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000117_0001
1-benzyl-3-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (1 g, 3.54 mmol, 1 equiv) in THF (70 mL) was added 2M LiHMDS (in THF) (2.3 mL, 4.60 mmol, 1.3 equiv) at -20 °C under N2. The resulting mixture was stirred 30 min at -20 °C. The reaction mixture was added MeI (653.6 mg, 4.60 mmol, 1.3 equiv) at -20 °C. The resulting mixture was stirred 30 min at -20 °C and then stirred 16 h at rt. The reaction mixture was quenched by water (50 mL) and extracted with EA (3 x 70 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 1-benzyl-3-methyl- 6-nitro-3,4-dihydroquinolin-2(1H)-one (0.3 g, 28.59%) as a white solid.
Figure imgf000118_0001
6-amino-1-benzyl-3-methyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-3-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (0.28 g, 0.944 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (527.7 mg, 9.44 mmol, 10 equiv) and NH4Cl (504.9 mg, 9.76 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 3-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (160 mg, 46.37%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O: 266.14 m/z, found 267.20 [M+H]+.
Figure imgf000118_0002
1-(tert-butyl)-3-(1-(3-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-benzyl-3-methyl-3,4-dihydroquinolin-2(1H)-one (150 mg, 0.563 mmol, 1 equiv) in CH3CN (5 mL) was added 2-isocyanato-2-methylpropane (167.48 mg, 1.689 mmol, 3 equiv) and KHCO3 (112.7 mg, 1.126 mmol, 2 equiv). The resulting mixture was stirred for overnight at 70 °C. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(3- cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (18.1 mg, 8.78%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2: 365.21 m/z, found 366.10 [M+H]+.1H NMR (400 MHz, Chloroform-d) δ 7.26 – 7.31 (m, 2H), 7.12 – 7.23 (m, 3H), 6.82 (d, J = 8.7 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 6.25 – 6.54 (m, 1H), 5.19 (d, J = 16.3 Hz, 1H), 5.05 (d, J = 16.0 Hz, 1H), 2.94 (d, J = 11.6 Hz, 1H), 2.58 – 2.81 (m, 2H), 1.33 (s, 9H), 1.30 (d, J = 6.2 Hz, 3H). Example 6: 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000119_0001
1-(3-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in dimethylformamide (10 mL) was added 1-(bromomethyl)-3-chlorobenzene (801.93 mg, 3.903 mmol, 1.5 equiv) and potassium methaneperoxoate potassium (724.41 mg, 5.204 mmol, 2.0 equiv) at rt. The mixture was stirred for overnight at rt. The reaction mixture was quenched by water (30 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine, dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 1-(3- chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (700 mg, 88.29%) as a white solid. MS (ESI): mass calcd. C16H13ClN2O3: for 316.06 m/z, found 317.10[M+H]+.
Figure imgf000119_0002
6-amino-1-(3-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(3-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (700 mg, 2.210 mmol, 1 equiv) in EtOH (5 mL)/H2O (0.5 mL) was added Fe (1.2 g, 22.1 mmol, 10 equiv) and NH4Cl (1.2 g, 22.100 mmol, 10 equiv) at rt. The mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water (30 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine, dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with PE/EA (2:1) to afford 6-amino-1-(3-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one (600 mg, 94.68%) as a brown solid. MS (ESI): mass calcd. C16H15ClN2O: for 286.09 m/z, found 287.10[M+H]+.
Figure imgf000120_0001
1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea A solution of 6-amino-1-(3-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one (50 mg, 0.174 mmol, 1 equiv) in DCM (3 mL) was added TEA (35.29 mg, 0.348 mmol, 2.0 equiv) and 2- isocyanato-2-methylpropane (20.74 mg, 0.209 mmol, 1.2 equiv) at 0 °C. The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water (20 mL). The resulting mixture was extracted with DCM (3 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (2:1) to afford 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (30.4 mg, 45.11%) as white solid. MS (ESI): mass calcd. C21H24ClN3O2 for : 385.16 m/z, found 386.1 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.24 – 7.40 (m, 4H), 7.15 – 7.18 (m, 1H), 6.96 – 7.01 (m, 1H), 6.75 – 6.80 (m, 1H), 5.92 (s, 1H), 5.10 (s, 2H), 2.85 – 2.92 (m, 2H), 2.61 – 2.70 (m, 2H), 1.27 (s, 9H). Example 7: 1-(4-benzyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea
Figure imgf000121_0001
4-benzyl-7-nitro-2H-1,4-benzoxazin-3-one To a solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (400 mg, 2.060 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (860.4 mg, 6.181 mmol, 3 equiv) and benzyl bromide (387.6 mg, 2.266 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 4-benzyl-7-nitro-2H-1,4-benzoxazin-3-one (500 mg, 72.97%) as a yellow solid. MS (ESI): mass calcd. for C15H12N2O4284.08 m/z, found 285.05 [M+H]+.
Figure imgf000121_0002
7-amino-4-benzyl-2H-1,4-benzoxazin-3-one To a solution of 4-benzyl-7-nitro-2H-1,4-benzoxazin-3-one (500 mg, 1.759 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (982.2 mg, 17.589 mmol, 10 equiv) and NH4Cl (940.8 mg, 17.589 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 7-amino-4-benzyl-2H-1,4-benzoxazin-3-one (200 mg, 44.72%) as a yellow solid. MS (ESI): mass calcd. for C15H14N2O2, 254.11 m/z, found 255.05 [M+H]+.
Figure imgf000122_0001
1-(4-benzyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea To a solution of 7-amino-4-benzyl-2H-1,4-benzoxazin-3-one (150 mg, 0.590 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (175.4 mg, 1.770 mmol, 3 equiv) and TEA (59.6 mg, 0.590 mmol, 1 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(4-benzyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (56.8 mg, 27.14%) as a white solid. MS (ESI): mass calcd. for C20H23N3O3, 353.17 m/z, found 354.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.21 (s, 1H), 7.17 – 7.41 (m, 7H), 6.86 (dd, J = 8.6, 2.1 Hz, 1H), 6.71 – 6.82 (m, 1H), 5.93 (s, 1H), 5.11 (s, 2H), 4.74 (d, J = 2.1 Hz, 2H), 1.26 (d, J = 2.1 Hz, 9H). Example 8: 1-(tert-butyl)-3-(1-(2-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000122_0002
1-(2-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1086.6 mg, 7.805 mmol, 3 equiv) and 1-(bromomethyl)-2- chlorobenzene (583.64 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-(2-chlorobenzyl)-6-nitro-3,4- dihydroquinolin-2(1H)-one (400 mg, 48.54%) as a white solid. MS (ESI): mass calcd. for C16H13ClN2O3: 316.06 m/z, found 317.00 [M+H]+.
Figure imgf000123_0001
6-amino-1-(2-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(2-chlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (400 mg, 1.263 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (705.3 mg, 12.63 mmol, 10 equiv) and NH4Cl (675.6 mg, 12.63 mmol, 10 equiv).The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-(2-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one (0.3 g, 82.87%) as a yellow solid. MS (ESI): mass calcd. for C16H15ClN2O: 286.09 m/z, found 287.00 [M+H]+.
Figure imgf000123_0002
1-(tert-butyl)-3-(1-(2-chlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-(2-chlorobenzyl)-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.349 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (103.9 mg, 1.048 mmol, 3 equiv) and TEA (105.8 mg, 1.048 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(2-chlorobenzyl)- 2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (39.7 mg, 29.40%) as a white solid. MS (ESI): mass calcd. for C21H24ClN3O2: 385.16 m/z, found 386.05 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.47 – 7.56 (m, 1H), 7.35 – 7.42 (m, 1H), 7.26 (d, J = 8.7 Hz, 2H), 6.92 – 7.01 (m, 2H), 6.47 – 6.62 (m, 1H), 5.91 (s, 1H), 5.07 (s, 2H), 2.88 – 2.96 (m, 2H), 2.63 – 2.72 (m, 2H), 1.25 (s, 9H). Example 9: tert-butyl (1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)carbamate Synthetic Scheme
Figure imgf000124_0001
tert-butyl (1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)carbamate To a solution of 6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.396 mmol, 1 equiv) in DCM (6 mL) was added Boc2O (129.7 mg, 0.594 mmol, 1.5 equiv) and DMAP (72.6 mg, 0.594 mmol, 1.5 equiv). The resulting mixture was stirred for 5 h at rt. The mixture was diuted with DCM (30 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 45% B in 9 min, 45% B; Wave Length: 254/220 nm; RT1(min): 8.6; Number Of Runs: 0) to afford tert-butyl (1- benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)carbamate (19.4 mg, 10.21%) as a yellow solid. MS (ESI): mass calcd. for C21H24N2O3: 352.18 m/z, found 353.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 9.21 (s, 1H), 7.39 – 7.16 (m, 6H), 7.10 (dd, J = 9.0, 2.4 Hz, 1H), 6.82 (d, J = 8.8 Hz, 1H), 5.11 (s, 2H), 2.88 (t, J = 7.2 Hz, 2H), 2.66 (dd, J = 8.7, 6.0 Hz, 2H), 1.45 (s, 9H). Example 10: 1-(1-benzyl-4,4-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000125_0001
4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one (0.4 g, 2.28 mmol, 1 equiv) in H2O (2 mL) and H2SO4 (9 mL) was added HNO3 (0.3 mL) at -10 °C. The reaction was stirred for 2 h at -10-0 °C. The reaction mixture was quenched by water. The mixture was acidified pH = 7 with NaHCO3 (aq.) and then extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (0.3 g, 59.76%) as a white solid. MS (ESI): mass calcd. for C11H12N2O3, 220.08 m/z, found 221.10 [M+H]+ .
Figure imgf000126_0001
1-benzyl-4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (280 mg, 1.270 mmol, 1 equiv) in CH3CN (10 mL) was added Cs2CO3 (828.3 mg, 2.542 mmol, 2.0 equiv) and (bromomethyl)benzene (238.9 mg, 1.397 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-benzyl-4,4-dimethyl-6-nitro-3,4- dihydroquinolin-2(1H)-one (300 mg, 76.14%) as a white solid. MS (ESI): mass calcd. for C18H18N2O3: 310.13 m/z, found 311.25 [M+H]+.
Figure imgf000126_0002
6-amino-1-benzyl-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-4,4-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (280 mg, 0.902 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (503.9 mg, 9.02 mmol, 10 equiv) and NH4Cl (482.5 mg, 9.02 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-benzyl-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 39.68%) as a yellow solid. MS (ESI): mass calcd. for C18H20N2O: 280.16 m/z, found 281.20 [M+H]+.
Figure imgf000127_0001
1-(1-benzyl-4,4-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.357 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (106.1 mg, 1.07 mmol, 3 equiv) and TEA (108.1 mg, 1.07 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep C18 OBD Column, 30*50 mm, 5μm 13nm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 45% B in 9 min, 45% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(1-benzyl-4,4-dimethyl-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (51.0 mg, 37.50%) as a white solid. MS (ESI): mass calcd. for C23H29N3O2: 379.23 m/z, found 380.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.16 (s, 1H),7.38 (s, 1H),7.20 - 7.31 (m, 5H),7.02 (s, 1H), 6.86 (s, 1H), 5.88 (s, 1H), 5.13 (s, 2H), 2.50 - 2.55 (s, 2H), 1.26 (s, 9H), 1.22 (s, 6H). Example 11: 1-(1-benzyl-3-(methoxymethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000128_0001
1-benzyl-3-(methoxymethyl)-6-nitro-3,4-dihydroquinolin-2-one To a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2-one (1.3 g, 4.605 mmol, 1 equiv) in THF (40 mL) was added 2M LDA (in THF) (3.5 mL, 0.74 g, 6.907 mmol, 1.5 equiv) at -78 °C under N2. The resulting mixture was stirred 30 min at -78 °C. The reaction mixture was added bromo(methoxy)methane (1.15 g, 9.203 mmol, 2.00 equiv) at -78 °C. The resulting mixture was stirred for 1 h at -78 °C and 16 h at rt. The reaction mixture was quenched by NH4Cl (aq.) and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-benzyl-3- (methoxymethyl)-6-nitro-3,4-dihydroquinolin-2-one (400 mg, 12.30%) as a yellow solid. MS (ESI): mass calcd. for C18H18N2O4, 326.13 m/z, found 327.05 [M+H]+.
Figure imgf000129_0001
6-amino-1-benzyl-3-(methoxymethyl)-3,4-dihydroquinolin-2-one To a solution of 1-benzyl-3-(methoxymethyl)-6-nitro-3,4-dihydroquinolin-2-one (300 mg, 0.919 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (513.3 mg, 9.193 mmol, 10 equiv) and NH4Cl (491.7 mg, 9.193 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-benzyl-3-(methoxymethyl)-3,4- dihydroquinolin-2-one (200 mg, 26.38%) as a yellow solid. MS (ESI): mass calcd. for C18H20N2O2, 296.15 m/z, found 297.10 [M+H]+.
Figure imgf000129_0002
1-(1-benzyl-3-(methoxymethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-3-(methoxymethyl)-3,4-dihydroquinolin-2-one (100 mg, 0.337 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (100.3 mg, 1.012 mmol, 3 equiv) and TEA (102.4 mg, 1.012 mmol, 3 equiv).The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(1-benzyl-3-(methoxymethyl)-2- oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (12.1 mg, 8.95%) as a white solid. MS (ESI): mass calcd. for C23H29N3O3, 395.22 m/z, found 396.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.19 (d, J = 5.5 Hz, 3H), 7.04 – 7.17 (m, 3H), 6.90 (d, J = 8.7 Hz, 1H), 6.66 (d, J = 9.1 Hz, 1H), 5.83 (s, 1H), 4.99 (s, 2H), 3.53 – 3.64 (m, 1H), 3.46 (t, J = 8.2 Hz, 1H), 3.20 (d, J = 1.7 Hz, 3H), 2.60 – 2.95 (m, 3H), 1.16 (d, J = 1.7 Hz, 9H). Example 12: 1-(1-benzyl-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert-butyl)ur ea Synthetic Scheme
Figure imgf000130_0001
2-(benzylamino)-5-nitrobenzonitrile To a solution of 2-fluoro-5-nitrobenzonitrile (5 g, 30.100 mmol, 1 equiv) in ACN (100 mL) was added benzylamine (4.84 g, 45.150 mmol, 1.5 equiv) and K2CO3 (8.32 g, 60.200 mmol, 2 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 2-(benzylamino)-5-nitrobenzonitrile (6.875 g, 90.18%) as a yellow solid. MS(ESI): mass calcd. for C14H11N3O2: 253.26 m/z, found 254.00[M+H]+.
Figure imgf000131_0001
2-(aminomethyl)-N-benzyl-4-nitroaniline To a solution of 2-(benzylamino)-5-nitrobenzonitrile (400 mg, 1.579 mmol, 1 equiv) in THF (20 mL) was added NaBH4 (238.99 mg, 6.316 mmol, 4 equiv) and Boron trifluoride diethyl etherate (896.66 mg, 6.316 mmol, 4 equiv). The reaction was stirred at 0 °C for 0.5h. The reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with DCM/MeOH (0-30%) to give 2-(aminomethyl)-N-benzyl-4-nitroaniline (41 mg, 10.09%) as a yellow solid. MS (ESI): mass calcd for C14H15N3O2: 257.29 m/z, found 258.10 [M+H]+.
Figure imgf000131_0002
1-benzyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a solution of 2-(aminomethyl)-N-benzyl-4-nitroaniline (260 mg, 1.011 mmol, 1 equiv) in THF (20 mL) was added CDI (163.86 mg, 1.011 mmol, 1 equiv) at 0 °C. The reaction was stirred at rt for 15 min. To the above mixture was added CDI (327.71 mg, 2.022 mmol, 2 equiv) dropwise at 0 °C. The resulting mixture was stirred for additional 15 min at rt. After the final reaction mixture was stirred for 18 h at 70 °C. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-6-nitro-3,4-dihydroquinazolin-2-one (198 mg, 69.17%) as a white solid. MS (ESI): mass calcd. for C15H13N3O3: 283.29 m/z, found 284.05 [M+H]+.
Figure imgf000132_0001
1-benzyl-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a solution of 1-benzyl-6-nitro-3,4-dihydroquinazolin-2-one (184 mg, 0.650 mmol, 1 equiv) in THF (20 mL) was added NaH (23.38 mg, 0.975 mmol, 1.5 equiv) at 0°C. The reaction was stirred at 0°C for 30 min. To the above mixture was added MeI (184.38 mg, 1.300 mmol, 2 equiv) dropwise at 0 °C. The final reaction mixture was stirred for 1h at rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-3-methyl-6-nitro-4H- quinazolin-2-one (122 mg, 63.18%) as a white solid. MS (ESI): mass calcd. for C16H15N3O3: 297.31 m/z, found 298.00 [M+H]+.
Figure imgf000132_0002
6-amino-1-benzyl-3-methyl-3,4-dihydroquinazolin-2(1H)-one To a solution of 1-benzyl-3-methyl-6-nitro-4H-quinazolin-2-one (122 mg, 0.410 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (10%, 122 mg) at rt. The mixture was hydrogenated at room temperature under 30 psi of hydrogen pressure for 1h. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 10 mL). The filtrate was concentrated under reduced pressure. This resulted in 6-amino-1-benzyl-3-methyl-4H-quinazolin-2-one (110 mg, 100.28%) as a white solid. MS (ESI): mass calcd. for C16H17N3O: 267.33 m/z, found 268.20 [M+H]+.
Figure imgf000133_0001
1-(1-benzyl-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-3-methyl-4H-quinazolin-2-one (100 mg, 0.374 mmol, 1 equiv) in DCM (10 mL, 157.306 mmol, 420.53 equiv) was added 2-isocyanato-2- methylpropane (370.82 mg, 3.740 mmol, 10 equiv) and Pyridine (295.89 mg, 3.740 mmol, 10 equiv). The reaction was stirred at rt for 3 days. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18, 30*150 mm, 5μm column (eluent: 36% to 56% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 1-(1-benzyl-3-methyl-2-oxo-4H-quinazolin- 6-yl)-3-tert-butylurea (23.2 mg, 16.92%) as a white solid. MS (ESI): mass calcd. for C21H26N4O2:366.47 m/z, found:367.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.90 – 8.21 (m, 1H), 7.18 – 7.39 (m, 6H), 6.80 – 7.00 (m, 1H), 6.44 – 6.68 (m, 1H), 5.72 – 5.99 (m, 1H), 4.90 – 5.20 (m, 2H), 4.28 – 4.51 (m, 2H), 2.80 – 3.01 (m, 3H), 1.20 – 1.31 (m, 9H). Example 13: (S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)u rea
Figure imgf000133_0002
Synthetic Scheme
Figure imgf000134_0001
(S)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (R)-1-phenylethanol (1 g, 8.186 mmol, 1 equiv) in DCM (30 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (1.57 g, 8.186 mmol, 1 equiv) and PPh3 (2.15 g, 8.186 mmol, 1 equiv). DEAD (1.43 g, 8.211 mmol, 1 equiv) was added the mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford (S)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (300 mg, 11.61%) as a yellow solid. MS (ESI): mass calcd. for C17H16N2O3, 296.12 m/z, found 297.15 [M+H]+.
Figure imgf000134_0002
(S)-6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (S)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (280 mg, 0.945 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford (S)-6-amino-1-(1- phenylethyl)-3,4-dihydroquinolin-2(1H)-one (220 mg, 86.81%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O, 266.14 m/z, found 267.15 [M+H]+.
Figure imgf000135_0001
(S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of (S)-6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.751 mmol, 1 equiv) in DCM (10.00 mL) was added 2-isocyanato-2-methylpropane (372.2 mg, 3.755 mmol, 5 equiv) and TEA (227.9 mg, 2.253 mmol, 3 equiv) stirred overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)- 1,2,3,4-tetrahydroquinolin-6-yl)urea (73.9 mg, 26.88%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2, 365.21 m/z, found 366.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.18 – 7.39 (m, 6H), 6.81 (d, J = 8.8 Hz, 1H), 6.42 – 6.52 (m, 1H), 6.15 (d, J = 8.0 Hz, 1H), 5.91 (s, 1H), 2.80 (d, J = 5.6 Hz, 2H), 2.54 – 2.70 (m, 2H), 1.67 (t, J = 5.7 Hz, 3H), 1.24 (dd, J = 4.1, 2.2 Hz, 9H). Example 14: (R)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)u
Figure imgf000135_0002
(R)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (S)-1-phenylethanol (1 g, 8.186 mmol, 1 equiv) in DCM (30 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (1.57 g, 8.186 mmol, 1 equiv) and PPh3 (2.15 g, 8.186 mmol, 1 equiv). DEAD (1.43 g, 8.211 mmol, 1 equiv) was added the mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford (R)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (300 mg, 11.61%) as a yellow solid. MS (ESI): mass calcd. for C17H16N2O3, 296.12 m/z, found 297.15 [M+H]+.
Figure imgf000136_0001
(R)-6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (R)-6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (280 mg, 0.945 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford (R)-6-amino-1-(1- phenylethyl)-3,4-dihydroquinolin-2(1H)-one (220 mg, 86.81%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O, 266.14 m/z, found 267.15 [M+H]+.
Figure imgf000136_0002
(R)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of (R)-6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.751 mmol, 1 equiv) in DCM (10.00 mL) was added 2-isocyanato-2-methylpropane (372.2 mg, 3.755 mmol, 5 equiv) and TEA (227.9 mg, 2.253 mmol, 3 equiv) stirred overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 3-tert-butyl-1-{2-oxo-1-[(1R)-1-phenylethyl]-3,4- dihydroquinolin-6-yl}urea (70 mg, 25.46%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2, 365.21 m/z, found 366.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.18 – 7.39 (m, 6H), 6.81 (d, J = 8.8 Hz, 1H), 6.42 – 6.52 (m, 1H), 6.15 (d, J = 8.0 Hz, 1H), 5.91 (s, 1H), 2.80 (d, J = 5.6 Hz, 2H), 2.54 – 2.70 (m, 2H), 1.67 (t, J = 5.7 Hz, 3H), 1.24 (dd, J = 4.1, 2.2 Hz, 9H). Example 15: 1-(1-benzyl-4-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea
Figure imgf000137_0001
4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 4-methyl-3,4-dihydroquinolin-2(1H)-one (0.4 g, 2.48 mmol, 1 equiv) in H2O (2 mL) and H2SO4 (9 mL) was added HNO3 (0.3 mL) at -10 °C. The reaction was stirred for 2 h at -10-0 °C. The reaction mixture was quenched by water. The mixture was acidified pH = 7 with NaHCO3 (aq.) and then extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (0.28 g, 54.79%) as a white solid. MS (ESI): mass calcd. for C10H10N2O3, 206.07 m/z, found 207.05 [M+H]+ .
Figure imgf000137_0002
1-benzyl-4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (260 mg, 1.260 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (522.0 mg, 3.782 mmol, 3 equiv) and (bromomethyl)benzene (237.1 mg, 1.386 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-benzyl-4-methyl-6-nitro-3,4- dihydroquinolin-2(1H)-one (200 mg, 53.62%) as a white solid. MS (ESI): mass calcd. for C17H16N2O3: 296.12 m/z, found 297.05 [M+H]+.
Figure imgf000138_0001
6-amino-1-benzyl-4-methyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-4-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (180 mg, 0.607 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (339.3 mg, 6.07 mmol, 10 equiv) and NH4Cl (324.7 mg, 6.07 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-benzyl-4-methyl-3,4-dihydroquinolin-2(1H)-one (120 mg, 74.07%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O: 266.14 m/z, found 267.15 [M+H]+.
Figure imgf000138_0002
1-(1-benzyl-4-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-4-methyl-3,4-dihydroquinolin-2(1H)-one (120 mg, 0.451 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (133.9 mg, 1.351 mmol, 3 equiv) and TEA (136.4 mg, 1.351 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(1-benzyl-4-methyl-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (47.9 mg, 44.35%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2: 365.21 m/z, found 366.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.21 – 7.32 (m, 6H), 6.99 (dd, J = 8.8, 2.4 Hz, 1H), 6.82 (d, J = 8.8 Hz, 1H), 5.90 (s, 1H), 5.05 – 5.17 (m, 2H), 2.90 – 3.13 (m, 1H), 2.68 – 2.86 (m, 1H), 2.33 – 2.50 (m, 1H), 1.26 (s, 9H), 1.18 (d, J = 6.9 Hz, 3H). Example 16: 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(4-hydroxy-2- methylbutan-2-yl)urea Synthetic Scheme
Figure imgf000139_0001
6-amino-1-benzyl-3,4-dihydroquinolin-2(1H)-one A solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2-one (400 mg, 1.417 mmol, 1 equiv) in EtOH (10 mL) and H2O (2 mL) was treated with Fe (791.29 mg, 14.170 mmol, 10 equiv) and NH4Cl (757.92 mg, 14.170 mmol, 10 equiv) for 2 h at 60 °C.The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM/MeOH (5%-11%) to afford 6-amino-1-benzyl-3,4- dihydroquinolin-2-one (320 mg, 89.51%) as a white solid. MS (ESI): mass calcd. for C16H16N2O, 252.13m/z, found 253.25 [M+H]+.
Figure imgf000140_0001
1-benzyl-6-isocyanato-3,4-dihydroquinolin-2(1H)-one To a stirred solution of 6-amino-1-benzyl-3,4-dihydroquinolin-2-one (180 mg, 0.713 mmol, 1 equiv) in anhydrous DCM (5 mL) was added TEA (216.57 mg, 2.139 mmol, 3 equiv) and Triphosgene (105.84 mg, 0.356 mmol, 0.5 equiv) at 0 °C, and stirred for 2 h at rt. After completion of reaction to afford 1-benzyl-6-isocyanato-3,4-dihydroquinolin-2-one as a crude. MS (ESI): mass calcd. for C17H14N2O2, 278.11m/z, found 279.05 [M+H]+
Figure imgf000140_0002
1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(4-hydroxy-2-methylbutan-2-yl)urea To a stirred solution of 1-benzyl-6-isocyanato-3,4-dihydroquinolin-2-one (crude) in anhydrous DCM (5 mL) was added 3-amino-3-methylbutan-1-ol (220.3 mg, 2.139 mmol, 3 equiv) was added at 0 °C. The resulting mixture was stirred for 1 h at rt. Desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The crude product was purified by HPLC with the following conditions Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 21% B to 41% B in 10 min, 41% B; Wave Length: 254/220 nm; RT1(min): 10; Number Of Runs: 0; to afford 1-(1-benzyl-2-oxo-3,4- dihydroquinolin-6-yl)-3-(4-hydroxy-2-methylbutan-2-yl)urea (23.3 mg, 16.47%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.39 – 7.12 (m, 6H), 6.96 (dd, J = 8.8, 2.5 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.10 (s, 2H), 4.39 (t, J = 4.9 Hz, 1H), 3.48 (td, J = 7.1, 4.7 Hz, 2H), 2.88 (dd, J = 8.8, 5.8 Hz, 2H), 2.65 (dd, J = 8.8, 5.8 Hz, 2H), 1.79 (t, J = 7.2 Hz, 2H), 1.25 (s, 6H). Example 17: 1-(tert-butyl)-3-(1-(4-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000141_0001
4-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1086.6 mg, 7.805 mmol, 3 equiv) and 4- (bromomethyl)benzonitrile (558.09 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 4-((6-nitro-2-oxo-3,4- dihydroquinolin-1(2H)-yl)methyl)benzonitrile (400 mg, 50.00%) as a white solid. MS (ESI): mass calcd. for C17H13N3O3: 307.10 m/z, found 308.15 [M+H]+.
Figure imgf000141_0002
4-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a solution of 4-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (300 mg, 0.976 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (545.1 mg, 9.76 mmol, 10 equiv) and NH4Cl (522.1 mg, 9.76 mmol, 10 equiv).The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 4-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (108 mg, 40.05%) as a yellow solid. MS (ESI): mass calcd. for C17H15N3O: 277.12 m/z, found 278.25 [M+H]+.
Figure imgf000142_0001
1-(tert-butyl)-3-(1-(4-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 4-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (80 mg, 0.288 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (85.8 mg, 0.865 mmol, 3 equiv) and TEA (87.4 mg, 0.865 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 45% B in 10.5 min; Wave Length: 220 nm; RT1(min): 12; Number Of Runs: 1) to afford 1-(tert-butyl)-3-(1-(4-cyanobenzyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea (38.9 mg, 36.02%) as a white solid. MS (ESI): mass calcd. for C22H24N4O: 376.19 m/z, found 377.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.75 – 7.81 (m, 2H), 7.40 (d, J = 8.2 Hz, 2H), 7.34 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.8, 2.5 Hz, 1H), 6.71 (d, J = 8.8 Hz, 1H), 5.92 (s, 1H), 5.18 (s, 2H), 2.86 – 2.94 (m, 2H), 2.63 – 2.71 (m, 2H), 1.26 (s, 9H). Example 18: 1-(tert-butyl)-3-(1-(3-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000142_0002
3-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (1086.6 mg, 7.805 mmol, 3 equiv) and 3- (bromomethyl)benzonitrile (558.09 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 3-((6-nitro-2-oxo-3,4- dihydroquinolin-1(2H)-yl)methyl)benzonitrile (350 mg, 43.75%) as a white solid. MS (ESI): mass calcd. for C17H13N3O3: 307.10 m/z, found 308.05 [M+H]+.
Figure imgf000143_0001
3-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a solution of 3-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (300 mg, 0.976 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (545.1 mg, 9.76 mmol, 10 equiv) and NH4Cl (522.1 mg, 9.76 mmol, 10 equiv).The resulting mixture was stirred for 2 h at 60 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 3-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (120 mg, 44.44%) as a yellow solid. MS (ESI): mass calcd. for C17H15N3O: 277.12 m/z, found 278.20 [M+H]+.
Figure imgf000143_0002
1-(tert-butyl)-3-(1-(3-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 3-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (80 mg, 0.288 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (85.8 mg, 0.865 mmol, 3 equiv) and TEA (87.4 mg, 0.865 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 25% B to 45% B in 10.5 min; Wave Length: 220 nm; RT1(min): 12; Number Of Runs: 1) to afford 1-(tert-butyl)-3-(1-(3-cyanobenzyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea (47.9 mg, 44.35%) as a white solid. MS (ESI): mass calcd. for C22H24N4O: 376.19 m/z, found 377.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.67 – 7.75 (m, 2H), 7.48 – 7.58 (m, 2H), 7.33 (d, J = 2.4 Hz, 1H), 6.98 (dd, J = 8.8, 2.5 Hz, 1H), 6.74 (d, J = 8.8 Hz, 1H), 5.92 (s, 1H), 5.14 (s, 2H), 2.86 – 2.94 (m, 2H), 2.64 – 2.72 (m, 2H), 1.26 (s, 9H). Example 1: 3-tert-butyl-1-{1-[(2-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea
Figure imgf000144_0001
1-[(2-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (2 g, 10.407 mmol, 1 equiv) and 1-(bromomethyl)-2-fluorobenzene (3147.60 mg, 16.651 mmol, 1.6 equiv) in dimethylformamide (20 mL) was added K2CO3 (4346.47 mg, 31.221 mmol, 3 equiv). The reaction was stirred at rt for 3h. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1- [(2-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (2.06 g, 65.92%) as a white solid. MS (ESI): mass calcd. for C16H13FN2O3: 300.09 m/z, found 301.05[M+H]+.
Figure imgf000145_0001
6-amino-1-[(2-fluorophenyl)methyl]-3,4-dihydroquinolin-2-one To a solution of 1-[(2-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (1 g, 3.330 mmol, 1 equiv) in EA (35 mL) was added Pd/C (1 g). The resulting mixture was stirred for 1 h at room temperature under hydrogen atmosphere. Desired product could be detected by LCMS. The resulting mixture was filtered, the filter cake was washed with EA (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(2-fluorophenyl)methyl]- 3,4-dihydroquinolin-2-one (990 mg, 109.98%) as a white solid. MS (ESI): mass calcd for C16H15FN2O: 270.12 m/z, found 271.10[M+H]+.
Figure imgf000145_0002
3-tert-butyl-1-{1-[(2-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea To a solution of 6-amino-1-[(2-fluorophenyl)methyl]-3,4-dihydroquinolin-2-one (980 mg, 3.626 mmol, 1 equiv) in DCM/toluene (10 mL, 2/1) was added 2-isocyanato-2- methylpropane (1077.9 mg, 10.873 mmol, 3.00 equiv) The reaction was stirred at 90 oC for 1h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using aXBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(2-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea (412.3 mg, 30.33%) as a white solid. LC/MS (ESI): mass calcd. for C21H24FN3O2:369.19 m/z, found:370.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.35 (m, 1H), 7.23 - 7.32 (m, 2H), 7.00 -7.09(m, 3H), 6.73 (m, 1H), 5.93 (s, 1H), 5.12 (s, 2H), 2.89 (m, 2H), 2.66 (m, 2H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ-118.31. Example 20: 1-(tert-butyl)-3-(1-(2-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000146_0001
2-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a stirred solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (400 mg, 2.081 mmol, 1 equiv) in DMF (10 mL) was added 2-(bromomethyl)benzonitrile (612.09 mg, 3.122 mmol, 1.5 equiv) and potassium methaneperoxoate potassium (869.29 mg, 6.243 mmol, 3 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. The resulting mixture was extracted with brine (1 x 30 mL). The combined organic layers were washed with EA (3 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile (447 mg, 70.0%) as a yellow solid. MS (ESI): mass calcd. for C17H13N3O3: 307.10 m/z, found 308.10 [M +H] +.
Figure imgf000146_0002
2-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile To a stirred solution of 2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile (400 mg, 1.302 mmol, 1 equiv) in ethyl alcohol (10 mL) and water (2 mL) was added iron (726.89 mg, 13.020 mmol, 10 equiv) and NH4Cl (696.24 mg, 13.020 mmol, 10 equiv) and stirred for 1 h at 60 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluted with PE/EA (2:1) to afford 2-((6- amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (280 mg, 77.57%) as a white solid. MS (ESI): mass calcd. for C17H15N3O: 277.12 m/z, found 278.05 [M +H] +.
Figure imgf000147_0001
1-(tert-butyl)-3-(1-(2-cyanobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea A solution of 2-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)benzonitrile (150 mg, 0.541 mmol, 1 equiv) in DCM (5 mL) was add TEA (164.20 mg, 1.623 mmol, 3 equiv) and 2-isocyanato-2-methylpropane (214.48 mg, 2.164 mmol, 4 equiv) at 0 °C. The resulting mixture was stirred for 1 h at rt. The resulting mixture was concentrated under reduced pressure. The crude product was purified by HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0; to afford 3-tert-butyl-1-{1-[(2- cyanophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea (69.2 mg, 33.91%) as a white solid. MS (ESI):mass calcd. for C22H24N4O2: 376.19 m/z, found 377.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.15 (s, 1H), 7.86 (dd, J = 7.7, 1.3 Hz, 1H), 7.61 (td, J = 7.7, 1.4 Hz, 1H), 7.44 (td, J = 7.6, 1.2 Hz, 1H), 7.37 (d, J = 2.4 Hz, 1H), 7.25 – 7.18 (m, 1H), 7.00 (dd, J = 8.7, 2.5 Hz, 1H), 6.69 (d, J = 8.8 Hz, 1H), 5.94 (s, 1H), 5.25 (s, 2H), 2.93 (dd, J = 8.7, 5.8 Hz, 2H), 2.67 (dd, J = 8.7, 5.8 Hz, 2H), 1.27 (s, 9H). Example 21: 1-(tert-butyl)-3-(1-(3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea
Figure imgf000147_0002
6-amino-1-[(3-methylphenyl)methyl]-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (8 mL) was added K2CO3 (1086.6 mg, 7.862 mmol, 3 equiv) and 1-(bromomethyl)-3- methylbenzene (529.6 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-[(3-methylphenyl)methyl]-6-nitro-3,4- dihydroquinolin-2-one (500 mg, 64.44%) as a yellow solid. MS (ESI): mass calcd. for C17H16N2O3296.12 m/z, found 297.00 [M+H]+.
Figure imgf000148_0001
6-amino-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one To a solution of 1-[(3-methylphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.687 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (942.2 mg, 16.873 mmol, 10 equiv) and NH4Cl (902.5 mg, 16.873 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-[(3- methylphenyl)methyl]-3,4-dihydroquinolin-2-one (400 mg, 88.46%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O 266.14 m/z, found 267.10 [M+H]+.
Figure imgf000148_0002
1-(tert-butyl)-3-(1-(3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-[(3-methylphenyl)methyl]-3,4-dihydroquinolin-2-one (400 mg, 1.502 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (446.6 mg, 4.506 mmol, 3 equiv) and TEA (455.9 mg, 4.505 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 38% B to 60% B in 10 min, 60% B; Wave Length: 254/220 nm; RT1(min): 10; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(3-methylbenzyl)- 2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (282.3 mg, 51.15%) as a white solid. MS (ESI): mass calcd. for C22H27N3O2, 365.21 m/z, found 366.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.17 (t, J = 7.7 Hz, 1H), 6.92 – 7.09 (m, 4H), 6.76 (d, J = 8.8 Hz, 1H), 5.92 (s, 1H), 5.05 (s, 2H), 2.87 (dd, J = 8.8, 5.8 Hz, 2H), 2.64 (dd, J = 8.7, 5.8 Hz, 2H), 2.25 (s, 3H), 1.27 (s, 9H). Example 22: 1-(tert-butyl)-3-(1-(3-methoxybenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-
Figure imgf000149_0001
1-[(3-methoxyphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (8 mL) was added K2CO3 (1078.7 mg, 7.862 mmol, 3 equiv) and 1-(bromomethyl)-3- methoxybenzene (575.4 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1-[(3-methoxyphenyl)methyl]-6-nitro-3,4- dihydroquinolin-2-one (500 mg, 60.71%) as a yellow solid. MS (ESI): mass calcd. for C17H16N2O4312.11 m/z, found 313.05 [M+H]+.
Figure imgf000150_0001
6-amino-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one To a solution of 1-[(3-methoxyphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.601 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (894.0 mg, 16.009 mmol, 10 equiv) and NH4Cl (856.3 mg, 16.009 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-[(3- methoxyphenyl)methyl]-3,4-dihydroquinolin-2-one (400 mg, 88.33%) as a yellow solid. MS (ESI): mass calcd. for C17H18N2O2282.14 m/z, found 283.10 [M+H]+.
Figure imgf000150_0002
1-(tert-butyl)-3-(1-(3-methoxybenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-[(3-methoxyphenyl)methyl]-3,4-dihydroquinolin-2-one (400 mg, 1.417 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (421.3 mg, 4.250 mmol, 3 equiv) and TEA (430.0 mg, 4.250 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 50% B in 9 min, 50% B; Wave Length: 254/220 nm; RT1(min): 8.6; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(3- methoxybenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (85.3 mg, 15.75%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3, 381.21 m/z, found 382.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.16 – 7.25 (m, 1H), 6.97 (dd, J = 8.7, 2.5 Hz, 1H), 6.71 – 6.82 (m, 4H), 5.92 (s, 1H), 5.06 (s, 2H), 3.70 (s, 3H), 2.87 (dd, J = 8.8, 5.8 Hz, 2H), 2.64 (dd, J = 8.7, 5.8 Hz, 2H), 1.26 (s, 9H). Example 23: 1-(tert-butyl)-3-(2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4- tetrahydroquinolin-6-yl)urea
Figure imgf000151_0001
6-nitro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (8 mL) was added K2CO3 (1086.6 mg, 7.862 mmol, 3 equiv) and 1-(bromomethyl)-3- (trifluoromethyl)benzene (684.1 mg, 2.862 mmol, 1.1 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS, The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (PE/EA=3:1r) to afford 6-nitro-1-{[3-(trifluoromethyl)phenyl]methyl}-3,4- dihydroquinolin-2-one (500 mg, 54.86%) as a yellow solid. MS (ESI): mass calcd. for C17H13F3N2O3350.09 m/z, found 350.95 [M+H]+.
Figure imgf000151_0002
6-amino-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-1-{[3-(trifluoromethyl)phenyl]methyl}-3,4-dihydroquinolin-2-one (500 mg, 1.427 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (797.1 mg, 14.274 mmol, 10 equiv) and NH4Cl (763.4 mg, 14.274 mmol, 10 equiv). The resulting mixture was stirred 30 min at 60 °C until the starting material was totally consumed by LCMS, The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (PE/EA=2:1) to afford 6-amino-1-{[3-(trifluoromethyl)phenyl]methyl}-3,4- dihydroquinolin-2-one (300 mg, 64.88%) as a yellow solid.MS (ESI): mass calcd. for C17H15F3N2O 320.11 m/z, found 321.05 [M+H]+.
Figure imgf000152_0001
1-(tert-butyl)-3-(2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-{[3-(trifluoromethyl)phenyl]methyl}-3,4-dihydroquinolin-2- one (300 mg, 0.937 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (278.5 mg, 2.810 mmol, 3 equiv) and TEA (284.3 mg, 2.810 mmol, 3 equiv).The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3*20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.The compound was separated by preparative HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 45% B in 10 min, 45% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 3- tert-butyl-1-(2-oxo-1-{[3-(trifluoromethyl)phenyl]methyl}-3,4-dihydroquinolin-6-yl)urea (125.4 mg, 31.80%) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O2, 419.18 m/z, found 419.95 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.45 – 7.63 (m, 4H), 7.34 (d, J = 2.4 Hz, 1H), 7.00 (dd, J = 8.7, 2.5 Hz, 1H), 6.80 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.20 (s, 2H), 2.89 (dd, J = 8.8, 5.7 Hz, 2H), 2.67 (dd, J = 8.6, 5.8 Hz, 2H), 1.27 (s, 9H). Example 24: 3-tert-butyl-1-{1-[(2,5-dichlorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6- yl}urea
Figure imgf000153_0001
1-[(2,5-dichlorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one To a stirred solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added 2-(bromomethyl)-1,4-dichlorobenzene (936.34 mg, 3.903 mmol, 1.5 equiv) and potassium carbonate (724.41 mg, 5.204 mmol, 2 equiv) at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 3 h at room temperature. The reaction was monitored by LCMS. The resulting mixture was quenched with water (20 mL) and extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 1-[(2,5-dichlorophenyl)methyl]-6-nitro-3,4- dihydroquinolin-2-one (500 mg, 54.72%) as a white solid. LC/MS: mass calcd. for C16H12Cl2N2O3: 350.02 m/z, found:350.90 [M+H] +
Figure imgf000153_0002
6-amino-1-[(2,5-dichlorophenyl)methyl]-3,4-dihydroquinolin-2-one To a stirred solution of 1-[(2,5-dichlorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.424 mmol, 1 equiv) in EtOH/H2O (10 mL, 10/1) was added Fe (795.11 mg, 14.240 mmol, 10 equiv) and NH4Cl (761.58 mg, 14.240 mmol, 10 equiv) at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 2 h at 60°C. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 6-amino-1-[(2,5-dichlorophenyl)methyl]-3,4- dihydroquinolin-2-one (280 mg, 61.23%) as a white solid. LC/MS: mass calcd. for C16H14Cl2N2O: 320.05 m/z, found:320.95 [M+H] +.
Figure imgf000154_0001
3-tert-butyl-1-{1-[(2,5-dichlorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea To a stirred solution of 6-amino-1-[(2,5-dichlorophenyl)methyl]-3,4-dihydroquinolin-2- one (140 mg, 0.436 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (129.63 mg, 1.308 mmol, 3 equiv) in DCM (10 mL) was added TEA (132.32 mg, 1.308 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 1 day at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by Prep-HPLC to afford 3-tert-butyl-1-{1-[(2,5-dichlorophenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea (20.8 mg, 11.34%) as a white solid. LC/MS: mass calcd for C21H23Cl2N3O2: 419.12 m/z, found:420.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.16 (s, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.40 - 7.37 (m, 2H), 7.01 - 6.95 (m, 2H), 6.60 (d, J = 8.4 Hz, 1H), 5.94 (s, 1H), 5.05 (s, 2H), 2.94 - 2.90 (m, 2H), 2.72 - 2.69 (m, 2H), 1.27 (s, 9H). Example 25: 1-(tert-butyl)-3-(1-(3,5-dichlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl) urea
Figure imgf000155_0001
1-(3,5-dichlorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a mixture of 6-nitro-3,4-dihydro-1H-quinolin-2-one (400 mg, 2.081 mmol, 1 equiv) and K2CO3 (869.29 mg, 6.243 mmol, 3 equiv) in DMF (20 mL) was added 1-(bromomethyl)-3,5- dichlorobenzene (749.07 mg, 3.122 mmol, 1.5 equiv). The reaction was stirred at rt for 2 hours. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[(3,5- dichlorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (647 mg, 88.51%) as a yellow solid. MS (ESI): mass calcd. for C16H12Cl2N2O3: 351.18m/z, found 350.95 [M+H]+.
Figure imgf000155_0002
6-amino-1-(3,5-dichlorobenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-[(3,5-dichlorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.424 mmol, 1 equiv) in MeOH/H2O (11 mL, 10/1) was added Fe (397.55 mg, 7.120 mmol, 5 equiv) and NH4Cl (380.79 mg, 7.120 mmol, 5 equiv). The reaction was stirred at 80 oC for 1 hours. After filtration, the filter cake was washed with EA (3 x 10 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The resulting mixture was concentrated under vacuum. This resulted in 6-amino-1-[(3,5-dichlorophenyl)methyl]-3,4- dihydroquinolin-2-one (418 mg, 91.40%) as a yellow solid. MS (ESI): mass calcd. for C16H14Cl2N2O: 321.20 m/z, found 320.95 [M+H]+.
Figure imgf000156_0001
1-(tert-butyl)-3-(1-(3,5-dichlorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-[(3,5-dichlorophenyl)methyl]-3,4-dihydroquinolin-2-one (100 mg, 0.311 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (92.59 mg, 0.933 mmol, 3 equiv) and TEA (94.51 mg, 0.933 mmol, 3 equiv) at 0 oC. The reaction was stirred at rt for 2 days. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column, 30*150 mm, 5μm column (eluent: 45% to 67% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(3,5-dichlorophenyl)methyl]-2-oxo- 3,4-dihydroquinolin-6-yl}urea (20 mg, 15.28%) as a white solid. MS (ESI): mass calcd. for C21H23Cl2N3O2:420.33 m/z, found:420.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.04 – 8.23 (m, 1H), 7.39 – 7.49 (m, 1H), 7.22 – 7.39 (m, 3H), 6.94 – 7.11 (m, 1H), 6.65 – 6.88 (m, 1H), 5.80 – 6.01 (m, 1H), 5.00 – 5.21 (m, 2H), 2.81 – 2.98 (m, 2H), 2.60 – 2.78 (m, 2H), 1.25 – 1.33 (m, 9H). Example 26: 1-(tert-butyl)-3-(1-(3-fluorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000156_0002
1-(3-fluorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (20 mL) was added K2CO3 (1086.62 mg, 7.806 mmol, 3 equiv) and (bromomethyl)-3- fluorobenzene (737.72 mg, 3.903 mmol, 1.5 equiv) at rt. The final reaction mixture was stirred for 1h at rt. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1- [(3-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (163 mg, 20.86%) as a yellow solid. MS (ESI): mass calcd. for C16H13FN2O3: 300.29 m/z, found 301.00 [M+H]+.
Figure imgf000157_0001
6-amino-1-(3-fluorobenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-[(3-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (155 mg, 0.516 mmol, 1 equiv) in MeOH/H2O (22 mL, 10/1) was added Fe (144.13 mg, 2.580 mmol, 5 equiv) and NH4Cl (138.05 mg, 2.580 mmol, 5 equiv). The reaction was stirred at 80 °C for 1 hour. The resulting mixture was filtered, the filter cake was washed with EA (3 x 20 mL). The filtrate was concentrated under reduced pressure. The residue was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(3-fluorophenyl)methyl]-3,4- dihydroquinolin-2-one hydrochloride (100 mg, 63.15%) as a yellow oil. MS (ESI): mass calcd. for C16H15FN2O: 270.31m/z, found 271.15 [M+H]+.
Figure imgf000157_0002
1-(tert-butyl)-3-(1-(3-fluorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-[(3-fluorophenyl)methyl]-3,4-dihydroquinolin-2-one (90 mg, 0.333 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (99.02 mg, 0.999 mmol, 3 equiv) and Pyridine (79.01 mg, 0.999 mmol, 3 equiv). The reaction was stirred at rt for 3 days. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(3-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin- 6-yl}urea (20.7 mg, 16.83%) as a white solid. MS (ESI): mass calcd. for C21H24FN3O2:369.44 m/z, found:370.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.96 – 8.19 (m, 1H), 7.30 – 7.40 (m, 2H), 6.93 – 7.19 (m, 4H), 6.69 – 6.81 (m, 1H), 5.80 – 6.00 (m, 1H), 4.98 – 5.29 (m, 2H), 2.81 – 2.94 (m, 2H), 2.60 – 2.71 (m, 2H), 1.19 – 1.29 (m, 9H).19F NMR (376 MHz, DMSO) δ (ppm): -113.2050. Example 27: 1-(tert-butyl)-3-(1-(4-fluorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea
Figure imgf000158_0001
1-(4-fluorobenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) and K2CO3 (1086.62 mg, 7.806 mmol, 3 equiv) in DMF (10 mL) was added 1-(bromomethyl)-4- fluorobenzene (737.78 mg, 3.903 mmol, 1.5 equiv). The reaction was stirred at rt for 2 hours. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[(4- fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (630 mg, 80.64%) as a yellow solid. MS(ESI): mass calcd. for C16H13FN2O3: 300.29 m/z, found 301.00 [M+H]+.
Figure imgf000158_0002
6-amino-1-(4-fluorobenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-[(4-fluorophenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (400 mg, 1.332 mmol, 1 equiv) in MeOH/H2O (22 mL, 10/1) was added Fe (371.94 mg, 6.660 mmol, 5 equiv) and NH4Cl (356.26 mg, 6.660 mmol, 5 equiv). The reaction was stirred for 1 hour at 80 oC. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The resulting mixture was concentrated under vacuum. This resulted in 6-amino-1-[(4- fluorophenyl)methyl]-3,4-dihydroquinolin-2-one (325 mg, 90.26%) as a yellow solid. MS (ESI): mass calcd. for C16H15FN2O: 270.31m/z, found 271.05 [M+H]+.
Figure imgf000159_0001
1-(tert-butyl)-3-(1-(4-fluorobenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-[(4-fluorophenyl)methyl]-3,4-dihydroquinolin-2-one (150 mg, 0.555 mmol, 1 equiv) in DCM (20 mL) was added 2-isocyanato-2-methylpropane (165.03 mg, 1.665 mmol, 3 equiv) and Pyridine (131.68 mg, 1.665 mmol, 3 equiv). The reaction was stirred at rt for 3 days. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(4-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin- 6-yl}urea (5.3 mg, 2.59%) as a white solid. MS (ESI): mass calcd. for C21H24FN3O2:369.44 m/z, found:370.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.11 – 7.40 (m, 3H), 6.96 – 7.11 (m, 3H), 6.81– 6.91 (m, 1H), 5.10– 5.22 (m, 2H), 2.82 – 3.03 (m, 2H), 2.59 – 2.82 (m, 2H), 1.29 – 1.41 (m, 9H). 19F NMR (376 MHz, DMSO) δ (ppm): -117.9957. Example 28: 1-(tert-butyl)-3-(2-oxo-1-(1-phenylpropyl)-1,2,3,4-tetrahydroquinolin-6- yl)urea
Figure imgf000160_0001
6-nitro-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (400 mg, 2.081 mmol, 1 equiv) in DMF (8 mL) was added K2CO3 (863.0 mg, 6.244 mmol, 3 equiv) and (1-bromopropyl)benzene (455.8 mg, 2.289 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-nitro-1-(1-phenylpropyl)-3,4-dihydroquinolin-2- one (150 mg, 22.24%) as a yellow solid. MS (ESI): mass calcd. for C18H18N2O3310.13 m/z, found 311.25 [M+H]+.
Figure imgf000160_0002
6-amino-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one (130 mg, 0.419 mmol, 1 equiv) in EtOH (8 mL) and H2O (2 mL) was added Fe (233.9 mg, 4.189 mmol, 10 equiv) and NH4Cl (224.0 mg, 4.189 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 60 °C. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 6-amino-1-(1-phenylpropyl)-3,4- dihydroquinolin-2-one (100 mg, 85.15%) as a yellow solid.MS (ESI): mass calcd. for C18H20N2O 280.16 m/z, found 281.10 [M+H]+.
Figure imgf000161_0001
1-(tert-butyl)-3-(2-oxo-1-(1-phenylpropyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a solution of 6-amino-1-(1-phenylpropyl)-3,4-dihydroquinolin-2-one (100 mg, 1.502 mmol, 1 equiv) in DCM (8 mL) was added 2-isocyanato-2-methylpropane (106.0 mg, 1.070 mmol, 3 equiv) and TEA (108.2 mg, 1.070 mmol, 3 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: Aeris PEPTIDE 5um XB-C18 Axia, 21.2 mm X 250 mm, 5 μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 39% B to 61% B in 10.5 min; Wave Length: 220 nm; RT1(min): 7.9; Number Of Runs: 2) to afford 1-(tert-butyl)-3-(2-oxo-1-(1-phenylpropyl)-1,2,3,4- tetrahydroquinolin-6-yl)urea (29.8 mg, 21.98%) as a white solid. MS (ESI): mass calcd. for C23H29N3O2, 379.23 m/z, found 380.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.20 – 7.41 (m, 6H), 6.81 (dd, J = 8.8, 2.5 Hz, 1H), 6.49 (d, J = 8.8 Hz, 1H), 5.95 (d, J = 13.1 Hz, 2H), 2.74 – 2.92 (m, 2H), 2.56 – 2.74 (m, 2H), 2.31 – 2.40 (m, 1H), 2.04 – 2.22 (m, 1H), 1.25 (s, 9H), 0.77 (t, J = 7.2 Hz, 3H). Example 29: 1-(1-benzyl-3,3-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000162_0001
N-(2-bromophenyl)pivalamide To a stirred solution of O-bromoaniline (3 g, 17.439 mmol, 1 equiv) in anhydrous DCM (50 mL) was added triethylamine (2.29 g, 22.671 mmol, 1.3 equiv) followed by 2,2- dimethylpropanoyl chloride (2.10 g, 17.439 mmol, 1 equiv) at 0 °C. The reaction mixture was stirred for 16 h at rt. After completion of reaction, the reaction mixture was quenched by addition of water (50 mL). The aqueous layer was extracted with DCM (50 mL). The combined organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude product which was further purified by silica gel column, eluted with PE / EA (3:1) to afford desired compound N-(2-bromophenyl)-2,2- dimethylpropanamide (2.7 g, 60.44%) as a yellow solid. MS (ESI): mass calcd. for C11H14BrNO: 255.03 m/z, found 256.05 [M +H]+.
Figure imgf000162_0002
3,3-dimethyl-3,4-dihydroquinolin-2(1H)-one To a stirred solution of N-(2-bromophenyl)-2,2-dimethylpropanamide (400 mg, 1.562 mmol, 1 equiv) in NMP (20 mL, 207.398 mmol, 132.81 equiv) was added Cs2CO3 (1.02 g, 3.124 mmol, 2 equiv), tris(4-methylphenyl)phosphane (95.06 mg, 0.312 mmol, 0.2 equiv), Pd(OAc)2 (35.06 mg, 0.156 mmol, 0.1 equiv) and TBHP (42.22 mg, 0.469 mmol, 0.3 equiv) at rt and stirred for overnight at 140 °C under N2. The reaction mixture was concentrated under reduced pressure to give crude product which was further purified by silica, gel column PE / EA (3:1) to afford desired compound 3,3-dimethyl-1,4-dihydroquinolin-2-one (0.2 g, 73.26%). MS (ESI): mass calcd. for C11H13NO: 175.10 m/z, found 176.25 [M +H]+.
Figure imgf000163_0001
3,3-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 3,3-dimethyl-3,4-dihydroquinolin-2(1H)-one (0.2 g, 1.14 mmol, 1 equiv) in H2O (2 mL) and H2SO4 (9 mL) was added HNO3 (0.2 mL) at -10 °C. The reaction was stirred for 2 h at -10-0 °C. The reaction mixture was quenched by water. The mixture was acidified pH = 7 with NaHCO3 (aq.) and then extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 3,3-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (0.15 g, 60.0%) as a white solid. MS (ESI): mass calcd. for C11H12N2O3, 220.08 m/z, found 221.05 [M+H]+ .
Figure imgf000163_0002
1-benzyl-3,3-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred solution of 3,3-dimethyl-6-nitro-1,4-dihydroquinolin-2-one (150 mg, 0.681 mmol, 1 equiv) in DMF (3 mL) was added K2CO3 (322.39 mg, 2.316 mmol, 3 equiv) and benzyl bromide (158.43 mg, 0.926 mmol, 1.2 equiv) at rt and stirred for overnight. The resulting mixture was extracted with EA (3 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 1-benzyl-3,3-dimethyl-6-nitro- 3,4-dihydroquinolin-2(1H)-one (150 mg, 71.09%) as a white solid. MS (ESI): mass calcd. for C18H18N2O3: 310.13 m/z, found 311.10 [M +H] +.
Figure imgf000164_0001
6-amino-1-benzyl-3,3-dimethyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-3,3-dimethyl-6-nitro-3,4-dihydroquinolin-2(1H)-one (130 mg, 0.419 mmol, 1 equiv) in 10 mL MeOH was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, then filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-benzyl-3,3-dimethyl-4H-quinolin-2-one (100 mg, 85.15%) as a white solid. MS (ESI): mass calcd. for C18H20N2O: 280.16 m/z, found 281.25 [M +H] +.
Figure imgf000164_0002
1-(1-benzyl-3,3-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a stirred solution of 6-amino-1-benzyl-3,3-dimethyl-4H-quinolin-2-one (50 mg, 0.178 mmol, 1 equiv) in anhydrous DCM (5 mL) was added TEA (54.14 mg, 0.534 mmol, 3 equiv) 2- isocyanato-2-methylpropane (70.72 mg, 0.712 mmol, 4 equiv) at 0 °C, and stirred for overnight at rt. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by HPLC with the following conditions Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 44% B to 60% B in 8.5 min; Wave Length: 220 nm; RT1(min): 6.9; Number Of Runs: 3; to afford 1-(1-benzyl- 3,3-dimethyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (5.6 mg, 8.26%) as a white solid. MS (ESI):mass calcd. for C23H29N3O2: 379.23 m/z, found 380.20 [M+H]+.1H NMR (300 MHz, Methanol-d4) δ 7.18 – 7.34 (m, 6H), 7.01 – 7.04 (m, 1H), 6.82 (d, J = 8.7 Hz, 1H), 5.16 (s, 2H), 2.84 (s, 2H), 1.36 (s, 9H), 1.23 (s, 6H). Example 30: 1-(tert-butyl)-3-(1-(2-hydroxy-1-phenylethyl)-2-oxo-1,2,3,4-tetrahydroquinoli n-6-yl)urea Synthetic Scheme
Figure imgf000165_0001
Into a 40 mL round-bottom flask was added 2-bromo-2-phenylethan-1-ol (3 g, 21.713 mmol, 1 equiv) and bromotrimethylsilane (3.66 g, 23.884 mmol, 1.1 equiv) at rt. The resulting mixture was stirred for additional overnight at rt. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE /EA (5:1) afford 2-bromo-2-phenylethanol (2.7 g, 61.85%) as a yellow semi-solid.
Figure imgf000165_0002
(2-bromo-2-phenylethoxy)(tert-butyl)dimethylsilane To a stirred solution of 2-bromo-2-phenylethanol (1.4 g, 6.963 mmol, 1 equiv) in anhydrous DCM (30 mL) was added t-butyldimethylchlorosilane (2.10 g, 13.926 mmol, 2 equiv) and Imidazole (1.42 g, 20.889 mmol, 3 equiv) at rt and stirred for 1 h. The reaction progress was monitored by TLC. The residue was purified by silica gel column chromatography, eluted with EA (35%-58%) to afford (2-bromo-2-phenylethoxy)(tert-butyl)dimethylsilane (1.2 g, 54.65%) as a yellow semi-solid.
Figure imgf000166_0001
1-(2-((tert-butyldimethylsilyl)oxy)-1-phenylethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred solution of (2-bromo-2-phenylethoxy)(tert-butyl)dimethylsilane (500 mg, 1.586 mmol, 1 equiv) in dimethylformamide (10 mL) was added 6-nitro-3,4-dihydro-1H- quinolin-2-one (274.25 mg, 1.427 mmol, 0.9 equiv) and Cs2CO3 (1549.92 mg, 4.758 mmol, 3 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. The resulting mixture was extracted with EA (3 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 1-(2-((tert- butyldimethylsilyl)oxy)-1-phenylethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (350 mg, 51.74%) as a yellow solid. MS (ESI): mass calcd. for C23H30N2O4Si: 426.20 m/z, found 427.15 [M +H] +.
Figure imgf000166_0002
6-amino-1-(2-((tert-butyldimethylsilyl)oxy)-1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(2-((tert-butyldimethylsilyl)oxy)-1-phenylethyl)-6-nitro-3,4- dihydroquinolin-2(1H)-one (80 mg, 0.188 mmol, 1 equiv) in 10 mL MeOH was added Pd/C (10%, 50 mg) under nitrogen atmosphere in a 50 mL. The mixture was hydrogenated at room temperature for 1 h under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-(2-((tert- butyldimethylsilyl)oxy)-1-phenylethyl)-3,4-dihydroquinolin-2(1H)-one (75 mg, 100.0%) as a white solid. MS (ESI): mass calcd. for C23H32N2O2S: 396.22 m/z, found 397.30 [M +H] +.
Figure imgf000167_0001
1-(tert-butyl)-3-(1-(2-((tert-butyldimethylsilyl)oxy)-1-phenylethyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea o a stirred solution of 6-amino-1-(2-((tert-butyldimethylsilyl)oxy)-1-phenylethyl)-3,4- dihydroquinolin-2(1H)-one (100 mg, 0.252 mmol, 1 equiv) in DCM (10 mL) was added 2- isocyanato-2-methylpropane (74.99 mg, 0.756 mmol, 3 equiv) and toluene (5 mL, 46.993 mmol, 186.38 equiv) at rt and stirred for overnight at 90 °C. The reaction progress was monitored by LCMS. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product. MS (ESI): mass calcd. for C28H41N3O3Si: 495.29 m/z, found 496.30 [M +H] +.
Figure imgf000167_0002
1-(tert-butyl)-3-(1-(2-hydroxy-1-phenylethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of 1-(tert-butyl)-3-(1-(2-((tert-butyldimethylsilyl)oxy)-1- phenylethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (crude) in tetrahydrofuran (5 mL) was added Triethylamine trihydrofluoride (300 mg) and stirred for 2 h at 60 °C. The reaction progress was monitored by LCMS. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by HPLC with the following conditions Column: Xselect CSH F-Phenyl OBD column, 19*250 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 45% B to 75% B in 7 min, 75% B; Wave Length: 254/220 nm; RT1(min): 6.4; Number Of Runs: 0 to afford 3-tert-butyl-1-[1-(2-hydroxy-1-phenylethyl)-2-oxo-3,4- dihydroquinolin-6-yl]urea (4 mg, 10.30%) as a white solid. MS (ESI):mass calcd. for C22H27N3O3: 381.21 m/z, found 382.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.33 (d, J = 4.0 Hz, 5H), 7.25 (q, J = 4.2 Hz, 1H), 6.87 (dd, J = 8.8, 2.5 Hz, 1H), 6.60 (d, J = 8.8 Hz, 1H), 5.89 (d, J = 13.5 Hz, 1H), 5.71 (d, J = 7.1 Hz, 1H), 5.01 (t, J = 5.5 Hz, 1H), 4.26 (dt, J = 11.4, 6.0 Hz, 1H), 4.07 (dt, J = 11.1, 6.1 Hz, 1H), 2.93 – 2.77 (m, 2H), 2.58 (q, J = 6.5 Hz, 2H), 1.26 (s, 9H). Example 31: 1-(1-benzyl-2-oxo-3H-indol-5-yl)-3-tert-butylurea Synthetic Scheme
Figure imgf000168_0001
1-benzyl-5-nitroindole-2,3-dione To a solution of 1H-indole-2,3-dione, 5-nitro (1 g, 5.205 mmol, 1 equiv) and benzyl bromide (979.24 mg, 5.726 mmol, 1.1 equiv) in dimethylformamide (10 mL) was added K2CO3 (1438.66 mg, 10.41 mmol, 2 equiv). The reaction was stirred at rt for 3h. Quenched with water (40 mL) and extracted with EA (3 x 40 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-5-nitroindole-2,3-dione (665 mg, 45.27%) as a white solid. MS (ESI): mass calcd for C15H10N2O4: 282.06 m/z, found 283.15[M+H]+.
Figure imgf000168_0002
benzyl-5-nitro-3H-indol-2-one To a solution of 1-benzyl-5-nitroindole-2,3-dione (500 mg, 1.771 mmol, 1 equiv) in Hydrazinium hydroxide solution (5 mL) was stirred at 120 °C for 3h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to 1-benzyl-5-nitro-3H-indol-2-one (437 mg, 91.96%) as a black solid. MS (ESI): mass calcd for C15H14N2O: 238.11 m/z, found 239.15[M+H]+.
Figure imgf000169_0001
1-(1-benzyl-2-oxo-3H-indol-5-yl)-3-tert-butylurea To a solution of 5-amino-1-benzyl-3H-indol-2-one (200 mg, 0.839 mmol, 1 equiv) and 2- isocyanato-2-methylpropane (124.2 mg, 1.253 mmol, 1.49 equiv) in DCM (2 mL) was added TEA (253.2 mg, 2.502 mmol, 2.98 equiv). The reaction was stirred at rt for 16h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 1-(1-benzyl-2-oxo-3H-indol-5-yl)-3-tert-butylurea (26.3 mg, 9.27%) as a white solid. LC/MS (ESI): mass calcd. for C20H23N3O2:337.18 m/z, found:338.00 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.08 (s, 1H), 7.42 (m, 1H), 7.23-7.34 (m, 5H), 7.02 (m, 1H), 6.70 (m, 1H), 5.88 (s, 1H), 4.84 (s, 2H), 3.63 (s, 2H), 1.27 (s, 9H). Example 32: 1-(1-benzyl-3-isopropyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)u rea Synthetic Scheme
Figure imgf000170_0001
3-isopropyl-3,4-dihydroquinolin-2(1H)-one To a solution of 3,4-dihydro-1H-quinolin-2-one (2 g, 13.589 mmol, 1 equiv) in THF (70 mL) was added 2M LDA (in THF) (17 mL) at -78 °C under N2. The resulting mixture was stirred 30 min at -78 °C. The reaction mixture was added 2-bromopropane (2.01 g, 16.343 mmol, 1.2 equiv) at -78 °C. The resulting mixture was stirred 30 min at -78 °C and then stirred 16 h at rt. The reaction mixture was quenched by water (50 mL) and extracted with EA (3 x 70 mL).The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 3-isopropyl-3,4-dihydro-1H-quinolin-2-one (1 g, 21.98%) as a white solid. MS (ESI): mass calcd. for C12H15NO, 189.12 m/z, found 190.30 [M+H]+.
Figure imgf000170_0002
3-isopropyl-7-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 3-isopropyl-3,4-dihydro-1H-quinolin-2-one (510 mg, 2.695 mmol, 1 equiv) in H2O (2 mL) and H2SO4 (9 mL) was added HNO3 (0.2 mL)at -10 °C. The resulting mixture was stirred at -10 °C until the starting material was totally consumed by LCMS. The reaction mixture was quenched by water. The mixture was acidified pH = 7 with NaHCO3 (aq.) and then extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 3-isopropyl- 7-nitro-3,4-dihydro-1H-quinolin-2-one (200 mg, 24.28%) as a white solid. MS (ESI): mass calcd. for C12H14N2O3, 234.10 m/z, found 235.05 [M+H]+ .
Figure imgf000171_0001
1-benzyl-3-isopropyl-7-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 3-isopropyl-7-nitro-3,4-dihydro-1H-quinolin-2-one (140 mg, 0.598 mmol, 1 equiv) in DMF (5 mL) was added K2CO3 (247.7 mg, 1.793 mmol, 3 equiv) and benzyl bromide (112.4 mg, 0.657 mmol, 1.1 equiv). The resulting mixture was stirred 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 1-benzyl-3-isopropyl-7-nitro-3,4-dihydroquinolin- 2(1H)-one (100 mg, 49.05%) as a yellow solid. MS (ESI): mass calcd. for C19H20N2O3324.15 m/z, found 325.05 [M+H]+.
Figure imgf000171_0002
7-amino-1-benzyl-3-isopropyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-3-isopropyl-7-nitro-3,4-dihydroquinolin-2(1H)-one (140 mg, 0.432 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 7-amino-1-benzyl-3- isopropyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 77.79%) as a yellow solid. MS (ESI): mass calcd. for C19H22N2O, 294.17 m/z, found 295.10 [M+H].
Figure imgf000172_0001
1-(1-benzyl-3-isopropyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 7-amino-1-benzyl-3-isopropyl-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.340 mmol, 1 equiv) in DCM (5 mL) was added TEA (103.1 mg, 1.019 mmol, 3 equiv) and 2- isocyanato-2-methylpropane (101.0 mg, 1.019 mmol, 3 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XSelect CSH Fluoro Phenyl, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 31% B to 46% B in 10.5 min; Wave Length: 220 nm; RT1(min): 11.2; Number Of Runs: 7) to afford 1-(1-benzyl-3-isopropyl-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert- butylurea (3.3 mg, 2.46%) as a white solid. MS (ESI): mass calcd. for C24H31N3O2, 393.24 m/z, found 394.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.13 – 7.37 (m, 6H), 6.96 (dd, J = 8.6, 2.4 Hz, 1H), 6.76 (d, J = 8.7 Hz, 1H), 5.93 (s, 1H), 5.09 (d, J = 3.5 Hz, 2H), 2.92 (dd, J = 15.7, 5.3 Hz, 1H), 2.77 (dd, J = 15.7, 8.1 Hz, 1H), 2.36 (q, J = 6.9 Hz, 1H), 1.98 (p, J = 6.4 Hz, 1H), 1.26 (s, 9H), 0.94 (dd, J = 17.0, 6.7 Hz, 6H). Example 33: 3-tert-butyl-1-{4-[(3-chlorophenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl}urea
Figure imgf000172_0002
4-[(3-chlorophenyl)methyl]-7-nitro-2H-1,4-benzoxazin-3-one To a solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.575 mmol, 1 equiv) and 1-(bromomethyl)-3-chlorobenzene (794 mg, 3.863 mmol, 1.5 equiv) in dimethylformamide (10 mL) was added Cs2CO3 (2.538 g, 7.766 mmol, 3.02 equiv). The reaction was stirred at rt for 3h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 4-[(3-chlorophenyl)methyl]-7-nitro-2H-1,4-benzoxazin-3-one (203 mg, 24.73%) as a white solid. MS (ESI): mass calcd for C15H11ClN2O4: 318.04 m/z, found 319.00[M+H]+.
Figure imgf000173_0001
7-amino-4-[(3E)-4-chloro-2-methylidenebut-3-en-1-yl]-2H-1,4-benzoxazin-3-one To a mixture of 4-[(3-chlorophenyl)methyl]-7-nitro-2H-1,4-benzoxazin-3-one (700 mg, 2.196 mmol, 1 equiv) and NH4Cl (1.17g, 21.960 mmol, 10 equiv) in ethyl alcohol/ water (10 mL, 10/1) was added Fe (1.23 g, 21.960 mmol, 10 equiv). The reaction was stirred at 60 °C for 6h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4-[(3E)- 4-chloro-2-methylidenebut-3-en-1-yl]-2H-1,4-benzoxazin-3-one (183 mg, 31.48%) as a white solid. MS (ESI): mass calcd for C15H13ClN2O2: 288.07m/z, found 289.00[M+H]+.
Figure imgf000173_0002
3-tert-butyl-1-{4-[(3-chlorophenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl}urea To a solution of 7-amino-4-[(3-chlorophenyl)methyl]-2H-1,4-benzoxazin-3-one (200 mg, 0.693 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (103.00 mg, 1.039 mmol, 1.5 equiv) in DCM (2 mL) was added TEA (210.29 mg, 2.079 mmol, 3 equiv). The reaction was stirred at rt for 3h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{4-[(3-chlorophenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl}urea (45.7 mg, 16.97%). LC/MS (ESI): mass calcd. for C20H22ClN3O3:387.13 m/z, found:387.95 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.29 – 7.41 (m, 3H), 7.22 (m, 2H), 6.86-6.85 (m, 2H), 5.94 (s, 1H), 5.11 (s, 2H), 4.76 (s, 2H), 1.26 (s, 9H). Example 34 and Example 35: (R)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro -2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000174_0001
7-nitro-4-(1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.575 mmol, 1 equiv) in DMF (20 mL) was added Cs2CO3 (1683.41 mg, 5.150 mmol, 2 equiv) and (1- bromoethyl)benzene (714.92 mg, 3.863 mmol, 1.5 equiv). The reaction was stirred at rt for 3 hours. Quenched with water (25 mL) and extracted with EA (3 x 25 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-nitro-4-(1- phenylethyl)-2H-1,4-benzoxazin-3-one (564 mg, 73.42%) as a yellow liquid. MS (ESI): mass calcd. for C16H14N2O4: 298.30m/z, found 299.00 [M+H]+.
Figure imgf000175_0001
7-amino-4-(1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (560 mg, 1.890 mmol, 1 equiv) in MeOH/H2O (22 mL, 10:1) was added Fe (527.68 mg, 9.450 mmol, 5 equiv) and NH4Cl (505.43 mg, 9.450 mmol, 5 equiv). The reaction was stirred at 80 oC for 1 hour. The reaction was quenched with water at rt. The resulting mixture was filtered, the filter cake was washed with EA (3 x 10 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 6-amino-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (382 mg, 75.89%) as a yellow oil. MS (ESI): mass calcd. for C16H16N2O2: 268.32m/z, found 269.15 [M+H]+.
Figure imgf000175_0002
1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of 6-amino-1-[(3-fluorophenyl)methyl]-3,4-dihydroquinolin-2-one (380 mg, 1.406 mmol, 1 equiv) in DCM (15 mL) was added 2-isocyanato-2-methylpropane (557.45 mg, 5.624 mmol, 4 equiv) and TEA (426.77 mg, 4.218 mmol, 3 equiv) at 0 oC. The reaction was stirred at rt for 1 days. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(3-fluorophenyl)methyl]-2-oxo-3,4-dihydroquinolin- 6-yl}urea (120 mg, 23.11%) .LC/MS (ESI): mass calcd. for C21H25N3O3:367.45 m/z, found:368.00 [M+H]+.
Figure imgf000176_0001
(R)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(3-oxo-4-(1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea A sample of 3-tert-butyl-1-[3-oxo-4-(1-phenylethyl)-2H-1,4-benzoxazin-7-yl]urea (120 mg) was separated by chiral-HPLC using a CHIRAL ART Amylose-SA, 2*25 cm, 5 μm column (eluent: 30% to 30% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer 3- tert-butyl-1-{3-oxo-4-[(1R)-1-phenylethyl]-2H-1,4-benzoxazin-7-yl}urea (37.1 mg) as a white solid. MS(ESI): mass calcd. for C21H25N3O3: 367.45, found: 368.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.99 – 8.31 (m, 1H), 7.11 – 7.44 (m, 6H), 6.43 – 6.67 (m, 2H), 6.04 – 6.24 (m, 1H), 5.80 – 5.98 (m, 1H), 4.55 – 4.80 (m, 2H), 1.67 – 1.80 (m, 3H), 1.22 – 1.31 (m, 9H). And to yield second enantiomer 3-tert-butyl-1-{3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4- benzoxazin-7-yl}urea (22.7 mg) as a white solid. MS(ESI): mass calcd. for C21H25N3O3: 367.45, found: 368.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.10 – 8.26 (m, 1H), 7.17 – 7.40 (m, 6H), 6.45 – 6.70 (m, 2H), 6.02 – 6.21 (m, 1H), 5.85 – 6.00 (m, 1H), 4.60 – 4.70 (m, 2H), 1.70 – 1.80 (m, 3H), 1.20 – 1.30 (m, 9H). Example 36 and Example 37: (R)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethyl)-3-oxo-3,4-di hydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethy l)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea
Figure imgf000176_0002
1-(1-bromoethyl)-3-chlorobenzene To a solution of 1-(3-chlorophenyl)ethanol (1.5 g, 9.578 mmol, 1 equiv) in Et2O (10 mL) was added PBr3 (5.19 g, 19.156 mmol, 2 equiv) at 0 oC. The reaction was stirred at rt for 2 hours. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. This resulted in 1-(1- bromoethyl)-3-chlorobenzene (762 mg, 36.24%) as a white oil. MS (ESI): mass calcd. for C8H8BrCl: 219.51 m/z, found 220.95 [M+H]+.
Figure imgf000177_0001
4-(1-(3-chlorophenyl)ethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.575 mmol, 1 equiv) in DMF (15 mL) was added Cs2CO3 (1683.4 mg, 5.151 mmol, 2 equiv) and 1-(1- bromoethyl)-3-chlorobenzene (565.2 mg, 2.575 mmol, 1 equiv). The final reaction mixture was stirred for 2 hours at rt. Quenched with water (25 mL) and extracted with EA (3 x 25 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 4-[1-(3-chlorophenyl)ethyl]-7-nitro-2H-1,4-benzoxazin-3-one (560 mg, 73.40%) as a yellow solid. MS (ESI): mass calcd. for C16H13ClN2O4: 332.74 m/z, found 333.00 [M+H]+.
Figure imgf000177_0002
7-amino-4-(1-(3-chlorophenyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 1-[1-(3-chlorophenyl)ethyl]-6-nitro-3,4-dihydroquinolin-2-one (560 mg, 1.693 mmol, 1 equiv) in MeOH/H2O (22 mL, 10:1) was added Fe (472.73 mg, 8.465 mmol, 5 equiv) and NH4Cl (452.80 mg, 8.465 mmol, 5 equiv). The reaction was stirred at 80 oC for 1 hour. The reaction was stirred for 1 hour at 80 oC. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[1-(3- chlorophenyl)ethyl]-3,4-dihydroquinolin-2-one (470 mg, 92.29%) as a yellow oil. MS (ESI): mass calcd. for C16H15ClN2O2: 302.76 m/z, found:303.00 [M+H]+.
Figure imgf000178_0001
1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea To a solution of 6-amino-1-[1-(3-chlorophenyl)ethyl]-3,4-dihydroquinolin-2-one (470 mg, 1.563 mmol, 1 equiv) in DCM (15 mL) was added 2-isocyanato-2-methylpropane (619.60 mg, 6.252 mmol, 4 equiv) and TEA (474.36 mg, 4.689 mmol, 3 equiv) at 0 oC. The reaction was stirred at rt for 1 day. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS, 30*150 mm, 5μm column (eluent: 49% to 69% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[1-(3-chlorophenyl)ethyl]-2-oxo-3,4- dihydroquinolin-6-yl}urea (470 mg, 74.8%) as a white solid. MS (ESI): mass calcd for C21H24ClN3O3:401.89 m/z, found:402.00 [M+H]+.
Figure imgf000178_0002
(R)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(1-(3-chlorophenyl)ethyl)-3-oxo- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-{4-[1-(3-chlorophenyl)ethyl]-3-oxo-2H-1,4-benzoxazin-7- yl}urea (200 mg) was separated by chiral-HPLC using a CHIRALPAK IG, 2*25 cm, 5 μm column (eluent: 30% to 30% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer example 3-tert-butyl-1-{4-[(1R)-1-(3-chlorophenyl)ethyl]-3-oxo-2H-1,4-benzoxazin- 7-yl}urea (48.0 mg) as a white solid. LC/MS: mass calcd. for C21H24ClN3O3: 401.89, found:402.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.10 – 8.30 (m, 1H), 7.30 – 7.44 (m, 3H), 7.20 – 7.30 (m, 2H), 6.51 – 6.72 (m, 2H), 5.99 – 6.16 (m, 1H), 5.81 – 5.99 (m, 1H), 4.59 – 4.78 (m, 2H), 1.68 – 1.78 (m, 3H), 1.24 – 1.29 (m, 9H). And to yield second enantiomer example 3-tert-butyl-1-{4-[(1S)-1-(3-chlorophenyl)ethyl]-3-oxo-2H-1,4-benzoxazin-7-yl}urea (26.2 mg) as a white solid. MS(ESI): mass calcd. for C21H24ClN3O3: 401.89, found:402.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.96 – 8.35 (m, 1H), 7.30 – 7.49 (m, 4H), 6.65 – 6.75 (m, 1H), 6.48 – 6.51 (m, 1H), 6.00 – 6.17 (m, 1H), 5.81 – 6.00 (m, 1H), 4.60 – 4.80 (m, 2H), 1.68 – 1.81 (m, 3H), 1.18 – 1.44 (m, 9H). Example 38: 1-(3-benzyl-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)-3-(tert-butyl)urea
Figure imgf000179_0001
3-benzyl-6-nitrobenzo[d]oxazol-2(3H)-one To a solution of 6-nitrobenzo[d]oxazol-2(3H)-one (300 mg, 1.666 mmol, 1 equiv) in CH3CN (6 mL) was added KI (276.49 mg, 1.666 mmol, 1 equiv), K2CO3 (788.35 mg, 5.664 mmol, 3.4 equiv) and benzyl bromide (112.4 mg, 0.657 mmol, 1.1 equiv). The resulting mixture was stirred for 5 h at 60 °C. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford 3-benzyl-6- nitrobenzo[d]oxazol-2(3H)-one (300 mg, 66.65%) as a yellow solid. MS (ESI): mass calcd. for C14H10N2O4270.06 m/z, found 271.05 [M+H]+.
Figure imgf000180_0001
6-amino-3-benzylbenzo[d]oxazol-2(3H)-one To a solution of 3-benzyl-6-nitrobenzo[d]oxazol-2(3H)-one (200 mg, 0.740 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (50 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 6-amino-3-benzylbenzo[d]oxazol-2(3H)-one (150 mg, 84.36%) as a white solid. MS (ESI): mass calcd. for C14H12N2O2, 240.09 m/z, found 241.20 [M+H]+.
Figure imgf000180_0002
1-(3-benzyl-2-oxo-2,3-dihydrobenzo[d]oxazol-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-3-benzylbenzo[d]oxazol-2(3H)-one (100 mg, 0.416 mmol, 1 equiv) in DCM (8 mL) was added TEA (126.3 mg, 1.249 mmol, 3 equiv) and 2-isocyanato-2- methylpropane (123.7 mg, 1.249 mmol, 3 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(3-benzyl-2-oxo-2,3-dihydrobenzo[d]oxazol-6- yl)-3-(tert-butyl)urea (12.8 mg, 9.03%) as a off-white solid. MS (ESI): mass calcd. for C19H21N3O3, 339.16 m/z, found 340.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.61 (d, J = 1.9 Hz, 1H), 7.27 – 7.39 (m, 5H), 7.01 (d, J = 8.5 Hz, 1H), 6.89 (dd, J = 8.5, 2.0 Hz, 1H), 5.95 (s, 1H), 4.99 (s, 2H), 1.27 (s, 9H). Example 39: 1-(tert-butyl)-3-(1-(2-fluoro-5-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahyd roquinolin-6-yl)urea
Figure imgf000181_0001
1-(2-fluoro-5-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydroquinolin-2(1H)-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added Cs2CO3 (1.695 g, 5.20 mmol, 2 equiv) and 2-(bromomethyl)-1-fluoro- 4-(trifluoromethyl)benzene (735.1 mg, 2.860 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 7-nitro-4-(3- (trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (533.4 mg, 55.66%) as a white solid. MS (ESI): mass calcd. for C17H12F4N2O3: 368.08 m/z, found 369.10 [M+H]+.
Figure imgf000181_0002
6-amino-1-(2-fluoro-5-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 7-nitro-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (200 mg, 0.543 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (50 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 6-amino-1-(2- fluoro-5-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one (130 mg, 71.0%) as a white solid. MS (ESI): mass calcd. for C17H14F4N2O, 338.10 m/z, found 339.05 [M+H]+.
Figure imgf000181_0003
1-(tert-butyl)-3-(1-(2-fluoro-5-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea To a stirred solution of 6-amino-1-(2-fluoro-5-(trifluoromethyl)benzyl)-3,4- dihydroquinolin-2(1H)-one (130 mg, 0.384 mmol, 1 equiv) in DCM (6 mL) & toluene (2 mL) was added 2-isocyanato-2-methylpropane (114.34 mg, 1.153 mmol, 3 equiv) stirred 16 h at 90 °C. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 45% B in 9 min, 45% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(2-fluoro-5- (trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (22.5 mg, 13.39%) as a white solid. MS (ESI): mass calcd. for C22H23F4N3O2, 437.17 m/z, found 438.15 [M+H]+ .1H NMR (300 MHz, DMSO-d6) δ 8.15 (s, 1H), 7.66 – 7.78 (m, 1H), 7.48 (d, J = 9.3 Hz, 1H), 7.29 – 7.41 (m, 2H), 7.03 (dd, J = 8.7, 2.5 Hz, 1H), 6.81 (d, J= 8.8 Hz, 1H), 5.94 (s, 1H), 5.18 (s, 2H), 2.81 – 2.96 (m, 2H), 2.60 – 2.71 (m, 2H), 1.26 (s, 9H). Example 40: 1-(tert-butyl)-3-(4-(3-methylbenzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazi n-7-yl)urea
Figure imgf000182_0001
4-(3-methylbenzyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.575 mmol, 1 equiv) in anhydrous dimethylformamide (10 mL) was added 1-(bromomethyl)-3- methylbenzene (619.59 mg, 3.348 mmol, 1.3 equiv) and Cs2CO3 (2.52 g, 7.725 mmol, 3 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. After completion of reaction, the resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (1 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA=3:1) to afford 4-[(3-methylphenyl)methyl]-7-nitro-2H-1,4- benzoxazin-3-one (750 mg, 97.63%) as a yellow solid. MS (ESI): mass calcd. for C16H14N2O4: 298.10 m/z, found 299.05 [M +H] +.
Figure imgf000183_0001
7-amino-4-(3-methylbenzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-[(3-methylphenyl)methyl]-7-nitro-2H-1,4-benzoxazin-3-one (300 mg, 1.006 mmol, 1 equiv) in 20 mL MeOH was added Pd/C (10%, 150mg) under nitrogen atmosphere in a 50 mL bar. The mixture was hydrogenated at room temperature for 30 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. This resulted in 7-amino-4-[(3-methylphenyl)methyl]-2H- 1,4-benzoxazin-3-one (250 mg, 92.65%) as a white solid. MS (ESI): mass calcd. for C16H16N2O2: 268.12 m/z, found 269.10 [M +H] +.
Figure imgf000183_0002
1-(tert-butyl)-3-(4-(3-methylb enzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a stirred solution of 7-amino-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin-3-one (50 mg, 0.186 mmol, 1 equiv) in DCM (5 mL) and Toluene (2.5 mL) was added 2-isocyanato-2- methylpropane (55.42 mg, 0.558 mmol, 3 equiv) at 90 °C and stirred for overnight. The reaction progress was monitored by TLC. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by HPLC with the following conditions Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0 to afford 3-tert-butyl-1-{4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4- benzoxazin-7-yl}urea (2.6 mg, 3.74%) as a white solid. MS (ESI):mass calcd. for C21H25N3O3: 367.19 m/z, found 368.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.19 (d, J = 6.2 Hz, 2H), 7.06 (d, J = 7.3 Hz, 3H), 6.85 (d, J = 8.6 Hz, 1H), 6.75 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.06 (s, 2H), 4.74 (s, 2H), 2.27 (s, 3H), 1.26 (s, 9H). Example 41 and Example 42: (R)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4] oxazin-7-yl)-3-(tert-butyl)urea and (S)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b ][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000184_0001
4-benzyl-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.441 mmol, 1 equiv) in DMF (10 mL) was added Cs2CO3 (941.99 mg, 2.882 mmol, 2 equiv) and benzyl bromide (369.72 mg, 2.162 mmol, 1.5 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 4-benzyl-2- methyl-7-nitro-2H-1,4-benzoxazin-3-one (377 mg, 87.70%) as a yellow solid. MS (ESI): mass calcd. for C16H14N2O4: 298.30m/z, found 299.00 [M+H]+.
Figure imgf000185_0001
7-amino-4-benzyl-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 6-nitro-1-(1-phenylethyl)-3,4-dihydroquinolin-2-one (560 mg, 1.890 mmol, 1 equiv) in MeOH/H2O (22 mL, 10:1) was added Fe (527.68 mg, 9.450 mmol, 5 equiv) and NH4Cl (505.43 mg, 9.450 mmol, 5 equiv). The reaction was stirred at 80 oC for 1 hour. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 10 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7- amino-4-benzyl-2-methyl-2H-1,4-benzoxazin-3-one (530 mg, 99.87%) as a yellow oil. MS (ESI): mass calcd. for C16H16N2O2: 268.32m/z, found 269.05 [M+H]+.
Figure imgf000185_0002
1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea To a solution of 7-amino-4-benzyl-2-methyl-2H-1,4-benzoxazin-3-one (540 mg, 2.013 mmol, 1 equiv) in DCM/Toluene (15 mL, 2:1) was added 2-isocyanato-2-methylpropane (798.04 mg, 8.052 mmol, 4 equiv). The reaction was stirred at 90 oC for 2 hours. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-(4-benzyl-2-methyl-3-oxo-2H-1,4- benzoxazin-7-yl)-3-tert-butylurea (498 mg, 67.34%) as a white solid. MS (ESI): mass calcd. for C21H25N3O3:367.45 m/z, found:368.10 [M+H]+.
Figure imgf000186_0001
(R)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert- butyl)urea and (S)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea A sample of 1-(4-benzyl-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl)-3-tert-butylurea (200 mg) was separted by chiral-HPLC using a CHIRALPAK IE, 2*25 cm, 5 μm column (eluent: 15% to 15% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer example 1- [(2R)-4-benzyl-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea (29.8 mg) as a white solid. MS(ESI): mass calcd. for C21H25N3O3: 367.45, found: 368.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.14 – 8.32 (m, 1H), 7.15 – 7.45 (m, 6H), 6.68 – 6.98 (m, 2H), 5.79 – 6.09 (m, 1H), 5.00 – 5.17 (m, 2H), 4.70 – 4.90 (m, 1H), 1.43 – 1.65 (m, 3H), 1.18 – 1.31 (m, 9H). And to yield second enantiomer example 1-[(2S)-4-benzyl-2-methyl-3-oxo-2H-1,4-benzoxazin- 7-yl]-3-tert-butylurea (25.5 mg) as a white solid. MS(ESI): mass calcd. for C21H25N3O3: 367.45, found: 368.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.08 – 8.31 (m, 1H), 7.30 – 7.39 (m, 2H), 7.18 – 7.30 (m, 4H), 6.71 – 6.92 (m, 2H), 5.75 – 6.07 (m, 1H), 5.01 – 5.19 (m, 2H), 4.72 – 4.87 (m, 1H), 1.42 – 1.53 (m, 3H), 1.21 – 1.33 (m, 9H). Example 43 and Example 44: (R)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-methyl-3-oxo-3,4-di hydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-met hyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea
Figure imgf000186_0002
4-(3-chlorobenzyl)-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.402 mmol, 1 equiv) in DMF (10 mL) was added Cs2CO3 (1569.98 mg, 4.804 mmol, 2 equiv) and 1- (bromomethyl)-3-chlorobenzene (740.30 mg, 3.603 mmol, 1.5 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 4- [(3-chlorophenyl)methyl]-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (790 mg, 98.85%) as a yellow solid. MS (ESI): mass calcd. for C16H13ClN2O4: 332.74 m/z, found 332.95 [M+H]+.
Figure imgf000187_0001
7-amino-4-(3-chlorobenzyl)-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-[(3-chlorophenyl)methyl]-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (570 mg, 1.713 mmol, 1 equiv) in EtOH (20 mL) was added SnCl2 (1641.36 mg, 8.565 mmol, 5 equiv). The reaction was stirred at 60 oC for 1 hour. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4-[(3-chlorophenyl)methyl]-2-methyl-2H-1,4- benzoxazin-3-one (500 mg, 96.41%) as a yellow solid. MS (ESI): mass calcd. for C16H15ClN2O2: 302.76m/z, found 302.95 [M+H]+.
Figure imgf000187_0002
1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea To a solution of 7-amino-4-[(3-chlorophenyl)methyl]-2-methyl-2H-1,4-benzoxazin-3-one (500 mg, 1.651 mmol, 1 equiv) in DCM/Toluene (25 mL, 2:1) was added 2-isocyanato-2- methylpropane (654.86 mg, 6.604 mmol, 4 equiv). The reaction was stirred at 90 oC for 2 hours. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-tert-butyl-1-{4- [(3-chlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl}urea (170 mg, 25.61%) as a white solid. LC/MS (ESI): mass calcd. For C21H24ClN3O3: 401.89 m/z, found:402.05 [M+H]+.
Figure imgf000188_0001
(R)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and (S)-1-(tert-butyl)-3-(4-(3-chlorobenzyl)-2-methyl-3-oxo- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-{2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4- benzoxazin-7-yl}urea (170 mg) was separated by chiral-HPLC using a CHIRALPAK IE, 2*25 cm, 5 μm column (eluent: 20% to 20% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer example 3-tert-butyl-1-[(2R)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4- benzoxazin-7-yl]urea (23 mg) as a white solid. MS(ESI): mass calcd. for C21H24ClN3O3: 401.89, found: 402.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.93 – 8.31 (m, 1H), 7.29 – 7.44 (m, 3H), 7.13 – 7.29 (m, 2H), 6.68 – 6.99 (m, 2H), 5.84 – 6.08 (m, 1H), 4.97 – 5.21 (m, 2H), 4.71 – 4.89 (m, 1H), 1.40 – 1.58 (m, 3H), 1.21 – 1.30 (m, 9H). And to yield second enantiomer example 3-tert-butyl-1-[(2S)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl]urea (24.5 mg) as a white solid. MS(ESI): mass calcd. for C21H24ClN3O3: 401.89, found: 402.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.00 – 8.32 (m, 1H), 7.30 – 7.49 (m, 3H), 7.19 – 7.30 (m, 2H), 6.70 – 6.92 (m, 2H), 5.81 – 6.06 (m, 1H), 5.00 – 5.21 (m, 2H), 4.66 – 4.91 (m, 1H), 1.43 – 1.54 (m, 3H), 1.25 – 1.30 (m, 9H). Example 45 and Example 45: 3-tert-butyl-1-{3-methyl-2-oxo-1-[(1S)-1-phenylethyl]-4H- quinazolin-6-yl}urea and 3-tert-butyl-1-{3-methyl-2-oxo-1-[(1R)-1-phenylethyl]-4H- quinazolin-6-yl}urea
Figure imgf000189_0001
3-methyl-6-nitro-1-(1-phenylethyl)-4H-quinazolin-2-one To a solution of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (600 mg, 2.896 mmol, 1 equiv) and (1-bromoethyl)benzene (800 mg, 4.323 mmol, 1.49 equiv) in dimethylformamide (30 mL) was added Cs2CO3 (2802.47 mg, 8.575 mmol, 2.96 equiv). The reaction was stirred at rt for 3h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-methyl-6-nitro- 1-(1-phenylethyl)-4H-quinazolin-2-one (765 mg, 84.85%) as a white solid. MS (ESI): mass calcd for C17H19N3O2: 311.13 m/z, found 312.20[M+H]+.
Figure imgf000189_0002
6-amino-3-methyl-1-(1-phenylethyl)-4H-quinazolin-2-one To a solution of 3-methyl-6-nitro-1-(1-phenylethyl)-4H-quinazolin-2-one (755 mg, 2.425 mmol, 1 equiv) in ethyl alcohol/ water (10 mL, 10/1) was added iron (1354.24 mg, 24.250 mmol, 10 equiv) and NH4Cl (1297.13 mg, 24.250 mmol, 10 equiv). The reaction was stirred at 80 °C for 3h. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to 6-amino-3-methyl-1- (1-phenylethyl)-4H-quinazolin-2-one (200 mg, 29.31%) as a white solid. MS (ESI): mass calcd forC17H19N3O: 281.15m/z, found 282.20 [M+H]+.
Figure imgf000190_0001
1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinazolin-6-yl)urea To a solution of 6-amino-3-methyl-1-(1-phenylethyl)-4H-quinazolin-2-one (190 mg, 0.675 mmol, 1 equiv) in DCM/toluene (5 mL2/1) was added2-isocyanato-2-methylpropane (220.64 mg, 2.226 mmol, 3.30 equiv) The reaction was stirred at 90°C for 3h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-tert-butyl-1-{3-methyl-2-oxo-1- [(1R)-1-phenylethyl]-4H-quinazolin-6-yl}urea (178 mg, 69.3%) as a white solidLC/MS (ESI): mass calcd. for C22H28N4O2:380.22 m/z, found:381.10 [M+H]+.
Figure imgf000190_0002
(S)-1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-phenylethyl)-1,2,3,4-tetrahydroquinazolin-6- yl)urea and (R)-1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-phenylethyl)-1,2,3,4- tetrahydroquinazolin-6-yl)urea. A sample of 1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-phenylethyl)-1,2,3,4- tetrahydroquinazolin-6-yl)urea (170 mg) was separated by chiral-HPLC using a (R, R)-WHELK- O1-Kromasil 2.11*25 cm, 5 μm column (eluent: MtBE(0.5% 2M NH3-MeOH)--HPLC and Hex(EtOH--HPLC) to yield first example cyclopentyl 3-tert-butyl-1-{3-methyl-2-oxo-1-[(1R)-1- phenylethyl]-4H-quinazolin-6-yl}urea (26.6 mg). LC/MS (ESI): mass calcd. for C22H28N4O2:380.22 m/z, found:381.05 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.26 – 7.37 (m, 5H), 6.80 (m, 1H), 6.37 (m, 1H), 5.92 (m, 2H), 4.32 (m, 2H), 2.92 (s, 3H), 1.74 (m, 3H), 1.19-1.25 (s, 10H). And to yield second enantiomer example cyclopentyl 3-tert-butyl-1- {3-methyl-2-oxo-1-[(1S)-1-phenylethyl]-4H-quinazolin-6-yl}urea (41.6 mg). LC/MS (ESI): mass calcd. for C22H28N4O2:380.22 m/z, found:381.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.26 – 7.37 (m, 5H), 6.80 (m, 1H), 6.37 (m, 1H), 5.92 (m, 2H), 4.32 (m, 2H), 2.92 (s, 3H), 1.74 (m, 3H), 1.19-1.25 (m, 10H). Example 46 and Example 52: (S)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3-methyl-2- oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea and (R)-1-(tert-butyl)-3-(1-(1-(4- chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea
Figure imgf000191_0001
1-(1-(3-chlorophenyl)ethyl)-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (500 mg, 2.413 mmol, 1 equiv) and Cs2CO3 (1.57 g, 4.826 mmol, 2 equiv) in DMF (10 mL) was added 1-(1- bromoethyl)-3-chlorobenzene (635.68 mg, 2.896 mmol, 1.2 equiv) in portions at 25°C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1-(1-(3- chlorophenyl)ethyl)-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one (640 mg, 76.70%) as a yellow solid. MS (ESI): mass calcd. for C17H16ClN3O3: 345.09 m/z, found 345.78 [M+H] +.
Figure imgf000191_0002
6-amino-1-(1-(3-chlorophenyl)ethyl)-3-methyl-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 1-(1-(3-chlorophenyl)ethyl)-3-methyl-6-nitro-3,4- dihydroquinazolin-2(1H)-one (630 mg, 1.822 mmol, 1 equiv) and Fe (1017.48 mg, 18.220 mmol, 10 equiv) in EtOH (8 mL) and H2O (0.8 mL) was added NH4Cl (974.57 mg, 18.220 mmol, 10 equiv) in portions at 70 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-amino-1-(1-(3-chlorophenyl)ethyl)-3-methyl-3,4- dihydroquinazolin-2(1H)-one (240 mg, 41.71%) as a yellow solid. MS (ESI): mass calcd. for C16H16ClN3O: 315.11 m/z, found 315.80 [M +H] +.
Figure imgf000192_0001
1-(tert-butyl)-3-(1-(1-(3-chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6 -yl)urea To a stirred mixture of 6-amino-1-(1-(3-chlorophenyl)ethyl)-3-methyl-3,4- dihydroquinazolin-2(1H)-one(242 mg, 0.766 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (151.93 mg, 1.532 mmol, 2 equiv) in DCM (5 mL) was added TEA (232.63 mg, 2.298 mmol, 3 equiv) at 25 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (30 mL). The resulting mixture was extracted with EA (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1-(tert-butyl)- 3-(1-(1-(3-chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea (110 mg, 34.60%) as a white solid. MS (ESI): mass calcd. for C22H27ClN4O2: 414.18 m/z, found 414.93[M +H] +
Figure imgf000192_0002
(S)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4- tetrahydroquinazolin-6-yl)urea and (R)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3- methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea The 1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3-methyl-2-oxo-1,2,3,4- tetrahydroquinazolin-6-yl)urea (110 mg) was separated by chiral-HPLC using a CHIRALPAK ID, 2*25 cm, 5 μm column (eluent: 10% to 10% (v/v) : EtOH--HPLC and Hex(0.5% 2M NH3- MeOH) to yield first enantiomer example (R)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3- methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea (24.4 mg, 7.57%) as a white solid. LC/MS: mass calcd for C22H27ClN4O2: 414.18, found: 415.00 [M+H]+.1H-NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.32 – 7.39 (m, 1H), 7.20 – 7.30 (m, 4H), 6.80 – 6.92 (m, 1H), 6.39 (d, J = 8.8 Hz, 1H), 5.75 – 5.96 (m, 2H), 4.32 (s, 2H), 2.92 (s, 3H), 1.74 (d, J = 7.1 Hz, 3H), 1.26 (s, 9H). And to yield second enantiomer example (S)-1-(tert-butyl)-3-(1-(1-(4-chlorophenyl)ethyl)-3-methyl- 2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea (16.4 mg, 5.19%) as a white soild. LC/MS: mass calcd. for C22H27ClN4O2: 414.18, found: 415.00 [M+H]+.1H-NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.20 – 7.42 (m, 5H), 6.81 – 6.91 (m, 1H), 6.38 (d, J = 8.8 Hz, 1H), 5.78 – 5.96 (m, 2H), 4.32 (s, 2H), 2.91 (s, 3H), 1.73 (d, J = 7.1 Hz, 3H), 1.25 (s, 9H). Example 47: 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazo lin-6-yl)urea
Figure imgf000193_0001
3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 3-methyl-1,4-dihydroquinazolin-2-one (1 g, 6.166 mmol, 1 equiv) in H2SO4 (30 mL) was added KNO3 (0.50 g, 4.933 mmol, 0.8 equiv) in portions at 0°C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 3-methyl-6- nitro-3,4-dihydroquinazolin-2(1H)-one (400 mg, 31.31%) as a white solid. MS (ESI): mass calcd. for C9H9N3O3: 207.06 m/z, found 208.19 [M+H] +.
Figure imgf000194_0001
1-(3-chlorobenzyl)-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one (300 mg, 1.448 mmol, 1 equiv) and 1-(bromomethyl)-3-chlorobenzene (357.03 mg, 1.738 mmol, 1.2 equiv) in DMF (5 mL) was added Cs2CO3 (1419.69 mg, 4.344 mmol, 3 equiv) in portions at 25 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1-(3-chlorobenzyl)-3-methyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one (400 mg, 83.27%) as a yellow solid. MS (ESI): mass calcd. for C16H14ClN3O3: 331.07 m/z, found 331.76 [M +H] +.
Figure imgf000194_0002
6-amino-1-(3-chlorobenzyl)-3-methyl-3,4-dihydroquinazolin-2(1H)-one To a stirred mixture of 1-(3-chlorobenzyl)-3-methyl-6-nitro-3,4-dihydroquinazolin- 2(1H)-one (395 mg, 1.191 mmol, 1 equiv) and Fe (664.90 mg, 11.910 mmol, 10 equiv) in EtOH (5 mL) and H2O (0.5 mL) was added NH4Cl (636.86 mg, 11.910 mmol, 10 equiv) in portions at 25 degrees C under air atmosphere. The resulting mixture was stirred for 2h at 60 degrees C. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-amino-1-(3-chlorobenzyl)-3- methyl-3,4-dihydroquinazolin-2(1H)-one (160 mg, 44.53%) as a yellow solid. MS (ESI): mass calcd. for C16H16ClN3O: 301.10 m/z, found 301.77[M +H] +.
Figure imgf000195_0001
1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea To a stirred mixture of 6-amino-1-(3-chlorobenzyl)-3-methyl-3,4-dihydroquinazolin-2(1H)- one (150 mg, 0.497 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (98.55 mg, 0.994 mmol, 2 equiv) in dichloromethane (1 mL) was added TEA (150.90 mg, 1.491 mmol, 3 equiv) dropwise at 0 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (30 mL). The resulting mixture was extracted with EA (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford the 1-(tert- butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)urea(31.9 mg, 15.65%) as a white solid. MS (ESI): mass calcd. for C21H25ClN4O2: 400.17 m/z, found 401.00[M +H]+.1H-NMR (300 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.22 – 7.40 (m, 4H), 7.18 (d, J = 7.4 Hz, 1H), 6.90 – 6.99 (m, 1H), 6.56 (d, J = 8.7 Hz, 1H), 5.91 (s, 1H), 5.02 (s, 2H), 4.42 (s, 2H), 2.95 (s, 3H), 1.26 (s, 9H). Example 48: 1-(tert-butyl)-3-(1-(2-fluoro-3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin
Figure imgf000195_0002
1-(2-fluoro-3-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 1.561 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-2-fluoro-3-methylbenzene (412.08 mg, 2.029 mmol, 1.3 equiv) and Cs2CO3 (1.53 g, 4.683 mmol, 3 equiv) at rt and stirred for overnight. After completion of reaction, the resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (1 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 1-(2-fluoro-3- methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (420 mg, 85.60%) as a yellow solid. MS (ESI): mass calcd. for C17H15FN2O3: 314.11 m/z, found 315.05 [M +H] +.
Figure imgf000196_0001
6-amino-1-(2-fluoro-3-methylbenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(2-fluoro-3-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.636 mmol, 1 equiv) in 20 mL MeOH was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-(2-fluoro-3-methylbenzyl)-3,4-dihydroquinolin-2(1H)-one (160 mg, 88.44%) as a white solid. MS (ESI): mass calcd. for C17H17FN2O: 284.13 m/z, found 285.10 [M +H] +.
Figure imgf000196_0002
1-(tert-butyl)-3-(1-(2-fluoro-3-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of 6-amino-1-[(2-fluoro-3-methylphenyl)methyl]-3,4- dihydroquinolin-2-one (80 mg, 0.281 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2- methylpropane (139.46 mg, 1.405 mmol, 5 equiv) and Et3N (85.41 mg, 0.843 mmol, 3 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by Prep-HPLC with the following conditions Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0 to afford 3-tert-butyl-1-{1-[(2-fluoro- 3-methylphenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea (29.0 mg, 26.78%) as a white solid. MS (ESI): mass calcd. for C22H26FN3O2: 383.20 m/z, found 384.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.34 (d, J = 2.4 Hz, 1H), 7.15 (t, J = 7.3 Hz, 1H), 6.92 – 7.04 (m, 2H), 6.76 – 6.88 (m, 1H), 6.71 (d, J = 8.7 Hz, 1H), 5.93 (s, 1H), 5.09 (s, 2H), 2.89 (dd, J = 8.7, 5.8 Hz, 2H), 2.65 (dd, J = 8.7, 5.8 Hz, 2H), 2.25 (d, J = 2.0 Hz, 3H), 1.27 (s, 9H). Example 49: 1-(tert-butyl)-3-(1-(2-fluoro-5-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin
Figure imgf000197_0001
1-(2-fluoro-5-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 1.561 mmol, 1 equiv) in DMF (5 mL) was added 2-(bromomethyl)-1-fluoro-4-methylbenzene (412.08 mg, 2.029 mmol, 1.3 equiv) and Cs2CO3 (1.53 g, 4.683 mmol, 3 equiv) at rt and stirred for overnight. After completion of reaction, the resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (1 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 1-(2-fluoro-5- methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (400 mg, 81.52%) as a yellow solid. MS (ESI): mass calcd. for C17H15FN2O: 314.11 m/z, found 315.05 [M +H] +.
Figure imgf000197_0002
6-amino-1-(2-fluoro-5-methylbenzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(2-fluoro-5-methylbenzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.636 mmol, 1 equiv) in 20 mL MeOH was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 30 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-(2-fluoro-5-methylbenzyl)-3,4-dihydroquinolin-2(1H)-one (160 mg, 88.40%) as a white solid. MS (ESI): mass calcd. for C17H17FN2O: 284.13 m/z, found 285.10 [M +H] +.
Figure imgf000198_0001
1-(tert-butyl)-3-(1-(2-fluoro-5-methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of 6-amino-1-[(2-fluoro-5-methylphenyl)methyl]-3,4- dihydroquinolin-2-one (80 mg, 0.281 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2- methylpropane (139.46 mg, 1.405 mmol, 5 equiv) and Et3N (85.41 mg, 0.843 mmol, 3 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by HPLC with the following conditions Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0 to afford 1-(tert-butyl)-3-(1-(2-fluoro-5- methylbenzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (8.1 mg, 7.49%) as a white solid. MS (ESI): mass calcd. for C22H26FN3O2: 383.20 m/z, found 384.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.34 (d, J = 2.4 Hz, 1H), 7.16 – 7.04 (m, 2H), 7.00 (dd, J = 8.7, 2.5 Hz, 1H), 6.81 (d, J = 7.5 Hz, 1H), 6.71 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.06 (s, 2H), 2.89 (t, J = 7.2 Hz, 2H), 2.66 (dd, J = 8.8, 5.8 Hz, 2H), 2.19 (s, 3H), 1.27 (s, 9H). Example 50: 1-(tert-butyl)-3-(3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H-benzo[b] [1,4]oxazin-7-yl)urea
Figure imgf000199_0001
7-nitro-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (500 mg, 2.575 mmol, 1 equiv) in DMF (10 mL) was added Cs2CO3 (1.678 g, 5.150 mmol, 2 equiv) and 1- (bromomethyl)-3-(trifluoromethyl)benzene (676.8 mg, 2.832 mmol, 1.1 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 7-nitro-4- (3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (473.8 mg, 52.23%) as a white solid. MS (ESI): mass calcd. for C16H11F3N2O4: 352.07 m/z, found 353.05 [M+H]+.
Figure imgf000199_0002
7-amino-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 0.852 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (100 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 7-amino-4-(3- (trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (263.6 mg, 96.0%) as a white solid. MS (ESI): mass calcd. for C16H13F3N2O2, 322.09 m/z, found 323.05 [M+H]+.
Figure imgf000199_0003
1-(tert-butyl)-3-(3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea To a stirred solution of 7-amino-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (90 mg, 0.279 mmol, 1 equiv) in DCM (6 mL) & toluene (2mL) was added 2- isocyanato-2-methylpropane (82.97 mg, 0.837 mmol, 3 equiv) stirred 16 h at 90 °C. The reaction mixture was concentrated under vacuum to yield a crude product which was directly purified Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea (2.1 mg, 1.7%) as a white solid. MS (ESI): mass calcd. for C21H22F3N3O3, 421.16 m/z, found 422.15 [M+H]+ . 1H NMR (300 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.49 – 7.70 (m, 4H), 7.21 (d, J = 2.3 Hz, 1H), 6.87 – 6.90 (m, 1H), 6.75 – 6.79 (m, 1H), 5.94 (s, 1H), 5.19 – 5.23 (m, 2H), 4.75 – 4.78 (m, 2H), 1.23 – 1.29 (m, 9H). Example 51: 1-(1-benzyl-3-(hydroxymethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000200_0001
1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylic acid In a 50-mL round bottom flask, to a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2- one (500 mg, 1.771 mmol, 1 equiv) in tetrahydrofuran (5 mL, 0.069 mmol, 0.04 equiv) was added dropwise 2M LDA (in THF) (1 mL, 1.948 mmol, 1.1 equiv) at -78 °C under N2. The reaction mixture was stirred at -78 °C for 30 min. The reaction was quenched with dry ice. The mixture was stirred for 10 min at rt. NH4Cl (aq.) was added into the mixture. The mixture was acidified pH = 5 with HCl (1N). The solution was extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified by silica gel column, eluted with DCM /MeOH (8:1) to afford 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylic acid (150 mg, 25.95%) as an off-white solid. MS (ESI): mass calcd. for C17H14N2O5, 326.09 m/z, found 327.05 [M+H]+.
Figure imgf000201_0001
methyl 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylate A solution of 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylic acid (500 mg, 1.532 mmol, 1 equiv) in methanol (10 mL) was added sulfuric acid (0.1 mL) at rt. The resulting mixture was stirred for 2 h at 70 °C. Quench with water (10 mL) and extract with ethyl acetate (3 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE /EA (3:1) to afford methyl 1-benzyl-6-nitro-2-oxo- 1,2,3,4-tetrahydroquinoline-3-carboxylate (150 mg, 28.76%) as white solid. MS (ESI): mass calcd. for C18H16N2O5, 340.11 m/z, found 341.05 [M+H]+.
Figure imgf000201_0002
methyl 6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylate To a solution of methyl 1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylate (200 mg, 0.588 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford methyl 6- amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylate (150 mg, 79.36%) as a yellow solid. MS (ESI): mass calcd. for C18H18N2O3, 310.13 m/z, found 311.15 [M+H]+.
Figure imgf000202_0001
methyl 1-benzyl-6-(3-(tert-butyl)ureido)-2-oxo-1,2,3,4-tetrahydroquinoline-3-carboxylate To a stirred solution of methyl 6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinoline-3- carboxylate (150 mg, 0.483 mmol, 1 equiv) in DCM (8 mL) was added 2-isocyanato-2- methylpropane (239.5 mg, 2.417 mmol, 5 equiv) and TEA (146.7 mg, 1.450 mmol, 3 equiv) stirred overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) to afford methyl 1-benzyl-6-[(tert- butylcarbamoyl)amino]-2-oxo-3,4-dihydroquinoline-3-carboxylate (70 mg, 31.95%) as a yellow solid. MS (ESI): mass calcd. for C23H27N3O4, 409.20 m/z, found 410.25 [M+H]+.
Figure imgf000202_0002
1-(1-benzyl-3-(hydroxymethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a stirred solution of methyl 1-benzyl-6-[(tert-butylcarbamoyl)amino]-2-oxo-3,4- dihydroquinoline-3-carboxylate (70 mg, 0.171 mmol, 1 equiv) in MeOH (5 mL) was added NaBH4 (64.6 mg, 1.709 mmol, 10 equiv) stirred 3 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-[1- benzyl-3-(hydroxymethyl)-2-oxo-3,4-dihydroquinolin-6-yl]-3-tert-butylurea (9.9 mg, 15.13%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3, 381.21 m/z, found 382.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.25 – 7.36 (m, 3H), 7.12 – 7.24 (m, 3H), 6.96 (dd, J = 8.6, 2.4 Hz, 1H), 6.74 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.08 (s, 2H), 4.78 (s, 1H), 3.68 – 3.83 (m, 1H), 3.59 (d, J = 10.8 Hz, 1H), 2.98 (dd, J = 15.4, 5.6 Hz, 1H), 2.75 – 2.90 (m, 1H), 2.65 – 2.75 (m, 1H), 1.26 (s, 9H). Example 53: 3-tert-butyl-1-{1-[(2-cyano-3-methylphenyl)methyl]-2-oxo-3,4-dihydroquinoli
Figure imgf000203_0001
2-methyl-6-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile To a stirred solution of 2-(bromomethyl)-6-methylbenzonitrile (400 mg, 1.904 mmol, 1 equiv) and 6-nitro-3,4-dihydro-1H-quinolin-2-one (548.88 mg, 2.856 mmol, 1.5 equiv) in DMF (15 mL) was added K2CO3 (662.69 mg, 4.760 mmol, 2.5 equiv) in portions at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 2 h at room temperature. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 2-methyl-6-[(6- nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile (400 mg, 65.38%) as a white solid. MS: mass calcd. for C18H15N3O3: 321.11 m/z, found 322.05 [M +H] +.
Figure imgf000203_0002
2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-6-methylbenzonitrile To a stirred solution of 2-methyl-6-[(6-nitro-2-oxo-3,4-dihydroquinolin-1- yl)methyl]benzonitrile (400 mg, 1.245 mmol, 1 equiv) in EtOH/H2O (11 mL, 10/1) was added Fe (695.16 mg, 12.450 mmol, 10 equiv) and NH4Cl (665.85 mg, 12.450 mmol, 10 equiv) in portions at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 2 h at 60°C. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The resulting mixture was concentrated under vacuum. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 2-[(6-amino-2- oxo-3,4-dihydroquinolin-1-yl)methyl]-6-methylbenzonitrile (300 mg, 82.72%) as a light yellow solid.MS: mass calcd. for C18H17N3O: 291.14 m/z, found 292.10 [M +H] +.
Figure imgf000204_0001
3-tert-butyl-1-{1-[(2-cyano-3-methylphenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea To a stirred solution of 2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-6- methylbenzonitrile (150 mg, 0.515 mmol, 1 equiv) in toluene/DCM (10 mL, 1/2) was added 2- isocyanato-2-methylpropane (153.11 mg, 1.545 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 3 h at 90°C. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 3-tert-butyl-1-{1-[(2-cyano-3-methylphenyl)methyl]-2-oxo-3,4- dihydroquinolin-6-yl}urea (10.3 mg, 5.10%) as a white solid. MS: mass calcd. For C23H26N4O2: 390.21 m/z, found 391.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.47 (t, J = 7.8 Hz, 1H), 7.35 – 7.33 (m, 2H),6.99 – 6.97 (m, 2H), 6.66 (d, J = 8.7 Hz, 1H), 5.92 (s, 1H), 5.21 (s, 2H), 2.91 (t, J = 7.8 Hz, 2H), 2.67 (m, 2H), 2.51 (s, 3H), 1.26 (s, 9H). Example 54: 1-(tert-butyl)-3-(1-(2-fluoro-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea
Figure imgf000205_0001
1-(2-fluoro-3-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.602 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (431.50 mg, 3.122 mmol, 2 equiv) and 1- (bromomethyl)-2-fluoro-3-(trifluoromethyl)benzene (601.86 mg, 2.341 mmol, 1.5 equiv). The resulting mixture was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 20 mL), brine (2 x 20 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 1- (2-fluoro-3-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (450 mg, 78.27%) as a yellow solid. MS (ESI): mass calcd. for C17H12F4N2O3: 368.08 m/z, found 369.05 [M+H]+.
Figure imgf000205_0002
6-amino-1-(2-fluoro-3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(2-fluoro-3-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin- 2(1H)-one (430 mg, 1.168 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (100 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 6- amino-1-(2-fluoro-3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin-2(1H)-one (370 mg, 93.67%) as a yellow solid. MS (ESI): mass calcd. for C17H14F4N2O: 338.10 m/z, found 339.15 [M+H]+.
Figure imgf000205_0003
1-(tert-butyl)-3-(1-(2-fluoro-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea To a solution of 6-amino-1-(2-fluoro-3-(trifluoromethyl)benzyl)-3,4-dihydroquinolin- 2(1H)-one (150 mg, 0.443 mmol, 1 equiv) in DCM (10 mL) was added TEA (179.47 mg, 1.772 mmol, 4 equiv) and 2-isocyanato-2-methylpropane (219.77 mg, 2.215 mmol, 5 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to afford 1-(tert-butyl)-3-(1-(2- fluoro-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (45.3 mg, 23.26%) as a white solid. MS (ESI): mass calcd. for C22H23F4N3O2: 437.17, found: 437.95 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.14 (s, 1H), 7.61 – 7.72 (m, 1H), 7.25 – 7.40 (m, 3H), 6.96 – 7.05 (m, 1H), 6.77 (d, J = 8.8 Hz, 1H), 5.93 (s, 1H), 5.18 (s, 2H), 2.85 – 2.95 (m, 2H), 2.61 – 2.71 (m, 2H), 1.26 (s, 9H). Example 55: 3-tert-butyl-1-(1-{[2-cyano-5-(trifluoromethyl) phenyl] methyl}-2-oxo-3,4- dihydroquinolin-6-yl) urea Synthetic Scheme
Figure imgf000206_0001
Figure imgf000207_0001
1-bromo-2-(bromomethyl)-4-(trifluoromethyl)benzene To a stirred solution of [2-bromo-5-(trifluoromethyl) phenyl] methanol (2 g, 7.842 mmol, 1 equiv) in DCM (20 mL) was added phosphorus tribromide (2.12 g, 7.842 mmol, 1 equiv) in portions at 0°C under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. This resulted in 1-bromo-2- (bromomethyl)-4-(trifluoromethyl) benzene (1.3 g, 52.14%) as a yellow oil. The crude product was used in the next step directly without further purification.
Figure imgf000207_0002
1-{[2-bromo-5-(trifluoromethyl) phenyl] methyl}-6-nitro-3,4-dihydroquinolin-2-one To a stirred solution of 1-bromo-2-(bromomethyl)-4-(trifluoromethyl) benzene (1.39 g, 4.372 mmol, 1.2 equiv) and 6-nitro-3,4-dihydro-1H-quinolin-2-one (700.16 mg, 3.643 mmol, 1.0 equiv) in DMF (15 mL) was added K2CO3 (1014.40 mg, 7.287 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1-{[2-bromo-5-(trifluoromethyl) phenyl] methyl}-6-nitro-3,4- dihydroquinolin-2-one (1.4 g, 89.76%) as a yellow oil. LC/MS: mass calcd. for C17H12BrF3N2O3: 428.00, found: 428.95 [M+H] +.
Figure imgf000208_0001
2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl) methyl]-4-(trifluoromethyl)benzonitrile To a stirred solution of 1-{[2-bromo-5-(trifluoromethyl) phenyl] methyl}-6-nitro-3,4- dihydroquinolin-2-one (600 mg, 1.398 mmol, 1 equiv) and Zn(CN)2 (196.98 mg, 1.678 mmol, 1.2 equiv) in DMF (10 mL) was added BrettPhos Pd G3 (253.45 mg, 0.280 mmol, 0.2 equiv) and BrettPhos (150.08 mg, 0.280 mmol, 0.2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at 120°C under nitrogen atmosphere. After cooling down to rt, the reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 2-[(6-nitro-2-oxo-3,4- dihydroquinolin-1-yl) methyl]-4-(trifluoromethyl) benzonitrile (100 mg, 19.06%) as a yellow oil. Desired product could be detected by TLC.
Figure imgf000208_0002
2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl) methyl]-4-(trifluoromethyl)benzonitrile To a stirred solution of 2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl) methyl]-4- (trifluoromethyl) benzonitrile (100 mg, 0.266 mmol, 1 equiv) in AcOH was added Zn (209.04 mg, 3.192 mmol, 12 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 2-[(6-amino-2-oxo-3,4- dihydroquinolin-1-yl) methyl]-4-(trifluoromethyl) benzonitrile (85 mg, 92.54%) as an off-white solid. LC/MS: mass calcd. for C18H14F3N3O: 345.11, found: 346.05 [M+H] +.
Figure imgf000209_0001
3-tert-butyl-1-(1-{[2-cyano-5-(trifluoromethyl) phenyl] methyl}-2-oxo-3,4-dihydroquinolin- 6-yl) urea To a stirred solution of 2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl) methyl]-4- (trifluoromethyl) benzonitrile (100 mg, 0.290 mmol, 1 equiv) in DCM/Toluene (10 mL, 2/1) was added 2-isocyanato-2-methylpropane (114.83 mg, 1.160 mmol, 4 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at 90 oC under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by Prep-HPLC with the following conditions to afford 3-tert-butyl-1-(1- {[2-cyano-5-(trifluoromethyl) phenyl] methyl}-2-oxo-3,4-dihydroquinolin-6-yl) urea (20.3 mg, 15.49%) as a white solid. MS (ESI): mass calcd. for C23H23F3N4O2: 444.2m/z, found 445.10 [M+H] +. 1H NMR (300 MHz, DMSO-d6) δ 8.09 – 8.19 (m, 2H), 7.86 (d, J = 8.1 Hz, 1H), 7.55 (s, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.8, 2.4 Hz, 1H), 6.77 (d, J = 8.7 Hz, 1H), 5.94 (s, 1H), 5.33 (s, 2H), 2.92 (t, J = 7.3 Hz, 2H), 2.67 (t, J = 7.2 Hz, 2H), 1.27 (s, 9H).19F NMR (282 MHz, DMSO) δ -62.00. Example 56: (S)-1-(tert-butyl)-3-(2-oxo-1-(1-(m-tolyl)ethyl)-1,2,3,4-tetrahydroquinolin-6- yl)urea
Figure imgf000209_0002
(S)-6-nitro-1-(1-(m-tolyl)ethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (1R)-1-(3-methylphenyl)ethanol (300 mg, 2.203 mmol, 1 equiv) in DCM (5 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (465.64 mg, 2.423 mmol, 1.1 equiv) and PPh3 (577.76 mg, 2.203 mmol, 1 equiv). DEAD (383.62 mg, 2.203 mmol, 1 equiv) was added the mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford (S)-6-nitro-1-(1-(m-tolyl)ethyl)-3,4-dihydroquinolin- 2(1H)-one (310 mg, 45.35%) as a yellow solid. MS (ESI): mass calcd. for C18H18N2O3: 310.353, found: 311.10 [M+H]+.
Figure imgf000210_0001
(S)-6-amino-1-(1-(m-tolyl)ethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of (S)-6-nitro-1-(1-(m-tolyl)ethyl)-3,4-dihydroquinolin-2(1H)-one (200 mg, 0.644 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford (S)-6-amino-1-(1-(m- tolyl)ethyl)-3,4-dihydroquinolin-2(1H)-one (170 mg, 94.44%) as a yellow solid. MS (ESI): mass calcd. For C18H20N2O, 280.16 m/z, found 281.15 [M+H]+.
Figure imgf000210_0002
(S)-1-(tert-butyl)-3-(2-oxo-1-(1-(m-tolyl)ethyl)-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred solution of (S)-6-amino-1-(1-(m-tolyl)ethyl)-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.357 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (176.79 mg, 1.785 mmol, 5 equiv) and TEA (144.37 mg, 1.428 mmol, 4 equiv) stirred overnight at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 42% B in 10 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.6; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3-(2-oxo-1-(1-(m-tolyl)ethyl)-1,2,3,4- tetrahydroquinolin-6-yl)urea (20.4 mg, 15.03%) as a white solid. MS (ESI): mass calcd. for C23H29N3O2: 379.23 m/z, found 380.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.32 (d, J = 2.5 Hz, 1H), 7.22 (t, J = 7.5 Hz, 1H), 7.01 – 7.11 (m, 3H), 6.78 – 6.87 (m, 1H), 6.44 – 6.53 (m, 1H), 6.06 – 6.20 (m, 1H), 5.93 (s, 1H), 2.73 – 2.88 (m, 2H), 2.52 – 2.73 (m, 2H), 2.29 (s, 3H), 1.66 (d, J = 7.2 Hz, 3H), 1.26 (s, 9H). Example 57: (S)-1-(tert-butyl)-3-(3-oxo-4-(1-(m-tolyl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]ox azin-7-yl)urea
Figure imgf000211_0001
(S)-7-nitro-4-(1-(m-tolyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (R)-1-(m-tolyl)ethan-1-ol (500 mg, 3.671 mmol, 1 equiv), 7-nitro-2H- benzo[b][1,4]oxazin-3(4H)-one (784.03 mg, 4.038 mmol, 1.1 equiv) and PPh3 (1.44 g, 5.506 mmol, 1.5 equiv) in DCM (20 mL ) was added DEAD (958.87 mg, 5.506 mmol, 1.5 equiv) under N2 at 0 °C. The reaction was stirred for 16 h at rt. The mixture was diuted with EtOAc (50 mL), washed with water (30 mL) and brine (30 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE /EA (3:1) to afford (S)-7-nitro-4-(1-(m-tolyl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one (620 mg, 54.07%) as a yellow oil. LC/MS: C17H16N2O4: 312.325, found: 313.10 [M+H]+.
Figure imgf000212_0001
(S)-7-amino-4-(1-(m-tolyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (S)-7-nitro-4-(1-(m-tolyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 0.961 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (50 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford (S)-7-amino-4-(1-(m- tolyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (200 mg, 73.75%) as a white solid. MS (ESI): mass calcd. for C17H18N2O2, 282.14 m/z, found 283.20 [M+H]+.
Figure imgf000212_0002
(S)-1-(tert-butyl)-3-(3-oxo-4-(1-(m-tolyl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea To a stirred solution of (S)-7-amino-4-(1-(m-tolyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)- one (200 mg, 0.708 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (351.1 mg, 3.542 mmol, 5 equiv) and TEA (215.0 mg, 2.125 mmol, 3 equiv) stirred 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 37% B to 57% B in 10 min; Wave Length: 220 nm; RT1(min): 9.5; Number Of Runs: 2) to afford (S)-1-(tert-butyl)-3-(3-oxo-4-(1-(m-tolyl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea (44.8 mg, 16.56%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3, 381.21 m/z, found 382.15 [M+H]+ .1H NMR (300 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.20 – 7.28 (m, 2H), 7.10 (dd, J = 14.1, 6.8 Hz, 3H), 6.62 (dd, J = 8.9, 2.4 Hz, 1H), 6.55 (d, J = 8.8 Hz, 1H), 6.09 (q, J = 7.2 Hz, 1H), 5.94 (s, 1H), 4.57 – 4.75 (m, 2H), 2.30 (s, 3H), 1.70 (d, J = 7.2 Hz, 3H), 1.25 (s, 9H). Example 58: 3-tert-butyl-1-{1-[(1S)-1-(2-fluorophenyl)ethyl]-2-oxo-3,4-dihydroquinolin-6-y l}ure
Figure imgf000213_0001
1-[(1S)-1-(2-fluorophenyl)ethyl]-6-nitro-3,4-dihydroquinolin-2-one To a solution of (1R)-1-(2-fluorophenyl)ethanol (500 mg, 3.567 mmol, 1 equiv) and 6- nitro-3,4-dihydro-1H-quinolin-2-one (685.57 mg, 3.567 mmol, 1 equiv) in DCM (20 mL) was added triphenylphosphine (935.71 mg, 3.567 mmol, 1 equiv). The reaction was stirred at rt under N2 for 0.5 hour. To the above mixture was added DEAD (621.29 mg, 3.567 mmol, 1 equiv) dropwise at 0 oC. The reaction was stirred at rt under N2 for 3h. Quenched with water (20 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[(1S)-1-(2-fluorophenyl)ethyl]-6-nitro-3,4- dihydroquinolin-2-one (723 mg, 64.48%) as a white solid. MS (ESI): mass calcd for C17H15FN2O3: 314.11 m/z, found 315.00[M+H]+.
Figure imgf000213_0002
6-amino-1-[(1S)-1-(2-fluorophenyl)ethyl]-3,4-dihydroquinolin-2-one To a solution of 1-[(1S)-1-(2-fluorophenyl)ethyl]-6-nitro-3,4-dihydroquinolin-2-one (710 mg, 2.259 mmol, 1 equiv) in methanol (20 mL) was added Pd/C (710 mg). The resulting mixture was stirred for 1 h at room temperature under hydrogen atmosphere. Desired product could be detected by LCMS. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1-(2- fluorophenyl)ethyl]-3,4-dihydroquinolin-2-one (420 mg, 65.39%) as a black liquid. MS (ESI): mass calcd for C17H17FN2O: 284.13m/z, found 285.10 [M+H]+.
Figure imgf000214_0001
3-tert-butyl-1-{1-[(1S)-1-(2-fluorophenyl)ethyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea To a solution of 6-amino-1-[(1S)-1-(2-fluorophenyl)ethyl]-3,4-dihydroquinolin-2-one (200 mg, 0.703 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (139.46 mg, 1.406 mmol, 2 equiv) in DCM (20 mL,) was added TEA (213.54 mg, 2.109 mmol, 3.00 equiv). The reaction was stirred at rt for 16h. Quenched with water (20 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column150 mm x 30 mm x 5 μm (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(1S)-1-(2-fluorophenyl)ethyl]-2-oxo-3,4- dihydroquinolin-6-yl}urea (21.2 mg, 7.85%) as a white solid. LC/MS (ESI): mass calcd. for C22H26FN3O2:383.20 m/z, found:384.00 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.46 – 7.55 (m, 1H), 7.21 – 7.33 (m, 2H), 7.17 (m, 1H), 7.06 (m, 1H), 6.93 (m, 1H), 6.81 (m, 1H), 6.00 (m, 1H), 5.89 (s, 1H), 2.69 – 2.79 (m, 2H), 2.40 – 2.50 (m, 2H), 1.76 (s, 3H), 1.26 (s, 9H). 19F NMR (376 MHz, DMSO) δ -116.16. Example 59: 1-(tert-butyl)-3-(1-(2-cyano-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydr oquinolin-6-yl)urea Synthetic Scheme
Figure imgf000215_0001
2-bromo-1-(bromomethyl)-3-(trifluoromethyl)benzene To a solution of [2-bromo-3-(trifluoromethyl)phenyl]methanol (1.5 g, 5.882 mmol, 1 equiv) in DCM (10 mL) was added phosphorus tribromide (3.18 g, 11.764 mmol, 2 equiv) at 0 oC. The reaction was stirred at rt for 2 hours. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. This resulted in 2-bromo-1-(bromomethyl)-3- (trifluoromethyl)benzene (1.83 g, 97.86%) as a yellow oil. MS (ESI): mass calcd. for C8H5Br2F3: 317.93m/z, found 318.90 [M+H]+.
Figure imgf000215_0002
1-(2-bromo-3-(trifluoromethyl)benzyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (426.14 mg, 2.217 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (617.40 mg, 4.434 mmol, 2 equiv) and 2-bromo-1- (bromomethyl)-3-(trifluoromethyl)benzene (705 mg, 2.217 mmol, 1 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-{[2-bromo-3-(trifluoromethyl)phenyl]methyl}-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 52.54%) as a white solid. MS(ESI): mass calcd. for C17H12BrF3N2O3: 429.19 m/z, found 428.95 [M+H]+.
Figure imgf000216_0001
2-((6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)-6-(trifluoromethyl)benzonitrile To a solution of 1-{[2-bromo-3-(trifluoromethyl)phenyl]methyl}-6-nitro-3,4- dihydroquinolin-2-one (250 mg, 0.582 mmol, 1 equiv) in DMF (10 mL) was added Zn(CN)2 (82.07 mg, 0.698 mmol, 1.2 equiv), Zn (19.04 mg, 0.291 mmol, 0.5 equiv), BrettPhos (62.53 mg, 0.116 mmol, 0.2 equiv) and BrettPhos Pd G3 (158.41 mg, 0.175 mmol, 0.3 equiv). The reaction was stirred at 120 °C under N2 for 1h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-6- (trifluoromethyl)benzonitrile (90 mg, 41.17%) as a white solid. MS (ESI): mass calcd. for C18H12F3N3O3: 375.31 m/z, found:370.15 [M+H]+.
Figure imgf000216_0002
2-((6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)methyl)-6-(trifluoromethyl)benzonitrile To a solution of 2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-6- (trifluoromethyl)benzonitrile (140 mg, 0.373 mmol, 1 equiv) in HOAc (10 mL) was added zinc (243.89 mg, 3.730 mmol, 10 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 2-[(6-amino-2-oxo-3,4- dihydroquinolin-1-yl)methyl]-6-(trifluoromethyl)benzonitrile (90 mg, 69.87%) as a white solid. MS (ESI): mass calcd. for C18H14F3N3O: 345.33m/z, found:368.10 [M+Na]+.
Figure imgf000217_0001
1-(tert-butyl)-3-(1-(2-cyano-3-(trifluoromethyl)benzyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea To a solution of 2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-6- (trifluoromethyl)benzonitrile (90 mg, 0.261 mmol, 1 equiv) in DCM (10 mL) was added 2- isocyanato-2-methylpropane (103.35 mg, 1.044 mmol, 4 equiv) and TEA (79.12 mg, 0.783 mmol, 3 equiv). The reaction was stirred at rt for 1 day. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 30% to 50% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-(1-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-2-oxo-3,4-dihydroquinolin-6-yl)urea (51 mg, 44.03%) as a white solid. MS (ESI): mass calcd. for C23H23F3N4O2: 444.46m/z, found:445.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.04 - 8.29 (m, 1H), 7.89 - 8.00 (m, 1H), 7.73 - 8.00 (m, 1H), 7.45 – 7.55 (m, 1H), 7.35 – 7.45 (m, 1H), 6.88 – 7.09 (m, 1H), 6.67 – 6.80 (m, 1H), 5.82 – 6.07 (m, 1H), 5.20 – 5.50 (m, 2H), 2.86 – 3.07 (m, 2H), 2.60 – 2.78 (m, 2H), 1.15 – 1.38 (m, 9H).19F NMR (376 MHz, DMSO) δ (ppm): -60.5758. Example 60: (S)-3-tert-butyl-1-(2-oxo-1-{1-[3-(trifluoromethyl)phenyl]ethyl}-3,4-dihydroqu inolin-6-yl)urea
Figure imgf000217_0002
6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one To a solution of (1R)-1-[3-(trifluoromethyl)phenyl]ethanol (500 mg, 2.629 mmol, 1 equiv) and 6-nitro-3,4-dihydro-1H-quinolin-2-one (505.28 mg, 2.629 mmol, 1 equiv) in DCM (20 mL) was added triphenylphosphine (689.64 mg, 2.629 mmol, 1 equiv). The reaction was stirred at rt under N2 for 0.5 hour. To the above mixture was addedDEAD (457.91 mg, 2.629 mmol, 1 equiv) dropwise at 0 oC. The reaction was stirred at rt under N2 for 3h. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4- dihydroquinolin-2-one (285 mg, 29.75%) as a white solid.MS (ESI): mass calcd. for C18H15F3N2O3: 364.10 m/z, found 365.11[M+H]+.
Figure imgf000218_0001
6-amino-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one To a solution of 6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one (285 mg, 0.782 mmol, 1 equiv) in methanol (3 mL) was added Pd/C (150 mg). The reaction was stirred at rt under H2 for 1h. The resulting mixture was stirred for 1 h at room temperature under hydrogen atmosphere. Desired product could be detected by LCMS. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 10 mL). The filtrate was concentrated under reduced pressure.The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2- one (225 mg, 86.03%) as a white solid. MS (ESI): mass calcd for C18H17F3N2O: 334.13 m/z, found 335.15 [M+H]+.
Figure imgf000218_0002
(S)-3-tert-butyl-1-(2-oxo-1-{1-[3-(trifluoromethyl)phenyl]ethyl}-3,4-dihydroquinolin-6- yl)urea To a solution of 6-amino-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-3,4-dihydroquinolin-2-one (200 mg, 0.598 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (177.90 mg, 1.794 mmol, 3 equiv) in DCM (3 mL) was added TEA (181.60 mg, 1.794 mmol, 3 equiv). The reaction was stirred at rt for 16h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound (S)-3-tert-butyl-1-(2-oxo-1-{1-[3-(trifluoromethyl)phenyl]ethyl}-3,4- dihydroquinolin-6-yl)urea (20.1 mg, 7.73%) as a white solid. LC/MS (ESI): mass calcd. for C23H26F3N3O2:433.20 m/z, found:432.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.10 (s, 1H), 7.52 – 7.65 (m, 4H), 7.33 (s, 1H), 6.90 (m, 1H), 6.52 (m, 1H), 6.08 (m, 1H), 5.92 (s, 1H), 2.79-2.89 (m, 2H), 2.52 – 2.66 (m, 2H), 1.74 (m, 3H), 1.26 (s, 9H). 19F NMR (282 MHz, DMSO) δ -60.93. Example 61: (S)-1-(tert-butyl)-3-(1-(1-(2-cyanophenyl)ethyl)-2-oxo-1,2,3,4-tetrahydroquino lin-6-yl)urea
Figure imgf000219_0001
(S)-1-(1-(2-bromophenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of (1R)-1-(2-bromophenyl)ethanol (1 g, 4.974 mmol, 1 equiv) in THF (10 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (1.05 g, 5.471 mmol, 1.1 equiv), PPh3 (1.30 g, 4.974 mmol, 1 equiv) at rt. The resulting mixture was stirred at 0 °C under a nitrogen atmosphere, which was added DEAD (0.87 g, 4.974 mmol, 1 equiv). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (PE/EA=3:1) to afford (S)-1-(1-(2- bromophenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (1.2 g, 64.30%) as a yellow oil. MS (ESI): mass calcd. for C17H15BrN2O3: 374.03, found: 375.05[M+H]+.
Figure imgf000220_0001
(S)-2-(1-(6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)ethyl)benzonitrile A solution of 1-[(1S)-1-(2-bromophenyl)ethyl]-6-nitro-3,4-dihydroquinolin-2-one (400 mg, 1.066 mmol, 1 equiv) in dimethylformamide (10 mL) was added Zn(CN)2 (250.35 mg, 2.132 mmol, 2.0 equiv) and Pd(PPh3)4 (123.19 mg, 0.107 mmol, 0.1 equiv) at rt. The resulting mixture was stirred for overnight at 110 °C. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with NaCl (3x5 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford (S)-2-(1-(6-nitro-2-oxo-3,4- dihydroquinolin-1(2H)-yl)ethyl)benzonitrile (78 mg, 22.77%) as an off-white solid. MS (ESI): mass calcd. for C18H15N3O3, 321.11 m/z, found 322.20 [M+H]+ .
Figure imgf000220_0002
(S)-2-(1-(6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)ethyl)benzonitrile solution of (S)-2-(1-(6-nitro-2-oxo-3,4-dihydroquinolin-1(2H)-yl)ethyl)benzonitrile (70 mg, 0.218 mmol, 1 equiv) in MeOH (5 mL)/H2O (0.5 mL) was added Fe (121.65 mg, 2.180 mmol, 10 equiv) and NH4Cl (116.52 mg, 2.180 mmol, 10 equiv) at rt. The resulting mixture was stirred for 1 h at 60 °C. The precipitated solids were collected by filtration and washed with MeOH (3 x 5 mL). The aqueous layer was extracted with DCM (3 x 5 mL). The organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (2:1) to afford (S)-2-(1-(6-amino-2-oxo-3,4- dihydroquinolin-1(2H)-yl)ethyl)benzonitrile (60 mg, 94.53%) as a yellow solid. MS (ESI): mass calcd. for C18H17N3O, 291.14 m/z, found 292.20 [M+H]+ .
Figure imgf000221_0001
(S)-1-(tert-butyl)-3-(1-(1-(2-cyanophenyl)ethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea A solution of (S)-2-(1-(6-amino-2-oxo-3,4-dihydroquinolin-1(2H)-yl)ethyl)benzonitrile (70 mg, 0.240 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (119.09 mg, 1.200 mmol, 5.0 equiv) and TEA (97.25 mg, 0.960 mmol, 4.0 equiv) at rt. The resulting mixture was stirred for overnight at TR. The resulting mixture was extracted with EA (5 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product 50 mg was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 30% B to 54% B in 7 min; Wave Length: 220 nm; RT1(min): 6.5; Number Of Runs: 0 ) to afford (S)-1-(tert-butyl)-3-(1-(1-(2-cyanophenyl)ethyl)- 2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (22.5 mg, 23.93%) as white solid. MS (ESI):mass calcd.C23H26N4O2 for :390.21 m/z, found 391.20 [M+H]+.1H NMR (400 MHz, Chloroform-d) δ 7.72 (d, J = 7.9 Hz, 1H), 7.56 – 7.66 (m, 2H), 7.21 – 7.32 (m, 1H), 7.18 – 7.20 (m, 1H),7.09 – 7.12(m,1H), 6.79 – 6.92 (m, 1H), 6.61 – 6.74 (m, 1H), 5.87 – 6.02 (m, 2H), 2.92 – 3.01 (m, 1H), 2.71 – 2.83 (m, 1H), 2.53 – 2.69 (m, 1H), 2.42 – 2.53 (m, 1H), 1.96 (d, J = 7.1 Hz, 3H), 1.35 – 1.40 (m, 9H). Example 62: (S)-1-(tert-butyl)-3-(3-oxo-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)urea
Figure imgf000221_0002
(S)-7-nitro-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (1R)-1-[3-(trifluoromethyl)phenyl]ethanol (500 mg, 2.629 mmol, 1 equiv), 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (510.47 mg, 2.629 mmol, 1 equiv) and PPh3 (1034.46 mg, 3.944 mmol, 1.5 equiv) in DCM (10 mL). The reaction was stirred at rt under N2 for 0.5 hour. To the above mixture was added DEAD (686.86 mg, 3.944 mmol, 1.5 equiv) dropwise at 0 oC. The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-nitro-4-[(1S)-1-[3- (trifluoromethyl)phenyl]ethyl]-2H-1,4-benzoxazin-3-one (550 mg, 57.11%) as a yellow solid. MS (ESI): mass calcd. for C17H13F3N2O4: 366.30m/z, found 367.00 [M+H]+.
Figure imgf000222_0001
(S)-7-amino-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-nitro-4-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-2H-1,4-benzoxazin- 3-one (500 mg, 1.365 mmol, 1 equiv) in MeOH/H2O (16.5 mL, 10:1) was added Fe (762.29 mg, 13.650 mmol, 10 equiv) and NH4Cl (730.15 mg, 13.650 mmol, 10 equiv). The reaction was stirred at 80 oC for 1 hour. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. This resulted in 7-amino-4-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]- 2H-1,4-benzoxazin-3-one (450 mg, 98.02%) as a yellow soild. MS (ESI): mass calcd. for C17H15F3N2O2: 336.31m/z, found 337.15 [M+H]+.
Figure imgf000222_0002
(S)-1-(tert-butyl)-3-(3-oxo-4-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of 7-amino-4-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]-2H-1,4- benzoxazin-3-one (150 mg, 0.446 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2- methylpropane (176.86 mg, 1.784 mmol, 4 equiv) and TEA (135.40 mg, 1.338 mmol, 3 equiv). The reaction was stirred at rt for 1 day. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 35% to 60% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{3-oxo-4-[(1S)-1-[3- (trifluoromethyl)phenyl]ethyl]-2H-1,4-benzoxazin-7-yl}urea (50.1 mg, 25.80%) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O3: 435.45 m/z, found:436.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.10 – 8.31 (m, 1H), 7.45 – 7.80 (m, 4H), 7.07 – 7.38 (m, 1H), 6.44 – 6.85 (m, 2H), 6.01 – 6.19 (m, 1H), 5.79 – 6.01 (m, 1H), 4.50 – 4.90 (m, 2H), 1.70 – 1.90 (m, 3H), 1.21 – 1.32 (m, 9H).19F NMR (376 MHz, DMSO) δ (ppm): -60.8904. Example 63: 5-benzyl-6-oxo-7,8-dihydro-5H-naphthalen-2-yl N-tert-butylcarbamate Synthetic Scheme
Figure imgf000223_0001
1-benzyl-6-methoxy-3,4-dihydroquinolin-2-one To a solution of 6-methoxy-3,4-dihydro-1H-quinolin-2-one (473.5 mg, 2.672 mmol, 1 equiv) and benzyl bromide (1142.56 mg, 6.680 mmol, 2.5 equiv) in dimethylformamide (25 mL) was added Cs2CO3 (2619.92 mg, 8.016 mmol, 3 equiv). The reaction was stirred at rt for 3h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give tert-butyl 1- benzyl-6-methoxy-3,4-dihydroquinolin-2-one (640 mg, 89.60%) as a white solid.MS (ESI): mass calcd. for C17H17NO2: 267.13 m/z, found 268.10[M+H]+.
Figure imgf000224_0001
1-benzyl-6-hydroxy-3,4-dihydroquinolin-2-one To a solution of 1-benzyl-6-methoxy-3,4-dihydroquinolin-2-one (620 mg, 2.319 mmol, 1 equiv) in DCM (8 mL) was added boron tribromide (7 mL) at -78 °C under nitrogen atmosphere. The reaction was stirred at -78 °C for 3h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-6-hydroxy-3,4-dihydroquinolin-2-one (312 mg, 53.11%) as a black solid. MS (ESI): mass calcd for C16H15NO2: 253.11 m/z, found 253.95[M+H]+.
Figure imgf000224_0002
5-benzyl-6-oxo-7,8-dihydro-5H-naphthalen-2-yl N-tert-butylcarbamate To a solution of 1-benzyl-6-hydroxy-3,4-dihydro-1H-naphthalen-2-one (132 mg, 0.523 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (103.32 mg, 1.042 mmol, 2 equiv) in DCM (2 mL) was added TEA (158.20 mg, 1.563 mmol, 3 equiv). The reaction was stirred at rt for 16h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 5-benzyl-6-oxo-7,8-dihydro-5H-naphthalen-2-yl N-tert-butylcarbamate (50.2 mg, 27.27%) as a white solid. LC/MS (ESI): mass calcd. for C21H24N2O3:352.18 m/z, found:353.05 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 7.50 (s, 1H), 7.22 – 7.37 (m, 5H), 6.97 (m, 1H), 6.76 – 6.91 (m, 2H), 5.13 (s, 2H), 2.89-3.01 (m, 2H), 2.69-2.73 (m, 2H), 1.25 (s, 9H). Example 64: 1-(tert-butyl)-3-(1-(cyclohexylmethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)u rea
Figure imgf000225_0001
1-(cyclohexylmethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred solution of 6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 1.561 mmol, 1.0 equiv) in DMF (15 mL) was added Cs2CO3 (1.53 g, 4.683 mmol, 3.0 equiv) and (bromomethyl)cyclohexane (276.44 mg, 1.561 mmol, 1 equiv) at rt and stirred for overnight. The reaction progress was monitored by LCMS. After completion of reaction, the resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with water (3 x2 0 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA =3:1) to afford 1-(cyclohexylmethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (310 mg, 68.87%) as a yellow solid. MS (ESI): mass calcd. for C16H20N2O3: 288.15 m/z, found 289.15 [M +H]+.
Figure imgf000225_0002
6-amino-1-(cyclohexylmethyl)-3,4-dihydroquinolin-2(1H)-one To a solution of 1-(cyclohexylmethyl)-6-nitro-3,4-dihydroquinolin-2-one (100 mg, 0.347 mmol, 1 equiv) in methanol (5 mL) was added Pd/C (30 mg) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. This resulted in 6-amino-1-(cyclohexylmethyl)-3,4-dihydroquinolin-2(1H)-one as a white solid. MS (ESI): mass calcd. for C16H22N2O: 258.17 m/z, found 259.30 [M +H]+.
Figure imgf000225_0003
1-(tert-butyl)-3-(1-(cyclohexylmethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea A solution of 6-amino-1-(cyclohexylmethyl)-3,4-dihydroquinolin-2(1H)-one (50 mg, 0.194 mmol, 1 equiv), triethylamine (58.75 mg, 0.582 mmol, 3 equiv) and 2-isocyanato-2- methylpropane (57.55 mg, 0.582 mmol, 3 equiv) in DCM (5 mL) was stirred for overnight at rt. The residue was purified by silica gel column chromatography (PE/EA = 2:1) to afford crude products. The residue was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 26% B to 50% B in 9 min, 50% B; Wave Length: 254/220 nm; RT1(min): 8.9;) to afford 1-(tert-butyl)-3-(1- (cyclohexylmethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (26.5 mg, 38.18%) as a white solid. MS (ESI):mass calcd. for C21H31N3O2: 357.24 m/z, found 358.25 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.14 (s, 1H), 7.28 (d, J = 2.4 Hz, 1H), 7.12 (dd, J = 8.8, 2.5 Hz, 1H), 6.99 (d, J = 8.8 Hz, 1H), 5.94 (s, 1H), 3.76 (d, J = 6.9 Hz, 2H), 2.77 (dd, J = 8.8, 5.8 Hz, 2H), 2.47 (d, J = 6.3 Hz, 2H), 1.55 - 1.64 (m, 6H), 1.28 (s, 9H), 1.09 (t, J = 9.5 Hz, 3H), 0.95 (t, J = 11.2 Hz, 2H). Example 65:1-(1-benzyl-7-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000226_0001
7-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 7-fluoro-3,4-dihydro-1H-quinolin-2-one (526 mg, 3.185 mmol, 1 equiv) in H2SO4 (3.2 mL) was added HNO3 (200.6 mg,3.185 mmol, 1.0 equiv)at 0 °C.The resulting mixture was stirred at 0 °C until the starting material was totally consumed by LCMS, the reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (PE/EA = 3:1) to afford 7-fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 35.30%) as a yellow solid. MS (ESI): mass calcd. for C9H7FN2O3, 210.04 m/z, found 211.10 [M+H]+.
Figure imgf000227_0001
1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 7-fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 1.427 mmol, 1 equiv) in DMF (8 mL) was added K2CO3 (591.8 mg, 4.282 mmol, 3 equiv) and benzyl bromide(488.3 mg, 2.855 mmol, 2 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS. The reaction mixture was quenched by water and extracted with EA (3*20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2-one (250 mg, 57.56%) as a white solid.MS (ESI): mass calcd. for C16H13FN2O3, 300.09 m/z, found 301.10 [M+H]+.
Figure imgf000227_0002
6-amino-1-benzyl-7-fluoro-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2-one (250 mg, 0.833 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The mixture was hydrogenated at room temperature for 50 min under hydrogen atmosphere using a hydrogen balloon, then filtered through a Celite pad and concentrated under reduced pressure to afford 6-amino-1-benzyl-7- fluoro-3,4-dihydroquinolin-2-one (220 mg, 86.79%) as a yellow solid. MS (ESI): mass calcd. for C16H15FN2O, 270.12 m/z, found 271.10 [M+H]+
Figure imgf000228_0001
1-(1-benzyl-7-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a stirred solution of 6-amino-1-benzyl-7-fluoro-3,4-dihydroquinolin-2-one (200 mg, 0.740 mmol, 1 equiv) in DCM (10.00 mL) was added 2-isocyanato-2-methylpropane (366.7 mg, 3.699 mmol, 5 equiv) and TEA (224.6 mg, 2.220 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid. The compound was separated by preparative HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 32% B to 52% B in 10 min; Wave Length: 254 nm; RT1(min): 10.2; Number Of Runs: 3) to afford 1-(1-benzyl-7- fluoro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (34.5 mg, 12.51%) as a white solid. MS (ESI): mass calcd. for C21H24FN3O2, 369.19 m/z, found 370.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 7.94 (dd, J = 5.7, 3.2 Hz, 2H), 7.27 – 7.37 (m, 2H), 7.18 – 7.27 (m, 3H), 6.79 (d, J = 13.3 Hz, 1H), 6.43 (s, 1H), 5.10 (s, 2H), 2.87 (t, J = 7.2 Hz, 2H), 2.66 (dd, J = 8.7, 5.8 Hz, 2H), 1.26 (s, 9H). Example 66: 1-(1-benzyl-7-chloro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea
Figure imgf000229_0001
Synthesis of 7-chloro-6-nitro-3,4-dihydro-1H-quinolin-2-one To a stirred solution of 7-chloro-3,4-dihydro-1H-quinolin-2-one (2 g, 11.012 mmol, 1 equiv) in H2SO4 was added KNO3 (1.11 g, 11.012 mmol, 1 equiv) at 0°C. The final reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS. The reaction was poured into the Water/Ice (100 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step directly without further purification. MS: mass calcd. for C9H7ClN2O3: 226.01 m/z, found 227.00 [M+H]+.
Figure imgf000229_0002
1-benzyl-7-chloro-6-nitro-3,4-dihydroquinolin-2-one To a stirred solution of 7-chloro-6-nitro-3,4-dihydro-1H-quinolin-2-one (1.8 g, 7.943 mmol, 1 equiv) in DMF (20 mL) was added benzyl bromide (2.72 g, 15.886 mmol, 2 equiv) and Cs2CO3 (5.19 g, 15.886 mmol, 2 equiv) at room temperature. The final reaction mixture was stirred for 3 h at room temperature. The reaction was monitored by LCMS. The resulting mixture was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 1-benzyl-7-chloro-6-nitro-3,4- dihydroquinolin-2-one (1.3 g, 51.67%) as an orange solid. MS: mass calcd. for C16H13ClN2O3: 316.06 m/z, found 317.15 [M+H]+.
Figure imgf000229_0003
6-amino-1-benzyl-7-chloro-3,4-dihydroquinolin-2-one To a stirred solution of 1-benzyl-7-chloro-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.579 mmol, 1 equiv) in EtOH/H2O (10 mL, 10/1) were added Fe (881.56 mg, 15.790 mmol, 10 equiv) and NH4Cl (844.38 mg, 15.790 mmol, 10 equiv) at room temperature. The final reaction mixture was stirred for 3 h at 60°C. The reaction was monitored by LCMS. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in 6-amino-1-benzyl-7-chloro- 3,4-dihydroquinolin-2-one (300 mg, 66.27%) as a yellow solid. The crude product was used in the next step directly without further purification. MS (ESI): mass calcd. for C16H15ClN2O: 286.09 m/z, found 287.05 [M+H]+.
Figure imgf000230_0001
1-(1-benzyl-7-chloro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea To a stirred solution of 6-amino-1-benzyl-7-chloro-3,4-dihydroquinolin-2-one (140 mg, 0.488 mmol, 1 equiv) in toluene/DCM (10 mL, 1/2) was added 2-isocyanato-2-methylpropane (145.19 mg, 1.464 mmol, 3 equiv) dropwise at room temperature under nitrogen atmosphere. The final reaction mixture was stirred for 3 h at 90°C. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by Prep-HPLC to afford 1-(1-benzyl-7-chloro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (5.0 mg, 2.65%) as a white solid. MS (ESI): mass calcd. for C21H24ClN3O2: 385.16 m/z, found 386.15 [M+H] +.1H NMR (400 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.74 (s, 1H), 7.32 – 7.22 (m, 5H), 6.92 (s, 1H), 6.80 (s, 1H), 5.12 (s, 2H), 2.91 (t, J = 7.2 Hz, 2H), 2.67 (t, J = 7.2 Hz, 2H), 1.28 (s, 9H). Example 67: 1-(5-benzyl-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000231_0001
ethyl (E)-3-(3-amino-6-chloropyridin-2-yl)acrylate A solution of 6-chloro-2-iodopyridin-3-amine (2 g, 7.860 mmol, 1 equiv), ethyl (2E)-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoate (1.77 g, 7.860 mmol, 1 equiv), sodium methaneperoxoate sodium (2.5 g, 23.580 mmol, 3 equiv) and Pd(dppf)Cl2 (575.11 mg, 0.786 mmol, 0.1 equiv) in water (3 mL) and 1,4-dioxane (30 mL) was stirred for overnight at 90 °C under N2 atmosphere. The mixture was allowed to cool down to rt.The reaction progress was monitored by LCMS. The resulting mixture was extracted with EA (3 x 60 mL). The combined organic layers were washed with brine (1x100 mL). After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA = 3:1) to afford ethyl (E)-3-(3-amino-6-chloropyridin-2-yl)acrylate (1.5 g, 84.20%) as a yellow solid. MS (ESI): mass calcd. for C10H11ClN2O2: 226.05 m/z, found 227.20 [M +H] +.
Figure imgf000231_0002
ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate A solution of ethyl (2E)-3-(3-amino-6-chloropyridin-2-yl)prop-2-enoate (1 g, 4.412 mmol, 1 equiv) in methanol (10 mL) and tetrahydrofuran (20 mL) was treated with cobalt (II) chloride (286.40 mg, 2.206 mmol, 0.5 equiv) for 30 min at 0 °C under nitrogen atmosphere followed by the addition of NaBH4 (834.51 mg, 22.060 mmol, 5 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 1 h at rt. The reaction was quenched with H2O at 0 °C. The resulting mixture was filtered. The filter cake was washed with EA (50 mL). The filtrate was concentrated under reduced pressure. This resulted in ethyl 3-(3-amino-6-chloropyridin-2- yl)propanoate (950 mg, 94.16%) as a yellow solid. MS (ESI): mass calcd. for C10H13ClN2O2: 228.07 m/z, found 229.15 [M +H] +.
Figure imgf000232_0001
6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate (900 mg, 3.936 mmol, 1 equiv) in HCl (2M, in EA) (30 mL) at rt and stirred for 2 h. The precipitated solids were collected by filtration and washed with diethyl ether (3 x 50 mL). This resulted in 6-chloro-3,4- dihydro-1H-1,5-naphthyridin-2-one (600 mg, 83.49%) as a white solid. MS (ESI): mass calcd. for C8H7ClN2O: 182.02 m/z, found 183.05 [M +H] +.
Figure imgf000232_0002
1-benzyl-6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of 6-chloro-3,4-dihydro-1H-1,5-naphthyridin-2-one (600 mg, 3.286 mmol, 1 equiv) in anhydrous dimethylformamide (15 mL, 0.027 mmol, 0.17 equiv) was added benzyl bromide (730.57 mg, 4.272 mmol, 1.3 equiv) and Cs2CO3 (3.22 g, 9.858 mmol, 3 equiv) at rt and stirred for 2 h. After completion of reaction, the resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA = (3:1) to afford 1-benzyl-6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one (680 mg, 75.88%) as a yellow solid. MS (ESI): mass calcd. for C15H13ClN2O: 272.07 m/z, found 273.05 [M +H] +.
Figure imgf000233_0001
tert-butyl (1-benzyl-2-oxo-1,2,3,4-tetrahydro-1,7-naphthyridin-6-yl)carbamate A solution of 1-benzyl-6-chloro-3,4-dihydro-1,5-naphthyridin-2-one (200 mg, 0.733 mmol, 1 equiv) in 1,4-dioxane (15 mL, 0.023 mmol, 0.31 equiv) was treated with tert-butyl carbamate (171.82 mg, 1.466 mmol, 2 equiv), Cs2CO3 (477.86 mg, 1.466 mmol, 2 equiv), EPHOS (39.22 mg, 0.073 mmol, 0.1 equiv) and EPhos Pd G4 (67.36 mg, 0.073 mmol, 0.1 equiv) for overnight at 90 °C under nitrogen atmosphere. The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford tert-butyl (1-benzyl-2-oxo-1,2,3,4- tetrahydro-1,7-naphthyridin-6-yl)carbamate (180 mg, 69.45%) as a white solid. MS (ESI): mass calcd. for C20H23N3O3: 353.17 m/z, found 354.25 [M +H] +.
Figure imgf000233_0002
6-amino-1-benzyl-3,4-dihydro-1,5-naphthyridin-2(1H)-one Into a 20 mL bar were added tert-butyl N-(5-benzyl-6-oxo-7,8-dihydro-1,5-naphthyridin- 2-yl)carbamate (80 mg, 0.226 mmol, 1 equiv) and HCl (10 mL, 10.000 mmol, 44.18 equiv) at rt and strried for 3 h. The crude product/ resulting mixture was used in the next step directly without further purification. This resulted in 6-amino-1-benzyl-3,4-dihydro-1,5-naphthyridin- 2(1H)-one (60 mg, 75%) as a white solid. MS (ESI): mass calcd. for C15H15N3O: 253.12 m/z, found 254.20 [M +H] +.
Figure imgf000233_0003
1-(5-benzyl-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)-3-(tert-butyl)urea A solution of 6-amino-1-benzyl-3,4-dihydro-1,5-naphthyridin-2-one (60 mg, 0.237 mmol, 1 eq uiv), TEA (71.91 mg, 0.711 mmol, 3 equiv) and 2-isocyanato-2-methylpropane (117.41 mg, 1.185 mmol, 5 equiv) in DCM (10 mL, 157.306 mmol, 664.11 equiv) was stirred for overnight at rt. The residue was purified by silica gel column chromatography, eluted with EA (2:1) to afford 1-(5-benzyl-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)-3-(tert-butyl)urea (43.1 mg, 51.44%) as a white solid. MS (ESI): mass calcd. for C20H24N4O2: 352.19 m/z, found 353.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.87 (s, 1H), 7.95 (s, 1H), 7.37 – 7.16 (m, 6H), 7.11 (d, J = 8.9 Hz, 1H), 5.10 (s, 2H), 2.99 (dd, J = 8.7, 6.2 Hz, 2H), 2.80 (dd, J = 8.8, 6.2 Hz, 2H), 1.31 (s, 9H). Example 68: 1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000234_0001
1-benzyl-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (500 mg, 2.616 mmol, 1 equiv) in DMF (20 mL) was added benzyl bromide (671.10 mg, 3.924 mmol, 1.5 equiv) and K2CO3 (728.31 mg, 5.232 mmol, 2 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-6-nitroquinoxalin-2-one (450 mg, 61.16%) as a yellow solid. MS (ESI): mass calcd. for C15H11N3O3: 281.27m/z, found 282.00 [M+H]+.
Figure imgf000235_0001
6-amino-1-benzylquinoxalin-2(1H)-one To a solution of 1-benzyl-6-nitroquinoxalin-2-one (250 mg, 0.889 mmol, 1 equiv) in MeOH/H2O (11 mL, 10:1) was added Fe (496.36 mg, 8.890 mmol, 10 equiv) and NH4Cl (475.43 mg, 8.890 mmol, 10 equiv). The reaction was stirred at 80 oC for 1 hour. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. This resulted in 6- amino-1-benzylquinoxalin-2-one (220 mg, 98.50%) as a yellow soild. MS (ESI): mass calcd. for C15H13N3O: 251.29 m/z, found 252.05 [M+H]+.
Figure imgf000235_0002
1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzylquinoxalin-2-one (220 mg, 0.875 mmol, 1 equiv) in DCM/Toluene (15 mL, 2:1) was added 2-isocyanato-2-methylpropane (520.74 mg, 5.250 mmol, 6 equiv). The reaction was stirred at 90 oC for 2 hours. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 26% to 45% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 1-(1-benzyl-2-oxoquinoxalin- 6-yl)-3-tert-butylurea (55 mg, 17.93%) as a yellow solid. MS (ESI): mass calcd. for C20H22N4O2: 350.42 m/z, found:351.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.40 – 8.52 (m, 1H), 8.25 – 7.38 (m, 1H), 7.83 – 8.01 (m, 1H), 7.20 – 7.50(m, 7H), 5.90 – 6.10 (m, 1H), 5.35 – 5.61 (m, 2H), 1.19 – 1.39 (m, 9H). Example 69: 1-(1-benzyl-8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea
Figure imgf000236_0001
8-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 8-fluoro-3,4-dihydro-1H-quinolin-2-one (500 mg, 3.027 mmol, 1 equiv) in H2SO4 (3 mL) was added HNO3 (190.7 mg, 3.027 mmol, 1.0 equiv)at -10 °C.The resulting mixture was stirred at -10 °C until the starting material was totally consumed by LCMS, The reaction mixture was quenched by water and adjusting PH with saturaed sodium bicarbonate aqueous solution and then extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (PE/EA=3:1) to afford 8-fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 46.66%) as a yellow solid. MS (ESI): mass calcd. for C9H7FN2O3, 210.04 m/z, found 211.10 [M+H]+
Figure imgf000236_0002
1-benzyl-8-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 8-fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (250 mg, 1.190 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (493.2 mg, 3.569 mmol, 3 equiv) and benzyl bromide (305.1 mg, 1.784 mmol, 1.5 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS. The mixture was diuted with EtOAc (100 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 4-benzyl-7-nitro-2H-1,4-benzoxazin-3-one (250 mg, 65.39%) as a yellow solid. MS (ESI): mass calcd. for C16H13FN2O3: 300.09 m/z, found 301.15 [M+H]+.
Figure imgf000237_0001
6-amino-1-benzyl-8-fluoro-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-8-fluoro-6-nitro-3,4-dihydroquinolin-2-one (250 mg, 0.833 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (30 mg). The reaction mixture was then stirred for 1 h under hydrogen atmosphere (balloon). The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure to afford 6-amino-1-benzyl-8-fluoro-3,4- dihydroquinolin-2-one (180 mg, 77.83%) as a yellow solid.MS (ESI): mass calcd. for C16H15FN2O, 270.12 m/z, found 271.10 [M+H]+.
Figure imgf000237_0002
1-(1-benzyl-8-fluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a stirred solution of 6-amino-1-benzyl-8-fluoro-3,4-dihydroquinolin-2-one (140 mg, 0.518 mmol, 1 equiv) in DCM (10.00 mL) was added 2-isocyanato-2-methylpropane (256.7 mg, 2.590 mmol, 5 equiv) and TEA (157.2 mg, 1.554 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water and extracted with EA (3*20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.The compound was separated by preparative HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 39% B to 58% B in 7 min; Wave Length: 254 nm; RT1(min): 6.4; Number Of Runs: 3) to afford 1-(1-benzyl-8- fluoro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (23.4 mg, 12.13%) as a white solid. MS (ESI): mass calcd. for C21H24FN3O2, 369.19 m/z, found 370.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.26 (t, J = 7.3 Hz, 2H), 7.08 – 7.21 (m, 4H), 6.87 (s, 1H), 6.01 (s, 1H), 5.11 (s, 2H), 2.86 (t, J = 6.7 Hz, 2H), 2.54 – 2.70 (m, 2H), 1.25 (s, 9H). Example 70: 1-(tert-butyl)-3-(3-methyl-2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4-tetrahyd roquinazolin-6-yl)urea
Figure imgf000238_0001
3-methyl-6-nitro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinazolin-2(1H)-one To a solution of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (120 mg, 0.579 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3-(trifluoromethyl)benzene (152.29 mg, 0.637 mmol, 1.1 equiv) and Cs2CO3 (378.58 mg, 1.158 mmol, 2 equiv). The reaction was stirred at rt for 1 hour. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-methyl-6-nitro- 1-{[3-(trifluoromethyl)phenyl]methyl}-4H-quinazolin-2-one (170 mg, 80.35%) as a yellow solid. MS (ESI): mass calcd. for C17H14F3N3O3: 365.31m/z, found 366.05 [M+H]+.
Figure imgf000238_0002
6-amino-3-methyl-1-(3-(trifluoromethyl)benzyl)-3,4-dihydroquinazolin-2(1H)-one To a solution of 3-methyl-6-nitro-1-{[3-(trifluoromethyl)phenyl]methyl}-4H-quinazolin-2-one (170 mg, 0.465 mmol, 1 equiv) in MeOH/H2O (11 mL, 10:1) was added Fe (259.88 mg, 4.650 mmol, 10 equiv) and NH4Cl (248.92 mg, 4.650 mmol, 10 equiv). The reaction was stirred for 1 hour at 80 oC. The reaction was quenched with water at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. This resulted in 6-amino-3-methyl-1-{[3-(trifluoromethyl)phenyl]methyl}-4H- quinazolin-2-one (150 mg, 96.15%) as a yellow oil. MS (ESI): mass calcd. for C17H16F3N3O: 335.33m/z, found 336.05 [M+H]+.
Figure imgf000239_0001
1-(tert-butyl)-3-(3-methyl-2-oxo-1-(3-(trifluoromethyl)benzyl)-1,2,3,4- tetrahydroquinazolin-6-yl)urea To a solution of 6-amino-3-methyl-1-{[3-(trifluoromethyl)phenyl]methyl}-4H-quinazolin-2-one (160 mg, 0.477 mmol, 1 equiv) in DCM/Toluene (15 mL, 2:1) was added 2-isocyanato-2- methylpropane (283.80 mg, 2.862 mmol, 6 equiv). The reaction was stirred at 90 oC for 2 hours. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-(3-methyl-2-oxo-1-{[3-(trifluoromethyl)phenyl]methyl}-4H- quinazolin-6-yl)urea (21.3 mg, 10.27%) as a white solid.MS (ESI): mass calcd. for C22H25F3N4O2: 434.46 m/z, found:435.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.91 – 8.20 (m, 1H), 7.43 – 7.66 (m, 4H), 7.17 – 7.36 (m, 1H), 6.84 – 7.02 (m, 1H), 6.45 – 6.62 (m, 1H), 5.76 – 6.00 (m, 1H), 4.94 – 5.20 (m, 2H), 4.30 – 4.48 (m, 2H), 2.85 – 3.01 (m, 3H), 1.17 – 1.33 (m, 9H).19F NMR (376 MHz, DMSO) δ (ppm): -60.9887. Experimental Procedures Example 71: 1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000240_0001
4-(3-(difluoromethyl)benzyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of 7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 1.545 mmol, 1 equiv) in DMF (15 mL) was added 1-(bromomethyl)-3-(difluoromethyl)benzene (376 mg, 1.700 mmol, 1.1 equiv) and Cs2CO3 (1.5 g, 4.635 mmol, 3 equiv) at rt and stirred for overnight. The reaction was quenched with water (10 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford 4-(3-(difluoromethyl)benzyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (220 mg, 42.59%).
Figure imgf000240_0002
7-amino-4-(3-(difluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-(3-(difluoromethyl)benzyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (100 mg, 0.299 mmol, 1 equiv) in MeOH (15 mL) was added Pd/C (50 mg) under nitrogen atmosphere. The resulting mixture was placed at rt under a hydrogen atmosphere until the starting material was totally consumed by LCMS. The mixture was filtered and concentrated under vacuum to afford The catalyst was removed by filtration through a pad of Celite and rinsed with MeOH. The filtrate was concentrated to afford 7-amino-4-(3-(difluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin- 3(4H)-on (57 mg, 62.62%) as a white solid.
Figure imgf000241_0001
1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)urea A solution of 7-amino-4-(3-(difluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (50 mg, 0.164 mmol, 1 equiv), TEA (49.88 mg) and 2-isocyanato-2-methylpropane (48.87 mg, 0.492 mmol, 3 equiv) in DCM (5 mL) was stirred for overnight at rt. The residue was purified by silica gel column chromatography, eluted with EA (35%-46%) to afford crude product. The residue was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart C18, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 61% B in 9 min, 61% B; Wave Length: 220/254 nm) to afford1-(tert- butyl)-3-(4-(3-(difluoromethyl)benzyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea (4 mg, 6.03%) as a white solid. MS (ESI): mass calcd. for C21H23F2N3O3: 403.17 m/z, found 404.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.44 – 7.51 (m, 4H), 7.21 (s, 1H), 7.01 (s, 1H), 6.82 – 6.88 (m, 1H), 6.76 (d, J = 9.0 Hz, 1H), 5.93 (s, 1H), 5.16 (s, 2H), 4.75 (s, 2H), 1.25 (s, 9H). The following examples in Table 3 were made according to the procedure for Example 71. Table 3
Figure imgf000241_0002
Figure imgf000242_0001
Figure imgf000243_0001
Examples 81 and 82: (S)-1-(tert-butyl)-3-(2-methyl-3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and (R)-1-(tert-butyl)-3-(2-methyl-3-oxo-4-(3- (trifluoromethyl)benzyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000244_0001
2-methyl-7-nitro-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (250 mg, 1.201 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3-(trifluoromethyl)benzene (315.77 mg, 1.321 mmol, 1.1 equiv) and Cs2CO3 (784.99 mg, 2.402 mmol, 2 equiv). The reaction was stirred at rt for 1 hours. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 2-methyl-7- nitro-4-{[3-(trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-3-one (390 mg, 88.66%) as a yellow solid.MS (ESI): mass calcd. for C17H13F3N2O4: 366.08 m/z, found 367.00 [M+H]+.
Figure imgf000245_0001
7-amino-2-methyl-4-(3-(trifluoromethyl)benzyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-4-{[3-(trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-3- one (390 mg, 1.065 mmol, 1 equiv) in MeOH/H2O (10:1 ,11 mL) was added Fe (594.59 mg, 10.650 mmol, 10 equiv) and NH4Cl (569.51 mg, 10.650 mmol, 10 equiv). The resulting mixture was filtered, and the filter cake was washed with EA (3 x 30 mL). The filtrate was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 7-amino-2-methyl-4-{[3-(trifluoromethyl)phenyl]methyl}-2H-1,4- benzoxazin-3-one (350 mg, 97.74%) as a yellow oil. MS (ESI): mass calcd. forC17H15F3N2O2: 336.11m/z, found 337.05 [M+H]+.
Figure imgf000245_0002
1-(tert-butyl)-3-(2-methyl-3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of 7-amino-2-methyl-4-{[3-(trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-3- one (190 mg, 0.565 mmol, 1 equiv) in DCM/Toluene (2:1, 15 mL) was added 2-isocyanato-2- methylpropane (336.03 mg, 3.390 mmol, 6 equiv). The reaction was stirred at 90℃ for 2 hours. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-(2-methyl-3-oxo-4-{[3- (trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-7-yl)urea (40 mg, 16.25%) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O3: 435.18 m/z, found:436.20 [M+H]+.
Figure imgf000246_0001
(S)-1-(tert-butyl)-3-(2-methyl-3-oxo-4-(3-(trifluoromethyl)benzyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and (R)-1-(tert-butyl)-3-(2-methyl-3-oxo-4-(3- (trifluoromethyl)benzyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-(2-methyl-3-oxo-4-{[3-(trifluoromethyl)phenyl]methyl}-2H-1,4- benzoxazin-7-yl)urea (40 mg) was separated by chiral-HPLC using a CHIRALPAK ID, 2*25 cm, 5 μm column (eluent: 10% to 10% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer example example 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-{[3- (trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-7-yl]urea (16.4 mg, 41.00%) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O3: 435.18 m/z, found:436.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.10 – 8.40 (m, 1H), 7.45 – 7.72 (m, 4H), 7.13 – 7.30 (m, 1H), 6.81 – 6.96 (m, 1H), 6.61 – 6.81 (m, 1H), 5.87 – 6.10 (m, 1H), 5.10 – 5.30 (m, 1H), 4.75 – 4.92 (m, 1H), 1.40 – 1.53 (m, 3H), 1.24 – 1.30 (m, 9H). 19F NMR (376 MHz, DMSO) δ(ppm): -61.0904.and to yield second enantiomer example 3-tert-butyl-1-[(2S)-2-methyl-3-oxo-4-{[3- (trifluoromethyl)phenyl]methyl}-2H-1,4-benzoxazin-7-yl]urea (14.4 mg, 36.00%) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O3: 435.18 m/z, found:436.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.10 – 8.35 (m, 1H), 7.50 – 7.71 (m, 4H), 7.13 – 7.30 (m, 1H), 6.82 – 6.99 (m, 1H), 6.70 – 6.81 (m, 1H), 5.85 – 6.02 (m, 1H), 5.09 – 5.27 (m, 2H), 4.72 – 4.92 (m, 1H), 1.40 – 1.56 (m, 3H), 1.24 – 1.30 (m, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -61.0913. The following examples in Table 4 were made according to the procedure for examples 81 and 82. Table 4
Figure imgf000246_0002
Figure imgf000247_0001
Figure imgf000248_0001
Example 89: 1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000249_0001
1-(3-chlorobenzyl)-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (400 mg, 2.093 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3-chlorobenzene (644.99 mg, 3.139 mmol, 1.5 equiv) and K2CO3 (582.65 mg, 4.186 mmol, 2 equiv). The reaction was stirred at rt for 1hour. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[(3-chlorophenyl)methyl]-6- nitroquinoxalin-2-one (510 mg, 77.19%) as a yellow solid. MS (ESI): mass calcd. for C15H10ClN3O3: 315.04m/z, found 316.05 [M+H]+.
Figure imgf000249_0002
6-amino-1-(3-chlorobenzyl)quinoxalin-2(1H)-one To a solution of 1-[(3-chlorophenyl)methyl]-6-nitroquinoxalin-2-one (400 mg, 1.267 mmol, 1 equiv) in MeOH/H2O (10:1, 11 mL) was added Fe (707.55 mg, 12.670 mmol, 10 equiv) and NH4Cl (677.71 mg, 12.670 mmol, 10 equiv). The reaction was stirred at 80 °C for 1 hour. The mixture was allowed to cool down to rt. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 x 20 mL). The filtrate was concentrated under reduced pressure. The mixture was diluted with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(3- chlorophenyl)methyl]quinoxalin-2-one (266 mg, 73.48%) as a yellow solid. MS (ESI): mass calcd. for C15H12ClN3O: 285.07m/z, found 286.00 [M+H]+.
Figure imgf000250_0001
1-(tert-butyl)-3-(1-(3-chlorobenzyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)urea To a solution of 6-amino-1-[(3-chlorophenyl)methyl]quinoxalin-2-one (130 mg, 0.455 mmol, 1 equiv) in DCM: Toluene (2:1 , 15 mL) was added 2-isocyanato-2-methylpropane (270.62 mg, 2.730 mmol, 6 equiv). The reaction was stirred at 90 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(3-chlorophenyl)methyl]-2-oxoquinoxalin-6-yl}urea (22.2 mg, 12.68%) as a yellow solid. MS (ESI): mass calcd. for C20H21ClN4O2: 384.14m/z, found:383.10 [M-H]-.1H NMR (400 MHz, DMSO-d6) δ 8.40 – 8.53 (m, 1H), 8.20 – 8.40 (m, 1H), 7.91 – 8.10 (m, 1H), 7.30 – 7.50 (m, 5H), 7.10– 7.27 (m, 1H), 5.80 – 6.20 (m, 1H), 5.30 – 5.60 (m, 2H), 1.21 – 1.40 (m, 9H). The following examples in Table 5 were made according to the procedure for example 89. Table 5
Figure imgf000250_0002
Figure imgf000251_0001
Figure imgf000252_0001
Example 98: (S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000253_0001
(S)-6-nitro-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (400 mg, 2.093 mmol, 1 equiv) in DCM (10 mL) was added (R)-1-phenylethan-1-ol (255.65 mg, 2.093 mmol, 1 equiv) and PPh3 (823.32 mg, 3.139 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (546.67 mg, 3.139 mmol, 1.5 equiv) in portions at 0℃. The resulting mixture was stirred for additional 1 hour at rt. Quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to 6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (108 mg, 17.48%) as a yellow solid. MS (ESI): mass calcd. for C16H13N3O3: 295.10m/z, found 296.00 [M+H]+.
Figure imgf000253_0002
(S)-6-amino-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (100 mg, 0.339 mmol, 1 equiv) in MeOH/H2O (10:1 , 11 mL) was added Fe (189.11 mg, 3.390 mmol, 10 equiv) and NH4Cl (181.14 mg, 3.390 mmol, 10 equiv).The reaction was stirred at 80 °C for 1 hour. The mixture was allowed to cool down to rt. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 x 20 mL). The filtrate was concentrated under reduced pressure. The mixture was diluted with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1- phenylethyl]quinoxalin-2-one (85 mg, 94.61%) as a yellow solid. MS (ESI): mass calcd. for C16H15N3O: 265.12m/z, found 266.15 [M+H]+.
Figure imgf000254_0001
(S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-1,2-dihydroquinoxalin-6-yl)urea To a solution of 6-amino-1-[(1S)-1-phenylethyl]quinoxalin-2-one (85 mg, 0.320 mmol, 1 equiv) in DCM: Toluene (2:1, 15 mL) was added 2-isocyanato-2-methylpropane (190.56 mg, 1.920 mmol, 6 equiv). The reaction was stirred at 90 °C under N2 for 3 hours. After cooling down to rt, the reaction was quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column, 30*150 mm, 5μm column (eluent: 34% to 56% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea (22.1 mg, 18.93%) as a yellow solid. MS (ESI): mass calcd. for C21H24N4O2: 364.19m/z, found:365.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.31 – 8.50 (m, 1H), 8.17 – 8.31 (m, 1H), 7.77 – 8.02 (m, 1H), 7.20 – 7.40 (m, 6H), 6.86 – 7.20 (m, 1H), 6.50 – 6.80 (m, 1H), 5.67 – 6.20 (m, 1H), 1.76 – 1.95 (m, 1H), 1.17 – 1.32 (m, 9H). The following examples in Table 6 were made according to the procedure for example 98. Table 6
Figure imgf000254_0002
Figure imgf000255_0001
Figure imgf000256_0001
Figure imgf000257_0001
Figure imgf000258_0001
Figure imgf000259_0001
Figure imgf000260_0001
Figure imgf000261_0001
Figure imgf000262_0001
Figure imgf000263_0001
Figure imgf000264_0001
Figure imgf000265_0001
Figure imgf000266_0001
Figure imgf000267_0001
Figure imgf000268_0001
Figure imgf000269_0001
Example 157: 3-tert-butyl-1-[(2R)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4- benzoxazin-7-yl]urea. Synthetic Scheme
Figure imgf000270_0001
(2R)-7-bromo-2-methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin-3-one To a stirred solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (400 mg, 1.652 mmol, 1 equiv) in DMF (4 mL) was added 1-(bromomethyl)-3-methylbenzene (458.70 mg, 2.478 mmol, 1.5 equiv) and Cs2CO3 (1080.10 mg, 3.304 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford (2R)-7-bromo-2-methyl-4-[(3- methylphenyl)methyl]-2H-1,4-benzoxazin-3-one (500 mg, 87.40%) as a white solid. LC/MS(ESI): mass calcd. for C17H16BrNO2: 345.04 m/z, found 346.00 [M+H, M+H+2]+.
Figure imgf000271_0001
(2R)-7-amino-2-methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin-3-one To a stirred solution of (2R)-7-bromo-2-methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin- 3-one (490 mg, 1.192 mmol, 1 equiv) in EtOH/H2O(4:1, 5 mL) was added N1,N2- dimethylcyclohexane-1,2-diamine (101.69 mg, 0.715 mmol, 0.6 equiv), CuSO4 (190.17 mg, 1.192 mmol, 1 equiv), sodium ascorbate (474.50 mg, 2.384 mmol, 2 equiv) and NaN3 (232.39 mg, 3.576 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at 80 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford (2R)-7-amino-2- methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin-3-one (150 mg, 44.59%) as a white solid. LC/MS(ESI): mass calcd. for C9H18N2O3: 282.14 m/z, found 283.10 [M+H]+.
Figure imgf000271_0002
3-tert-butyl-1-[(2R)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7- yl]urea To a stirred solution of (2R)-7-amino-2-methyl-4-[(3-methylphenyl)methyl]-2H-1,4-benzoxazin- 3-one (140 mg, 0.496 mmol, 1 equiv) in DCM / Toluene(2:1, 2mL) was added 2-isocyanato-2- methylpropane (147.47 mg, 1.488 mmol, 3 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 3-tert- butyl-1-[(2R)-2-methyl-4-[(3-methylphenyl)methyl]-3-oxo-2H-1,4-benzoxazin-7-yl]urea (41.1 mg, 21.37%) as a white solid. LC/MS(ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.23 – 7.18 (m, 2H), 7.06 – 6.99 (m, 3H), 6.84 (d, J = 8.7 Hz, 1H), 6.76 – 6.72 (m, 1H), 5.93 (s, 1H), 5.05 (m, 2H), 4.84 – 4.78 (m, 1H), 2.26 (s, 3H), 1.48 (d, J = 6.6 Hz, 3H), 1.26 (s, 9H). The following examples in Table 7 were made according to the procedure for example 157. Table 7
Figure imgf000272_0001
Figure imgf000273_0001
Figure imgf000274_0001
Figure imgf000275_0001
Figure imgf000276_0001
Figure imgf000277_0001
Figure imgf000278_0004
Figure imgf000278_0005
Figure imgf000278_0003
Figure imgf000278_0002
Example 179: 3-tert-butyl-1-{3-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea Synthetic Scheme
Figure imgf000278_0001
Figure imgf000279_0001
(S)-3-methyl-6-nitro-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 3-methyl-6-nitro-1H-quinoxalin-2-one (600 mg, 2.924 mmol, 1 equiv) and 1- phenyl-ethanol (357.26 mg, 2.924 mmol, 1 equiv) in DCM (10 mL) was added PPh3 (1150.55 mg, 4.386 mmol, 1.5 equiv). After the reaction was stirred at rt for 30 min under N2, DEAD (509.30 mg, 2.924 mmol, 1 equiv) was added at 0 oC. The reaction was stirred at rt under N2 for 3 h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-methyl-6-nitro- 1-[(1S)-1-phenylethyl]quinoxalin-2-one (217 mg, 23.99%) as a white solid. MS (ESI): mass calcd. for C17H15N3O3: 309.11m/z, found 310.05 [M+H]+
Figure imgf000279_0002
6-amino-3-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one To a solution of 3-methyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (200 mg, 0.647 mmol, 1 equiv) in methanol (8 mL) and water (0.8 mL) was added iron (361.08 mg, 6.470 mmol, 10 equiv) and NH4Cl (345.85 mg, 6.470 mmol, 10 equiv). The reaction was stirred at 60 °C for 3 h. After cooling down to rt, the reaction was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-3-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (95 mg, 52.60%) as a white solid. MS (ESI): mass calcd. for C17H17N3O: 279.14 m/z, found 280.10 [M+H]+
Figure imgf000279_0003
3-tert-butyl-1-{3-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea To a solution of 6-amino-3-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (85 mg, 0.304 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (60.33 mg, 0.608 mmol, 2 equiv) in DCM (2 mL) was added TEA (92.37 mg, 0.912 mmol, 3 equiv). The reaction was stirred at rt for 6h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{3-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea (20.1 mg, 17.37%) as a white solid. LC/MS (ESI): mass calcd. for C22H26N4O2:378.21 m/z, found:379.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.33 (s, 1H), 7.79 (s, 1H), 7.16-7.40 (m, 6H), 7.00 (s, 1H), 6.64 (s, 1H), 6.00 (s, 1H), 2.47-2.51 (m, 3H), 1.86-1.88 (d, J = 6 Hz, 3H), 1.27 (s, 9H). The following examples in Table 8 were made according to the procedure for example 179. Table 8
Figure imgf000280_0001
Figure imgf000281_0002
Example 183: 1-(tert-butyl)-3-((R)-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000281_0001
(R)-7-bromo-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of(2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (270 mg, 1.115 mmol, 1 equiv) in DCM (10 mL) was added 1-[(1R)-1-bromoethyl]-3-chlorobenzene (9.07 mg, 0.041 mmol, 1 equiv) and PPh3 (438.83 mg, 1.672 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5 hour. To the above mixture was added DEAD (291.37 mg, 1.672 mmol, 1.5 equiv) in portions at 0°C. The resulting mixture was stirred for additional 1 hour at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-4-[(1S)-1-(3- chlorophenyl)ethyl]-2-methyl-2H-1,4-benzoxazin-3-one (194 mg, 45.69%) as a white oil. MS (ESI): mass calcd. for C17H15BrClNO2: 379.00m/z, found 380.95,382.95 [M+H, M+H+2]+.
Figure imgf000282_0001
(R)-7-amino-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-4-[(1S)-1-(3-chlorophenyl)ethyl]-2-methyl-2H-1,4-benzoxazin-3- one (180 mg, 0.473 mmol, 1 equiv) in EtOH/H2O (4:1, 10 mL) was added (1R,2R)-N1,N2- dimethylcyclohexane-1,2-diamine (40.36 mg, 0.284 mmol, 0.6 equiv), CuSO4 (75.47 mg, 0.473 mmol, 1 equiv), azidosodium (92.22 mg, 1.419 mmol, 3 equiv) and sodium ascorbate (188.30 mg, 0.946 mmol, 2 equiv). The reaction was stirred at 80 °C under N2 for 1 day. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-[(1S)-1-(3-chlorophenyl)ethyl]-2-methyl-2H-1,4-benzoxazin-3-one (110 mg, 73.43%) as a yellow oil. MS (ESI): mass calcd. for C17H17ClN2O2: 316.10m/z, found 317.15 [M+H]+.
Figure imgf000282_0002
1-(tert-butyl)-3-((R)-4-((S)-1-(3-chlorophenyl)ethyl)-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-[(1S)-1-(3-chlorophenyl)ethyl]-2-methyl-2H-1,4-benzoxazin-3- one (100 mg, 0.316 mmol, 1 equiv) in DCM/Toluene (2:1, 12 mL) was added 2-isocyanato-2- methylpropane (312.93 mg, 3.160 mmol, 10 equiv). The reaction was stirred at 90 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS, 30*150 mm, 5μm column (eluent: 53% to 66% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-4-[(1S)-1-(3- chlorophenyl)ethyl]-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (27.5 mg, 20.95%) as a white solid. MS (ESI): mass calcd. for C22H26ClN3O3: 415.17 m/z, found:416.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.07 – 8.37 (m, 1H), 7.30 – 7.60 (m, 3H), 7.00 – 7.30 (m, 2H), 6.52 – 6.74 (m, 2H), 5.83 – 6.10 (m, 2H), 4.55 – 4.88 (m, 1H), 1.66 – 1.85 (m, 3H), 1.40 – 1.53 (m, 3H), 1.22 – 1.30 (m, 9H). The following examples in Table 9 were made according to the procedure for example 183. Table 9
Figure imgf000283_0001
Figure imgf000284_0002
Figure imgf000284_0001
Figure imgf000284_0003
Figure imgf000284_0004
Figure imgf000285_0001
Figure imgf000286_0002
Example 191: 1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(1-methylcyclobutyl)urea Synthetic Scheme
Figure imgf000286_0001
benzyl-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (3 g, 15.695 mmol, 1 equiv) and benzyl bromide (5368.79 mg, 31.390 mmol, 2 equiv) in dimethylformamide (100 mL) was added Cs2CO3 (15388.46 mg, 47.085 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-6-nitroquinoxalin-2-one (3.1 g, 70.22%) as a yellow solid. MS (ESI): mass calcd. for C15H11N3O3: 281.08 m/z, found 282.10 [M+H]+
Figure imgf000287_0001
6-amino-1-benzylquinoxalin-2(1H)-one To a solution of 1-benzyl-6-nitroquinoxalin-2-one (3.1 g, 11.021 mmol, 1 equiv) in methanol (80 mL) and water (8 mL) was added iron (6.15 g, 110.210 mmol, 10 equiv) and NH4Cl (5.90 g, 110.210 mmol, 10 equiv). The reaction was stirred at 60 °C for 6 h. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6- amino-1-benzylquinoxalin-2-one (1 g, 36.11%) as a red solid. MS (ESI): mass calcd. for C15H13N3O: 251.11 m/z, found 252.10 [M+H]+
Figure imgf000287_0002
1-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(1-methylcyclobutyl)urea To a solution of 6-amino-1-benzylquinoxalin-2-one (100 mg, 0.398 mmol, 1 equiv) in DCM (3 mL) was added triphosgene (47.23 mg, 0.159 mmol, 0.4 equiv). The reaction was stirred at 0 °C under N2 for 1h. The reaction was added TEA (201.35 mg, 1.990 mmol, 5 equiv) and stirred at rt under N2 for 1h. The reaction was added 1-methylcyclobutan-1-amine (67.77 mg, 0.796 mmol, 2 equiv) and stirred at rt under N2 for 1h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 21.2 mm X 250 mm, 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1-(1-benzyl-2-oxoquinoxalin-6-yl)-3- (1-methylcyclobutyl)urea (32.5 mg, 22.48%) as a white solid. LC/MS (ESI): mass calcd. for C21H22N4O2:362.17 m/z, found:363.00 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.33 (s, 1H), 7.99 (s, 1H), 7.42-7.43 (m, 1H), 7.20-7.39 (m, 6H), 6.32 (s, 1H), 5.45 (s, 2H), 2.24- 2.28 (m, 2H), 1.68-1.96 (m, 4H), 1.40 (s, 3H). The following examples in Table 10 were made according to the procedure for example 191. Table 10
Figure imgf000288_0001
Figure imgf000289_0001
Example 200: (S)-1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)- 1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000290_0001
(S)-6-isocyanato-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a stirred solution of (S)-6-amino-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (70 mg, 0.200 mmol, 1 equiv) in DCM (5 mL) was added TEA (81 mg, 0.802 mmol, 4 equiv) and triphosgene (24 mg, 0.080 mmol, 0.4 equiv) at 0 °C under a nitrogen atmosphere, the reaction was stirred 1 h at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (5 mL) and extracted with DCM (3 x 10 mL). The combined organic extracts were concentrated under vacuum to yield a 70 mg crude (S)- 6-isocyanato-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one as a yellow solid.
Figure imgf000290_0002
(S)-1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6- yl)urea To a stirred solution of (S)-6-isocyanato-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin- 2(1H)-one (70 mg, 0.187 mmol, 1 equiv) in DCM (5 mL) was added TEA (57 mg, 0.560 mmol, 3 equiv) and 1-methylcyclobutanamine hydrochloride (27 mg, 0.224 mmol, 1.2 equiv) at 0 °C under a nitrogen atmosphere, the reaction was stirred 1 h at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford (S)-1-(1- methylcyclobutyl)-3-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6- yl)urea (9.2 mg, 10.67%) as a yellow solid. MS (ESI): mass calcd. for C23H23F3N4O3, 460.17 m/z, found 461.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.24 (s, 1H), 7.95 (d, J = 2.5 Hz, 1H), 7.47 (t, J = 8.3 Hz, 1H), 7.29 (d, J = 8.2 Hz, 4H), 7.11 (s, 1H), 6.57 (s, 1H), 6.31 (s, 1H), 2.28 (q, J = 9.5 Hz, 2H), 1.89 (d, J = 7.0 Hz, 4H), 1.76 (dd, J = 9.2, 6.0 Hz, 3H), 1.39 (s, 3H). The following examples in Table 11 were made according to the procedure for example 200. Table 11
Figure imgf000291_0001
Figure imgf000292_0001
Figure imgf000293_0001
Figure imgf000294_0001
Figure imgf000295_0001
Figure imgf000296_0001
Figure imgf000297_0003
Figure imgf000297_0004
Figure imgf000297_0002
Figure imgf000297_0001
Example 226: 1-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-7-yl]-3-(1-methylcyclobutyl)urea Synthetic Scheme
Figure imgf000298_0001
1-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-7- yl]-3-(1-methylcyclobutyl)urea To a solution of (2R)-7-amino-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (100 mg, 0.296 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (35.19 mg, 0.118 mmol, 0.4 equiv) at 0°C under N2 atmosphere. The resulting mixture was stirred for 1 h at 25 °C. Then TEA (90.00 mg, 0.888 mmol, 3 equiv) was added into above system at 0 °C. The resulting mixture was stirred for 1 h at 25 °C. To the above mixture was added 1- methylcyclobutan-1-amine (50.49 mg, 0.592 mmol, 2 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD 30*150 mm, 5μm column (eluent: 42% to 60% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 1-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-7-yl]-3-(1-methylcyclobutyl)urea (32.5 mg, 24.40%) as a white solid. MS (ESI): mass calcd. for C22H23F3N4O3:448.17 m/z, found:449.15 [M+H]+. 1H NMR (400 MHz, DMSO- d6) δ 8.24 (s, 1H), 8.03 – 8.12 (m, 1H), 7.81 (d, J = 7.7 Hz, 1H), 7.53 (d, J = 8.0 Hz, 1H), 7.25 (d, J = 2.0 Hz, 1H), 6.76 – 6.86 (m, 2H), 6.23 (s, 1H), 5.24 (s, 2H), 4.78 – 4.88 (m, 1H), 2.22 – 2.30 (m, 2H), 1.82 – 1.93 (m, 2H), 1.69 – 1.82 (m, 2H), 1.47 (d, J = 6.7 Hz, 3H), 1.38 (s, 3H).19F NMR (376 MHz, DMSO) δ -66.54. The following examples in Table 12 were made according to the procedure for example 226. Table 12
Figure imgf000299_0001
Figure imgf000300_0001
Figure imgf000301_0001
Figure imgf000302_0001
Figure imgf000303_0002
Example 236: (R)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(4-hydroxy-2- methylbutan-2-yl)urea Synthetic Scheme
Figure imgf000303_0001
Figure imgf000304_0001
methyl (R)-2-(5-bromo-2-nitrophenoxy)propanoate To a solution of 2-amino-5-nitrophenol (900 mg, 0.065 mmol, 1 equiv) in THF (15 mL) was added methyl (2S)-2-hydroxypropanoate (649.68 mg, 6.240 mmol, 1.5 equiv) and PPh3 (1636.87 mg, 6.240 mmol, 1.5 equiv). The resulting mixture was maintained under nitrogen and stirred at rt for 1 h. Then DEAD (1086.85 mg, 6.240 mmol, 1.5 equiv) was added at 0 oC. The resulting mixture was maintained under nitrogen and stirred at 80°C for 3 h. After cooling down to rt, the reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0- 20% ethyl acetate/petroleum ether) to afford the methyl (2R)-2-(5-bromo-2- nitrophenoxy)propanoate (1.12 g, 88.53%) as a yellow oil. LC/MS: mass calcd. for C10H10BrNO5: 302.97, found: 304.09, 306.09 [M+H, M+H+2]+.
Figure imgf000304_0002
(R)-7-bromo-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-(5-bromo-2-nitrophenoxy)propanoate (1.1 g, 3.63 mmol, 1 equiv) in MeOH (15 mL) and H2O (1.5 mL) was added Fe (2.03 g, 36.3mmol, 10 equiv) and NH4Cl (1.96 g, 36.3 mmol, 10 equiv). The resulting mixture was maintained under nitrogen and stirred at 70 °C for 3 h. After cooling down to rt, the reaction was quenched with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the methyl (2R)-2-(5- bromo-2-nitrophenoxy)propanoate (800 mg, 91.53%) as a yellow oil. LC/MS: mass calcd. for C9H8BrNO2: 240.97, found: 241.07, 243.97 [M+H, M+H+2]+.
Figure imgf000305_0001
(R)-4-benzyl-7-bromo-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (2 g, 8.262 mmol, 1 equiv) and benzyl bromide (2826.22 mg, 16.524 mmol, 2 equiv) in dimethylformamide (50 mL) was added Cs2CO3 (8100.74 mg, 24.786 mmol, 3 equiv). The reaction was stirred at 100 °C under N2 for 3 h. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-benzyl-7-bromo-2-methyl-2H-1,4-benzoxazin-3- one (2.35 g, 85.62%) as a white solid. LC/MS (ESI): mass calcd. for C16H14BrNO2: 331.02 m/z, found:332.00, 334.00 [M+H, M+H+2]+.
Figure imgf000305_0002
(R)-7-amino-4-benzyl-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-benzyl-7-bromo-2-methyl-2H-1,4-benzoxazin-3-one (2.35 g, 5.916 mmol, 1 equiv) in ethyl alcohol (60 mL) and water (15 mL) was added N1,N2- dimethylcyclohexane-1,2-diamine (0.50 g, 3.550 mmol, 0.6 equiv), CuSO4 (0.94 g, 5.916 mmol, 1 equiv) and NaN3 (1.15 g, 17.75 mmol, 3 equiv). The reaction was stirred at 80 °C under N2 for 3 h. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-benzyl-2-methyl-2H-1,4-benzoxazin-3-one (1.3 g, 81.89%) as a white solid. LC/MS (ESI): mass calcd. for C16H16N2O2: 268.12 m/z, found:269.15 [M+H]+.
Figure imgf000306_0001
(R)-1-(4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(4-hydroxy-2- methylbutan-2-yl)urea To a solution of (2R)-7-amino-4-benzyl-2-methyl-2H-1,4-benzoxazin-3-one (100 mg, 0.373 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (44.24 mg, 0.149 mmol, 0.4 equiv). The reaction was stirred at 0 °C under N2 for 1 h. The reaction was added TEA (188.57 mg, 1.865 mmol, 5 equiv) and stirred at rt under N2 for 1 h. The reaction was added 3-amino-3- methylbutan-1-ol (153.80 mg, 1.492 mmol, 4 equiv) and stirred at rt under N2 for 1 h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep Phenyl OBD Column 19*150 mm, 5μm (eluent: 27% to 49% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 1-[(2R)-4-benzyl-2-methyl-3- oxo-2H-1,4-benzoxazin-7-yl]-3-(4-hydroxy-2-methylbutan-2-yl)urea (36.2 mg, 24.36%) as a white solid. LC/MS (ESI): mass calcd. for C22H27N3O4:397.20 m/z, found:398.00 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.27 (s, 1H), 7.26-7.38 (m, 2H), 7.20-7.25 (m, 4H), 6.75-6.86 (m, 2H), 5.95 (s, 1H), 5.09 (s, 2H), 4.79-4.81 (m, 1H), 4.37-4.41 (m, 1H), 3.44-3.50 (m, 2H), 1.78- 1.80 (t, J = 6 Hz, 2H), 1.46-1.48 (m, 3H), 1.25 (s, 6H). The following examples in Table 13 were made according to the procedure for example 236. Table 13
Figure imgf000306_0002
Figure imgf000307_0001
Example 239: 1-(2-hydroxy-2-methylpropyl)-3-(2-oxo-1-(3-(trifluoromethoxy)benzyl)-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000308_0001
6-nitro-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a mixture of 6-nitroquinoxalin-2(1H)-one (1.8 g, 9.417 mmol, 1.2 equiv) and 1-(bromomethyl)- 3-(trifluoromethoxy)benzene (2.00 g, 7.848 mmol, 1 equiv) in DMF (30 mL) was stirred and added K2CO3 (2.18 g, 15.695 mmol, 2 equiv). The reaction was stirred at rt for 1 h. The reaction mixture was quenched by ice water (30 mL) and extracted with EA (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford 6-nitro-1-(3- (trifluoromethoxy)benzyl)quinoxalin-2(1H)-one (1.7 g, 59.31%) as a yellow solid. MS (ESI): mass calcd. for C16H10F3N3O4, 365.06 m/z, found 366.00 [M+H]+.
Figure imgf000308_0002
6-amino-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a mixture of 6-nitro-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one (1.0 g, 2.738 mmol, 1 equiv) in EtOH (25 mL) and water (5 mL). Fe (1.53 g, 27.380 mmol, 10 equiv) and NH4Cl (1.46 g, 27.380 mmol, 10 equiv) was added into the mixture. The resulting solution was stirred at 60 °C for 1 h. The reaction was quenched with water (30 mL) at rt. After filtration, the filter cake was washed with EA (3 x 20 mL). The filtrate was added water (20 mL) and extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 6-amino-1-(3- (trifluoromethoxy)benzyl)quinoxalin-2(1H)-one (700 mg, 75.51%) as a yellow solid. MS (ESI): mass calcd. for C16H12F3N3O2, 335.09 m/z, found 336.05 [M+H]+.
Figure imgf000309_0001
1-(2-hydroxy-2-methylpropyl)-3-(2-oxo-1-(3-(trifluoromethoxy)benzyl)-1,2- dihydroquinoxalin-6-yl)urea To a mixture of 6-amino-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one (70 mg, 0.209 mmol, 1 equiv) in DCM (6 mL) was stirred under nitrogen atmosphere and added triphosgene (19 mg, 0.063 mmol, 0.3 equiv) at 0 °C. The reaction was stirred at rt for 1 h. TEA (53 mg, 0.522 mmol, 2.5 equiv) was added into the mixture at 0°C. The reaction was stirred at rt for 1 h.1-amino- 2-methylpropan-2-ol (23 mg, 0.251 mmol, 1.2 equiv) was added into the mixture at 0 °C. The reaction was stirred at rt for 1 h. The reaction mixture was quenched by 1N hydrochloric acid (5 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 1-(2-hydroxy-2- methylpropyl)-3-(2-oxo-1-(3-(trifluoromethoxy)benzyl)-1,2-dihydroquinoxalin-6-yl)urea (26.1 mg, 27.71%) as a yellow solid. MS (ESI): mass calcd. for C21H21F3N4O4, 450.15 m/z, found 451.00 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.33 (s, 1H), 8.01 (d, J = 2.4 Hz, 1H), 7.41 – 7.53 (m, 2H), 7.32 – 7.41 (m, 2H), 7.21 – 7.32 (m, 2H), 6.18 (t, J = 5.7 Hz, 1H), 5.49 (s, 2H), 4.55 (s, 1H), 3.05 (d, J = 5.7 Hz, 2H), 1.09 (s, 6H). The following examples in Table 14 were made according to the procedure for example 239. Table 14
Figure imgf000310_0001
Figure imgf000311_0002
Example 246: 1-(tert-butyl)-3-(1-(1-(3-cyclobutylphenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000311_0001
1-(3-cyclobutylphenyl)ethan-1-one In a 100-mL round bottom flask, to a solution of 1-bromo-3-cyclobutylbenzene (500 mg, 2.369 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added dropwise n-BuLi (2.5 M, 1.1 mL, 2.843 mmol, 1.2 equiv) at -78 °C under N2 atmosphere. The reaction mixture was stirred at -78 °C for 30 mins. N-methoxy-N-methylacetamide (292 mg, 2.843 mmol, 1.2 equiv) was added into the solution and the mixture was stirred for another 1 h. The reaction was quenched with water/sat. NH4Cl (5 mL), and then the mixture was extracted with ether/EtOAc (2 x 15 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with PE/EA (5/1) to afford 1-(3-cyclobutylphenyl)ethan-1-one (280 mg, 67.96%) as a white solid. MS (ESI): mass calcd. for C12H14O, 174.10 m/z, found 175.20 [M+H]+.
Figure imgf000312_0001
1-(3-cyclobutylphenyl)ethan-1-ol A solution of 1-(3-cyclobutylphenyl)ethanone (280 mg, 1.607 mmol, 1 equiv) in MeOH (5 mL) was added NaBH4 (152 mg, 4.018 mmol, 2.5 equiv) at rt. The resulting mixture was stirred for 1h at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford 1-(3-cyclobutylphenyl)ethan-1-ol (200 mg, 70.61%) as a white solid. MS: mass calcd. for C12H16O: 176.12, found: 159.15 [M-OH]+.
Figure imgf000312_0002
1-(1-bromoethyl)-3-cyclobutylbenzene A solution of 1-(3-cyclobutylphenyl)ethan-1-ol (40 mg, 0.227 mmol, 1 equiv) in DCM (5 mL) was added phosphorus tribromide (154 mg, 0.568 mmol, 2.5 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by ice-water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product 1-(1-bromoethyl)-3- cyclobutylbenzene (40 mg, 73.70%) was used in the next step directly without further purification.
Figure imgf000313_0001
1-(1-(3-cyclobutylphenyl)ethyl)-6-nitroquinoxalin-2(1H)-one A solution of 1-(1-bromoethyl)-3-cyclobutylbenzene (150 mg, 0.627 mmol, 1 equiv) in dimethylformamide (5 mL) was added 6-nitroquinoxalin-2(1H)-one (239 mg, 1.254 mmol, 2.0 equiv) and K2CO3 (260 mg, 1.881 mmol, 3.0 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL)The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with NaCl (aq., 3 x 5 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (1/3) to afford 1-(1-(3-cyclobutylphenyl)ethyl)-6-nitroquinoxalin-2(1H)-one (50 mg, 23.00%) as a light yellow solid. MS (ESI): mass calcd. C20H19N3O3 for: 349.14 m/z, found 350.20 [M+H]+.
Figure imgf000313_0002
6-amino-1-(1-(3-cyclobutylphenyl)ethyl)quinoxalin-2(1H)-one A solution of 1-(1-(3-cyclobutylphenyl)ethyl)-6-nitroquinoxalin-2(1H)-one (60 mg, 0.172 mmol, 1 equiv) in EtOH (5 mL)/H2O (0.5 mL) was added Fe (96 mg, 1.720 mmol, 10 equiv) and NH4Cl (92 mg, 1.720 mmol, 10 equiv) at rt. The resulting mixture was stirred for 1 h at 60 °C. The mixture was allowed to cool down to rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford 6-amino-1-(1-(3-cyclobutylphenyl)ethyl)quinoxalin- 2(1H)-one (45 mg, 82.04%) as a light yellow solid. MS: mass calcd. for C20H21N3O: 319.17, found: 320.20 [M+H]+.
Figure imgf000313_0003
1-(tert-butyl)-3-(1-(1-(3-cyclobutylphenyl)ethyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)urea A solution of 6-amino-1-(1-(3-cyclobutylphenyl)ethyl)quinoxalin-2(1H)-one (40 mg, 0.125 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (37 mg, 0.375 mmol, 3.0 equiv) and triethylamine (51 mg, 0.500 mmol, 4.0 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL). The resulting mixture was extracted with EA (5 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product 30 mg was purified by Prep-HPLC with the following conditions (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0) to affored 3-tert-butyl-1-{1-[1-(3- cyclobutylphenyl)ethyl]-2-oxoquinoxalin-6-yl}urea (7.8 mg, 14.70%) as a yellow solid. MS (ESI): mass calcd. C25H30N4O2 for: 418.24 m/z, found 419.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.39 (s, 1H), 8.27 (s, 1H), 7.92 (d, J = 2.6 Hz, 1H), 7.21 – 7.32 (m, 2H), 6.98 – 7.21 (m, 4H), 6.55 – 6.59 (m, 1H), 6.03 (s, 1H), 3.45 – 3.51 (m, 1H), 2.16 – 2.30 (m, 2H), 1.98 – 2.09 (m, 2H), 1.90 – 1.98 (m, 1H), 1.82 – 1.89 (m, 3H), 1.72 –1.81 (m, 1H), 1.30 (s, 9H). The following examples in Table 15 were made according to the procedure for example 246. Table 15
Figure imgf000314_0001
Figure imgf000315_0001
Figure imgf000316_0001
Figure imgf000317_0002
Example 255: 1-((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclobutyl)urea Synthetic Scheme
Figure imgf000317_0001
1-((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(1- methylcyclobutyl)urea To a stirred mixture of (R)-7-amino-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (100 mg, 0.354 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (42.04 mg, 0.142 mmol, 0.4 equiv) at 0 degrees C under nitrogen atmosphere. After 30 min, the TEA (107.52 mg, 1.062 mmol, 3 equiv) was added at 0 degrees C. After stirring for 30 min at 0 degrees C, the 1-methylcyclobutan-1-amine (60.32 mg, 0.708 mmol, 2 equiv) and TEA (107.52 mg, 1.062 mmol, 3 equiv) was added to the mixture at 0 degrees C. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 20 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10 mmol /L NH4HCO3) to afford the 1-((R)-2-methyl-3-oxo- 4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclobutyl)urea (63.0 mg, 45.21%) as a white solid. MS (ESI): mass calcd. for C23H27N3O3: 393.21 m/z, found 394.05[M +H] +.1H-NMR (300 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.31 – 7.42 (m, 2H), 7.20 – 7.31 (m, 4H), 6.55 – 6.69 (m, 2H), 6.22 (s, 1H), 6.03 – 6.16 (m, 1H), 4.69 – 4.82 (m, 1H), 2.17 – 2.32 (m, 2H), 1.81 – 1.93 (m, 2H), 1.65 – 1.80 (m, 5H), 1.44 (d, J = 6.8 Hz, 3H), 1.36 (s, 3H). The following examples in Table 16 were made according to the procedure for example 255. Table 16
Figure imgf000318_0001
Figure imgf000319_0001
Figure imgf000320_0001
Figure imgf000321_0002
Figure imgf000321_0001
Example 266: 3-tert-butyl-1-{1-[(2-cyano-5-methylphenyl)methyl]-2-oxo-3,4- dihydroquinolin-6-yl}urea. Synthetic Scheme
Figure imgf000322_0001
1-bromo-2-(bromomethyl)-4-methylbenzene To a stirred solution of (2-bromo-5-methylphenyl) methanol (600 mg, 2.984 mmol, 1 equiv) in DCM (6 mL) was added mg, 8.952 mmol, 3 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 3 h at room temperature under nitrogen atmosphere.The reaction was monitored by LCMS. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 1-bromo-2-(bromomethyl)-4- methylbenzene (300 mg, 38.09%) as a yellow solid. LC/MS (ESI): mass calcd. for C8H8Br2: 263.90 m/z, found: 265.05, 267.05 [M+H, M+H+2]+.
Figure imgf000323_0001
1-[(2-bromo-5-methylphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one To a stirred solution of 1-bromo-2-(bromomethyl)-4-methylbenzene (280 mg, 1.061 mmol, 1 equiv) in DMF (3 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (203.85 mg, 1.061 mmol, 1 equiv) and Cs2CO3 (691.24 mg, 2.122 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (20 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 1-[(2-bromo-5-methylphenyl)methyl]- 6-nitro-3,4-dihydroquinolin-2-one (100 mg, 25.12%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H15BrN2O3: 374.03 m/z, found 375.00, 377.00 [M+H, M+H+2]+.
Figure imgf000323_0002
4-methyl-2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile To a stirred solution of 1-[(2-bromo-5-methylphenyl)methyl]-6-nitro-3,4-dihydroquinolin-2-one (150 mg, 0.400 mmol, 1 equiv) and Zn(CN)2 (31.36 mg, 0.480 mmol, 1.2 equiv) in DMF(2 mL) was added BrettPhos Pd G3 (72.48 mg, 0.080 mmol, 0.2 equiv), BrettPhos (42.92 mg, 0.080 mmol, 0.2 equiv) and Zn (3.92 mg, 0.060 mmol, 0.15 equiv) in portions at room temperature under nitrogen atmosphere.The resulting mixture was stirred for 3 h at 120 °C under nitrogen atmosphere.The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 4-methyl-2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl)methyl]benzonitrile (50 mg, 38.92%) as a yellow solid. LC/MS (ESI): mass calcd. for C18H15N3O3:321.11 m/z, found 322.10 [M+H]+.
Figure imgf000324_0001
2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-4-methylbenzonitrile To a stirred solution of 4-methyl-2-[(6-nitro-2-oxo-3,4-dihydroquinolin-1-yl) methyl] benzonitrile (70 mg, 0.218 mmol, 1 equiv) in HOAc (1 mL) was added Zn (142.42 mg, 2.180 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-4-methylbenzonitrile (60 mg, 94.53%) as a yellow solid. LC/MS (ESI): mass calcd. for C18H17N3O:291.14 m/z, found 292.10 [M+H]+
Figure imgf000324_0002
3-tert-butyl-1-{1-[(2-cyano-5-methylphenyl)methyl]-2-oxo-3,4-dihydroquinolin-6-yl}urea To a stirred solution of 2-[(6-amino-2-oxo-3,4-dihydroquinolin-1-yl)methyl]-4- methylbenzonitrile (50 mg, 0.172 mmol, 1 equiv) in DCM/Toluene(2:1,1 mL) was added 2- isocyanato-2-methylpropane (51.04 mg, 0.516 mmol, 3 equiv) in portions at room temperature. The final reaction mixture was stirred overnight at 90 °C. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 3-tert-butyl-1-{1-[(2-cyano-5-methylphenyl)methyl]- 2-oxo-3,4-dihydroquinolin-6-yl}urea (6.7 mg, 9.68%) as a white solid. LC/MS (ESI): mass calcd. for C23H26N4O2:390.20 m/z, found 391.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.74 (d, J = 8.1 Hz, 1H), 7.34 (s, 1H), 7.26 (d, J = 8.1 Hz, 1H), 7.00 (d, J = 11.4 Hz, 2H), 6.66 (d, J = 9.0 Hz, 1H), 5.93 (s, 1H), 5.19 (s, 2H), 2.95 – 2.90 (m, 2H), 2.73 – 2.65 (m, 2H), 2.31 (s, 3H), 1.26 (s, 9H). Example 267: 1-(1-benzyl-5-chloro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000325_0001
5-chloro-3,4-dihydroquinolin-2(1H)-one To a solution of 3-chloro-2-iodoaniline (1.5 g, 5.918 mmol, 1 equiv), AIBN (388.71 mg, 2.367 mmol, 0.4 equiv) and tributyltin (2583.73 mg, 8.877 mmol, 1.5 equiv) in DMSO (10 mL) was added ethyl acrylate (2369.91 mg, 23.672 mmol, 4 equiv). The reaction was stirred at 120 °C under N2 for 3 h. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-chloro-3,4-dihydro-1H-quinolin-2-one (348 mg, 32.38%) as a white solid. LC/MS: mass calcd. for C9H8ClNO: 181.03, found: 182.05 [M+H]+.
Figure imgf000326_0001
5-chloro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 5-chloro-3,4-dihydro-1H-quinolin-2-one (376 mg, 2.070 mmol, 1 equiv) in sulfuric acid (4 mL) was added KNO3 (167.84 mg, 1.660 mmol, 0.80 equiv). The reaction was stirred at rt for 1 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-chloro-6-nitro-3,4-dihydro-1H-quinolin-2-one (96 mg, 20.46%) as a white solid. LC/MS: mass calcd. for C9H7ClN2O3: 226.01, found: 226.85 [M+H]+.
Figure imgf000326_0002
benzyl-5-chloro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 5-chloro-6-nitro-3,4-dihydro-1H-quinolin-2-one (151 mg, 0.666 mmol, 1 equiv) in dimethylformamide (3 mL) was added Cs2CO3 (653.31 mg, 1.998 mmol, 3 equiv) and (bromomethyl)benzene (653.31 mg, 1.998 mmol, 3 equiv). The reaction was stirred at rt for 1 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-5-chloro-6- nitro-3,4-dihydroquinolin-2-one (199 mg, 94.29%) as a white solid. LC/MS (ESI): mass calcd. for C16H13ClN2O3: 316.06 m/z, found:317.05 [M+H]+.
Figure imgf000327_0002
6-amino-1-benzyl-5-chloro-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-5-chloro-6-nitro-3,4-dihydroquinolin-2-one (100 mg, 0.316 mmol, 1 equiv) in ethyl alcohol (3 mL) was added iron (177 mg, 3.169 mmol, 10.04 equiv), NH4Cl (167 mg, 3.122 mmol, 9.89 equiv) and water (0.3 mL). The reaction was stirred at 60 °C for 3 h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 6-amino-1-benzyl-5-chloro-3,4-dihydroquinolin-2-one (74.5 mg, 82.29%) as a white solid. LC/MS (ESI): mass calcd. for C16H15ClN2O: 286.09 m/z, found:287.05 [M+H]+.
Figure imgf000327_0001
1-(1-benzyl-5-chloro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-5-chloro-3,4-dihydroquinolin-2-one (70 mg, 0.244 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (48.29 mg, 0.487 mmol, 2.00 equiv) in DCM (1 mL) was added TEA (73.93 mg, 0.731 mmol, 2.99 equiv). The reaction was stirred at rt for 3 h. Quenched with water (3 mL) and extracted with DCM (3 x 3 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column,50 mm x 30 mm x 5 μm (eluent: 36% to 56% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1- (1-benzyl-5-chloro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (23.4 mg, 24.79%) as a white solid. LC/MS (ESI): mass calcd. for C21H24ClN3O2:385.16 m/z, found:386.00 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.78-7.85 (m, 2H), 7.26-7.34 (m, 2H), 7.18-7.26 (m, 3H), 6.87- 6.91 (m, 2H), 5.13 (s, 2H), 3.04-3.05 (m, 2H), 2.68-2.71 (m, 2H), 1.26 (s, 9H). Example 268 and Example 269: 1-(tert-butyl)-3-((S)-2-methyl-3-oxo-4-((S)-1-phenylethyl)- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4- ((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000328_0001
2-methyl-7-nitro-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (350 mg, 1.681 mmol, 1 equiv) in DCM (10 mL) was added (R)-1-phenylethan-1-ol (205.40 mg, 1.681 mmol, 1 equiv) and PPh3 (661.48 mg, 2.522 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5 h. To the above mixture was added DEAD (439.21 mg, 2.522 mmol, 1.5 equiv) dropwise at 0℃. The resulting mixture was stirred for additional 1 hour at rt. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 2-methyl-7-nitro-4-[(1S)-1-phenylethyl]-2H-1,4- benzoxazin-3-one (270 mg, 51.42%) as a yellow solid. MS (ESI): mass calcd. for C17H16N2O4: 312.11 m/z, found 313.15 [M+H]+.
Figure imgf000329_0001
7-amino-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 2-methyl-7-nitro-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (270 mg, 0.864 mmol, 1 equiv) in MeOH/H2O (10:1 ,11 mL) was added Fe (482.77 mg, 8.640 mmol, 10 equiv) and NH4Cl (462.41 mg, 8.640 mmol, 10 equiv). The reaction was stirred at 80℃ for 1 hour. The resulting mixture was filtered, and the filter cake was washed with EA (3 x 30 mL). The filtrate was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 7-amino-2-methyl-4-[(1S)-1-phenylethyl]-2H- 1,4-benzoxazin-3-one (200 mg, 81.96%) as a yellow oil. MS (ESI): mass calcd. for C17H18N2O2: 282.14 m/z, found 283.00 [M+H]+.
Figure imgf000329_0002
1-(tert-butyl)-3-(2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin- 7-yl)urea To a solution of 7-amino-2-methyl-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (130 mg, 0.460 mmol, 1 equiv) in DCM/Toluene (2:1, 15 mL) was added 2-isocyanato-2-methylpropane (273.86 mg, 2.760 mmol, 6 equiv). The reaction was stirred at 90℃ for 2 hours. After cooling to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column, 30*150 mm, 5μm column (eluent: 43% to 65% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4- benzoxazin-7-yl}urea (40 mg, 22.77%) as a white solid.MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found:382.20 [M+H]+.
Figure imgf000330_0001
1-(tert-butyl)-3-((S)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1- phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-{2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-7- yl}urea (40 mg) was separated by chiral-HPLC using a CHIRALPAK IE, 2*25 cm, 5 μm column (eluent: 10% to 10% (v/v) Hex(0.5% 2M NH3-MeOH) and EtOH) to yield first enantiomer example 3-tert-butyl-1-[(2S)-2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-7- yl]urea (17.6 mg, 44.00%) as a white solid. MS(ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found: 382.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.11 – 8.29 (m, 1H), 7.25 – 7.45 (m, 6H), 6.55 – 6.68 (m, 1H), 6.35 – 6.55 (m, 1H), 6.11 – 6.26 (m, 1H), 5.85 – 6.03 (m, 1H), 4.63 – 4.79 (m, 1H), 1.64 – 1.71 (m, 3H), 1.43 – 1.59 (m, 3H), 1.25 – 1.26 (m, 9H).and to yield second enantiomer example 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4- benzoxazin-7-yl]urea (17 mg, 42.50%) as a white solid. MS(ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found: 382.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.84 – 8.20 (m, 1H), 7.32 – 7.46 (m, 2H), 7.10 – 7.32 (m, 4H), 6.49 – 6.65 (m, 2H), 6.02 – 6.18 (m, 1H), 5.76 – 5.97 (m, 1H), 4.63 – 4.79 (m, 1H), 1.73 – 1.81 (m, 3H), 1.40 – 1.55 (m, 3H), 1.26 – 1.29 (m, 9H). Example 270: 1-(1-benzyl-7-cyano-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea. Synthetic Scheme
Figure imgf000331_0001
1-benzyl-6-nitro-2-oxo-3,4-dihydroquinoline-7-carbonitrile To a stirred solution of 1-benzyl-7-chloro-6-nitro-3,4-dihydroquinolin-2-one (500 mg, 1.579 mmol, 1 equiv) in DMF (5 mL) was added Zn(CN)2 (222.43 mg, 1.895 mmol, 1.2 equiv), BrettPhos Pd G3 (286.20 mg, 0.316 mmol, 0.2 equiv), BrettPhos (169.47 mg, 0.316 mmol, 0.2 equiv) and Zn (15.48 mg, 0.237 mmol, 0.15 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at 120 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 1-benzyl-6-nitro-2-oxo-3,4-dihydroquinoline-7-carbonitrile (200 mg, 41.23%) as a white solid. LC/MS (ESI): mass calcd. for C17H13N3O3: 307.10 m/z, found 308.05 [M+H]+ .
Figure imgf000331_0002
6-amino-1-benzyl-2-oxo-3,4-dihydroquinoline-7-carbonitrile To a stirred solution of 1-benzyl-6-nitro-2-oxo-3,4-dihydroquinoline-7-carbonitrile (200 mg, 0.651 mmol, 1 equiv) in AcOH (2 mL) was added Zn (425.50 mg, 6.510 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-1-benzyl-2-oxo-3,4- dihydroquinoline-7-carbonitrile (80 mg, 44.44%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H15N3O: 277.12 m/z, found 278.05 [M+H] +.
Figure imgf000332_0001
1-(1-benzyl-7-cyano-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea To a stirred solution of 6-amino-1-benzyl-2-oxo-3,4-dihydroquinoline-7-carbonitrile (70 mg, 0.252 mmol, 1 equiv) in DCM/Toluene (2:1,1 mL) was added 2-isocyanato-2-methylpropane (75.07 mg, 0.756 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 90 °C under air atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 26% to 45% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 1-(1-benzyl-7-cyano-2-oxo- 3,4-dihydroquinolin-6-yl)-3-tert-butylurea (3.9 mg, 4.01%) as a white solid. LC/MS (ESI): mass calcd. for C22H24N4O2: 376.19 m/z, found 377.15 [M+H]+.1H NMR (300 MHz, Methanol-d4) δ 7.82 (s, 1H), 7.35 – 7.31 (m, 2H), 7.24 (t, J = 7.1 Hz, 3H), 7.11 (s, 1H), 5.20 (s, 2H), 3.08 – 3.01 (m, 2H), 2.80 – 2.75 (m, 2H), 1.35 (s, 9H). Example 271: 1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-(3-(trifluoromethyl)phenyl)ethyl)- 1,2,3,4-tetrahydroquinazolin-6-yl)urea Synthetic Scheme
Figure imgf000333_0001
3-methyl-6-nitro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)-one To a solution of 3-methyl-6-nitro-1,4-dihydroquinazolin-2-one (200 mg, 0.966 mmol, 1 equiv) in DMF (5mL) was added K2CO3 (0.40 g, 2.890 mmol, 3 equiv) and 1-(1-bromoethyl)-3-(trifluoromethyl)benzene (269 mg, 1.06 mmol, 1.1 equiv). The resulting mixture was placed at rt and stirred overnight. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by silica gel column to afford 3-methyl-6-nitro-1-(1-(3- (trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)-one (100 mg, 27.31%) as a yellow solid.
Figure imgf000333_0002
6-amino-3-methyl-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)-one To a solution of 3-methyl-6-nitro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)- one (100 mg, 0.264 mmol, 1 equiv) in MeOH (10 mL) was added Pd/C (28.0 mg, 10%, 0.026 mmol, 0.1 equiv). The resulting mixture was placed at rt under a hydrogen atmosphere until the starting material was totally consumed by LCMS. The mixture was filtered and concentrated under vacuum to afford 6-amino- 3-methyl-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin-2(1H)-one (60 mg, 65.15%) as a yellow solid. MS (ESI): mass calcd. for C18H18F3N3O, 349.14 m/z, found 350.10 [M+H]+.
Figure imgf000334_0001
1-(tert-butyl)-3-(3-methyl-2-oxo-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-1,2,3,4- tetrahydroquinazolin-6-yl)urea To a stirred solution of 6-amino-3-methyl-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydroquinazolin- 2(1H)-one (100 mg, 0.286 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2-methylpropane (142 mg, 1.431 mmol, 5 equiv) and TEA (87 mg, 0.859 mmol, 3 equiv) stirred overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid.The compound was separated by preparative HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) to afford 1-(tert- butyl)-3-(3-methyl-2-oxo-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-1,2,3,4-tetrahydroquinazolin-6-yl)urea (12.6 mg, 9.78%) as a white solid. MS (ESI): mass calcd. for C23H27F3N4O2, 448.21 m/z, found 449.05 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.58 (s, 4H), 7.30 (s, 1H), 6.88 (d, J = 8.7 Hz, 1H), 6.40 (d, J = 8.8 Hz, 1H), 5.83 – 5.96 (m, 2H), 4.32 (s, 2H), 2.91 (s, 3H), 1.78 (d, J = 7.0 Hz, 3H), 1.26 (s, 9H). Example 272: 1-(1-benzyl-3-methyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000335_0001
3-methyl-6-nitroquinoxalin-2(1H)-one To a solution of 2-amino-4-nitroaniline (2 g, 13.060 mmol, 1 equiv) in ethyl alcohol (20 mL) was added ethyl pyruvate (1.82 g, 15.674 mmol, 1.20 equiv). The reaction was stirred at rt for 3 h. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-methyl-6-nitro- 1H-quinoxalin-2-one (1.78 g, 66.43%) as a yellow solid. MS (ESI): mass calcd. for C9H7N3O3: 205.05m/z, found 206.05 [M+H]+
Figure imgf000335_0002
benzyl-3-methyl-6-nitroquinoxalin-2(1H)-one To a solution of 3-methyl-6-nitro-1H-quinoxalin-2-one (400 mg, 1.950 mmol, 1 equiv) and benzyl bromide (500.17 mg, 2.925 mmol, 1.5 equiv) in dimethylformamide (5 mL) was added K2CO3 (814.22 mg, 5.850 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-3-methyl-6- nitroquinoxalin-2-one (212 mg, 36.82%) as a white solid. MS (ESI): mass calcd. for C16H13N3O3: 295.10 m/z, found 296.10 [M+H]+
Figure imgf000336_0001
6-amino-1-benzyl-3-methylquinoxalin-2(1H)-one To a solution of 1-benzyl-3-methyl-6-nitroquinoxalin-2-one (129 mg, 0.437 mmol, 1 equiv) in water (0.2 mL) and methanol (2 mL) was added iron (243.96 mg, 4.370 mmol, 10 equiv) and NH4Cl (233.67 mg, 4.370 mmol, 10 equiv). The reaction was stirred at 60 °C for 3 h. After cooling down to rt, the reaction was quenched with water (5 mL) and extracted with EA (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 6-amino-1-benzyl-3-methylquinoxalin-2-one (54.6 mg, 47.11%) as a white solid. LC/MS (ESI): mass calcd. for C16H15N3O: 265.12 m/z, found:266.15 [M+H]+.
Figure imgf000336_0002
1-(1-benzyl-3-methyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-3-methylquinoxalin-2-one (55 mg, 0.207 mmol, 1 equiv) in DCM (1 mL)/toluene (0.5 mL) was added 2-isocyanato-2-methylpropane (61.65 mg, 0.621 mmol, 3 equiv). The reaction was stirred at 90 °C for 3 h. After cooling down to rt, the reaction was quenched with water (5 mL) and extracted with DCM (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1-(1-benzyl-3- methyl-2-oxoquinoxalin-6-yl)-3-tert-butylurea (15.3 mg, 20.22%) as a yellow solid. LC/MS (ESI): mass calcd. for C21H24N4O2:364.19 m/z, found:365.051H NMR (400 MHz, DMSO-d6) δ 8.38 (s, 1H), 7.84 (s, 1H), 7.23-7.40 (m, 7H), 6.01 (s, 1H), 5.45 (s, 2H), 2.49 (s, 3H), 1.29 (s, 9H). Example 273: 3-tert-butyl-1-{1-[(3-chlorophenyl)methyl]-3-methyl-2-oxoquinoxalin-6- yl}urea. Synthetic Scheme
Figure imgf000337_0001
1-[(3-chlorophenyl) methyl]-3-methyl-6-nitroquinoxalin-2-one To a stirred solution of 3-methyl-6-nitro-1H-quinoxalin-2-one (400 mg, 1.950 mmol, 1 equiv) and 1-(bromomethyl)-3-chlorobenzene (600.90 mg, 2.925 mmol, 1.5 equiv) in DMF (5 mL) was added K2CO3 (542.81 mg, 3.900 mmol, 2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at room temperature under nitrogen atmosphere. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 1-[(3-chlorophenyl) methyl]-3-methyl-6-nitroquinoxalin-2-one (300 mg, 46.67%) as a yellow solid. LC/MS (ESI): mass calcd. for C16H12ClN3O3: 329.06 m/z, found 330.00 [M+H]+ .
Figure imgf000338_0001
6-amino-1-[(3-chlorophenyl) methyl]-3-methylquinoxalin-2-one To a stirred solution of 1-[(3-chlorophenyl) methyl]-3-methyl-6-nitroquinoxalin-2-one (300 mg, 0.910 mmol, 1 equiv) in MeOH / H2O (10:1, 4 mL) were added NH4Cl (486.66 mg, 9.100 mmol, 10 equiv) and Fe (508.08 mg, 9.100 mmol, 10 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at 60 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with CH2Cl2 / MeOH (10:1) to afford 6-amino-1- [(3-chlorophenyl) methyl]-3-methylquinoxalin-2-one (240 mg, 88.00%) as a white solid. LC/MS (ESI): mass calcd. for C16H14ClN3O: 299.08 m/z, found 300.00 [M+H]+.
Figure imgf000338_0002
3-tert-butyl-1-{1-[(3-chlorophenyl) methyl]-3-methyl-2-oxoquinoxalin-6-yl} urea To a stirred solution of 6-amino-1-[(3-chlorophenyl) methyl]-3-methylquinoxalin-2-one (100 mg, 0.334 mmol, 1 equiv) in DCM/Toluene (2:1,1 mL) was added 2-isocyanato-2- methylpropane (99.21 mg, 1.002 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a Xselect CSH OBD C18 150 mm x 30 mm x 5 μm column (eluent: 37% to 62% (v/v) CH3CN and H2O with 0.1%FA) to afford 3-tert-butyl-1-{1-[(3-chlorophenyl) methyl]-3-methyl-2-oxoquinoxalin-6-yl}urea (22.4 mg, 16.17%) as a yellow solid. LC/MS (ESI): mass calcd. for C21H23ClN4O2: 398.2 m/z, found 399.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.40 (s, 1H), 7.84 (d, J = 2.1 Hz, 1H), 7.38 – 7.28 (m, 6H), 6.02 (s, 1H), 5.45 (s, 2H), 2.49 (s, 3H) 1.30 (s, 9H). Example 274: 1-(tert-butyl)-3-(6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea Synthetic Scheme
Figure imgf000339_0001
6-chloro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of 6-chloro-3,4-dihydro-1H-1,5-naphthyridin-2-one (300 mg, 1.643 mmol, 1 equiv) in anhydrous dimethylformamide (5 mL) was added 1-(bromomethyl)-3- (trifluoromethyl)benzene (432 mg, 1.807 mmol, 1.1 equiv) and Cs2CO3 (1.6 g, 4.929 mmol, 3 equiv) at rt and stirred for 2 h. After completion of reaction, the reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford 6-chloro-1-(3- (trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (390 mg, 69.67%) as a light- yellow solid.
Figure imgf000340_0001
tert-butyl (6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2- yl)carbamate A solution of 6-chloro-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (390 mg, 1.145 mmol, 1 equiv) in 1,4-dioxane (5 mL) was treated with tert-butyl carbamate (269 mg, 2.290 mmol, 2 equiv), Cs2CO3 (1.11 g, 3.435 mmol, 3 equiv), EPHOS (61 mg, 0.115 mmol, 0.1 equiv) and EPhos Pd G4 (105 mg, 0.115 mmol, 0.1 equiv) for overnight at 100 °C under nitrogen atmosphere. The reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford tert-butyl (6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate (0.5 g, 96.01%) .
Figure imgf000340_0002
6-amino-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of tert-butyl (6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate (350 mg, 0.831 mmol, 1 equiv) in HCl (4N in EA, 10 mL) at rt. The reaction was stirred for 2 h at rt. The resulting mixture was concentrated under reduced pressure to afford 6-amino-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (300 mg, 112.42%) as a white solid.
Figure imgf000341_0001
1-(tert-butyl)-3-(6-oxo-5-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2- yl)urea A solution of 6-amino-1-(3-(trifluoromethyl)benzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (50 mg, 0.156 mmol, 1 equiv), TEA (79 mg, 0.780 mmol, 5 equiv) and 2-isocyanato-2-methylpropane (77 mg, 0.780 mmol, 5 equiv) in DCM (5 mL) was stirred for overnight at rt. The mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with EA (46%-57%) to afford crude products. The residue was purified by reversed-phase flash chromatography with the following conditions Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 40% B to 65% B in 9 min, 65% B; Wave Length: 254/220 nm; RT1(min): 8.0; Number Of Runs: 0 to afford 3-tert-butyl-1-(6-oxo-5-{[3-(trifluoromethyl)phenyl]methyl}-7,8- dihydro-1,5-naphthyridin-2-yl)urea (19.6 mg, 29.91%) as a white solid. MS (ESI):mass calcd. for C21H23F3N4O2: 420.18 m/z, found 421.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.89 (s, 1H), 7.91 (s, 1H), 7.66 – 7.58 (m, 2H), 7.54 (t, J = 7.6 Hz, 1H), 7.48 (d, J = 7.7 Hz, 1H), 7.32 (d, J = 8.9 Hz, 1H), 7.13 (d, J = 8.9 Hz, 1H), 5.18 (s, 2H), 2.96 – 3.04 (m, 2H), 2.81 (dd, J = 8.8, 6.2 Hz, 2H), 1.30 (s, 9H). Example 275: (R)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3- (tert-butyl)urea Synthetic Scheme
Figure imgf000341_0002
Figure imgf000342_0001
2-(1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)acetonitrile 3-In a 100-mL round bottom flask, to a solution of 1-benzyl-6-nitro-3,4-dihydroquinolin-2-one (300 mg, 1.063 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added dropwise LDA (2M, in THF, 0.63 mL, 1.276 mmol, 1.2 equiv) at -78 °C under N2 atmosphere. The reaction mixture was stirred at -78 °C for 20 mins. Then a solution of 2-bromoacetonitrile (153 mg, 1.276 mmol, 1.2 equiv) in 3 mL THF was added dropwise and the mixture was stirred for another 30 mins. The reaction was quenched with NH4Cl (aq., 10 mL), and then the mixture were extracted with EtOAc (2 x 15 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 2-(1-benzyl 6-nitro-2-oxo-1,2,3,4-tetrahydroquinolin-3- yl)acetonitrile (180 mg, 52.71%) as a white solid.
Figure imgf000342_0002
2-(6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)acetonitrile A solution of 2-(1-benzyl-6-nitro-2-oxo-3,4-dihydroquinolin-3-yl)acetonitrile (180 mg, 0.560 mmol, 1 equiv) in EtOH (5 mL) and H2O (0.5 mL) was added Fe (313 mg, 5.600 mmol, 10 equiv) and NH4Cl (300 mg, 5.600 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at 80 °C. The reaction was quenched with H2O (10 mL). The resulting mixture was extracted with EA (10 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 2-(6-amino-1-benzyl-2-oxo-1,2,3,4- tetrahydroquinolin-3-yl)acetonitrile (100 mg, 61.27%) as a brown solid. MS (ESI): mass calcd. for C18H17N3O, 291.14 m/z, found 292.15 [M+H]+.
Figure imgf000343_0001
(R)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea A solution of 2-(6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)acetonitrile (70 mg, 0.240 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (72 mg, 0.720 mmol, 3.0 equiv) and TEA (122 mg, 1.200 mmol, 5.0 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction was quenched with H2O (10 mL). The resulting mixture was extracted with EA (10 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column (PE/EA=2:1) and separated by Prep-Chiral-HPLC with the following conditions ( Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH-- HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 12 min; Wave Length: 220/254 nm; RT1(min): 6.534; RT2(min): 10.398; Sample Solvent: EtOH--HPLC; Injection Volume: 1.8 mL; Number Of Runs: 4 ) to affored (R)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea (18.8 mg, 19.39%) as an off-white solid. MS (ESI): mass calcd. C23H26N4O2 for: 390.21 m/z, found 391.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.35 (m, 3H), 7.24 – 7.28 (m, 2H), 7.19 – 7.21 (m,1H), 7.01 – 7.06 (m, 1H), 6.79 – 6.86 (m, 1H), 5.97 (s, 1H), 5.03 – 5.20 (m, 2H), 3.05 – 3.15 (m, 1H), 2.83 – 3.02 (m, 4H), 1.26 (s, 9H). And to afford (S)-1-(1-benzyl-3-(cyanomethyl)-2- oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (11.9 mg, 12.66%) as an off-white solid. MS (ESI): mass calcd. C23H26N4O2 for : 390.21m/z, found 391.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.35 (m, 3H), 7.24 – 7.28 (m, 2H), 7.19 – 7.21 (m,1H), 7.01 – 7.06 (m, 1H), 6.79 – 6.86 (m, 1H), 5.97 (s, 1H), 5.03 – 5.20 (m, 2H), 3.05 – 3.15 (m, 1H), 2.83 – 3.02 (m, 4H), 1.26 (s, 9H). Example 276: (S)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3- (tert-butyl)urea
Figure imgf000343_0002
(S)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea A solution of 2-(6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinolin-3-yl)acetonitrile (70 mg, 0.240 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (72 mg, 0.720 mmol, 3.0 equiv) and TEA (122 mg, 1.200 mmol, 5.0 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction was quenched with H2O (10 mL). The resulting mixture was extracted with EA (10 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column (PE/EA=2:1) and separated by Prep-Chiral-HPLC with the following conditions ( Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH-- HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 12 min; Wave Length: 220/254 nm; RT1(min): 6.534; RT2(min): 10.398; Sample Solvent: EtOH--HPLC; Injection Volume: 1.8 mL; Number Of Runs: 4 ) to affored (R)-1-(1-benzyl-3-(cyanomethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea (18.8 mg, 19.39%) as an off-white solid. MS (ESI): mass calcd. C23H26N4O2 for: 390.21 m/z, found 391.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.35 (m, 3H), 7.24 – 7.28 (m, 2H), 7.19 – 7.21 (m,1H), 7.01 – 7.06 (m, 1H), 6.79 – 6.86 (m, 1H), 5.97 (s, 1H), 5.03 – 5.20 (m, 2H), 3.05 – 3.15 (m, 1H), 2.83 – 3.02 (m, 4H), 1.26 (s, 9H). And to afford (S)-1-(1-benzyl-3-(cyanomethyl)-2- oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (11.9 mg, 12.66%) as an off-white solid. MS (ESI): mass calcd. C23H26N4O2 for : 390.21m/z, found 391.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.35 (m, 3H), 7.24 – 7.28 (m, 2H), 7.19 – 7.21 (m,1H), 7.01 – 7.06 (m, 1H), 6.79 – 6.86 (m, 1H), 5.97 (s, 1H), 5.03 – 5.20 (m, 2H), 3.05 – 3.15 (m, 1H), 2.83 – 3.02 (m, 4H), 1.26 (s, 9H). Example 277: (R)-1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea Synthetic Scheme
Figure imgf000344_0001
Figure imgf000345_0001
2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of 2-amino-5-nitrophenol (1 g, 6.488 mmol, 1 equiv) in anhydrous dimethylformamide (30 mL) was added K2CO3 (2.71 g, 19.464 mmol, 3 equiv) and methyl 2- bromobutanoate (1.41 g, 7.786 mmol, 1.2 equiv) at 0 °C and stirred for 30 min and the mixture is stirred for overnight at 90 °C. The reaction mixture was quenched by addition of water (15 mL). The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to give 2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (1.3 g, 90.17%) as a yellow solid.
Figure imgf000345_0002
4-benzyl-2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of To a stirred solution of 2-amino-5-nitrophenol (0.5 g, 2.25 mmol, 1 equiv) in anhydrous dimethylformamide (6 mL) was added Cs2CO3 (1.5 g, 4.50 mmol, 2 equiv) and BnBr (460 mg, 2.70 mmol, 1.2 equiv) at rt and stirred for 16 h. The reaction mixture was quenched by addition of water (15 mL). The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to give 4-benzyl-2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin- 3(4H)-one (0.5 g, 71.43%) as a yellow solid.
Figure imgf000345_0003
7-amino-4-benzyl-2-ethyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-benzyl-2-ethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (200 mg, 0.640 mmol, 1 equiv) in 15 mL MeOH was added Pd/C (10%, 0.1 g) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 1 h under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. This resulted in 7-7-amino-4-benzyl-2-ethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (180 mg, 99.56%) as a white solid.
Figure imgf000346_0001
(R) -1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert- butyl)urea A solution of 7-amino-4-benzyl-2-ethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.531 mmol, 1 equiv), TEA (269 mg, 2.655 mmol, 5 equiv) and 2-isocyanato-2-methylpropane (263 mg, 2.655 mmol, 5 equiv) in DCM (10 mL) was stirred for overnight at rt. The reaction mixture was quenched by addition of water (15 mL). The aqueous layer was extracted with DCM (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (2:1) to give crude product. The residue was purified by Chiral HPLC with the following conditions Column: CHIRALPAK ID, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 15% B to 15% B in 16 min; Wave Length: 220/254 nm; RT1(min): 9.531; RT2(min): 12.768; Sample Solvent: EtOH--HPLC; Injection Volume: 0.8 mL; Number Of Runs: 6 to afford (R)-1-(4-benzyl- 2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (15.5 mg, 7.64%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.20 (d, J = 2.8 Hz, 1H), 7.22 – 7.33 (m, 6H), 6.85 (dd, J = 8.7, 2.8 Hz, 1H), 6.76 – 6.66 (m, 1H), 5.94 (d, J = 2.8 Hz, 1H), 5.10 (t, J = 3.5 Hz, 2H), 4.63 – 4.65 (m, 1H), 1.78 – 1.91 (m, 2H), 1.26 (s, 9H), 1.04 (t, J = 7.4 Hz, 3H). And (S)-1-(4-benzyl-2-ethyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (13.4 mg, 6.60%) as a white solid.MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.20 (d, J = 2.8 Hz, 1H), 7.22 – 7.33 (m, 6H), 6.85 (dd, J = 8.7, 2.8 Hz, 1H), 6.76 – 6.66 (m, 1H), 5.94 (d, J = 2.8 Hz, 1H), 5.10 (t, J = 3.5 Hz, 2H), 4.63 – 4.65 (m, 1H), 1.78 – 1.91 (m, 2H), 1.26 (s, 9H), 1.04 (t, J = 7.4 Hz, 3H). Example 278: (S)-1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea
Figure imgf000347_0001
(S)-1-(4-benzyl-2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea A solution of 7-amino-4-benzyl-2-ethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.531 mmol, 1 equiv), TEA (269 mg, 2.655 mmol, 5 equiv) and 2-isocyanato-2-methylpropane (263 mg, 2.655 mmol, 5 equiv) in DCM (10 mL) was stirred for overnight at rt. The reaction mixture was quenched by addition of water (15 mL). The aqueous layer was extracted with DCM (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (2:1) to give crude product. The residue was purified by Chiral HPLC with the following conditions Column: CHIRALPAK ID, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 15% B to 15% B in 16 min; Wave Length: 220/254 nm; RT1(min): 9.531; RT2(min): 12.768; Sample Solvent: EtOH--HPLC; Injection Volume: 0.8 mL; Number Of Runs: 6 to afford (R)-1-(4-benzyl- 2-ethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (15.5 mg, 7.64%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.20 (d, J = 2.8 Hz, 1H), 7.22 – 7.33 (m, 6H), 6.85 (dd, J = 8.7, 2.8 Hz, 1H), 6.76 – 6.66 (m, 1H), 5.94 (d, J = 2.8 Hz, 1H), 5.10 (t, J = 3.5 Hz, 2H), 4.63 – 4.65 (m, 1H), 1.78 – 1.91 (m, 2H), 1.26 (s, 9H), 1.04 (t, J = 7.4 Hz, 3H). And (S)-1-(4-benzyl-2-ethyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (13.4 mg, 6.60%) as a white solid.MS (ESI): mass calcd. for C22H27N3O3: 381.21 m/z, found 382.20 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.20 (d, J = 2.8 Hz, 1H), 7.22 – 7.33 (m, 6H), 6.85 (dd, J = 8.7, 2.8 Hz, 1H), 6.76 – 6.66 (m, 1H), 5.94 (d, J = 2.8 Hz, 1H), 5.10 (t, J = 3.5 Hz, 2H), 4.63 – 4.65 (m, 1H), 1.78 – 1.91 (m, 2H), 1.26 (s, 9H), 1.04 (t, J = 7.4 Hz, 3H). Example 279: 1-(1-benzyl-5-cyano-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000348_0001
1-benzyl-6-nitro-2-oxo-1,2,3,4-tetrahydroquinoline-5-carbonitrile To a solution of 1-benzyl-5-chloro-6-nitro-3,4-dihydroquinolin-2-one (180 mg, 0.568 mmol, 1 equiv) in dimethylformamide (2 mL) was added zinc (5.57 mg, 0.085 mmol, 0.15 equiv), zinc cyanide (80.07 mg, 0.682 mmol, 1.2 equiv), Brettphos Pd G3 (103.03 mg, 0.114 mmol, 0.2 equiv) and Brettphos (61.01 mg, 0.114 mmol, 0.2 equiv). The reaction was stirred at 120 °C under N2 for 3 h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-6-nitro-2-oxo-3,4-dihydroquinoline-5-carbonitrile (77 mg, 44.09%) as a white solid. LC/MS (ESI): mass calcd. for C17H13N3O3: 307.10 m/z, found:308.10 [M+H]+.
Figure imgf000349_0001
6-amino-1-benzyl-2-oxo-1,2,3,4-tetrahydroquinoline-5-carbonitrile To a solution of 1-benzyl-6-nitro-2-oxo-3,4-dihydroquinoline-5-carbonitrile (77 mg, 0.251 mmol, 1 equiv) in ethyl alcohol (20 mL) and water (2 mL) was added iron (140 mg, 2.507 mmol, 10.01 equiv) and NH4Cl (133 mg, 2.486 mmol, 9.92 equiv). The reaction was stirred at 60 °C for 3 h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-benzyl-2-oxo-3,4- dihydroquinoline-5-carbonitrile (22 mg, 31.66%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H15N3O: 277.12 m/z, found:278.10 [M+H]+.
Figure imgf000349_0002
1-(1-benzyl-5-cyano-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-2-oxo-3,4-dihydroquinoline-5-carbonitrile (80 mg, 0.288 mmol, 1 equiv) in DCM (4 mL) was added erbumine (42.20 mg, 0.576 mmol, 2 equiv). The reaction was stirred at 0 °C under N2 for 1 h. The reaction was added TEA (87.57 mg, 0.864 mmol, 3 equiv) and stirred at rt under N2 for 1 h. The reaction was added 2-methylpropan-2- amine (42.20 mg, 0.576 mmol, 2 equiv) and stirred at rt under N2 for 1 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a Aeris PEPTIDE 5um XB-C18 Axia 21.2 mm X 250 mm, 5 μm column (eluent: 43% to 57% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1- (1-benzyl-5-cyano-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (12.5 mg, 11.48%) as a white solid. LC/MS (ESI): mass calcd. for C22H24N4O2:376.19 m/z, found:377.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.71-7.74 (m, 1H), 7.11-7.36 (m, 6H), 6.80 (s, 1H), 5.14 (s, 2H), 3.06-3.11 (m, 2H), 2.72-2.77 (m, 2H), 1.27 (s, 9H). Example 280: (R)-1-(1-benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6- yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000350_0001
ethyl (R)-2-((6-chloro-3-nitropyridin-2-yl)oxy)propanoate To a solution of 2,6-dichloro-3-nitropyridine (2.0 g, 10.364 mmol, 1 equiv) in tetrahydrofuran (20 mL) was added 60% NaH (0.5 g, 12.437 mmol, 1.2 equiv) at 0 °C. The mixture was stirred for 15 min. ethyl lactate (1.47 g, 12.437 mmol, 1.2 equiv) was added and the mixture was allowed to warm to RT and stirred for 2 h. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with NaCl (20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (5/1) to afford ethyl (R)-2-((6-chloro-3-nitropyridin-2-yl)oxy)propanoate (2.0 g, 70.26%) as a white solid. MS (ESI): mass calcd. for C10H11ClN2O5, 274.04 m/z, found 275.05 [M+H]+.
Figure imgf000351_0001
(R)-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of ethyl (R)-2-((6-chloro-3-nitropyridin-2-yl)oxy)propanoate (500 mg, 1.820 mmol, 1 equiv) in EtOH (10 mL) and H2O (1 mL) was added Fe (2.0 g, 36.400 mmol, 20 equiv) and NH4Cl (1.9 g, 36.400 mmol, 20 equiv) at rt. The resulting mixture was stirred for overnight at 80 °C. The reaction was quenched with water (10 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The residue was purified by silica gel column chromatography, eluted with PE/EA (3:1) to afford (R)-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (300 mg, 82.97%) as a brown solid. MS (ESI): mass calcd. for C8H7ClN2O2, 198.02 m/z, found 199.10 [M+H]+.
Figure imgf000351_0002
(R)-1-benzyl-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of (R)-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (300 mg, 1.510 mmol, 1 equiv) in DMF (5 mL) was added BnBr (775 mg, 4.530 mmol, 3.0 equiv) and Cs2CO3 (2.0 g, 6.040 mmol, 4.0 equiv) at rt. The resulting mixture was stirred for 2 h at rt. The reaction was quenched with water (20 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with NaCl (aq., 3 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford (R)-1-benzyl-6-chloro-3- methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (300 mg, 68.79%) as a white solid. MS (ESI): mass calcd. for C15H13ClN2O2, 288.07 m/z, found 289.15 [M+H]+.
Figure imgf000352_0001
tert-butyl (R)-(1-benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6- yl)carbamate A solution of (R)-1-benzyl-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (150 mg, 0.520 mmol, 1 equiv) in dioxane (5 mL) was added tert-butyl carbamate (73 mg, 0.624 mmol, 1.2 equiv), Cs2CO3 (338 mg, 1.040 mmol, 2.0 equiv), Ephos (55.5 mg, 0.104 mmol, 0.2 equiv) and Ephos Pd G4 (48.0 mg, 0.052 mmol, 0.1 equiv) at rt. The resulting mixture was stirred for overnight at 90 °C. The resulting was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford tert-butyl (R)-(1- benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)carbamate (110 mg, 57.32%) as an off-white solid. MS (ESI): mass calcd. for C20H23N3O4, 369.17 m/z, found 370.10 [M+H]+.
Figure imgf000352_0002
(R)-6-amino-1-benzyl-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of tert-butyl (R)-(1-benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 6-yl)carbamate (100 mg, 0.271 mmol, 1 equiv) in 4N HCl (in EA, 10 mL) at rt. The resulting mixture was stirred for 2 h at rt. The resulting mixture was concentrated under reduced pressure to afford (R)-6-amino-1-benzyl-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (70 mg, 96.02%) was used in the next step directly without further purification. MS (ESI): mass calcd. for C15H15N3O2, 269.12 m/z, found 270.20 [M+H]+.
Figure imgf000353_0001
(R)-1-(1-benzyl-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)-3-(tert- butyl)urea A solution of (R)-6-amino-1-benzyl-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (80 mg, 0.297 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (294 mg, 2.970 mmol, 10 equiv) and TEA (300 mg, 2.970 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction was quenched with water (10 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The residue was purified by silica gel column chromatography, eluted with PE/EA (1:1) and C18 column to afford (R)-1-(1-benzyl-3-methyl-2-oxo-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazin-6-yl)-3-(tert-butyl)urea (28.0 mg, 25.51%) as a white solid. MS (ESI): mass calcd.C20H24N4O3 for: 368.18m/z, found 369.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 7.36 – 7.43 (m, 1H), 7.29 – 7.35 (m, 2H), 7.22 – 7.29 (m, 3H), 7.16 – 7.22 (m, 1H), 7.08 – 7.12 (m, 1H),4.79 – 5.19 (m, 3H), 1.53 (d, J = 6.8 Hz, 3H), 1.29 (s, 9H). Example 281: 1-(tert-butyl)-3-(5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea Synthetic Scheme
Figure imgf000354_0001
ethyl (E)-3-(3-amino-6-chloropyridin-2-yl)acrylate A solution of 6-chloro-2-iodopyridin-3-amine (2.3 g, 9.039 mmol, 1 equiv) ethyl (2E)-3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)prop-2-enoate (2.0 g, 9.039 mmol, 1 equiv), Na2CO3 (2.9 g, 27.117 mmol, 3 equiv) and Pd(dppf)Cl2 (0.66 g, 0.904 mmol, 0.1 equiv) in water (3 mL) and 1,4-dioxane (30 mL) was stirred for overnight at 90 °C under N2 atmosphere. The mixture was allowed to cool down to rt. The reaction mixture was quenched by water (50 mL) and extracted with EA (3 x 100 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford ethyl (E)-3-(3-amino-6-chloropyridin-2-yl)acrylate (1.5 g, 73.22%) as a yellow solid.
Figure imgf000354_0002
ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate A solution of ethyl (E)-3-(3-amino-6-chloropyridin-2-yl)acrylate (1.4 g, 6.177 mmol, 1 equiv) in methanol (10 mL) and tetrahydrofuran (20 mL) was treated with cobalt(II) chloride (0.40 g, 3.088 mmol, 0.5 equiv) for 30 min at 0 °C under nitrogen atmosphere followed by the addition of NaBH4 (1.17 g, 30.885 mmol, 5 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 1 h at rt. The reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (2:1) to afford ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate (1.2 g, 84.96%) as a white solid. MS (ESI): mass calcd. for C10H13ClN2O2, 228.07 m/z, found 229.10 [M+H]+.
Figure imgf000355_0001
6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of ethyl 3-(3-amino-6-chloropyridin-2-yl)propanoate (1.2 g, 5.248 mmol, 1 equiv) in EA (10 mL), HCl (4N, in EA, 20 mL) was added at rt. The reaction stirred for 2 h. The precipitated solids were collected by filtration and washed with Et2O (3 x 50 mL). This resulted in6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one (1.08 g, 112.71%) as a white solid. MS (ESI): mass calcd. for C8H7ClN2O, 182.02 m/z, found 183.10 [M+H]+.
Figure imgf000355_0002
6-chloro-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of 6-chloro-3,4-dihydro-1,5-naphthyridin-2(1H)-one (300 mg, 1.643 mmol, 1 equiv) in anhydrous DMF (20 mL) was added 1-(bromomethyl)-3-chlorobenzene (371 mg, 1.807 mmol, 1.1 equiv) and Cs2CO3 (1.6 g, 4.929 mmol, 3 equiv) at rt and stirred for 2 h. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford 6-chloro-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (293 mg, 58.06%) as a light-yellow solid. MS (ESI): mass calcd. for C15H12Cl2N2O, 306.03 m/z, found 307.00 [M+H]+.
Figure imgf000355_0003
tert-butyl (5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)carbamate To a stirred solution of 6-chloro-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (210 mg, 0.684 mmol, 1 equiv) in anhydrous toluene (15 mL) was added Cs2CO3 (670 mg, 2.052 mmol, 3 equiv) tert-butyl carbamate (160 mg, 1.368 mmol, 2 equiv), XantPhos (40 mg, 0.068 mmol, 0.1 equiv) Pd2(dba)3 (63 mg, 0.068 mmol, 0.1 equiv) at 90 °C and stirred for overnight. After completion of reaction, the reaction mixture was concentrated under reduced pressure to give crude product which was further purified by column chromatography (PE/EA=2:1) afford desired compound tert-butyl (5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2- yl)carbamate (204 mg, 76.93%) as a yellow semi-solid. MS (ESI): mass calcd. for C20H22ClN3O3, 387.13 m/z, found 388.20 [M+H]+.
Figure imgf000356_0001
6-amino-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred solution of tert-butyl (5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin- 2-yl)carbamate (220 mg, 0.567 mmol, 1 equiv) in EA (5 mL), HCl (4N, in EA, 10 mL) was added at rt. The reaction stirred for 2 h. The resulting mixture was concentrated under reduced pressure to afford 6-amino-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (250 mg, 153.17%) as a white solid. MS (ESI): mass calcd. for C15H14ClN3O, 287.08 m/z, found 288.20 [M+H]+.
Figure imgf000356_0002
1-(tert-butyl)-3-(5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea A solution of 6-amino-1-(3-chlorobenzyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (100 mg, 0.348 mmol, 1 equiv), TEA (176 mg, 1.740 mmol, 5 equiv) and 2-isocyanato-2-methylpropane (172 mg, 1.740 mmol, 5 equiv) in DCM (10 mL) was stirred for overnight at rt. The reaction was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with EA (55%-62%) to afford crude products. The residue was purified by reversed-phase flash chromatography with the following conditions Column: Xselect CSH C18 OBD Column 30*150mm 5μm, n; Mobile Phase A: Water(0.1%FA), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 35% B to 55% B in 10 min; Wave Length: 254/220 nm; RT1(min): 9.8; Number Of Runs: 3 to) afford 1-(tert-butyl)-3-(5-(3-chlorobenzyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea (26.2 mg, 19.44%) as a white solid. MS (ESI): mass calcd. for C20H23ClN4O2: 386.15 m/z, found 387.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.81 (s, 1H), 7.94 (s, 1H), 7.11 – 7.36 (m, 6H), 5.08 (s, 2H), 2.97 (dd, J = 8.8, 6.2 Hz, 2H), 2.78 (dd, J = 8.7, 6.2 Hz, 2H), 1.30 (s, 9H). Example 282: (S)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8- tetrahydro-1,5-naphthyridin-2-yl)urea Synthetic Scheme
Figure imgf000357_0001
6-chloro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a solution of 6-chloro-3,4-dihydro-1H-1,5-naphthyridin-2-one (400 mg, 2.190 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (908 mg, 6.571 mmol, 3 equiv) and 1-(1-bromoethyl)-3- (trifluoromethyl)benzene (831 mg, 3.286 mmol, 1 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS. The mixture was diuted with EtOAc (50 mL), washed with water (3 x 30 mL), brine (2 x 30 mL) and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0- 100% ethyl acetate/petroleum ether) to afford 6-chloro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)- 3,4-dihydro-1,5-naphthyridin-2(1H)-one (300 mg, 38.29%) as a yellow solid. MS (ESI): mass calcd. for C17H14ClF3N2O 354.07 m/z, found 355.05 [M+H]+.
Figure imgf000358_0001
tert-butyl (6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate To a solution of 6-chloro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5-naphthyridin- 2(1H)-one (300 mg, 0.846 mmol, 1 equiv) in dioxane (10 mL) was added tert-butyl carbamate (198.1 mg, 1.691 mmol, 2 equiv), Ephos (45.2 mg, 0.085 mmol, 0.1 equiv), EPhos Pd G4 (77.6 mg, 0.085 mmol, 0.1 equiv) and Cs2CO3 (826.5 mg, 2.537 mmol, 3 equiv). The resulting mixture was placed at 100 °C under a nitrogen atmosphere and stirred overnight until the starting material was totally consumed by LCMS, The reaction mixture was quenched by water and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford tert-butyl (6- oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)carbamate (200 mg, 54.31%) as a yellow solid. MS (ESI): mass calcd. for C22H24F3N3O3, 435.18 m/z, found 436.15 [M+H]+.
Figure imgf000358_0002
6-amino-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a solution of tert-butyl (6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate (20 mg, 0.046 mmol, 1 equiv) in HCl (4N, in EA, 2 mL). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS. The reaction mixture was concentrated under vacuum to yield 6-amino-1-{1-[3- (trifluoromethyl)phenyl]ethyl}-3,4-dihydro-1,5-naphthyridin-2-one (100 mg, 61.75%) as a yellow solid. MS (ESI): mass calcd. for C17H16F3N3O, 335.12 m/z, found 336.05 [M+H]+.
Figure imgf000359_0001
(S)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea To a stirred solution of 6-amino-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (100 mg, 0.298 mmol, 1 equiv) in DCM (10.00 mL) was added 2- isocyanato-2-methylpropane (147.8 mg, 1.491 mmol, 5 equiv) and TEA (90.5 mg, 0.895 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid. The compound was separated by Chiral-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro- 1,5-naphthyridin-2-yl)urea (11.7 mg, 8.86%) as a white solid. MS (ESI): mass calcd. for C22H25F3N4O2, 434.19 m/z, found 435.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 7.96 (s, 1H), 7.52 – 7.67 (m, 4H), 6.96 – 7.07 (m, 2H), 6.08 (d, J = 7.3 Hz, 1H), 2.61 – 3.06 (m, 4H), 1.74 (d, J = 7.1 Hz, 3H), 1.30 (s, 9H). And another isomer (R)-1-(tert-butyl)-3-(6-oxo-5-(1- (3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (9.1 mg, 6.96%) as a white solid. MS (ESI): mass calcd. for C22H25F3N4O2, 434.19 m/z, found 435.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 7.96 (s, 1H), 7.52 – 7.67 (m, 4H), 6.96 – 7.07 (m, 2H), 6.08 (d, J = 7.3 Hz, 1H), 2.61 – 3.06 (m, 4H), 1.74 (d, J = 7.1 Hz, 3H), 1.30 (s, 9H). Example 283: (R)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8- tetrahydro-1,5-naphthyridin-2-yl)urea
Figure imgf000360_0001
(R)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea To a stirred solution of 6-amino-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (100 mg, 0.298 mmol, 1 equiv) in DCM (10.00 mL) was added 2- isocyanato-2-methylpropane (147.8 mg, 1.491 mmol, 5 equiv) and TEA (90.5 mg, 0.895 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 100 mg crude product as a white solid. The compound was separated by Chiral-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3-(6-oxo-5-(1-(3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro- 1,5-naphthyridin-2-yl)urea (11.7 mg, 8.86%) as a white solid. MS (ESI): mass calcd. for C22H25F3N4O2, 434.19 m/z, found 435.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 7.96 (s, 1H), 7.52 – 7.67 (m, 4H), 6.96 – 7.07 (m, 2H), 6.08 (d, J = 7.3 Hz, 1H), 2.61 – 3.06 (m, 4H), 1.74 (d, J = 7.1 Hz, 3H), 1.30 (s, 9H). And another isomer (R)-1-(tert-butyl)-3-(6-oxo-5-(1- (3-(trifluoromethyl)phenyl)ethyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (9.1 mg, 6.96%) as a white solid. MS (ESI): mass calcd. for C22H25F3N4O2, 434.19 m/z, found 435.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 7.96 (s, 1H), 7.52 – 7.67 (m, 4H), 6.96 – 7.07 (m, 2H), 6.08 (d, J = 7.3 Hz, 1H), 2.61 – 3.06 (m, 4H), 1.74 (d, J = 7.1 Hz, 3H), 1.30 (s, 9H). Example 284: 3-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-1-phenylurea
Figure imgf000360_0002
3-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-1-phenylurea To a stirred solution of 6-amino-1-benzyl-3,4-dihydroquinolin-2-one (150 mg, 0.594 mmol, 1 equiv) in DCM (2 mL) was added triphosgene (59.98 mg, 0.202 mmol, 0.34 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at 0°C under nitrogen atmosphere. To the above mixture was added triethylamine (180.47 mg, 1.782 mmol, 3 equiv) in portions at room temperature. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added aniline (55.36 mg, 0.594 mmol, 1 equiv) in portions at 0 °C. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 26% to 45% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 3-(1-benzyl-2-oxo-3,4- dihydroquinolin-6-yl)-1-phenylurea (37.5 mg, 16.92%) as a white solid. LC/MS (ESI): mass calcd. for C23H21N3O2:371.2 m/z, found:372.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.55 (s, 1H), 7.43 – 7.41 (m, 3H), 7.31 – 7.21 (m, 8H), 7.01 – 6.89 (m, 1H), 6.84 (d, J = 8.7 Hz, 1H), 5.12 (s, 2H), 2.93 (m, 2H), 2.69 (m, 2H). Example 285: N-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide
Figure imgf000361_0001
N-(1-benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide To a stirred solution of 3,3-dimethylbutanamide (35.15 mg, 0.305 mmol, 1.1 equiv) and HATU (137.13 mg, 0.360 mmol, 1.3 equiv) in DMF (1 mL) were added 6-amino-1-benzyl-3,4- dihydroquinolin-2-one (70 mg, 0.277 mmol, 1 equiv) and DIEA (107.57 mg, 0.831 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2 h at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford N-(1- benzyl-2-oxo-3,4-dihydroquinolin-6-yl)-3,3-dimethylbutanamide (47.6 mg, 48.80%) as a white solid.LC/MS (ESI): mass calcd. for C22H26N2O2:350.2 m/z, found:351.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 9.69 (s, 1H), 7.51 (d, J = 2.1 Hz, 1H), 7.33 – 7.19 (m, 6H), 6.84 (d, J = 8.7 Hz, 1H), 5.11 (s, 2H), 2.92 – 2.88 (m, 2H), 2.69 – 2.64 (m, 2H), 2.13 (s, 2H), 0.99 (s, 9H). Example 286: (S)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-6-yl)urea Synthetic Scheme
Figure imgf000362_0001
(S)-6-chloro-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of (S)-6-chloro-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (100 mg, 0.503 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-3-chlorobenzene (155 mg, 0.754 mmol, 1.5 equiv) and Cs2CO3 (492 mg, 1.509 mmol, 3.0 equiv) at rt. The resulting mixture was stirred for 2h at rt. The reaction mixture was quenched by ice water (5 mL) and extracted with EA (3 x 5 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford (S)-6-chloro-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin- 2(3H)-one (150 mg, 92.19%) as a white solid. MS (ESI): mass calcd. for C15H12Cl2N2O2, 322.03 m/z, found 323.00 [M+H]+.
Figure imgf000363_0001
tert-butyl (S)-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)carbamate A solution of (S)-6-chloro-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)- one (150 mg, 0.464 mmol, 1 equiv) in toluene (5 mL) was added tert-butyl carbamate (70 mg, 0.603 mmol, 1.3 equiv), Cs2CO3 (453 mg, 1.392 mmol, 3.0 equiv), Ephos (49 mg, 0.093 mmol, 0.2 equiv) and EPhos Pd G4 (42 mg, 0.046 mmol, 0.1 equiv) at rt. The resulting mixture was stirred for overnight at 90 °C under N2. After cooled to room temperature the reaction mixture was quenched by water (5 mL) and extracted with EA (3 x 5 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford tert-butyl (S)-(1-(3- chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)carbamate (160 mg, 85.35%) as yellow oil. MS (ESI): mass calcd. for C20H22ClN3O4, 403.13 m/z, found 404.20 [M+H]+.
Figure imgf000363_0002
(S)-6-amino-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A solution of tert-butyl (S)-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)carbamate (150 mg, 0.371 mmol, 1 equiv) in EA (5 mL), 2N HCl (in EA, 5 mL) was added into the reaction at rt. The resulting mixture was stirred for 2 h at rt. The resulting mixture was concentrated under reduced pressure. The crude product (S)-6-amino-1-(3- chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (80 mg, 70.91%) was used in the next step directly without further purification. MS (ESI): mass calcd. for C15H14ClN3O2, 303.08 m/z, found 304.15 [M+H]+.
Figure imgf000364_0001
(S)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)urea A solution of (S)-6-amino-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)- one (80 mg, 0.263 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (261 mg, 2.630 mmol, 10 equiv) and TEA (266 mg, 2.630 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude was purified by flash chromatography with PE/EA (1:1) and Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 7 min, 60% B; Wave Length: 254/220 nm; RT1(min): 6.7; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl- 2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)urea (19.6 mg, 17.86%) as a white solid. MS (ESI): mass calcd. C20H23ClN4O3 for: 402.15m/z, found 403.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 7.29 – 7.41 (m, 4H), 7.17 – 7.26 (m, 2H), 7.09 (s, 1H), 5.02 – 5.16(m, 3H), 1.53 (d, J = 6.8 Hz, 3H), 1.28 (s, 9H). Example 287: (R)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-6-yl)urea
Figure imgf000364_0002
(R)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)urea A solution of (R)-6-amino-1-(3-chlorobenzyl)-3-methyl-1H-pyrido[2,3-b][1,4]oxazin-2(3H)- one (60 mg, 0.198 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (195 mg, 1.980 mmol, 10 equiv) and TEA (199 mg, 1.980 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude was purified by flash chromatography with PE/EA (1:1) and Prep-HPLC ( Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 7 min, 60% B; Wave Length: 254/220 nm; RT1(min): 6.7; Number Of Runs: 0) to afford (R)-1-(tert-butyl)-3-(1-(3-chlorobenzyl)-3-methyl- 2-oxo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)urea (12.1 mg, 14.76%) as a white solid. MS (ESI): mass calcd. C20H23ClN4O3 for: 402.15m/z, found 403.05[M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 7.29 – 7.41 (m, 4H), 7.17 – 7.26 (m, 2H), 7.09 (s, 1H), 5.02 – 5.16(m, 3H), 1.53 (d, J = 6.8 Hz, 3H), 1.28 (s, 9H). Example 288: (S)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000365_0001
2-(methoxymethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of 2-amino-5-nitrophenol (300 mg, 1.946 mmol, 1 equiv) in dimethylformamide (5 mL) was added Cs2CO3 (1.9 g, 5.838 mmol, 3.0 equiv) and methyl 2-bromo-3-methoxypropanoate (460.2 mg, 2.335 mmol, 1.2 equiv) at rt. The resulting mixture was stirred for overnight at 90 °C. The reaction mixture was quenched by water (5 mL) and extracted with EA (3 x 10 mL). The combined organic layers were washed with NaCl (20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 2-(methoxymethyl)-7-nitro-2H- benzo[b][1,4]oxazin-3(4H)-one (200 mg, 43.14%) as a white solid. MS (ESI): mass calcd. for C10H10N2O5238.06 m/z, found 239.10 [M+H]+.
Figure imgf000366_0001
4-benzyl-2-(methoxymethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of 2-(methoxymethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.630 mmol, 1 equiv) in dimethylformamide (5 mL) was added BnBr (323.1 mg, 1.890 mmol, 3.0 equiv) and Cs2CO3 (615.5 mg, 1.890 mmol, 3.0 equiv) at rt. The resulting mixture was stirred for 2h at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic layers were washed with NaCl (10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 4-benzyl-2-(methoxymethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)- one (130 mg, 62.88%) as a white solid. MS (ESI): mass calcd. for C17H16N2O5, 328.11 m/z, found 329.05 [M+H]+.
Figure imgf000366_0002
7-amino-4-benzyl-2-(methoxymethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-benzyl-2-(methoxymethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (130 mg, 0.396 mmol, 1 equiv) in methanol (5 mL) was added Pd/C (10%, 421.3 mg, 3.960 mmol, 10 equiv) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 30 min under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. The crude product 7-amino-4-benzyl-2-(methoxymethyl)- 2H-benzo[b][1,4]oxazin-3(4H)-one (100 mg, 84.65%) was used in the next step directly without further purification. MS (ESI): mass calcd. for C17H18N2O3, 298.13 m/z, found 353.10 [M+H]+.
Figure imgf000367_0001
(S)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea A solution of 7-amino-4-benzyl-2-(methoxymethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (70 mg, 0.235 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (232.6 mg, 2.350 mmol, 10 equiv) and Et3N (237.4 mg, 2.350 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with NaCl (20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product 40 mg was purified by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 25% B to 25% B in 23 min; Wave Length: 220/254 nm; RT1(min): 13.384; RT2(min): 18.251; Sample Solvent: EtOH--HPLC; Injection Volume: 0.6 mL; Number Of Runs: 7) to afford (S)-1-(4-benzyl-2-(methoxymethyl)-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.0 mg, 21.32%) as a white solid. MS (ESI): mass calcd. C22H27N3O4 for: 397.20m/z, found 398.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.42 (m, 2H), 7.19 – 7.29 (m, 4H), 6.71 – 6.88 (m, 1H), 6.57 – 6.71 (m, 1H), 5.94 (s, 1H), 5.10 – 5.31 (m, 1H), 4.89 – 5.10 (m, 2H), 3.71 – 3.81 (m, 2H), 3.30 (s, 3H), 1.25 (s, 9H). And to afford (R)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.5 mg, 21.91%) as a white solid. MS (ESI): mass calcd. C22H27N3O4 for : 397.20m/z, found 398.15[M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.42 (m, 2H), 7.19 – 7.29 (m, 4H), 6.71 – 6.88 (m, 1H), 6.57 – 6.71 (m, 1H), 5.94 (s, 1H), 5.10 – 5.31 (m, 1H), 4.89 – 5.10 (m, 2H), 3.71 – 3.81 (m, 2H), 3.30 (s, 3H), 1.25 (s, 9H). Example 289: (R)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea
Figure imgf000368_0001
(R)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea A solution of 7-amino-4-benzyl-2-(methoxymethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (70 mg, 0.235 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (232.6 mg, 2.350 mmol, 10 equiv) and Et3N (237.4 mg, 2.350 mmol, 10 equiv) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with NaCl (20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product 40 mg was purified by Prep-Chiral-HPLC with the following conditions (Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 25% B to 25% B in 23 min; Wave Length: 220/254 nm; RT1(min): 13.384; RT2(min): 18.251; Sample Solvent: EtOH--HPLC; Injection Volume: 0.6 mL; Number Of Runs: 7) to afford (S)-1-(4-benzyl-2-(methoxymethyl)-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.0 mg, 21.32%) as a white solid. MS (ESI): mass calcd. C22H27N3O4 for: 397.20m/z, found 398.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.29 – 7.42 (m, 2H), 7.19 – 7.29 (m, 4H), 6.71 – 6.88 (m, 1H), 6.57 – 6.71 (m, 1H), 5.94 (s, 1H), 5.10 – 5.31 (m, 1H), 4.89 – 5.10 (m, 2H), 3.71 – 3.81 (m, 2H), 3.30 (s, 3H), 1.25 (s, 9H). And to afford (R)-1-(4-benzyl-2-(methoxymethyl)-3-oxo-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (20.5 mg, 21.91%) as a white solid. MS (ESI): mass calcd. C22H27N3O4 for : 397.20m/z, found 398.15[M+H]+. 1H NMR (400 MHz, DMSO- d6) δ 8.19 (s, 1H), 7.29 – 7.42 (m, 2H), 7.19 – 7.29 (m, 4H), 6.71 – 6.88 (m, 1H), 6.57 – 6.71 (m, 1H), 5.94 (s, 1H), 5.10 – 5.31 (m, 1H), 4.89 – 5.10 (m, 2H), 3.71 – 3.81 (m, 2H), 3.30 (s, 3H), 1.25 (s, 9H). Example 290: (S)-1-(tert-butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea Synthetic Scheme
Figure imgf000369_0001
1-(1-bromoethyl)-3-chlorobenzene To a solution of 1-(3-chlorophenyl)ethan-1-ol (500 mg, 3.193 mmol, 1 equiv) in DCM (15 mL) was added PBr3 (950.6 mg, 3.512 mmol, 1.1 equiv) at 0 °C. The resulting mixture was placed at 0°C and stirred overnight until the starting material was totally consumed by TLC. The reaction mixture was quenched by ice water (10 mL) and extracted with EA (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford 1-(1- bromoethyl)-3-chlorobenzene (400 mg,57.08%) as a yellow oil.
Figure imgf000369_0002
6-chloro-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a solution of 1-(1-bromoethyl)-3-chlorobenzene (249.5 mg, 1.367 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (566.6 mg, 4.100 mmol, 3 equiv) and 1-(1-bromoethyl)-3- chlorobenzene (300 mg, 1.367 mmol, 1 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS. The reaction mixture was quenched by ice water (10 mL) and extracted with EA (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford 6-chloro-1-(1-(3- chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (200 mg, 45.26%) as a yellow solid.MS (ESI): mass calcd. for C16H14Cl2N2O 320.05 m/z, found 321.05 [M+H]+.
Figure imgf000370_0001
tert-butyl (5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2- yl)carbamate To a solution of 6-chloro-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (200 mg, 0.623 mmol, 1 equiv) in toluene (15 mL) was added tert-butyl carbamate (87.5 mg, 0.747 mmol, 1.2 equiv), Xantphos (36.0 mg, 0.062 mmol, 0.1 equiv), Pd2(dba)3 (57.0 mg, 0.062 mmol, 0.1 equiv) and Cs2CO3 (608.6 mg, 1.868 mmol, 3 equiv). The resulting mixture was placed at 90°C under a nitrogen atmosphere and stirred overnight until the starting material was totally consumed by LCMS. After cooled to room temperature the reaction mixture was quenched by ice water (10 mL) and extracted with EA (3 x 20 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA to afford tert-butyl (5-(1-(3-chlorophenyl)ethyl)-6-oxo- 5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)carbamate (140 mg, 53.72%) as a yellow solid.MS (ESI): mass calcd. for C21H24ClN3O3, 401.15 m/z, found 402.10 [M+H]+.
Figure imgf000370_0002
6-amino-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a solution of tert-butyl (5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)carbamate (20 mg, 0.050 mmol, 1 equiv) in HCl (in EA) (2 mL). The resulting mixture was placed at rtand stirred overnight. The reaction mixture was concentrated under vacuum to yield 6-amino-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (80 mg, 60.87%) as a yellow solid.MS (ESI): mass calcd. for C16H16ClN3O, 301.10 m/z, found 302.05 [M+H]+.
Figure imgf000371_0001
(S)-1-(tert-butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea To a stirred solution of 6-amino-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin- 2(1H)-one (80 mg, 0.265 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2- methylpropane (131.4 mg, 1.325 mmol, 5 equiv) and TEA (80.4 mg, 0.795 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude was purified by flash chromatography with PE/EA and Prep-HPLC ( Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) and Chiral Prep-HPLC (Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 13 min; Wave Length: 220/254 nm; RT1(min): 9.187; RT2(min): 10.914; Sample Solvent: EtOH--HPLC; Injection Volume: 1.2 mL; Number Of Runs: 4) to afford (S)-1-(tert-butyl)-3-(5-(1-(3- chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (3.6 mg, 3.38%) as a white solid. MS (ESI): mass calcd. for C21H25ClN4O2, 400.17 m/z, found 435.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.88 (s, 1H), 8.00 (s, 1H), 7.28 – 7.42 (m, 3H), 7.23 (d, J = 7.5Hz, 1H), 6.99 (q, J = 9.0 Hz, 2H), 6.07 (q, J = 7.1 Hz, 1H), 2.90 – 3.02 (m, 2H), 2.76 – 2.82 (m, 1H), 2.63 – 2.70 (m, 1H), 1.68 (d, J = 7.2 Hz, 3H), 1.30 (s, 9H). And another isomer to afford (R)-1-(tert- butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (5.2 mg, 4.89%) as a white solid. MS (ESI): mass calcd. for C21H25ClN4O2, 400.17 m/z, found 435.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.88 (s, 1H), 8.00 (s, 1H), 7.28 – 7.42 (m, 3H), 7.23 (d, J = 7.5Hz, 1H), 6.99 (q, J = 9.0 Hz, 2H), 6.07 (q, J = 7.1 Hz, 1H), 2.90 – 3.02 (m, 2H), 2.76 – 2.82 (m, 1H), 2.63 – 2.70 (m, 1H), 1.68 (d, J = 7.2 Hz, 3H), 1.30 (s, 9H). Example 291: (R)-1-(tert-butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro- 1,5-naphthyridin-2-yl)urea
Figure imgf000372_0001
(R)-1-(tert-butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5- naphthyridin-2-yl)urea To a stirred solution of 6-amino-1-(1-(3-chlorophenyl)ethyl)-3,4-dihydro-1,5-naphthyridin- 2(1H)-one (80 mg, 0.265 mmol, 1 equiv) in DCM (10 mL) was added 2-isocyanato-2- methylpropane (131.4 mg, 1.325 mmol, 5 equiv) and TEA (80.4 mg, 0.795 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude was purified by flash chromatography with PE/EA and Prep-HPLC ( Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) and Chiral Prep-HPLC (Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: 50% B to 50% B in 13 min; Wave Length: 220/254 nm; RT1(min): 9.187; RT2(min): 10.914; Sample Solvent: EtOH--HPLC; Injection Volume: 1.2 mL; Number Of Runs: 4) to afford (S)-1-(tert-butyl)-3-(5-(1-(3- chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (3.6 mg, 3.38%) as a white solid. MS (ESI): mass calcd. for C21H25ClN4O2, 400.17 m/z, found 435.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.88 (s, 1H), 8.00 (s, 1H), 7.28 – 7.42 (m, 3H), 7.23 (d, J = 7.5Hz, 1H), 6.99 (q, J = 9.0 Hz, 2H), 6.07 (q, J = 7.1 Hz, 1H), 2.90 – 3.02 (m, 2H), 2.76 – 2.82 (m, 1H), 2.63 – 2.70 (m, 1H), 1.68 (d, J = 7.2 Hz, 3H), 1.30 (s, 9H). And another isomer to afford (R)-1-(tert- butyl)-3-(5-(1-(3-chlorophenyl)ethyl)-6-oxo-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (5.2 mg, 4.89%) as a white solid. MS (ESI): mass calcd. for C21H25ClN4O2, 400.17 m/z, found 435.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.88 (s, 1H), 8.00 (s, 1H), 7.28 – 7.42 (m, 3H), 7.23 (d, J = 7.5Hz, 1H), 6.99 (q, J = 9.0 Hz, 2H), 6.07 (q, J = 7.1 Hz, 1H), 2.90 – 3.02 (m, 2H), 2.76 – 2.82 (m, 1H), 2.63 – 2.70 (m, 1H), 1.68 (d, J = 7.2 Hz, 3H), 1.30 (s, 9H). Example 292: 1-(4-benzyl-2,2-dimethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea Synthetic Scheme
Figure imgf000373_0001
4-benzyl-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of 2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 1.350 mmol, 1 equiv) in anhydrous DMF (10 mL) was added K2CO (373 mg, 2.700 mmol, 2 equiv) and BnBr (346 mg, 2.025 mmol, 1.5 equiv) at rt. The reaction mixture was stirred for 1.5 h at rt. After completion of reaction, the reaction mixture was quenched by addition of water (15 mL). The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate filtered and concentrated under reduced pressure. The residue was purified by silica gel column, eluted with PE/EA (3:1) to give 4-benzyl-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (420 mg, 99.60%) as a yellow oil. MS (ESI): mass calcd. for C17H16N2O4: 312.11, found: 313.20 [M+H]+.
Figure imgf000374_0001
7-amino-4-benzyl-2,2-dimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 4-benzyl-2,2-dimethyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (420 mg, 1.345 mmol, 1 equiv) in MeOH (15 mL) was added Pd/C under N2 atmosphere. The reaction was stirred at room temperature for 2.5 h under H2 atmosphere. The reaction was filtered. The filtrate was concentrated to give 7-amino-4-benzyl-2,2-dimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (340 mg, 87.11%) as a white solid. MS (ESI): mass calcd. for C17H18N2O2: 282.14, found: 283.10[M+H]+.
Figure imgf000374_0002
1-(4-benzyl-2,2-dimethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea To a mixture of 7-amino-4-benzyl-2,2-dimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (110 mg, 0.390 mmol, 1 equiv) and TEA (236 mg, 2.340 mmol, 6 equiv) in DCM (5 mL) was added 2- isocyanato-2-methylpropane (232 mg, 2.340 mmol, 6 equiv) at 0°C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified by silica gel column (PE/EA 1:1) and Prep- HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm) to give 1-(4-benzyl-2,2-dimethyl-3-oxo-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (21.7 mg, 14.53%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3, 381.21 m/z, found 382.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.31 – 7.35 (m, 2H), 7.20 – 7.26 (m, 4H), 6.83 (d, J = 8.7 Hz, 1H), 6.72 (dd, J = 8.7, 2.3 Hz, 1H), 5.94 (s, 1H), 5.08 (s, 2H), 1.46 (s, 6H), 1.26 (s, 9H). Example 293: 1-(tert-butyl)-3-(4-(1-(5-chloro-2-cyanophenyl)ethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000375_0001
1-bromo-2-(1-bromoethyl)-4-chlorobenzene To a mixture of 1-(2-bromo-5-chlorophenyl)ethan-1-ol (300 mg, 1.047 mmol, 1 equiv) in DCM (5 mL) was added PBr3 (567 mg, 2.094 mmol, 2 equiv) at 0 °C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by ice water (10 mL) and the pH value of the aqueous phase was adjusted to 8 with NaHCO3 (aq.). The mixture was extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified by silica gel column, eluted with PE/EA (20:1) to give 1- bromo-2-(1-bromoethyl)-4-chlorobenzene (218 mg, 69.76%) as yellow oil.
Figure imgf000376_0001
4-(1-(2-bromo-5-chlorophenyl)ethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a mixture of 7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (87 mg, 0.447 mmol, 1 equiv) and K2CO3 (187 mg, 1.340 mmol, 3 equiv) in DMF (5 mL). 1-Bromo-2-(1-bromoethyl)-4- chlorobenzene (200 mg, 0.670 mmol, 1.5 equiv) was added into the solution. The resulting solution was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum The residue was purified by silica gel column, eluted with PE/EA (3:1) to afford 4-(1-(2-bromo-5-chlorophenyl)ethyl)-7-nitro-2H-benzo[b][1,4]oxazin- 3(4H)-one (150 mg, 81.55%) as a yellow solid.
Figure imgf000376_0002
4-chloro-2-(1-(7-nitro-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)benzonitrile A solution of 4-(1-(2-bromo-5-chlorophenyl)ethyl)-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.360 mmol, 1 equiv) in DMF (3 mL) was added Zn(CN)2 (106 mg, 0.90 mmol, 2.5 equiv) and Pd(PPh3)4 (42 mg, 0.036 mmol, 0.1 equiv) at rt. The resulting mixture was maintained under nitrogen and stirred at 90 °C for overnight. After cooling down to rt, the reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography eluted with PE/EA (2:1) to afford 4-chloro-2-(1-(7-nitro-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)benzonitrile (100 mg, 77.52%). LC/MS: mass calcd. for C17H12ClN3O4: 357.05, found: 358.00 [M+H]+.
Figure imgf000377_0001
2-(1-(7-amino-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)-4-chlorobenzonitrile To a mixture of 4-chloro-2-(1-(7-nitro-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4- yl)ethyl)benzonitrile (50 mg, 0.140 mmol, 1 equiv) in EtOH (10 mL) and water (2 mL). Fe (78 mg, 1.400 mmol, 10 equiv) and NH4Cl (74.76 mg, 1.400 mmol, 10 equiv) were added into the mixture. The resulting solution was stirred at 60 °C for 1 h. The resulting mixture was concentrated under vacuum. The residue was purified by flash chromatography with PE/EA (1:1) to afford 2- (1-(7-amino-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)-4-chlorobenzonitrile (33 mg, 72.04%) as a yellow solid. LC/MS: mass calcd. for C17H14ClN3O2, 327.08 m/z, found 328.15 [M+H]+.
Figure imgf000377_0002
1-(tert-butyl)-3-(4-(1-(5-chloro-2-cyanophenyl)ethyl)-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a mixture of 2-(1-(7-amino-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)-4- chlorobenzonitrile (33 mg, 0.101 mmol, 1 equiv) and TEA (21 mg, 0.202 mmol, 2 equiv) in DCM (4 mL) was added 2-isocyanato-2-methylpropane (30 mg, 0.303 mmol, 3 equiv) at 0 °C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column with PE/EA (1:1) and Prep-HPLC ( Column: XBridge Prep Phenyl OBD Column, 19*150 mm, 5μm; Mobile Phase A: Water(50 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 35% B to 57% B in 10 min, 57% B; Wave Length: 254/220 nm) to give 1-(tert-butyl)-3-(4-(1-(5- chloro-2-cyanophenyl)ethyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea (4.7 mg, 10.92%) as a white solid. LC/MS: mass calcd. for C22H23ClN4O3, 426.15 m/z, found 427.05 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.21 (s, 1H), 7.79 – 7.84 (m, 2H), 7.58 (dd, J = 8.3, 2.0 Hz, 1H), 7.13 (d, J = 2.3 Hz, 1H), 6.88 (d, J = 8.9 Hz, 1H), 6.77 (dd, J = 8.8, 2.4 Hz, 1H), 5.91 – 5.93 (m, 2H), 4.54 (s, 2H), 1.82 (d, J = 7.0 Hz, 3H), 1.26 (s, 9H). Example 294: 1-(3-amino-1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000378_0001
1-benzyl-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (3 g, 15.695 mmol, 1 equiv) in DMF (50 mL) was added K2CO3 (6.51 g, 47.104 mmol, 3 equiv) and BnBr (4.03 g, 23.562 mmol, 1.5 equiv). The resulting mixture was placed at rt and stirred overnight until the starting material was totally consumed by LCMS, The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 50 mL). The combined organic layers were washed with NaCl (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 1-benzyl-6- nitroquinoxalin-2(1H)-one (3 g, 65.14%) as a yellow solid.MS (ESI): mass calcd. for C15H11N3O3 281.08 m/z, found 282.05 [M+H]+.
Figure imgf000379_0001
3-amino-1-benzyl-6-nitroquinoxalin-2(1H)-one An oven-dried Schlenk tube was charged with 1-benzyl-6-nitroquinoxalin-2(1H)-one (1 g, 3.555 mmol, 1.0 equiv), CAN (2.93 g, 5.325 mmol, 1.5 equiv) and a magnetic stirring bar, and then was purged with nitrogen for three times. EtOAc (36 mL) and H2O (128.1 mg, 7.111 mmol, 2.0 equiv) were added in turn via respective syringes, and the mixture was stirred at 20 °C in water bath. Then, TMSN3 (1.23 g, 10.676 mmol, 3.0 equiv) was added in turn via a syringe, and the mixture was stirred under the previous conditions until the substrate was consumed (monitored by TLC, about 10 hour). The mixture was added with CH2Cl2 (200 mL) and saturated aqueous Na2CO3 (200 mL). The organic layer was isolated and the remaining aqueous phase was further extracted with CH2Cl2 (150 mL × 2). The combined organic phase was washed with 0.3 M HCl (100 mL × 2), and adjusted pH to 8 by the addition of saturated aqueous Na2CO3, followed by extraction with CH2Cl2 (150 mL × 3). The received organic phase was washed with saturated brine (200 mL). The organic layer was dried over Na2SO4, and then filtered. The filtrate was evaporated under reduced pressure, and the resulting residue was purified by column chromatography on neutral aluminum oxide (petroleum ether/ethyl acetate, in some cases, 5-10 vol % triethylamine is helpful.) to afford 3-amino-1-benzyl-6-nitroquinoxalin-2(1H)-one (300 mg, 24.48%) as a yellow solid.MS (ESI): mass calcd. for C15H12N4O3, 296.09 m/z, found 297.10 [M+H]+ .
Figure imgf000379_0002
tert-butyl (4-benzyl-7-nitro-3-oxo-3,4-dihydroquinoxalin-2-yl)carbamate In a 100 mL round bottom flask, to a solution of 3-amino-1-benzyl-6-nitroquinoxalin-2(1H)- one (300 mg, 1.013 mmol, 1 equiv) in THF (10 mL) was added 60% NaH (60.7 mg, 1.519 mmol, 1.5 equiv) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at 0°C for 0.5 h . Then a solution of Boc2O (331.4 mg, 1.519 mmol, 1.5 equiv) in 5 mL THF was added dropwise and the mixture was stirred for another 120 mins. The reaction was quenched with water/sat. NH4Cl (40 mL), and then the mixture was extracted with ether/EtOAc (2 × 30mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0- 100% ethyl acetate/petroleum ether) to afford tert-butyl (4-benzyl-7-nitro-3-oxo-3,4- dihydroquinoxalin-2-yl)carbamate (200 mg, 49.45%) as a yellow solid.MS (ESI): mass calcd. for C20H20N4O5, 396.14 m/z, found 397.10 [M+H]+ .
Figure imgf000380_0001
tert-butyl (7-amino-4-benzyl-3-oxo-3,4-dihydroquinoxalin-2-yl)carbamate To a solution of tert-butyl (4-benzyl-7-nitro-3-oxo-3,4-dihydroquinoxalin-2-yl)carbamate (200 mg, 0.505 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (281.7 mg, 5.045 mmol, 10 equiv) and NH4Cl (269.8 mg, 5.045 mmol, 10 equiv).The resulting mixture was stirred for overnight at 60 °C. The reaction progress was monitored by LCMS, the reaction mixture was quenched by water and extracted with EA (3 × 20 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford tert-butyl (7-amino-4-benzyl-3-oxo-3,4-dihydroquinoxalin-2-yl)carbamate (100 mg, 46.83%) as a yellow solid.MS (ESI): mass calcd. for C20H22N4O3, 366.17 m/z, found 367.10 [M+H]+.
Figure imgf000380_0002
tert-butyl (4-benzyl-7-(3-(tert-butyl)ureido)-3-oxo-3,4-dihydroquinoxalin-2-yl)carbamate To a stirred solution of tert-butyl (7-amino-4-benzyl-3-oxo-3,4-dihydroquinoxalin-2- yl)carbamate (90 mg, 0.246 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2- methylpropane (121.7 mg, 1.228 mmol, 5 equiv) and TEA (74.5 mg, 0.737 mmol, 3 equiv) stirred overnight at rt. The reaction progress was monitored by LCMS. The reaction mixture was quenched by water and extracted with EA (3 × 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford tert-butyl (4-benzyl-7-(3-(tert-butyl)ureido)-3-oxo-3,4- dihydroquinoxalin-2-yl)carbamate (30 mg, 25.68%) as a yellow solid.MS (ESI): mass calcd. for C25H31N5O4, 465.24 m/z, found 466.15 [M+H]+.
Figure imgf000381_0001
1-(3-amino-1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea To a stirred solution of tert-butyl (4-benzyl-7-(3-(tert-butyl)ureido)-3-oxo-3,4- dihydroquinoxalin-2-yl)carbamate (30 mg, 0.064 mmol, 1 equiv)in HCl (in EA) (10 mL) stirred 2 h at rt. The reaction mixture was concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-100% ethyl acetate/petroleum ether) to afford 30 mg crude product as a white solid. The compound was separated by preparative HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 45% B to 73% B in 9 min, 73% B; Wave Length: 254/220 nm; RT1(min): 9.6; Number Of Runs: 0) to afford 1-(3-amino-1- benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(tert-butyl)urea (9.3 mg, 29.89%) as a yellow solid.MS (ESI): mass calcd. for C20H23N5O2, 365.19 m/z, found 366.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.55 (br, 2H), 8.43 (s, 1H), 7.69 (d, J = 2.3 Hz, 1H), 7.21 – 7.37 (m, 5H), 7.15 (d, J = 9.1 Hz, 1H), 7.04 (dd, J = 9.0, 2.4 Hz, 1H), 6.03 (s, 1H), 5.44 (s, 2H), 1.28 (s, 9H). Example 295: 1-(tert-butyl)-3-(3-ethyl-2-oxo-1-(3-(trifluoromethoxy)benzyl)-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000382_0001
3-ethyl-6-nitroquinoxalin-2(1H)-one To a solution of 2-amino-4-nitroaniline (2 g, 13.060 mmol, 1 equiv) in ethyl alcohol (30 mL) was added ethyl 2-oxobutanoate (2.039 g, 15.667 mmol, 1.20 equiv). The reaction was stirred at 80 °C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give tert-butyl 3-ethyl-6-nitro-1H-quinoxalin-2-one (2.5 g, 87.33%) as a red solid. MS (ESI): mass calcd. for C10H9N3O3: 219.06 m/z, found 220.10 [M+H]+ .
Figure imgf000383_0001
3-ethyl-6-nitro-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a solution of 3-ethyl-6-nitro-1H-quinoxalin-2-one (700 mg, 3.193 mmol, 1 equiv) and 1- (bromomethyl)-3-(trifluoromethoxy)benzene (2443.30 mg, 9.579 mmol, 3 equiv) in DMF (10 mL) was added K2CO3 (1333.70 mg, 9.579 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-ethyl-6-nitro-1-{[3- (trifluoromethoxy)phenyl]methyl}quinoxalin-2-one (819 mg, 65.20%) as a red solid. LC/MS: mass calcd. for C18H14F3N3O4: 393.09, found: 394.05 [M+H]+.
Figure imgf000383_0002
6-amino-3-ethyl-1-(3-(trifluoromethoxy)benzyl)quinoxalin-2(1H)-one To a solution of 3-ethyl-6-nitro-1-{[3-(trifluoromethoxy)phenyl]methyl}quinoxalin-2-one (400 mg, 1.017 mmol, 1 equiv) in methanol (15 mL) and water (1.5 mL) was added iron (567.93 mg, 10.170 mmol, 10 equiv) and NH4Cl (543.98 mg, 10.170 mmol, 10 equiv). The reaction was stirred at 60 °C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-3-ethyl-1-{[3- (trifluoromethoxy)phenyl]methyl}quinoxalin-2-one (300 mg, 81.19%) as a white solid. LC/MS (ESI): mass calcd. for C18H16F3N3O2: 363.12 m/z, found:364.05 [M+H]+.
Figure imgf000384_0001
1-(tert-butyl)-3-(3-ethyl-2-oxo-1-(3-(trifluoromethoxy)benzyl)-1,2-dihydroquinoxalin-6- yl)urea To a solution of 6-amino-3-ethyl-1-{[3-(trifluoromethoxy)phenyl]methyl}quinoxalin-2-one (100 mg, 0.275 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (81.85 mg, 0.825 mmol, 3 equiv) in DCM (3 mL) was added TEA (83.55 mg, 0.825 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm(eluent: 17% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-(3-ethyl-2-oxo-1-{[3- (trifluoromethoxy)phenyl]methyl}quinoxalin-6-yl)urea (25.1 mg, 19.69%) as a white solid. LC/MS (ESI): mass calcd. for C23H25F3N4O3:462.19 m/z, found:461.00 [M-H]-.19F NMR (282 MHz, DMSO) δ -56.69.1H NMR (300 MHz, DMSO-d6) δ 8.40 (s, 1H), 7.96 (m, 1H), 7.45 (m, 1H), 7.18 – 7.38 (m, 5H), 6.03 (s, 1H), 5.50 (s, 2H), 2.89 (m, 2H), 1.26-1.30 (m, 12H). Example 296: 1-(tert-butyl)-3-(3-oxo-4-(2,2,2-trifluoro-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000385_0001
5-bromo-2-((2,2,2-trifluoro-1-phenylethyl)amino)phenol To a solution of 2-amino-5-bromophenol (1 g, 5.318 mmol, 1 equiv) and trifluoroacetophenone (0.93 g, 5.318 mmol, 1 equiv) in DCM (20 mL) was added trimethylaluminium (1.15 g, 15.954 mmol, 3 equiv) under N2. The reaction was stirred at rt under N2 for 15h. To the above mixture was added BH3-Me2S (10.64 mL, 10.636 mmol, 2 equiv, 1.0 M in THF). The resulting mixture was stirred for additional 2h at rt. The reaction was quenched by dropwise addition of 20% aqueos NaOH and the vaqueous layer was extracted with DCM (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-bromo-2-[(2,2,2- trifluoro-1-phenylethyl)amino]phenol (1.5 g, 81.48%) as an orange oil. MS (ESI): mass calcd. for C14H11BrF3NO: 345.00 m/z, found 346.05, 348.05 [M+H, M+H+2]+.
Figure imgf000385_0002
ethyl 2-(5-bromo-2-((2,2,2-trifluoro-1-phenylethyl)amino)phenoxy)acetate To a solution of 5-bromo-2-[(2,2,2-trifluoro-1-phenylethyl)amino]phenol (400 mg, 1.156 mmol, 1 equiv) in DMF/H2O(10:1, 11 mL) was added ethyl bromoacetate (231.58 mg, 1.387 mmol, 1.2 equiv) and K2CO3 (321.74 mg, 2.312 mmol, 2 equiv). The reaction was stirred at 80 °C for 1h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give ethyl 2-{5-bromo-2-[(2,2,2-trifluoro-1-phenylethyl)amino]phenoxy}acetate (400 mg, 80.08%) as an orange solid. MS (ESI): mass calcd. for C18H17BrF3NO3: 431.03 m/z, found 432.05, 434.05 [M+H, M+H+2]+ .
Figure imgf000386_0001
2-(5-bromo-2-((2,2,2-trifluoro-1-phenylethyl)amino)phenoxy)acetic acid To a solution ofethyl 2-{5-bromo-2-[(2,2,2-trifluoro-1-phenylethyl)amino]phenoxy}acetate (240 mg, 0.555 mmol, 1 equiv) in THF/EtOH/H2O(1:1:1 , 15 mL) was added LiOH (66.49 mg, 2.775 mmol, 5 equiv). The reaction was stirred at rt for 1h. The mixture was acidified to pH = 5 with 1N HCl. Diluted with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 5-bromo- 2-[(2,2,2-trifluoro-1-phenylethyl)amino]phenoxyacetic acid (200 mg, 89.12%) as a white oil. MS (ESI): mass calcd. for C16H13BrF3NO3: 403.00 m/z, found:404.00, 406.00 [M+H, M+H+2]+.
Figure imgf000386_0002
7-bromo-4-(2,2,2-trifluoro-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 5-bromo-2-[(2,2,2-trifluoro-1-phenylethyl)amino]phenoxyacetic acid (200 mg, 0.495 mmol, 1 equiv) in EA (10 mL) was added T3P (3 mL, 50% in EA). The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-bromo-4- (2,2,2-trifluoro-1-phenylethyl)-2H-1,4-benzoxazin-3-one (140 mg, 73.27%) as a white solid. MS (ESI): mass calcd. for C16H11BrF3NO2: 384.99 m/z, found 385.90, 387.90 [M+H, M+H+2]+.
Figure imgf000387_0001
7-amino-4-(2,2,2-trifluoro-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of 7-bromo-4-(2,2,2-trifluoro-1-phenylethyl)-2H-1,4-benzoxazin-3-one (140 mg, 0.363 mmol, 1 equiv) in EtOH/ H2O (4:1, 10 mL) was added (1R,2R)-N1,N2- dimethylcyclohexane-1,2-diamine (30.94 mg, 0.218 mmol, 0.6 equiv), CuSO4 (57.86 mg, 0.363 mmol, 1 equiv), azidosodium (70.71 mg, 1.089 mmol, 3 equiv) and sodium ascorbate (144.37 mg, 0.726 mmol, 2 equiv). The reaction was stirred at 80°C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4- (2,2,2-trifluoro-1-phenylethyl)-2H-1,4-benzoxazin-3-one (70 mg, 59.91%) as a yellow solid. MS (ESI): mass calcd. for C16H13F3N2O2: 322.09 m/z, found 323.05 [M+H]+.
Figure imgf000387_0002
1-(tert-butyl)-3-(3-oxo-4-(2,2,2-trifluoro-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of 7-amino-4-(2,2,2-trifluoro-1-phenylethyl)-2H-1,4-benzoxazin-3-one (90 mg, 0.279 mmol, 1 equiv) in DCM/Toluene (2:1, 7.5 mL) was added 2-isocyanato-2-methylpropane (166.10 mg, 1.674 mmol, 6 equiv).The reaction was stirred at 90 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column, 30*150 mm, 5μm column (eluent: 45% to 67% (v/v) CH3CN and Water (10 mmol/L NH4HCO3)) to afford the title compound 3-tert-butyl-1-[3-oxo-4-(2,2,2-trifluoro-1-phenylethyl)- 2H-1,4-benzoxazin-7-yl]urea (22.3 mg, 18.95%) as a white solid. MS (ESI): mass calcd. for C21H22F3N3O3: 421.16 m/z, found 422.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.03 – 8.30 (m, 1H), 7.37 – 7.60 (m, 5H), 7.15 – 7.30 (m, 1H), 6.78 – 6.96 (m, 1H), 6.56 – 6.78 (m, 2H), 5.80 – 6.06 (m, 1H), 4.68 – 4.88 (m, 2H), 1.20 – 1.39 (m, 9H).19F NMR (376 MHz, DMSO) δ(ppm): - 63.7904. Example 297: (S)-1-(tert-butyl)-3-(1-(1-(3-methoxyphenyl)ethyl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea Synthetic Scheme
Figure imgf000388_0001
(S)-1-(1-(3-methoxyphenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of (1R)-1-(3-methoxyphenyl)ethanol (238 mg, 1.561 mmol, 1 equiv) in DCM (6 mL) was added 6-nitro-3,4-dihydro-1H-quinolin-2-one (300 mg, 1.561 mmol, 1 equiv), triphenylphosphine (409 mg, 1.561 mmol, 1 equiv) and DEAD (272 mg, 1.561 mmol, 1 equiv) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford (S)- 1-(1-(3-methoxyphenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (160 mg, 21.86%) as a yellow solid. MS (ESI): mass calcd. for C18H18N2O4, 326.13 m/z, found 327.10 [M+H]+.
Figure imgf000389_0001
(S)-6-amino-1-(1-(3-methoxyphenyl)ethyl)-3,4-dihydroquinolin-2(1H)-one To a mixture of (S)-1-(1-(3-methoxyphenyl)ethyl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (160 mg, 0.490 mmol, 1 equiv) in methanol (20 mL) was added Pd/C (10%) under nitrogen atmosphere. The mixture was hydrogenated at room temperature for 1 h under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. This resulted in (S)-6-amino-1-(1-(3-methoxyphenyl)ethyl)-3,4-dihydroquinolin-2(1H)-one (100 mg, 68.82%) as a white soild. MS (ESI): mass calcd. for C18H20N2O2, 296.15 m/z, found 297.15 [M+H]+.
Figure imgf000389_0002
(S)-1-(tert-butyl)-3-(1-(1-(3-methoxyphenyl)ethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6- yl)urea To a mixture of (S)-6-amino-1-(1-(3-methoxyphenyl)ethyl)-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.337 mmol, 1 equiv) and TEA (68 mg, 0.674 mmol, 2 equiv) in DCM (5 mL) was added 2- isocyanato-2-methylpropane (100 mg, 1.011 mmol, 3 equiv) at 0 °C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude was purified by flash chromatography with PE/EA (1:1) and Prep-HPLC ( Column: XBridge Shield RP18 OBD Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 37% B to 59% B in 10 min, 59% B; Wave Length: 254/220 nm ) to give (S)-1-(tert- butyl)-3-(1-(1-(3-methoxyphenyl)ethyl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (31.1 mg, 23.24%) as a white solid. MS (ESI): mass calcd. for C23H29N3O3, 395.22 m/z, found 396.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.32 (d, J = 2.4 Hz, 1H), 7.25 (t, J = 7.9 Hz, 1H), 6.88 – 6.74 (m, 4H), 6.49 (d, J = 8.8 Hz, 1H), 6.12 (q, J = 7.1 Hz, 1H), 5.91 (s, 1H), 3.72 (s, 3H), 2.76 – 2.84 (m, 2H), 2.56 – 2.63 (m, 2H), 1.66 (d, J = 7.1 Hz, 3H), 1.25 (s, 9H). Example 298: 1-(tert-butyl)-3-(1-(chroman-4-yl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea Synthetic Scheme
Figure imgf000390_0001
N-(2-bromo-4-nitrophenyl)chroman-4-amine To a stirred mixture of 2-bromo-1-fluoro-4-nitrobenzene (1.5 g, 6.818 mmol, 1 equiv) and 3,4- dihydro-2H-1-benzopyran-4-amine (1.02 g, 6.818 mmol, 1 equiv) in DMF (20 mL) was added Cs2CO3 (4.44 g, 13.636 mmol, 2 equiv) at 25°C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford N-(2-bromo-4- nitrophenyl)chroman-4-amine (900 mg, 37.8%) as a white solid. MS (ESI): mass calcd. for C15H13BrN2O3: 348.00 m/z, found 349.00 [M+H] +.
Figure imgf000391_0001
ethyl (E)-3-(2-(chroman-4-ylamino)-5-nitrophenyl)acrylate To a stirred mixture of N-(2-bromo-4-nitrophenyl)chroman-4-amine (465 mg, 1.332 mmol, 1 equiv) and ethyl (2E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoate (451.60 mg, 1.998 mmol, 1.5 equiv) in dioxane (15 mL) and H2O (1.5 mL) was added K2CO3 (552.13 mg, 3.996 mmol, 3 equiv) and Pd(dppf)Cl2 (97.44 mg, 0.133 mmol, 0.1 equiv) at 25°C under air atmosphere. The resulting mixture was stirred for 2h at 90 degrees C under nitrogen. Desired product could be detected by LCMS. The mixture was diluted with water (30 mL). The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford ethyl (E)-3-(2-(chroman-4-ylamino)-5-nitrophenyl)acrylate (480 mg, 97.84%) as a white solid. MS (ESI): mass calcd. for C20H20N2O5: 368.14 m/z, found 369.15 [M+H] +.
Figure imgf000391_0002
ethyl 3-(2-(chroman-4-ylamino)-5-nitrophenyl)propanoate To a stirred mixture of ethyl (E)-3-(2-(chroman-4-ylamino)-5-nitrophenyl)acrylate (400 mg, 1.086 mmol, 1 equiv) in EtOH (5 mL) was added RuCl(PPh3)2 (399.05 mg, 0.434 mmol, 0.40 equiv) at 25 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature under hydrogen. Desired product could be detected by LCMS. The resulting mixture was filtered, and the filter cake was washed with EtOH (4 x 10 mL). The filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford ethyl 3-(2-(chroman-4-ylamino)-5-nitrophenyl)propanoate (190 mg, 47.24%) as a white solid. MS (ESI): mass calcd. for C30H42N4O4: 370.15 m/z, found 371.15 [M +H] +.
Figure imgf000392_0001
3-(2-(chroman-4-ylamino)-5-nitrophenyl)propanoic acid To a stirred mixture of ethyl 3-(2-(chroman-4-ylamino)-5-nitrophenyl)propanoate (160 mg, 0.432 mmol, 1 equiv) in H2O (1 mL), EtOH (1 mL) and THF (1 mL) was added LiOH (31.04 mg, 1.296 mmol, 3 equiv) at 0 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was basified to PH = 5 with 2N HCl and diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 3-(2-(chroman-4-ylamino)-5- nitrophenyl)propanoic acid (120 mg, 81.15%) as a white solid. MS (ESI): mass calcd. for C18H18N2O5: 342.12 m/z, found 341.00 [M - H] -.
Figure imgf000392_0002
1-(chroman-4-yl)-6-nitro-3,4-dihydroquinolin-2(1H)-one The 3-(2-(chroman-4-ylamino)-5-nitrophenyl)propanoic acid (110 mg, 0.321 mmol, 1 equiv) was dissolved in T3P (2 mL) and EA (2 mL) at 0 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford 1-(chroman-4-yl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (100 mg, 95.96%) as a white solid. MS (ESI): mass calcd. for C18H16N2O4: 324.11 m/z, found 325.05[M +H] +.
Figure imgf000393_0001
6-amino-1-(chroman-4-yl)-3,4-dihydroquinolin-2(1H)-one To a stirred mixture of 1-(chroman-4-yl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (100 mg, 0.308 mmol, 1 equiv) in MeOH (5 mL) was added Fe (172.18 mg, 3.080 mmol, 10 equiv) and NH4Cl (164.92 mg, 3.080 mmol, 10 equiv) at room temperature under air atmosphere. The resulting mixture was stirred for 2h at 80 degrees C. Desired product could be detected by LCMS. After cooling down to room temperature, the mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the 6-amino-1-(chroman-4-yl)- 3,4-dihydroquinolin-2(1H)-one (50 mg, 55.09%) as a white solid. MS (ESI): mass calcd. for C18H18N2O2: 294.14 m/z, found 295.05[M + H] +.
Figure imgf000393_0002
1-(tert-butyl)-3-(1-(chroman-4-yl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred mixture of 6-amino-1-(chroman-4-yl)-3,4-dihydroquinolin-2(1H)-one (50 mg, 0.170 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (84.20 mg, 0.850 mmol, 5 equiv) in DCM (2 mL) and toluene (0.5 mL) was added TEA (85.94 mg, 0.850 mmol, 5 equiv) dropwise at room temperature under air atmosphere. The resulting mixture was stirred for 2h at 90 degrees C. Desired product could be detected by LCMS. After cooling down to room temperature, the mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5μm column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10 mmol /L NH4HCO3) to afford the 1-(tert-butyl)-3-(1-(chroman-4-yl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (7.2 mg, 10.72%) as a white solid. MS (ESI): mass calcd. for C23H27N3O3: 393.21 m/z, found 394.05[M +H] +. 1H-NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.34 (d, J = 2.5 Hz, 1H), 7.04 – 7.15 (m, 1H), 6.71 – 6.90 (m, 4H), 6.08 – 6.68 (m, 2H), 5.92 (s, 1H), 4.39 (d, J = 10.8 Hz, 1H), 4.19 – 4.33 (m, 1H), 2.78 – 2.89 (m, 2H), 2.58 – 2.69 (m, 3H), 1.92 (s, 1H), 1.24 (s, 9H). Example 299: (S)-N-(2-oxo-1-(1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-1,2- dihydroquinoxalin-6-yl)-3-(3-(trifluoromethyl)phenyl)propanamide
Figure imgf000394_0001
(S)-N-(2-oxo-1-(1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-1,2-dihydroquinoxalin-6-yl)-3-(3- (trifluoromethyl)phenyl)propanamide To a solution of 3-[3-(trifluoromethyl)phenyl]propanoic acid (43.07 mg, 0.197 mmol, 1.1 equiv) in DMF (5 mL) was added 6-amino-1-[(1S)-1-[6-(trifluoromethyl)pyridin-2-yl]ethyl]quinoxalin- 2-one (60 mg, 0.179 mmol, 1 equiv), HATU (88.72 mg, 0.233 mmol, 1.3 equiv) and DIEA (69.59 mg, 0.537 mmol, 3 equiv) at 0 °C. The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound N-{2-oxo-1-[(1S)-1-[6- (trifluoromethyl)pyridin-2-yl]ethyl]quinoxalin-6-yl}-3-[3-(trifluoromethyl)phenyl]propanamide (28.4 mg, 29.61%) as a yellow solid. MS (ESI): mass calcd. for C26H20F6N4O2: 534.15 m/z, found:535.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.25 (s, 1H), 8.10 – 8.20 (m, 1H), 7.95 – 8.10 (m, 1H), 7.77 – 7.86 (m, 1H), 7.50 – 7.70 (m, 6H), 7.00 – 7.50 (m, 1H), 6.55 (s, 1H), 2.97 – 3.11 (m, 2H), 2.62 – 2.77 (m, 2H), 1.90 – 1.98 (m, 3H).19F NMR (376 MHz, DMSO) δ(ppm): -60.9926, -66.5643. Example 300: (S)-4,4-dimethyl-N-(2-oxo-1-(1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-1,2- dihydroquinoxalin-6-yl)pentanamide Synthetic Scheme
Figure imgf000395_0001
(S)-4,4-dimethyl-N-(2-oxo-1-(1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-1,2- dihydroquinoxalin-6-yl)pentanamide To a solution of 4,4-dimethyl valeric acid (25.70 mg, 0.197 mmol, 1.1 equiv) in DMF (5 mL) was added 6-amino-1-[(1S)-1-[6-(trifluoromethyl)pyridin-2-yl]ethyl]quinoxalin-2-one (60 mg, 0.179 mmol, 1 equiv), HATU (88.72 mg, 0.233 mmol, 1.3 equiv) and DIEA (69.59 mg, 0.537 mmol, 3 equiv) at 0°C. The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XSelect CSH Fluoro Phenyl, 30*150 mm, 5μm column (eluent: 33% to 56% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 4,4-dimethyl-N-{2-oxo-1-[(1S)-1-[6- (trifluoromethyl)pyridin-2-yl]ethyl]quinoxalin-6-yl}pentanamide (31.9 mg, 39.81%) as a yellow solid. MS (ESI): mass calcd. for C23H25F3N4O2: 446.19 m/z, found:447.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 10.12 (s, 1H), 8.12 – 8.30 (m, 2H), 7.93 – 8.10 (m, 1H), 7.70 – 7.85 (m, 1H), 7.50 – 7.70 (m, 2H), 6.98 – 7.45 (m, 1H), 6.30 – 6.64 (m, 1H), 2.15 – 2.37 (m, 2H), 1.81 – 2.00 (m, 3H), 1.40 – 1.60 (m, 2H), 0.90 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -66.5505. Example 301: 1-(1-benzyl-2-oxo-3-(trifluoromethyl)-1,2,3,4-tetrahydroquinolin-6-yl)-3- (tert-butyl)urea Synthetic Scheme
Figure imgf000396_0001
6-bromo-3-(trifluoromethyl)quinolin-2(1H)-one 6-bromoquinolin-2(1H)-one (1.0 g, 4.46 mmol, 1.0 equiv), Mn(OAc)3.2H2O (3.59 g, 17.84 mmol, 4.0 equiv) and sodium trifluoromethanesulfinate (1.57 g, 13.87 mmol, 3.0 equiv) were dissolved in glacial acetic acid (25 mL), and the reaction was stirred at 25 °C for 24 hours. Water (100 mL) was added to the system, the mixture was extracted with ethyl acetate (100 mL × 2), the organic phases were combined, washed with saturated saline (100 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE/EA=3:1) to obtain 6-bromo-3- (trifluoromethyl)quinolin-2(1H)-one (0.5 g, 38.46%) as a yellow solid. MS (ESI): mass calcd. for C10H5BrF3NO, 290.95 m/z, found 292.15 [M+H]+.
Figure imgf000396_0002
1-benzyl-6-bromo-3-(trifluoromethyl)quinolin-2(1H)-one A solution of 6-bromo-3-(trifluoromethyl)quinolin-2(1H)-one (480 mg, 1.64 mmol, 1 equiv) in DMF (5 mL) was added BnBr (0.8 g, 4.90 mmol, 3.0 equiv) and Cs2CO3 (2.1 g, 6.560 mmol, 4.0 equiv) at rt. The resulting mixture was stirred for 2 h at rt. The reaction was quenched with water (20 mL). The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with NaCl (aq., 3 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 1-benzyl-6-bromo-3- (trifluoromethyl)quinolin-2(1H)-one (400 mg, 63.89%%) as a white solid. MS (ESI): mass calcd. for C17H11BrF3NO, 381.00 m/z, found 382.15 [M+H]+.
Figure imgf000397_0001
tert-butyl (1-benzyl-2-oxo-3-(trifluoromethyl)-1,2-dihydroquinolin-6-yl)carbamate A solution of 1-benzyl-6-bromo-3-(trifluoromethyl)quinolin-2(1H)-one (0.2 g, 0.524 mmol, 1 equiv) in toluene (5 mL) was treated with tert-butyl carbamate (123 mg, 1.04 mmol, 2 equiv), Cs2CO3 (0.5 g, 1.57 mmol, 3 equiv), XantPhos (30 mg, 0.052 mmol, 0.1 equiv) and Pd2(dba)3 (48 mg, 0.052 mmol, 0.1 equiv) for overnight at 90 °C under nitrogen atmosphere. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford tert- butyl (1-benzyl-2-oxo-3-(trifluoromethyl)-1,2-dihydroquinolin-6-yl)carbamate (120 mg, 54.79%). MS (ESI): mass calcd. for C22H21F3N2O3, 418.15.00 m/z, found 419.10 [M+H]+.
Figure imgf000397_0002
tert-butyl (1-benzyl-2-oxo-3-(trifluoromethyl)-1,2,3,4-tetrahydroquinolin-6-yl)carbamate To a solution of tert-butyl (1-benzyl-2-oxo-3-(trifluoromethyl)-1,2-dihydroquinolin-6- yl)carbamate (90 mg, 0.215 mmol, 1 equiv) in EtOH (5 mL) and HOAc (1 mL) was added Pd/C (50 mg) under nitrogen atmosphere. The resulting mixture was placed at 60 °C under a hydrogen atmosphere (50 bar) until the starting material was totally consumed by LCMS. The mixture was filtered and concentrated under vacuum to afford tert-butyl (1-benzyl-2-oxo-3-(trifluoromethyl)- 1,2,3,4-tetrahydroquinolin-6-yl)carbamate (90 mg crude) as a white solid.
Figure imgf000398_0001
6-amino-1-benzyl-3-(trifluoromethyl)-3,4-dihydroquinolin-2(1H)-one To a stirred solution of tert-butyl (1-benzyl-2-oxo-3-(trifluoromethyl)-1,2,3,4-tetrahydroquinolin- 6-yl)carbamate (90 mg crude) in HCl (4N in EA, 3 mL) at rt. The reaction was stirred for 2 h at rt. The resulting mixture was concentrated under reduced pressure to afford 6-amino-1-benzyl-3- (trifluoromethyl)-3,4-dihydroquinolin-2(1H)-one (80 mg crude) as a white solid.
Figure imgf000398_0002
1-(1-benzyl-2-oxo-3-(trifluoromethyl)-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea A solution of 6-amino-1-benzyl-3-(trifluoromethyl)-3,4-dihydroquinolin-2(1H)-one (80 mg crude), TEA (80 mg) and 2-isocyanato-2-methylpropane (80 mg) in DCM (5 mL) was stirred for overnight at rt. The mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluted with EA (46%-57%) and C18 column to afford 1-(1-benzyl- 2-oxo-3-(trifluoromethyl)-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea (2.7 mg) as a white solid. MS (ESI): mass calcd. for C22H24F3N3O2: 419.18 m/z, found 420.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.39 (d, J = 2.5 Hz, 1H), 7.28 – 7.37 (m, 2H), 7.18 – 7.28 (m, 3H), 7.05 (dd, J = 8.8, 2.5 Hz, 1H), 6.83 (d, J = 8.8 Hz, 1H), 5.96 (s, 1H), 5.08 – 5.18(m, 2H), 4.01 – 4.12 (m, 1H), 3.06 – 3.23 (m, 2H), 1.26 (s, 9H). Example 302: 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000399_0001
2-(benzylamino)-5-nitrobenzonitrile A solution of 2-fluoro-5-nitrobenzonitrile (2.0 g, 12.040 mmol, 1 equiv) in NMP (20 mL) was added benzylamine (1.94 g, 18.060 mmol, 1.5 equiv) and DIEA (3.11 g, 24.080 mmol, 2.0 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20mL). The combined organic layers were washed with NaCl (3 x 20mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 2-(benzylamino)-5-nitrobenzonitrile (2.2 g, 72.15%) as an off-white solid. LC/MS: mass calcd. for C14H11N3O2: 253.09, found: 254.10 [M+H]+.
Figure imgf000399_0002
2-(aminomethyl)-N-benzyl-4-nitroaniline A solution of 2-(benzylamino)-5-nitrobenzonitrile (500 mg, 1.974 mmol, 1 equiv) in methanol (5 mL) was added borane (54.61 mg, 3.948 mmol, 2.0 equiv) at 0°C. The resulting mixture was stirred for overnight at 60 °C. The reaction mixture was quenched by water (5 mL) and extracted with EA (3 x 10mL). The combined organic layers were washed with NaCl (3 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 2- (aminomethyl)-N-benzyl-4-nitroaniline (300 mg, 59.06%) as colorless oil. LC/MS: mass calcd. for C14H15N3O2: 257.12, found: 258.10[M+H]+.
Figure imgf000400_0001
1-benzyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one A solution of 2-(aminomethyl)-N-benzyl-4-nitroaniline (130 mg, 0.505 mmol, 1 equiv) in DCM (3 mL) was added triphosgene (45.0 mg, 0.151 mmol, 0.3 equiv) and triethylamine (127.8 mg, 1.262 mmol, 2.5 equiv). The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (5 mL) and extracted with EA (3 x 10 mL). The combined organic layers were washed with NaCl (3 x 10mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (3/1) to afford 1-benzyl-6-nitro-3,4-dihydroquinazolin- 2(1H)-one (37 mg, 25.85%) as a white solid. LC/MS: mass calcd. for C15H13N3O3: 283.10, found: 284.05 [M+H]+.
Figure imgf000400_0002
6-amino-1-benzyl-3,4-dihydroquinazolin-2(1H)-one A solution of 1-benzyl-6-nitro-3,4-dihydroquinazolin-2(1H)-one (32 mg, 0.113 mmol, 1 equiv) in EtOH (3 mL)/H2O (0.3 mL) was added Fe (63.1 mg, 1.130 mmol, 10 equiv) and NH4Cl (60.4 mg, 1.130 mmol, 10 equiv). The resulting mixture was stirred for 1h at 60 °C. The reaction mixture was quenched by water (5 mL) and extracted with EA (3 x 5mL). The combined organic layers were washed with NaCl (3 x 5mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (1/1) to afford 6-amino-1-benzyl-3,4-dihydroquinazolin- 2(1H)-one (20 mg, 69.90%) as an off-white solid. LC/MS: mass calcd. for C15H15N3O: 253.12, found: 254.20 [M+H]+.
Figure imgf000401_0001
1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert-butyl)urea A solution of 6-amino-1-benzyl-3,4-dihydroquinazolin-2(1H)-one (20 mg, 0.079 mmol, 1 equiv) in DCM (26.8 mg, 0.316 mmol, 4.0 equiv) was added 2-isocyanato-2-methylpropane (23.5 mg, 0.237 mmol, 3.0 equiv) and TEA (3 mL) at rt. The resulting mixture was stirred for overnight at rt. The reaction mixture was quenched by water (5 mL) and extracted with EA (5 mL x 3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product 30 mg was purified by Prep-HPLC with the following conditions ( Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 17% B to 42% B in 9 min, 42% B; Wave Length: 254/220 nm; RT1(min): 8.9; Number Of Runs: 0 ) to affored 1-(1-benzyl-2-oxo-1,2,3,4-tetrahydroquinazolin-6-yl)-3-(tert- butyl)urea (3.0 mg, 10.77%) as a white solid. MS (ESI): mass calcd. C20H24N4O2 for: 352.19 m/z, found 353.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.27 – 7.35 (m, 3H),7.24 – 7.19(m, 3H), 7.09 (s, 1H), 6.89 (dd, J = 8.8, 2.5 Hz, 1H), 6.57 (d, J = 8.8 Hz, 1H), 5.90 (s, 1H), 5.00 (s, 2H), 4.33 (s, 2H), 1.25 (s, 9H). Example 303: cyclopentyl ((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)carbamate Synthetic Scheme
Figure imgf000402_0001
Figure imgf000402_0002
(R)-7-bromo-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred mixture of (R)-7-bromo-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one (1 g, 4.131 mmol, 1 equiv) and (R)-1-phenylethan-1-ol (555.14 mg, 4.544 mmol, 1.1 equiv) in DCM (20 mL) was added PPh3 (1.63 g, 6.197 mmol, 1.5 equiv) at 25°C under air atmosphere. The resulting mixture was stirred at room temperature under nitrogen. After 30 min, the DEAD (719.44 mg, 4.131 mmol, 1 equiv) was added to the mixture. The resulting mixture was stirred for 2h at room temperature under nitrogen. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the (R)-7-bromo-2-methyl-4- ((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (1 g, 69.92%) as a white solid. MS (ESI): mass calcd. for C17H16BrNO2: 345.04 m/z, found 346.05 [M+H] +.
Figure imgf000402_0003
(R)-7-amino-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred mixture of (R)-7-bromo-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (1 g, 2.432 mmol, 1 equiv) in EtOH (8 mL) and H2O (2 mL) was added NaN3 (452.37 mg, 7.296 mmol, 3 equiv), N1,N2-dimethylcyclohexane-1,2-diamine (207.54 mg, 1.459 mmol, 0.6 equiv) , CuSO4 (388.10 mg, 2.432 mmol, 1 equiv) and sodium ascorbate (968.37 mg, 4.864 mmol, 2 equiv) at 25°C under air atmosphere. The resulting mixture was stirred for 2h at 80 degrees C under nitrogen. Desired product could be detected by LCMS. After cooling down to room temperature, the mixture was diluted with water (30 mL). The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the (R)-7- amino-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 43.70%) as a white solid. MS (ESI): mass calcd. for C17H18N2O2: 282.14 m/z, found 283.15 [M+H]+.
Figure imgf000403_0001
cyclopentyl ((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin- 7-yl)carbamate To a stirred mixture of (R)-7-amino-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (80 mg, 0.283 mmol, 1 equiv) and cyclopentyl carbonochloridate (126.31 mg, 0.849 mmol, 3 equiv) in DCM (3 mL) was added pyridine (67.24 mg, 0.849 mmol, 3 equiv) dropwise at 0 degrees C under air atmosphere. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5μm column (eluent: 51% to 73% (v/v) CH3CN and H2O with 10 mmol /L NH4HCO3) to afford the cyclopentyl ((R)-2- methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)carbamate (14.3 mg, 12.73%) as a white solid. MS (ESI): mass calcd. for C23H26N2O4: 394.19 m/z, found 395.10 [M+H]+.1H-NMR (300 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.31 – 7.41 (m, 2H), 7.20 – 7.31 (m, 3H), 7.20 (d, J = 2.4 Hz, 1H), 6.81 – 6.91 (m, 1H), 6.67 (d, J = 8.9 Hz, 1H), 6.03 – 6.16 (m, 1H), 5.00 – 5.10 (m, 1H), 4.70 – 4.82 (m, 1H), 1.79 – 1.94 (m, 2H), 1.71 – 1.78 (m, 3H), 1.49 – 1.70 (m, 6H), 1.45 (d, J = 6.8 Hz, 3H). Example 304: cyclopentyl N-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethyl)pyridin-2- yl]methyl}-2H-1,4-benzoxazin-7-yl]carbamate Synthetic Scheme
Figure imgf000404_0001
(2R)-7-bromo-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-3- one To a solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (500 mg, 2.066 mmol, 1 equiv) in DCM (10 mL) was added [6-(trifluoromethyl)pyridin-2-yl]methanol (439.02 mg, 2.479 mmol, 1.2 equiv) and PPh3 (812.65 mg, 3.099 mmol, 1.5 equiv). After the mixture was stirred at rt for 1 h, then DEAD (539.58 mg, 3.099 mmol, 1.5 equiv) was added at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-7-bromo-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (410 mg, 49.48%) as a white solid. MS(ESI): mass calcd. for C16H12BrF3N2O2:400.00 m/z, found:401.00, 403.00 [M+H, M+H+2]+.
Figure imgf000405_0001
(2R)-7-amino-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-3- one To a solution of (2R)-7-bromo-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (600 mg, 1.287 mmol, 1 equiv) in EtOH (10 mL) and H2O (1 mL) were added N1,N2-dimethylcyclohexane-1,2-diamine (109.84 mg, 0.772 mmol, 0.6 equiv) and CuSO4 (205.41 mg, 1.287 mmol, 1 equiv), sodium ascorbate (512.53 mg, 2.574 mmol, 2 equiv). The reaction was stirred at 80 °C for 2 h. The mixture was allowed to cool down to room temperature. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-2-methyl-4- {[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-3-one (290 mg, 66.80%) as a white solid. MS (ESI): mass calcd. for C16H14F2N3O2:337.10 m/z, found:338.10 [M+H]+.
Figure imgf000405_0002
cyclopentyl N-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-7-yl]carbamate To a solution of (2R)-7-amino-2-methyl-4-{[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (80 mg, 0.237 mmol, 1 equiv) in DCM (5 mL) was added cyclopentyl carbonochloridate (52.86 mg, 0.355 mmol, 1.5 equiv) and pyridine (56.28 mg, 0.711 mmol, 3 equiv) at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (30 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C1830*150 mm, 5μm column (eluent: 26% to 42% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound cyclopentyl N-[(2R)-2-methyl-3-oxo-4- {[6-(trifluoromethyl)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-7-yl]carbamate (24.7 mg, 23.14%) as a white solid. MS (ESI): mass calcd. for C22H22F3N3O4:449.16 m/z, found:450.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 9.54 (s, 1H), 8.03 – 8.11 (m, 1H), 7.81 (d, J = 7.7 Hz, 1H), 7.54 (d, J = 8.0 Hz, 1H), 7.22 (s, 1H), 7.01 (d, J = 9.4 Hz, 1H), 6.89 (d, J = 8.8 Hz, 1H), 5.25 (s, 2H), 5.04-5.09 (m, 1H), 4.80 – 4.90 (m, 1H), 1.80 – 1.88 (m, 3H), 1.61 – 1.70 (m, 5H), 1.47 (d, J = 6.7 Hz, 3H).19F NMR (376 MHz, DMSO) δ -66.56. Example 305: 1-[4-benzyl-2-(fluoromethyl)-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea Synthetic Scheme
Figure imgf000406_0001
methyl 2-bromo-3-[(tert-butyldiphenylsilyl)oxy]propanoate To a stirred solution of methyl 2-bromo-3-hydroxypropanoate (3 g, 16.393 mmol, 1 equiv) in DCM (30 mL) was added TBDPSCl (6.76 g, 24.590 mmol, 1.5 equiv), TEA (4.98 g, 49.179 mmol, 3 equiv) and DMAP (0.20 g, 1.639 mmol, 0.1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (150 mL) and extracted with DCM (3 x 150 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford methyl 2-bromo-3-[(tert- butyldiphenylsilyl)oxy] propanoate (6.5 g, 94.1%) as a yellow oil. LC/MS (ESI): mass calcd. for C20H25BrO3Si: 420.1 m/z, found:421.1, 423.1 [M+H, M+H+2]+.
Figure imgf000407_0001
2-{[(tert-butyldiphenylsilyl)oxy]methyl}-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one To a stirred solution of methyl 2-bromo-3-[(tert-butyldiphenylsilyl)oxy]propanoate (8 g, 18.984 mmol, 1 equiv) in DMF(80 mL) was added 2-amino-5-nitrophenol (2.93 g, 18.984 mmol, 1 equiv) and K2CO3 (5.25 g, 37.968 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 80 °C under air atmosphere. The reaction was monitored by LCMS. After cooling down to rt, the reaction was quenched with water (400 mL) and extracted with EA (3 x 300 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 2-{[(tert- butyldiphenylsilyl)oxy]methyl}-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (2.8 g, 31.88%) as a yellow solid. LC/MS (ESI): mass calcd. for C25H26N2O5Si:462.2 m/z, found:461.2 [M+H]+.
Figure imgf000407_0002
4-benzyl-2-{[(tert-butyldiphenylsilyl)oxy]methyl}-7-nitro-2H-1,4-benzoxazin-3-one To a stirred solution of 2-{[(tert-butyldiphenylsilyl)oxy]methyl}-7-nitro-2,4-dihydro-1,4- benzoxazin-3-one (1.2 g, 2.594 mmol, 1 equiv) and benzyl alcohol (0.28 g, 2.594 mmol, 1 equiv) in DCM was added PPh3 (0.68 g, 2.594 mmol, 1 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (0.68 g, 3.891 mmol, 1.5 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 2 h at room temperature. The reaction was monitored by LCMS. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 4-benzyl-2-{[(tert-butyldiphenylsilyl)oxy]methyl}-7-nitro-2H-1,4- benzoxazin-3-one (800 mg, 55.80%) as a yellow oil. LC/MS (ESI): mass calcd. for C32H32N2O5Si:552.2 m/z, found:551.3 [M+H]+.
Figure imgf000408_0002
4-benzyl-2-(hydroxymethyl)-7-nitro-2H-1,4-benzoxazin-3-one To a stirred solution of 4-benzyl-2-{[(tert-butyldiphenylsilyl)oxy]methyl}-7-nitro-2H-1,4- benzoxazin-3-one (800 mg, 1.447 mmol, 1 equiv) in THF (8 mL) was added Et3N.3HF (350.02 mg, 2.171 mmol, 1.5 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (40 mL) and extracted with EA (3 x 40 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 4-benzyl-2-(hydroxymethyl)-7-nitro-2H-1,4-benzoxazin-3-one (320 mg, 70.34%) as a yellow oil. LC/MS (ESI): mass calcd. for C16H14N2O5:314.1 m/z, found:315.2 [M+H]+.
Figure imgf000408_0001
4-benzyl-2-(fluoromethyl)-7-nitro-2H-1,4-benzoxazin-3-one To a stirred solution of 4-benzyl-2-(hydroxymethyl)-7-nitro-2H-1,4-benzoxazin-3-one (320 mg, 1.018 mmol, 1 equiv) in DCM (3 mL) was added DAST (328.23 mg, 2.036 mmol, 2 equiv) dropwise at 0 °C under air atmosphere. The resulting mixture was stirred for 3 h at 0 °C under air atmosphere. The reaction was monitored by LCMS. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 4-benzyl-2-(fluoromethyl)-7-nitro-2H-1,4-benzoxazin-3-one (130 mg, 40.37%) as a yellow oil. LC/MS (ESI): mass calcd. for C16H13FN2O4:316.1 m/z, found:317.20 [M+H]+.
Figure imgf000409_0001
7-amino-4-benzyl-2-(fluoromethyl)-2H-1,4-benzoxazin-3-one To a stirred solution of 4-benzyl-2-(fluoromethyl)-7-nitro-2H-1,4-benzoxazin-3-one (130 mg, 0.411 mmol, 1 equiv) in AcOH (1.5 mL) was added Zn (268.72 mg, 4.110 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 7-amino-4-benzyl-2- (fluoromethyl)-2H-1,4-benzoxazin-3-one (130 mg, 110.47%) as a yellow solid. LC/MS (ESI): mass calcd. for C16H15FN2O2:286.1 m/z, found:287.20 [M+H]+.
Figure imgf000409_0002
1-[4-benzyl-2-(fluoromethyl)-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea To a stirred solution of 7-amino-4-benzyl-2-(fluoromethyl)-2H-1,4-benzoxazin-3-one (120 mg, 0.419 mmol, 1 equiv) in DCM/Toluene (2:1, 1.5 mL) was added 2-isocyanato-2-methylpropane (124.65 mg, 1.257 mmol, 3 equiv) dropwise at room temperature under air atmosphere. The resulting mixture was stirred overnight at 90 °C under air atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using YMC-Actus Tiart ExRS C18150 mm x 30 mm x 5 μm column (eluent: 23% to 40% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford 1-[4-benzyl-2- (fluoromethyl)-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea (13.7 mg, 8.25%) as a white solid. LC/MS (ESI): mass calcd. for C21H24FN3O3:385.2 m/z, found:386.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.35 – 7.21 (m, 6H), 6.83 (d, J = 8.8 Hz, 1H), 6.74 (d, J = 8.8 Hz, 1H), 5.97 (s, 1H), 5.21 – 4.81 (m, 5H), 1.26 (s, 9H).19F NMR (376 MHz, DMSO-d6) δ - 231.52. Example 306 and Example 307: 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(6- (trifluoromethyl)pyrazin-2-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1- (tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)-1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000410_0001
1-(6-(trifluoromethyl)pyrazin-2-yl)ethan-1-one To a solution of 2-chloro-6-(trifluoromethyl)pyrazine (1 g, 5.479 mmol, 1 equiv) and bis(triphenylphosphine) palladium chloride (0.38 g, 0.548 mmol, 0.1 equiv) in dioxane (10 mL) was added tributyl(1-ethoxyethenyl)stannane (2.97 g, 8.219 mmol, 1.5 equiv) under N2. The reaction was stirred at 85 °C under N2 for 3h. The mixture was allowed to cool down to rt, and to the above mixture was added 1 N HCl (5 mL, 16.456 mmol). The resulting mixture was stirred for additional 1h at rt. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[6-(trifluoromethyl)pyrazin-2-yl]ethanone (640 mg, 60.21%) as a yellow oil. MS (ESI): mass calcd. for C7H5F3N2O: 190.04 m/z, found 191.00[M+H]+.
Figure imgf000411_0001
1-(6-(trifluoromethyl)pyrazin-2-yl)ethan-1-ol To a solution of 1-[6-(trifluoromethyl)pyrazin-2-yl]ethanone (600 mg, 3.156 mmol, 1 equiv) in MeOH (10 mL) was added NaBH4 (358.15 mg, 9.468 mmol, 3 equiv) at 0 °C. The reaction was stirred at rt for 1h. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[6-(trifluoromethyl)pyrazin-2-yl]ethanol (210 mg, 34.63%) as a yellow oil. MS (ESI): mass calcd. for C7H7F3N2O: 192.05 m/z, found 193.15 [M+H]+.
Figure imgf000411_0002
(2R)-7-bromo-2-methyl-4-(1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of 1-[6-(trifluoromethyl)pyrazin-2-yl]ethanol (200 mg, 1.041 mmol, 1 equiv) in DCM (10 mL) was added (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (251.97 mg, 1.041 mmol, 1 equiv) and PPh3 (409.53 mg, 1.561 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (271.92 mg, 1.561 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-2-methyl-4-{1-[6-(trifluoromethyl)pyrazin- 2-yl]ethyl}-2H-1,4-benzoxazin-3-one (162 mg, 37.39%) as a yellow oil. MS (ESI): mass calcd. for C16H13BrF3N3O2: 415.01 m/z, found:415.90, 417.90 [M+H, M+H+2]+.
Figure imgf000412_0001
(2R)-7-amino-2-methyl-4-(1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-4-{1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl}-2H-1,4- benzoxazin-3-one (150 mg, 0.360 mmol, 1 equiv) in EtOH/H2O(4:1, 10 mL) was added (1R,2R)- N1,N2-dimethylcyclohexane-1,2-diamine (30.76 mg, 0.216 mmol, 0.6 equiv), azidosodium (70.29 mg, 1.080 mmol, 3 equiv), CuSO4(57.52 mg, 0.360 mmol, 1 equiv) and sodium ascorbate (143.52 mg, 0.720 mmol, 2 equiv). The reaction was stirred at 80°C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-2-methyl-4-{1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl}-2H-1,4-benzoxazin-3-one (100 mg, 78.75%) as a yellow oil. MS (ESI): mass calcd. for C16H15F3N4O2: 352.11 m/z, found 353.10 [M+H]+.
Figure imgf000412_0002
1-(tert-butyl)-3-((2R)-2-methyl-3-oxo-4-(1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-2-methyl-4-{1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl}-2H-1,4- benzoxazin-3-one (100 mg, 0.284 mmol, 1 equiv) in DCM/Toluene(2:1, 15 mL) was added 2- isocyanato-2-methylpropane (281.37 mg, 2.840 mmol, 10 equiv). The reaction was stirred at 90 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with DCM (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 41% to 63% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-2-methyl- 3-oxo-4-{1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl}-2H-1,4-benzoxazin-7-yl]urea (40 mg, 31.21%) as a white solid. MS (ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found 452.00 [M+H]+.
Figure imgf000413_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)- 1-(6-(trifluoromethyl)pyrazin-2-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-{1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl}- 2H-1,4-benzoxazin-7-yl]urea (40 mg) was separated by chiral-HPLC using a CHIRALPAK IE, 2*25 cm, 5 μm column (eluent: 90% to 90% (v/v) HEX: DCM=3: 1(0.3% IPAmine) and IPA) to yield first example 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-[(1S)-1-[6-(trifluoromethyl)pyrazin-2- yl]ethyl]-2H-1,4-benzoxazin-7-yl]urea (16.3 mg, 40.75%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 452.05 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 9.10 (s, 1H), 9.00 (s, 1H), 8.25 (s, 1H), 7.25 (d, J = 2.3 Hz, 1H), 6.77 – 6.90 (m, 2H), 6.00 – 6.05 (m, 1H), 5.93 – 5.99 (m, 1H), 4.67 – 4.70 (m, 1H), 1.75 – 1.85 (m, 3H), 1.36 – 1.41 (m, 3H), 1.28 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -66.4059. And to yield second example 3-tert-butyl-1-[(2R)-2- methyl-3-oxo-4-[(1R)-1-[6-(trifluoromethyl)pyrazin-2-yl]ethyl]-2H-1,4-benzoxazin-7-yl]urea (15 mg, 37.50%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 452.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 9.08 (s, 1H), 8.98 (s, 1H), 8.25 (s, 1H), 7.24 (d, J = 2.4 Hz, 1H), 6.89 – 7.00 (m, 1H), 6.74 – 6.89 (m, 1H), 5.87 – 6.10 (m, 2H), 4.60 – 4.75 (m, 1H), 1.80 – 1.87 (m, 3H), 1.36 – 1.44 (m, 3H), 1.28 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -66.3907. Example 308: N-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-2,2-dimethylcyclopentane-1- carboxamide Synthetic Scheme
Figure imgf000414_0001
(Z)-6-(hydroxymethylene)-2,2-dimethylcyclohexan-1-one A solution of sodium methoxide (406 mg, 7.528 mmol, 1.9 equiv) in toluene (8 mL) was treated with 2,2-dimethylcyclohexan-1-one (500 mg, 3.962 mmol, 1 equiv).The solution was stirred at rt for 30 min. Ethyl formate (733 mg, 9.905 mmol, 2.5 equiv) was added into the reaction. The solution was stirred at rt for 12 h. The mixture was acidified to pH 7 with 2N HCl. The reaction mixture was quenched by ice water (20 mL) and extracted with DCM (3 x 20 mL).The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to yield a crude product which was directly purified by flash chromatography (0-60% EA/PE) to afford (Z)-6-(hydroxymethylene)-2,2-dimethylcyclohexan-1-one (300 mg, 49.10%) as white solid. MS (ESI): mass calcd. for C9H14O2, 154.10 m/z, found 153.00 [M-H]-.
Figure imgf000414_0002
2,2-dimethylcyclopentane-1-carboxylic acid A solution of (Z)-6-(hydroxymethylene)-2,2-dimethylcyclohexan-1-one (300 mg, 0.065 mmol, 1 equiv) in 2-methyl-2-propanol (6 mL) was treated with hydrogen peroxide (0.6 mL, 0.235 mmol, 3.63 equiv). The solution was stirred at rt for overnight. The mixture was acidified to pH 5 with NaHCO3 (aq.). The reaction mixture was quenched by ice water (20 mL) and extracted with Et2O (3 x 20 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to yield a crude product 2,2-dimethylcyclopentane-1-carboxylic acid (70 mg, 25.36%) as white oil. MS (ESI): mass calcd. for C8H14O2, 142.10 m/z, found 141.00 [M- H]-.
Figure imgf000415_0001
N-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6-yl)-2,2-dimethylcyclopentane-1-carboxamide To a mixture of 2,2-dimethylcyclopentane-1-carboxylic acid (70 mg, 0.492 mmol, 1 equiv) and 6- amino-1-benzylquinoxalin-2-one (123 mg, 0.492 mmol, 1 equiv) in pyridine (5 mL) was stirred ,and added EDCI (189 mg, 0.984 mmol, 2 equiv). The reaction was stirred at rt for 4 h. The resulting mixture was concentrated under vacuum. The crude product was purified by reverse phase flash with the following conditions to afford N-(1-benzyl-2-oxo-1,2-dihydroquinoxalin-6- yl)-2,2-dimethylcyclopentane-1-carboxamide (21.4 mg, 11.55%) as yellow solid. MS (ESI): mass calcd. for C23H25N3O2, 375.19 m/z, found 376.10 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 9.93 (s, 1H), 8.35 (s, 1H), 8.22 (d, J = 2.4 Hz, 1H), 7.71 (dd, J = 9.1, 2.5 Hz, 1H), 7.43 (d, J = 9.0 Hz, 1H), 7.29 – 7.37 (m, 2H), 7.25 – 7.27 (m, 3H), 5.47 (s, 2H), 2.46 (t, J = 7.7 Hz, 1H), 1.96 – 2.03 (m, 1H), 1.80 – 1.89 (m, 1H), 1.70 – 1.75 (m, 1H), 1.60 – 1.64 (m, 1H), 1.51 – 1.56 (m, 1H), 1.36 – 1.45 (m, 1H), 1.10 (s, 3H), 0.91 (s, 3H). Example 309: 1-(tert-butyl)-3-(1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)urea Synthetic Scheme
Figure imgf000415_0002
Figure imgf000416_0001
N-(2-bromo-4-nitrophenyl)-2,3-dihydro-1H-inden-1-amine To a stirred mixture of 2-bromo-1-fluoro-4-nitrobenzene (4.3 g, 19.546 mmol, 1 equiv) and 2,3- dihydro-1H-inden-1-amine (2.60 g, 19.546 mmol, 1 equiv) in DMF (50 mL) was added Cs2CO3 (15.92 g, 48.865 mmol, 2.5 equiv) in portions at 25°C under air atmosphere. The resulting mixture was stirred for 2 h at 80 °C. Desired product could be detected by LCMS. The mixture was diluted with water (200 mL). The resulting mixture was extracted with EA (3 x 200 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford N-(2-bromo-4-nitrophenyl)-2,3- dihydro-1H-inden-1-amine (3.6 g, 55.28%) as a white solid. LC/MS (ESI): mass calcd. for C15H13BrN2O2: 332.02 m/z, found 334.90 [M+H, M+H+2] +.
Figure imgf000416_0002
ethyl (E)-3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)acrylate To a stirred mixture of N-(2-bromo-4-nitrophenyl)-2,3-dihydro-1H-inden-1-amine (1.5 g, 4.502 mmol, 1 equiv) and ethyl (2E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoate (1.53 g, 6.753 mmol, 1.5 equiv) in dioxane (15 mL) and H2O (1.5 mL) was added K2CO3 (1.56 g, 11.255 mmol, 2.5 equiv) and Pd(dppf)Cl2 (329.42 mg, 0.450 mmol, 0.1 equiv) in portions at 25°C under nitrogen atmosphere. The resulting mixture was stirred for 2 h at 90 °C under N2. Desired product could be detected by LCMS. After cooling down to rt, the mixture was diluted with water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford ethyl (E)-3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5- nitrophenyl)acrylate (630 mg, 39.71%) as a white solid. LC/MS (ESI): mass calcd. for C20H20N2O4: 352.14 m/z, found 353.15 [M+H] +.
Figure imgf000417_0001
ethyl 3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoate To a stirred mixture of ethyl (E)-3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5- nitrophenyl)acrylate (610 mg, 1.731 mmol, 1 equiv) and CoCl2 (112.37 mg, 0.866 mmol, 0.5 equiv) in MeOH (0.5 mL) and THF (10 mL) was added NaBH4 (130.97 mg, 3.462 mmol, 2 equiv) in portions at 0 °C under N2 atmosphere. The resulting mixture was stirred for 2 h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (0- 100%) to afford ethyl 3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoate (200 mg, 35.89%) as a white solid. LC/MS (ESI): mass calcd. for C18H18N2O4: 354.16 m/z, found 355.15 [M+H] +.
Figure imgf000417_0002
3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoic acid To a stirred mixture of ethyl 3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoate (210 mg, 0.593 mmol, 1 equiv) in THF (2 mL), EtOH (2 mL) and H2O (2 mL) was added LiOH (42.57 mg, 1.779 mmol, 3 equiv) at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature. Desired product could be detected by LCMS. The mixture was basified to PH = 5 with 2N HCl and diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford 3-(2- ((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoic acid (180 mg, 93.08%) as a white solid. LC/MS (ESI): mass calcd. for C25H34N4O2: 326.13 m/z, found 327.15 [M+H] +.
Figure imgf000418_0001
1-(2,3-dihydro-1H-inden-1-yl)-6-nitro-3,4-dihydroquinolin-2(1H)-one To a stirred mixture of 3-(2-((2,3-dihydro-1H-inden-1-yl)amino)-5-nitrophenyl)propanoic acid (170 mg, 0.521 mmol, 1 equiv) in T3P (1 mL) at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the 1-(2,3-dihydro-1H-inden-1-yl)-6-nitro-3,4- dihydroquinolin-2(1H)-one (150 mg, 93.39%) as a white solid. LC/MS (ESI): mass calcd. for C18H16N2O3: 308.12 m/z, found 309.05[M+H] +.
Figure imgf000418_0002
6-amino-1-(2,3-dihydro-1H-inden-1-yl)-3,4-dihydroquinolin-2(1H)-one To a stirred mixture of 1-(2,3-dihydro-1H-inden-1-yl)-6-nitro-3,4-dihydroquinolin-2(1H)-one (170 mg, 0.551 mmol, 1 equiv) in MeOH (2 mL) and H2O (0.2 mL) was added Fe (307.90 mg, 5.510 mmol, 10 equiv) and NH4Cl (294.91 mg, 5.510 mmol, 10 equiv) in portions at room temperature nder air atmosphere. The resulting mixture was stirred for 2 h at 70 °C. Desired product could be detected by LCMS. After cooling down to rt, the mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-20% ethyl acetate/petroleum ether) to afford the 6-amino-1-(2,3-dihydro-1H-inden-1-yl)-3,4-dihydroquinolin-2(1H)-one (120 mg, 78.19%) as a yellow solid. LC/MS (ESI): mass calcd. for C18H18N2O: 278.14 m/z, found 279.15[M+H] +.
Figure imgf000419_0001
1-(tert-butyl)-3-(1-(2,3-dihydro-1H-inden-1-yl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea To a stirred mixture of 6-amino-1-(2,3-dihydro-1H-inden-1-yl)-3,4-dihydroquinolin-2(1H)-one (110 mg, 0.395 mmol, 1 equiv) in DCM/Toluene (2:1, 2mL) was added 2-isocyanato-2- methylpropane (117.53 mg, 1.185 mmol, 3 equiv) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2h at 90°C. Desired product could be detected by LCMS. After cooling down to rt, the mixture was diluted with water (10 mL). The resulting mixture was extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5μm column (eluent: 35% to 65% (v/v) CH3CN and H2O with 10 mmol /L NH4HCO3) to afford the 1-(tert-butyl)-3-(1-(2,3- dihydro-1H-inden-1-yl)-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)urea (31.6 mg, 21.16%) as a white solid. LC/MS (ESI): mass calcd. for C23H27N3O2: 377.21 m/z, found 378.15[M +H] +.1H NMR (300 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.32 – 7.29 (m, 2H), 7.20 (t, J = 7.3 Hz, 1H), 7.10 (t, J = 7.3 Hz, 1H), 6.94 (d, J = 7.5 Hz, 1H), 6.82 (d, J = 7.5 Hz, 1H), 6.36 (s, 2H), 5.92 (m, 1H), 3.15 – 3.11 (m, 1H), 2.98 – 2.94 (m, 1H), 2.85 – 2.80 (m, 2H), 2.57 – 2.55 (m, 2H), 2.43 – 2.27 (m, 2H), 1.25 (s, 9H). Example 310 and Example 311: 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(2- (trifluoromethyl)pyrimidin-4-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)-1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000420_0001
1-(2-(trifluoromethyl)pyrimidin-4-yl)ethan-1-one To a solution of 4-chloro-2-(trifluoromethyl)pyrimidine (1 g, 5.479 mmol, 1 equiv) and bis(triphenylphosphine) palladium chloride (0.38 g, 0.548 mmol, 0.1 equiv) in dioxane (10 mL) was added tributyl(1-ethoxyethenyl)stannane (2.97 g, 8.219 mmol, 1.5 equiv) under N2. The reaction was stirred at 85 °C under N2 for 3h. The mixture was allowed to cool down to rt., and to the above mixture was added 1 N HCl (5 mL). The resulting mixture was stirred for additional 1h at rt. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[2- (trifluoromethyl)pyrimidin-4-yl]ethanone (500 mg, 47.04%) as a yellow oil. MS (ESI): mass calcd. for C7H5F3N2O: 190.04 m/z, found 191.45 [M+H]+
Figure imgf000420_0002
1-(2-(trifluoromethyl)pyrimidin-4-yl)ethan-1-ol To a solution of 1-[2-(trifluoromethyl)pyrimidin-4-yl]ethanone (640 mg, 3.366 mmol, 1 equiv) in MeOH (10 mL) was added NaBH4 (382.03 mg, 10.098 mmol, 3 equiv) at 0 °C. The reaction was stirred at rt for 1h. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 1-[2-(trifluoromethyl)pyrimidin-4-yl]ethanol (400 mg, 58.75%) as a yellow oil. LCMS showed the desired product was generated and the reaction was not worked up. MS (ESI): mass calcd. for C7H7F3N2O: 192.05 m/z, found 193.00 [M+H]+.
Figure imgf000421_0001
(2R)-7-bromo-2-methyl-4-(1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of 1-[2-(trifluoromethyl)pyrimidin-4-yl]ethanol (270 mg, 1.405 mmol, 1 equiv) in DCM (10 mL) was added (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (340.16 mg, 1.405 mmol, 1 equiv) and PPh3 (552.87 mg, 2.107 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (367.09 mg, 2.107 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, water in ACN, 65% to 75% gradient in 10 min; detector, UV 254 nm. This resulted in (2R)-7-bromo-2- methyl-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}-2H-1,4-benzoxazin-3-one (110 mg, 18.81%) as a yellow oil. MS (ESI): mass calcd. for C16H13BrF3N3O2: 415.01 m/z, found:415.90,417.90 [M+H, M+H+2]+.
Figure imgf000421_0002
(2R)-7-amino-2-methyl-4-(1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}-2H-1,4- benzoxazin-3-one (180 mg, 0.432 mmol, 1 equiv) in EtOH/H2O (4:1, 10 mL) was added (1R,2R)- N1,N2-dimethylcyclohexane-1,2-diamine (36.91 mg, 0.259 mmol, 0.6 equiv), CuSO4 (69.02 mg, 0.432 mmol, 1 equiv), sodium ascorbate (172.23 mg, 0.864 mmol, 2 equiv) and NaN3 (84.35 mg, 1.296 mmol, 3 equiv). The reaction was stirred at 80°C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino- 2-methyl-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}-2H-1,4-benzoxazin-3-one (70 mg, 45.94%) as a yellow oil. MS (ESI): mass calcd. for C16H15F3N4O2: 352.12 m/z, found 353.00 [M+H]+.
Figure imgf000422_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-2-methyl-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}-2H-1,4- benzoxazin-3-one (70 mg, 0.199 mmol, 1 equiv) in DCM/Toluene (2:1, 7.5 mL) was added 2- isocyanato-2-methylpropane (196.96 mg, 1.990 mmol, 10 equiv). The reaction was stirred at 90 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Tiart C18 ExRS Column, 30*150 mm, 5μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-2- methyl-3-oxo-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}-2H-1,4-benzoxazin-7-yl]urea (40 mg, 44.59%) as a white solid. MS (ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found 452.15 [M+H]+.
Figure imgf000423_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)- 1-(2-(trifluoromethyl)pyrimidin-4-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-{1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl}- 2H-1,4-benzoxazin-7-yl]urea (40 mg) was separated by chiral-HPLC using a CHIRALPAK IE, 2*25 cm, 5 μm column (eluent: 10% to 10% (v/v) HEX: DCM=3: 1(0.5% 2M NH3-MeOH) to yield first example 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-[(1S)-1-[2-(trifluoromethyl)pyrimidin- 4-yl]ethyl]-2H-1,4-benzoxazin-7-yl]urea (12.8 mg, 32.00%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 474.15 [M+Na]+. 1H NMR (400 MHz, DMSO-d6) δ 8.81 - 9.10 (m, 1H), 8.26 (s, 1H), 7.59 – 7.90 (m, 1H), 7.16 – 7.34 (m, 1H), 6.58 – 7.03 (m, 2H), 5.78 – 6.01 (m, 2H), 4.65 – 4.80 (m, 1H), 1.78 (d, J = 6.9 Hz, 3H), 1.39 (d, J = 6.7 Hz, 3H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -69.1379. And to yield second example 3-tert-butyl- 1-[(2R)-2-methyl-3-oxo-4-[(1R)-1-[2-(trifluoromethyl)pyrimidin-4-yl]ethyl]-2H-1,4-benzoxazin- 7-yl]urea (13.7 mg, 34.25%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 452.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.90 - 9.10 (m, 1H), 8.26 (s, 1H), 7.63 – 7.94 (m, 1H), 7.15 – 7.36 (m, 1H), 6.65 – 7.00 (m, 2H), 5.98 (m, 1H), 5.80 – 5.95 (m, 1H), 4.68 – 4.72 (m, 1H), 1.70 – 1.86 (m, 3H), 1.33 – 1.45 (m, 3H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -69.1481. Example 312: 1-(tert-butyl)-3-(1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000424_0001
1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethan-1-ol To a stirring solution 2-fluoro-3-(trifluoromethoxy)benzaldehyde (500 mg, 2.403 mmol, 1 equiv) in tetrahydrofuran (10 mL) was added a solution of methylmagnesium bromide (2 M in THF, 1.8 mL, 3.604 mmol, 1.5 equiv) at 0 °C. The mixture was stirred at room temperature for 5 h. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude was purified by silica gel column with PE/EA (4:1) to afford 1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethan-1-ol (500 mg, 92.84%) as a yellow solid.
Figure imgf000424_0002
1-(1-bromoethyl)-2-fluoro-3-(trifluoromethoxy)benzene To a stirring solution 1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethan-1-ol (100 mg, 0.446 mmol, 1 equiv) in DCM (2 mL) was added a solution of CBr4 (163 mg, 0.491 mmol, 1.1 equiv) in 0 °C and PPh3 (129 mg, 0.491 mmol, 1.1 equiv) at 0 °C . The mixture was stirred at room temperature for 5 h. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum. The crude was purified by silica gel column with PE/EA (5:1) to afford 1-(1-bromoethyl)-2-fluoro-3-(trifluoromethoxy)benzene (100 mg, 78.09%) as a yellow solid.
Figure imgf000425_0001
1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)-6-nitroquinoxalin-2(1H)-one To a stirring solution 6-nitroquinoxalin-2(1H)-one (67 mg, 0.348 mmol, 1 equiv) in DMF (5 mL) was added K2CO3 (146 mg, 1.044 mmol, 3 equiv) and 1-(1-bromoethyl)-2-fluoro-3- (trifluoromethoxy)benzene (100 mg, 0.348 mmol, 1 equiv). The mixture was stirred at room temperature for 5 h. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude was purified by silica gel column with PE/EA (3:1) to afford 1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)-6-nitroquinoxalin-2(1H)-one (40 mg, 28.90%) as a yellow solid. MS (ESI): mass calcd. for C17H11F4N3O4: 397.07 found: 398.10 [M+H]+.
Figure imgf000425_0002
6-amino-1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a stirring solution 1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)-6-nitroquinoxalin-2(1H)- one (40 mg, 0.101 mmol, 1 equiv) in EtOH (2 mL) and H2O (0.5 mL) was added Fe (27 mg, 0.479 mmol, 4.74 equiv) and NH4Cl (27 mg, 0.500 mmol, 4.95 equiv). The mixture was stirred for 2 h at 60 °C. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude was purified by silica gel column with PE/EA (1:1) to afford 6-amino-1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (30 mg, 81.12%) as a yellow solid. MS (ESI): mass calcd. for C17H13F4N3O2: 367.07 found: 368.10 [M+H]+.
Figure imgf000425_0003
1-(tert-butyl)-3-(1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea To a stirring solution 6-amino-1-(1-(2-fluoro-3-(trifluoromethoxy)phenyl)ethyl)quinoxalin- 2(1H)-one (30 mg, 0.082 mmol, 1 equiv) in DCM (2 mL) was added 2-isocyanato-2- methylpropane (30 mg, 0.303 mmol, 3.71 equiv) and TEA (40 mg, 0.395 mmol, 4.84 equiv). The mixture was stirred at room temperature for 16 h. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The combined organic extracts were washed with brine (10 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude was purified by silica gel column with PE/EA (1:1) and Prep-HPLC (Column: XBridge Prep OBD C18, 30*150 mm, 5.0 μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B:ACN; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 7min ; Wave Length: 254/220 nm; RT1(min): 10.7; Number Of Runs: 3) to afford 1-(tert-butyl)-3-(1-(1-(2-fluoro-3- (trifluoromethoxy)phenyl)ethyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)urea (9.5 mg, 24.79%) as a yellow solid. MS (ESI): mass calcd. for C22H22F4N4O3: 466.15 found: 467.00[M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.14 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.73 (t, J = 7.4 Hz, 1H), 7.44 – 7.54 (m,3H), 7.37 (t, J = 8.2 Hz, 1H), 6.54 (s, 1H), 6.04 (s, 1H), 1.89 (d, J = 7.0 Hz, 3H), 1.29 (s, 9H). Example 313: (S)-1-(tert-butyl)-3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000426_0001
Figure imgf000427_0001
1-(3-(2,2,2-trifluoroethoxy)phenyl)ethan-1-one To a stirred solution of 3-hydroxyacetophenone (1.5 g, 11.017 mmol, 1 equiv) was added dimethylformamide (15 mL) followed by catalytic amount of 2,2,2-trifluoroethyl trifluoromethanesulfonate (3.84 g, 16.526 mmol, 1.5 equiv) and at room temperature. The reaction mixture was stirred at room temperature for a period of 3h. After completion of reaction, the reaction mixture was quenched by water (30 mL) and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford desired 1-(3-(2,2,2-trifluoroethoxy)phenyl)ethan-1- one (1.5 g, 62.40%) . MS (ESI): mass calcd. for C10H9F3O2: 218.175 m/z, found: 219.15 [M+H]+.
Figure imgf000427_0002
1-(3-(2,2,2-trifluoroethoxy)phenyl)ethan-1-ol To a solution of 1-(3-(2,2,2-trifluoroethoxy)phenyl)ethan-1-one (2.2 g, 10.249 mmol, 1 equiv) in methanol (30 mL) was added NaBH4 (0.77 g, 20.498 mmol, 2 equiv)) at 0 °C. The mixture was stirred for 30 min. The reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 100 mL). Then, the combined organic phase was washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (1:1) to afford 1-(3-(2,2,2- trifluoroethoxy)phenyl)ethan-1-ol (1.23 g, 54.68%) as a yellow oil.
Figure imgf000427_0003
1-(1-bromoethyl)-3-(2,2,2-trifluoroethoxy)benzene To a solution of 1-(3-(2,2,2-trifluoroethoxy)phenyl)ethan-1-ol (1.0 g, 4.542 mmol, 1 equiv) in methylene chloride (20 mL) the mixture was 0 °C then added phosphorus tribromide (6.15 g, 22.710 mmol, 5.0 equiv) slowly, the mixture was warmed to room temperature stirred at room temperature for 3 h. The reaction mixture was quenched by ice-water and extracted with EA (3 x 50 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 1-(1-bromoethyl)-3-(2,2,2- trifluoroethoxy)benzene (725 mg, 56.39%) as a yellow oil.
Figure imgf000428_0001
6-nitro-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 1-(1-bromoethyl)-3-(2,2,2-trifluoroethoxy)benzene (430 mg, 1.519 mmol, 1 equiv) in dimethylformamide (10 mL) was added potassium methaneperoxoate potassium (634 mg, 4.557 mmol, 3 equiv) at room temperature. Then 6-nitro-1H-quinoxalin-2-one (290 mg, 1.519 mmol, 1 equiv) was added at room temperature. The reaction was stirred at room temperature for 6h. The reaction mixture was quenched by water and extracted with EA (3 x 100 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (1:1) to afford 6-nitro-1-(1-(3-(2,2,2- trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (380 mg, 63.60%). MS (ESI): mass calcd. for C18H14F3N3O4: 393.322 m/z, found:394.20 [M+H]+.
Figure imgf000428_0002
6-amino-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a stirred solution of6-nitro-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (50 mg, 0.127 mmol, 1 equiv) in methanol (10 mL) and water (2 mL) was added iron (71 mg, 1.27 mmol, 10 equiv) followed by catalytic amount of NH4Cl (67 mg, 1.27 mmol, 10 equiv) at room temperature. The reaction mixture was stirred at 60 °C for a period of 0.5 h. The reaction mixture was quenched by water (20 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (1:1) to afford desired 6-amino-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (30 mg, 64.93%). MS (ESI): mass calcd. for C18H16F3N3O2: 363.34 m/z, found:364.20 [M+H]+.
Figure imgf000429_0001
(S)-1-(tert-butyl)-3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a stirring solution 6-amino-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (200 mg, 0.550 mmol, 1 equiv) in DCM (5 mL) was added a solution of 2-isocyanato-2- methylpropane (272 mg, 2.750 mmol, 5 equiv) and TEA (557 mg, 5.500 mmol, 10 equiv). The mixture was stirred at room temperature for 16 h. The mixture was concentrated to dryness under reduced pressure to provide the crude product, which was purified by C18 column (eluent: CH3CN and H2O), Prep-HPLC (Column: XBridge Prep OBD C18, 30*150 mm, 5.0 μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B:ACN; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 7min) and Chiral-HPLC (Column: (R, R)-WHELK-01-Kromasil 5*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: isocratic 30; Wave Length: 220/254 nm; RT1(min): 21.321; RT2(min): 28.591; Injection Volume: 0.9 mL; Number Of Runs: 5) to afford (S)-1-(tert-butyl)-3-(2-oxo-1- (1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea (8.2 mg, 3.21%) as a yellow solid. MS (ESI): mass calcd. for C23H25F3N4O3: 462.18 m/z, found: 463.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.26 (s, 1H), 7.93 (d, J = 2.5 Hz, 1H), 7.23 – 7.36 (m, 2H), 7.05 (br, 1H), 6.94 – 7.01 (m, 2H), 6.90 (d, J = 7.8 Hz, 1H), 6.57 (br, 1H), 6.03 (s, 1H), 4.76 (q, J = 8.9 Hz, 2H), 1.87 (d, J = 7.1 Hz, 3H), 1.28 (s, 9H). And another isomer (R)-1-(tert-butyl)- 3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea as a yellow solid (6 mg, 2.35%) as a yellow solid. MS (ESI): mass calcd. for C23H25F3N4O3: 462.18 m/z, found: 463.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.26 (s, 1H), 7.93 (d, J = 2.5 Hz, 1H), 7.23 – 7.36 (m, 2H), 7.05 (br, 1H), 6.94 – 7.01 (m, 2H), 6.90 (d, J = 7.8 Hz, 1H), 6.57 (br, 1H), 6.03 (s, 1H), 4.76 (q, J = 8.9 Hz, 2H), 1.87 (d, J = 7.1 Hz, 3H), 1.28 (s, 9H). Example 314: (R)-1-(tert-butyl)-3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea
Figure imgf000430_0001
(R)-1-(tert-butyl)-3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a stirring solution 6-amino-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (200 mg, 0.550 mmol, 1 equiv) in DCM (5 mL) was added a solution of 2-isocyanato-2- methylpropane (272 mg, 2.750 mmol, 5 equiv) and TEA (557 mg, 5.500 mmol, 10 equiv). The mixture was stirred at room temperature for 16 h. The mixture was concentrated to dryness under reduced pressure to provide the crude product, which was purified by C18 column (eluent: CH3CN and H2O), Prep-HPLC (Column: XBridge Prep OBD C18, 30*150 mm, 5.0 μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B:ACN; Flow rate: 60 mL/min; Gradient: 30% B to 60% B in 7min) and Chiral-HPLC (Column: (R, R)-WHELK-01-Kromasil 5*25 cm, 5 μm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)--HPLC, Mobile Phase B: EtOH--HPLC; Flow rate: 20 mL/min; Gradient: isocratic 30; Wave Length: 220/254 nm; RT1(min): 21.321; RT2(min): 28.591; Injection Volume: 0.9 mL; Number Of Runs: 5) to afford (S)-1-(tert-butyl)-3-(2-oxo-1- (1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea (8.2 mg, 3.21%) as a yellow solid. MS (ESI): mass calcd. for C23H25F3N4O3: 462.18 m/z, found: 463.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.26 (s, 1H), 7.93 (d, J = 2.5 Hz, 1H), 7.23 – 7.36 (m, 2H), 7.05 (br, 1H), 6.94 – 7.01 (m, 2H), 6.90 (d, J = 7.8 Hz, 1H), 6.57 (br, 1H), 6.03 (s, 1H), 4.76 (q, J = 8.9 Hz, 2H), 1.87 (d, J = 7.1 Hz, 3H), 1.28 (s, 9H). And another isomer (R)-1-(tert-butyl)- 3-(2-oxo-1-(1-(3-(2,2,2-trifluoroethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea as a yellow solid (6 mg, 2.35%) as a yellow solid. MS (ESI): mass calcd. for C23H25F3N4O3: 462.18 m/z, found: 463.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.26 (s, 1H), 7.93 (d, J = 2.5 Hz, 1H), 7.23 – 7.36 (m, 2H), 7.05 (br, 1H), 6.94 – 7.01 (m, 2H), 6.90 (d, J = 7.8 Hz, 1H), 6.57 (br, 1H), 6.03 (s, 1H), 4.76 (q, J = 8.9 Hz, 2H), 1.87 (d, J = 7.1 Hz, 3H), 1.28 (s, 9H). Example 315: (S)-N-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin- 6-yl)cyclopentanecarboxamide
Figure imgf000431_0001
(S)-N-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6- yl)cyclopentanecarboxamide To a stirring solution 6-amino-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (40 mg, 0.115 mmol, 1 equiv) in pyridine (3 mL) was added a solution of cyclopentanecarboxylic acid (16.99 mg, 0.150 mmol, 1.3 equiv) and EDCI (33 mg, 0.173 mmol, 1.5 equiv). The mixture was stirred at room temperature for 16 h. The mixture was concentrated and purified by C18 column (eluent: CH3CN and H2O) to afford (S)-N-(2-oxo-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)cyclopentanecarboxamide (32.3 mg, 19.32%) as a yellow solid. MS (ESI): mass calcd. for : C23H22F3N3O3: 445.44 m/z, found: 446.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 10.07 (s, 1H), 8.27 (s, 1H), 8.19 (d, J = 2.5 Hz, 1H), 7.59 (d, J = 9.1 Hz, 1H), 7.47 (t, J = 8.1 Hz, 1H), 7.30 (d, J = 4.9 Hz, 4H), 7.17 (s, 1H), 6.58 (s, 1H), 2.74 – 2.79 (m, 1H), 1.83 – 1.88 (m, 5H), 1.56 –1.68 (m, 4H), 1.45 – 1.53 (m, 2H). Example 316 and Example 317: 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(4- (trifluoromethyl)pyrimidin-2-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)-1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000432_0001
2-(1-ethoxyvinyl)-4-(trifluoromethyl)pyrimidine To a solution of 2-chloro-4-(trifluoromethyl)pyrimidine (5 g, 27.393 mmol, 1 equiv) in dioxane (70 mL) was added tributyl(1-ethoxyethenyl)stannane (14.84 g, 41.090 mmol, 1.5 equiv) and bis(triphenylphosphane) dichloropalladium (1.92 g, 2.739 mmol, 0.1 equiv). The resulting mixture was maintained under nitrogen and stirred at 90 °C for 3 h. After cooling down to rt, the reaction was quenched with water (150 mL). The resulting mixture was extracted with ethyl acetate (3 x 150 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the 2-(1-ethoxyvinyl)-4-(trifluoromethyl)pyrimidine (5 g, 83.66%) as a yellow solid.LC/MS: mass calcd. for C8H8F3N2O: 218.07 m/z, found: 219.00 [M+H]+.
Figure imgf000432_0002
1-(4-(trifluoromethyl)pyrimidin-2-yl)ethan-1-one To a solution of 2-(1-ethoxyvinyl)-4-(trifluoromethyl)pyrimidine (1.5 g, 6.875 mmol, 1 equiv) in dioxane (70 mL) was added 1N HCl (15 mL). The resulting mixture was maintained under air and stirred at rt for 3 h. The reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the 1-(4- (trifluoromethyl)pyrimidin-2-yl)ethan-1-one (800 mg, 61.20%) as a yellow oil.LC/MS: mass calcd. for C7H5F3N2O: 190.04 m/z, found: 191.10 [M+H]+.
Figure imgf000433_0001
1-(4-(trifluoromethyl)pyrimidin-2-yl)ethan-1-ol To a solution of 1-(4-(trifluoromethyl)pyrimidin-2-yl)ethan-1-one (2.2 g, 11.571 mmol, 1 equiv) in MeOH (24 mL) was added NaBH4 (656.62 mg, 17.357 mmol, 1.5 equiv) at 0°C. The resulting mixture was maintained under air and stirred at rt for 3 h. The reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the 1-(4-(trifluoromethyl)pyrimidin-2-yl)ethan-1-ol (1 g, 44.98%) as a yellow oil. LC/MS: mass calcd. for C7H7F3N2O: 192.05 m/z, found: 193.00 [M+H]+.
Figure imgf000433_0002
(2R)-7-bromo-2-methyl-4-(1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of 1-(4-(trifluoromethyl)pyrimidin-2-yl)ethan-1-ol (300 mg, 1.561 mmol, 1 equiv) in THF (5 mL) was added (R)-7-bromo-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one (377.96 mg, 1.561 mmol, 1 equiv) and PPh3 (614.30 mg, 2.341 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DIAD (473.58 mg, 2.341 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred for additional 1h at rt. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the (2R)-7-bromo-2-methyl-4-(1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one (550 mg, 84.64%) as a yellow oil. LC/MS: mass calcd. for C16H13BrF3N3O2: 415.01 m/z, found: 416.00, 418.00 [M+H, M+H+2]+.
Figure imgf000434_0001
(2R)-7-amino-2-methyl-4-(1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of 6-nitro-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]quinoxalin-2-one (540 mg, 1.122 mmol, 1 equiv) in EtOH (4 mL) and H2O (1 mL) was added (1R,2R)-N1,N2- dimethylcyclohexane-1,2-diamine (95.77 mg, 0.673 mmol, 0.6 equiv), CuSO4 (179.10 mg, 1.122 mmol, 1 equiv), sodium ascorbate (446.88 mg, 2.244 mmol, 2 equiv) and sodium azide (218.86 mg, 3.366 mmol, 3 equiv). The reaction was stirred at 80 °C for 1h. After cooling down to rt, the filtrate was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino- 2-methyl-4-(1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (360 mg, 91.06%) as a yellow solid. MS (ESI): mass calcd. for C16H15F3N4O2: 352.11 m/z, found 352.95 [M+H]+.
Figure imgf000434_0002
1-(tert-butyl)-3-((2R)-2-methyl-3-oxo-4-(1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-2-methyl-4-{1-[4-(trifluoromethyl)pyrimidin-2-yl]ethyl}-2H-1,4- benzoxazin-3-one (280 mg, 0.795 mmol, 1 equiv) in DCM (10 mL) was added TEA (482.53 mg, 4.770 mmol, 6 equiv) and 2-isocyanato-2-methylpropane (787.85 mg, 7.950 mmol, 10 equiv). The reaction was stirred at rt under N2 for 3h.Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP 18 OBD Column, 30*150 mm, 5μm column (eluent: 39% to 69% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-tert-butyl-1-[(2R)-2- methyl-3-oxo-4-{1-[4-(trifluoromethyl)pyrimidin-2-yl]ethyl}-2H-1,4-benzoxazin-7-yl]urea (80 mg, 22.30%) as a white solid. LC/MS (ESI): mass calcd. for C21H24F3N5O3:451.18 m/z, found:452.15 [M+H]+.
Figure imgf000435_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea and 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)- 1-(4-(trifluoromethyl)pyrimidin-2-yl)ethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea A sample of 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-{1-[4-(trifluoromethyl)pyrimidin-2-yl]ethyl}- 2H-1,4-benzoxazin-7-yl]urea (80 mg) was separated by chiral-HPLC using a CHIRALPAK IF 2*25 cm, 5 μm column (eluent: 25% to 25% (v/v) EtOH and Hex(0.5% 2M NH3-MeOH) to yield first enantiomer example 3-tert-butyl-1-[(2R)-2-methyl-3-oxo-4-[(1S)-1-[4- (trifluoromethyl)pyrimidin-2-yl]ethyl]-2H-1,4-benzoxazin-7-yl]urea (29.9 mg, 37.38%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 452.05 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 9.16 (d, J = 5.0 Hz, 1H), 8.25 (s, 1H), 7.91 (d, J = 5.1 Hz, 1H), 7.23 (d, J = 2.3 Hz, 1H), 6.88 – 7.00 (m, 1H), 6.70 – 6.88 (m, 1H), 5.96 (s, 1H), 5.81 – 5.13 (m, 1H), 4.61 (q, J = 6.7 Hz, 1H), 1.82 (d, J = 6.9 Hz, 3H), 1.34 (d, J = 6.7 Hz, 3H), 1.28 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -68.7349. and to yield second enantiomer example 3-tert-butyl-1-[(2R)-2- methyl-3-oxo-4-[(1R)-1-[4-(trifluoromethyl)pyrimidin-2-yl]ethyl]-2H-1,4-benzoxazin-7-yl]urea (25.9 mg, 32.38%) as a white solid. MS(ESI): mass calcd. for C21H24F3N5O3: 451.18 m/z, found: 452.05 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 9.16 (d, J = 5.0 Hz, 1H), 8.22 (s, 1H), 7.92 (d, J = 5.1 Hz, 1H), 7.23 (d, J = 2.2 Hz, 1H), 6.45 – 7.00 (m, 2H), 5.98 – 6.06 (m, 1H), 5.95 (s, 1H), 4.33 – 4.72 (m, 1H), 1.81 (d, J = 7.0 Hz, 3H), 1.36 (d, J = 6.8 Hz, 3H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -68.7105. Example 318: (S)-N-(2-oxo-1-(1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-1,2- dihydroquinoxalin-6-yl)-3-(3-(trifluoromethyl)phenyl)propanamide Synthetic Scheme
Figure imgf000436_0001
2-(1-ethoxyvinyl)-6-(trifluoromethoxy)pyridine To a solution of 2-chloro-6-(trifluoromethoxy)pyridine (1 g, 5.062 mmol, 1 equiv) and Pd(PPh3)2Cl2 (0.36 g, 0.506 mmol, 0.1 equiv) in dioxane (10 mL) was added tributyl(1- ethoxyethenyl)stannane (2.74 g, 7.593 mmol, 1.5 equiv) under N2. The reaction was stirred at 85 °C under N2 for 1h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 2-(1-ethoxyethenyl)-6- (trifluoromethoxy)pyridine (1 g, 84.71%) as a white oil. MS (ESI): mass calcd. for C10H10F3NO2: 233.07 m/z, found 233.95 [M+H]+.
Figure imgf000436_0002
1-(6-(trifluoromethoxy)pyridin-2-yl)ethan-1-one To a solution of 2-(1-ethoxyethenyl)-6-(trifluoromethoxy)pyridine (1 g, 4.288 mmol, 1 equiv) in dioxane (10 mL) was added 2N HCl (5 mL) .The reaction was stirred at rt for 1 day. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[6-(trifluoromethoxy)pyridin-2- yl]ethanone (0.75 g, 75.26%) as a white oil. MS (ESI): mass calcd. for C8H6F3NO2: 205.04 m/z, found 205.95 [M+H]+.
Figure imgf000437_0001
(R)-1-(6-(trifluoromethoxy)pyridin-2-yl)ethan-1-ol To a solution of (3aS)-1-methyl-3,3-diphenyl-hexahydropyrrolo[1,2-c][1,3,2]oxazaborole (162.14 mg, 0.585 mmol, 0.2 equiv) in THF (10 mL) was added BH3-Me2S (333.26 mg, 4.387 mmol, 1.5 equiv) and 1-[6-(trifluoromethoxy)pyridin-2-yl]ethanone (600 mg, 2.925 mmol, 1 equiv) at 0 °C. The reaction was stirred at 0 °C for 1h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (1R)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethanol (260 mg, 42.91%) as a white oil. MS (ESI): mass calcd. for C8H8F3NO2: 207.05 m/z, found:208.10 [M+H]+.
Figure imgf000437_0002
(S)-6-nitro-1-(1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)quinoxalin-2(1H)-one To a solution of (1R)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethanol (240 mg, 1.159 mmol, 1 equiv) in DCM (10 mL) was added 6-nitro-1H-quinoxalin-2-one (221.46 mg, 1.159 mmol, 1 equiv) and PPh3 (455.83 mg, 1.739 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5 h. To the above mixture was added DEAD (302.66 mg, 1.739 mmol, 1.5 equiv) at 0 °C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-nitro-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]quinoxalin-2-one (100 mg, 22.70%) as a yellow oil. MS (ESI): mass calcd. for C16H11F3N4O4: 380.07 m/z, found 380.95 [M+H]+.
Figure imgf000438_0001
(S)-6-amino-1-(1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)quinoxalin-2(1H)-one To a solution of 6-nitro-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]quinoxalin-2-one (1 g, 2.630 mmol, 1 equiv) in MeOH/H2O (10:1, 11 mL) was added Fe (1.47 g, 26.300 mmol, 10 equiv) and NH4Cl (1.41 g, 26.300 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. the resulting mixture was filtered, the filter cake was washed with MeOH (3x20 mL). The filtrate was concentrated under reduced pressure. Diluted with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2- yl]ethyl]quinoxalin-2-one (460 mg, 49.94%) as a yellow solid. MS (ESI): mass calcd. for C16H13F3N4O2: 350.10 m/z, found 351.00 [M+H]+.
Figure imgf000438_0002
(S)-N-(2-oxo-1-(1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-1,2-dihydroquinoxalin-6-yl)-3- (3-(trifluoromethyl)phenyl)propanamide To a solution of 6-amino-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]quinoxalin-2-one (90 mg, 0.257 mmol, 1 equiv) in DMF (10 mL) was added 3-[3-(trifluoromethyl)phenyl]propanoic acid (61.66 mg, 0.283 mmol, 1.1 equiv), HATU (127.00 mg, 0.334 mmol, 1.3 equiv) and DIEA (99.62 mg, 0.771 mmol, 3 equiv) at 0 °C. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 43% to 73% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound N-{2-oxo-1-[(1S)-1-[6-(trifluoromethoxy)pyridin-2- yl]ethyl]quinoxalin-6-yl}-3-[3-(trifluoromethyl)phenyl]propanamide (62.6 mg, 44.26%) as a yellow solid.LC/MS (ESI): mass calcd. for C26H20F6N4O3:550.14 m/z, found:551.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.14 (s, 1H), 8.24 (s, 1H), 8.14 (d, J = 2.5 Hz, 1H), 7.85 – 8.09 (m, 1H), 7.46 – 7.80 (m, 6H), 7.39 – 7.46 (m, 1H), 7.09 – 7.20 (m, 1H), 6.22 – 6.74 (m, 1H), 2.94 – 3.11 (m, 2H), 2.60 – 2.78 (m, 2H), 1.89 (d, J = 7.0 Hz, 3H).19F NMR (376 MHz, DMSO) δ(ppm): -55.3657, -60.9601, -61.0139. Example 319: (S)-1-(1-(1-(3-chlorophenyl)ethyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(1- methylcyclobutyl)urea Synthetic Scheme
Figure imgf000439_0001
(S)-1-(1-(3-chlorophenyl)ethyl)-6-nitroquinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (852 mg, 4.457 mmol, 1 equiv) in DCM (10 mL) was added (1R)-1-(3-chlorophenyl)ethanol (698.06 mg, 4.457 mmol, 1 equiv) and PPh3 (1753.68 mg, 6.685 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (1164.41 mg, 6.685 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitroquinoxalin-2-one (200 mg, 13.47%) as a yellow oil. MS (ESI): mass calcd. for C16H12ClN3O3: 329.06 m/z, found 329.95 [M+H]+.
Figure imgf000440_0001
(S)-6-amino-1-(1-(3-chlorophenyl)ethyl)quinoxalin-2(1H)-one To a solution of 1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitroquinoxalin-2-one (190 mg, 0.576 mmol, 1 equiv) in MeOH/H2O (10:1, 11 mL) was added Fe (321.79 mg, 5.760 mmol, 10 equiv) and NH4Cl (308.22 mg, 5.760 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. After cooling down to rt, the resulting mixture was filtered, the filter cake was washed with MeOH (3 x 20 mL). The filtrate was concentrated under reduced pressure. Diluted with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1-(3-chlorophenyl)ethyl]quinoxalin-2-one (100 mg, 57.90%) as a yellow solid. MS (ESI): mass calcd. for C16H14ClN3O: 299.08 m/z, found 300.00 [M+H]+.
Figure imgf000440_0002
(S)-1-(1-(1-(3-chlorophenyl)ethyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)-3-(1- methylcyclobutyl)urea To a solution of 6-amino-1-[(1S)-1-(3-chlorophenyl)ethyl]quinoxalin-2-one (90 mg, 0.300 mmol, 1 equiv) in DCM (10 mL) was added triphosgene (35.64 mg, 0.120 mmol, 0.4 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. Then, to the above mixture was added TEA (91.15 mg, 0.900 mmol, 3 equiv) under N2 at 0 °C. The resulting mixture was stirred for additional 1h at rt. Then, 1-methylcyclobutan-1-amine (51.13 mg, 0.600 mmol, 2 equiv) was added. The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP 18 OBD Column, 30*150 mm, 5μm column (eluent: 36% to 66% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 1-{1-[(1S)-1-(3-chlorophenyl)ethyl]-2- oxoquinoxalin-6-yl}-3-(1-methylcyclobutyl)urea (27.7 mg, 22.45%) as a yellow solid.LC/MS (ESI): mass calcd. for C22H23ClN4O2:410.15 m/z, found:411.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.25 (s, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.27 – 7.41 (m, 4H), 7.21 (d, J = 6.9 Hz, 1H), 6.93 – 7.16 (m, 1H), 6.55 (s, 1H), 6.33 (s, 1H), 2.18 – 2.33 (m, 2H), 1.86 – 1.94 (m, 5H), 1.70 – 1.82 (m, 2H), 1.39 (s, 3H). Example 320: (S)-1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(3-(trifluoromethyl)phenyl)ethyl)- 1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000441_0001
(S)-6-nitro-1-(1-(3-(trifluoromethyl)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 6-nitro-1H-quinoxalin-2-one (704 mg, 3.683 mmol, 1 equiv) in DCM (10 mL) was added (1R)-1-[3-(trifluoromethyl)phenyl]ethanol (700.39 mg, 3.683 mmol, 1 equiv) and PPh3 (1449.05 mg, 5.524 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (962.14 mg, 5.524 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]quinoxalin-2-one (200 mg, 14.35%) as a yellow oil. MS (ESI): mass calcd. for C17H12F3N3O3: 363.08 m/z, found 364.00 [M+H]+.
Figure imgf000442_0001
(S)-6-amino-1-(1-(3-(trifluoromethyl)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 6-nitro-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]quinoxalin-2-one (190 mg, 0.523 mmol, 1 equiv) in MeOH/H2O (10:1, 11 mL) was added Fe (292.06 mg, 5.230 mmol, 10 equiv) and NH4Cl (279.75 mg, 5.230 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. After cooling down to rt, the resulting mixture was filtered, the filter cake was washed with MeOH (3x20 mL). The filtrate was concentrated under reduced pressure. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1-[3- (trifluoromethyl)phenyl]ethyl]quinoxalin-2-one (96 mg, 55.07%) as a yellow solid. MS (ESI): mass calcd. for C17H14F3N3O: 333.11 m/z, found 334.00 [M+H]+.
Figure imgf000442_0002
(S)-1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(3-(trifluoromethyl)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a solution of 6-amino-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]quinoxalin-2-one (80 mg, 0.240 mmol, 1 equiv) in DCM (10 mL) was added triphosgene (28.49 mg, 0.096 mmol, 0.4 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. Then, to the above mixture was added TEA (72.86 mg, 0.720 mmol, 3 equiv) under N2 at 0 °C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added 1-methylcyclobutan-1-amine (40.87 mg, 0.480 mmol, 2 equiv). The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP 18 OBD Column, 30*150 mm, 5μm column (eluent: 36% to 66% (v/v) CH3CN and Water(10mmol/L NH4HCO3+0.1%NH3.H2O) to afford the title compound 3-(1- methylcyclobutyl)-1-{2-oxo-1-[(1S)-1-[3-(trifluoromethyl)phenyl]ethyl]quinoxalin-6-yl}urea (22.2 mg, 20.81%) as a yellow solid. MS (ESI): mass calcd. for C23H23F3N4O2: 444.18 m/z, found:445.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.24 (s, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.50 – 7.70 (m, 4H), 7.35 (s, 1H), 6.90 – 7.27 (m, 1H), 6.40 – 6.78 (m, 1H), 6.33 (s, 1H), 2.21 – 2.33 (m, 2H), 1.85 – 1.97 (m, 5H), 1.71 – 1.80 (m, 2H), 1.88 – 1.72 (m, 2H), 1.39 (s, 3H).19F NMR (376 MHz, DMSO) δ(ppm): -60.8769. Example 321: (R)-1-(2-methyl-3-oxo-4-((6-(trifluoromethoxy)pyridin-2-yl)methyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea Synthetic Scheme
Figure imgf000443_0001
(6-(trifluoromethoxy)pyridin-2-yl)methanol To a solution of 2-chloro-6-(trifluoromethoxy)pyridine (800 mg, 4.050 mmol, 1 equiv) in dioxane (8.0 mL) was added (tripropylstannyl)methanol (1694.91 mg, 6.075 mmol, 1.5 equiv) and Pd(PPh3)4 (467.99 mg, 0.405 mmol, 0.1 equiv). The reaction was stirred at 90 °C under N2 for 12 h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give [6-(trifluoromethoxy)pyridin-2-yl]methanol (500 mg, 63.93%) as a white solid. MS (ESI): mass calcd. for C7H6F3NO2: 193.04 m/z, found 194.05 [M+H]+ .
Figure imgf000444_0001
(R)-7-bromo-2-methyl-4-((6-(trifluoromethoxy)pyridin-2-yl)methyl)-2H- benzo[b][1,4]oxazin-3(4H)-one A solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (451.24 mg, 1.864 mmol, 1.2 equiv) in DCM (5.0 mL) was treated with [6-(trifluoromethoxy)pyridin-2-yl]methanol (300 mg, 1.553 mmol, 1 equiv) and PPh3 (611.17 mg, 2.329 mmol, 1.5 equiv) for 1h at rt under nitrogen atmosphere followed by the addition of DEAD (270.53 mg, 1.553 mmol, 1 equiv) dropwise at 0°C. The resulting mixture was stirred for 2h at rt under N2 atmosphere. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-6-fluoro-2- methyl-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (195 mg, 46.41%) as a white solid. MS (ESI): mass calcd. for C16H12BrF3N2O3: 416.00 m/z, found 416.95, 418.95 [M+H, M+H+2]+ .
Figure imgf000444_0002
(R)-7-amino-2-methyl-4-((6-(trifluoromethoxy)pyridin-2-yl)methyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-4-{[6-(trifluoromethoxy)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (180 mg, 0.373 mmol, 1 equiv) in EtOH (3 mL)/H2O (0.3 mL) was added N1,N2-dimethylcyclohexane-1,2-diamine (31.86 mg, 0.224 mmol, 0.6 equiv), CuSO4(59.58 mg, 0.373 mmol, 1 equiv), sodium ascorbate (148.66 mg, 0.746 mmol, 2 equiv) and NaN3 (72.80 mg, 1.119 mmol, 3 equiv). The reaction was stirred at 90 °C under N2 for 2 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-2-methyl-4-{[6- (trifluoromethoxy)pyridin-2-yl]methyl}-2H-1,4-benzoxazin-3-one (91 mg, 69.00%) as a white solid. LC/MS (ESI): mass calcd. for C16H14F3N3O3: 353.10 m/z, found:354.00 [M+H]+.
Figure imgf000445_0001
(R)-1-(2-methyl-3-oxo-4-((6-(trifluoromethoxy)pyridin-2-yl)methyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea To a solution of (2R)-7-amino-2-methyl-4-{[6-(trifluoromethoxy)pyridin-2-yl]methyl}-2H-1,4- benzoxazin-3-one (85 mg, 0.241 mmol, 1 equiv) in DCM (3 mL) was added Triphosgene (71.39 mg, 0.241 mmol, 1 equiv). The reaction was stirred at 0 °C under N2 for 1 h. The reaction was added TEA (194.77 mg, 1.928 mmol, 8 equiv) and stirred at 0 °C under N2 for 1 h. The reaction was added 1-methyl-cyclopentanamine (95.44 mg, 0.964 mmol, 4 equiv) and stirred at rt under N2 for 1 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1-[(2R)-2-methyl-3-oxo-4-{[6-(trifluoromethoxy)pyridin-2- yl]methyl}-2H-1,4-benzoxazin-7-yl]-3-(1-methylcyclopentyl)urea (35.8 mg, 31.05%) as a white solid. LC/MS (ESI): mass calcd. for C23H25F3N4O4:478.18 m/z, found:479.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.21 (s, 1H), 7.96-7.99 (m, 1H), 7.29-7.32 (m, 1H), 7.22-7.23 (m, 1H), 7.15-7.17 (m, 1H), 6.82-6.85 (m, 1H), 6.74-7.78 (m, 1H), 6.02 (s, 1H), 5.14 (s, 2H), 4.73-7.79 (m, 1H), 1.87-1.89 (m, 2H), 1.56-1.62 (m, 4H), 1.44-1.54 (m, 5H), 1.34 (s, 3H).19F NMR (282 MHz, DMSO) δ-55.06. Example 322: 1-((R)-2-methyl-3-oxo-4-((S)-1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea Synthetic Scheme
Figure imgf000446_0001
1-((R)-2-methyl-3-oxo-4-((S)-1-(6-(trifluoromethyl)pyridin-2-yl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea To a solution of (2R)-7-amino-2-methyl-4-[(1S)-1-[6-(trifluoromethyl)pyridin-2-yl]ethyl]-2H- 1,4-benzoxazin-3-one (70 mg, 0.199 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (23.65 mg, 0.080 mmol, 0.4 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. Then, to the above mixture was added TEA (60.49 mg, 0.597 mmol, 3 equiv) under N2 at 0 °C. The resulting mixture was stirred for additional 1h at rt. Then, 1-methyl-cyclopentanamine (39.52 mg, 0.398 mmol, 2 equiv) was added. The reaction was stirred at rt for 1h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP 18 OBD Column, 30*150 mm, 5μm column (eluent: 48% to 78% (v/v) CH3CN and H2O (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 1-[(2R)-2- methyl-3-oxo-4-[(1S)-1-[6-(trifluoromethyl)pyridin-2-yl]ethyl]-2H-1,4-benzoxazin-7-yl]-3-(1- methylcyclopentyl)urea (16.7 mg, 17.59%) as a yellow solid. LC/MS (ESI): mass calcd. for C24H27F3N4O3:476.20 m/z, found:477.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.97 – 8.11 (m, 1H), 7.70 – 7.90 (m, 1H), 7.52 – 7.67 (m, 1H), 7.23 (d, J = 1.9 Hz, 1H), 6.63 – 6.80 (m, 2H), 5.95 – 6.15 (m, 2H), 4.63 – 4.80 (m, 1H), 1.84 – 1.99 (m, 2H), 1.74 – 1.84 (m, 3H), 1.57 – 1.69 (m, 4H), 1.47 – 1.54 (m, 2H), 1.37 – 1.42 (m, 3H), 1.33 (s, 3H).19F NMR (282 MHz, DMSO) δ(ppm): -66.560. Example 323: 1-((R)-2-methyl-3-oxo-4-((S)-1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea Synthetic Scheme
Figure imgf000447_0001
(R)-7-bromo-2-methyl-4-((S)-1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (1R)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethanol (300 mg, 1.448 mmol, 1 equiv) in DCM (10 mL) was added (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (350.57 mg, 1.448 mmol, 1 equiv) and PPh3 (569.78 mg, 2.172 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (378.32 mg, 2.172 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give(2R)-7-bromo-2-methyl-4-[(1S)-1-[6- (trifluoromethoxy)pyridin-2-yl]ethyl]-2H-1,4-benzoxazin-3-one (220 mg, 35.23%) as a yellow solid. LC/MS: mass calcd. for C17H14BrF3N2O3: 430.01, found: 430.95, 432.95 [M+H, M+H+2]+.
Figure imgf000447_0002
(R)-7-amino-2-methyl-4-((S)-1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-4-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]-2H- 1,4-benzoxazin-3-one (210 mg, 0.487 mmol, 1 equiv) in EtOH/H2O (4:1, 10 mL) was added (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (41.56 mg, 0.292 mmol, 0.6 equiv), CuSO4 (77.73 mg, 0.487 mmol, 1 equiv), azidosodium (94.98 mg, 1.461 mmol, 3 equiv) and sodium ascorbate (193.94 mg, 0.974 mmol, 2 equiv). The reaction was stirred at 80°C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-7-amino-2-methyl-4-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]-2H- 1,4-benzoxazin-3-one (130 mg, 72.67%) as a yellow solid. MS (ESI): mass calcd. for C17H16F3N3O3: 367.11 m/z, found 368.05 [M+H]+.
Figure imgf000448_0001
1-((R)-2-methyl-3-oxo-4-((S)-1-(6-(trifluoromethoxy)pyridin-2-yl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(1-methylcyclopentyl)urea To a solution of (2R)-7-amino-2-methyl-4-[(1S)-1-[6-(trifluoromethoxy)pyridin-2-yl]ethyl]-2H- 1,4-benzoxazin-3-one (60 mg, 0.163 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (19.39 mg, 0.065 mmol, 0.4 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (49.59 mg, 0.489 mmol, 3 equiv) under N2 at 0 °C. The resulting mixture was stirred for additional 1h at rt. Then, 1-methyl-cyclopentanamine (32.40 mg, 0.326 mmol, 2 equiv) was added. The reaction was stirred at rt for 1h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column30*150 mmm column (eluent: 30% to 55% (v/v) CH3CN and Water (10 mmol/L NH4HCO3)) to afford the title compound 1-[(2R)-2-methyl-3-oxo-4-[(1S)-1- [6-(trifluoromethoxy)pyridin-2-yl]ethyl]-2H-1,4-benzoxazin-7-yl]-3-(1-methylcyclopentyl)urea (23.1 mg, 28.72%) as a white solid. LC/MS (ESI): mass calcd. for C24H27F3N4O4: 492.20 m/z, found:493.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.19 (s, 1H), 8.00 (t, J = 7.9 Hz, 1H), 7.35 (d, J = 7.7 Hz, 1H), 7.10 – 7.20 (m, 2H), 6.60 – 6.78 (m, 2H), 5.90 – 6.11 (m, 2H), 4.55 – 4.80 (m, 1H), 1.81 – 2.00 (m, 2H), 1.75 (d, J = 7.1 Hz, 3H), 1.45 – 1.70 (m, 6H), 1.38 – 1.45 (s, 3H), 1.25 – 1.38 (m, 3H).19F NMR (282 MHz, DMSO) δ (ppm): -55.093 Example 324: 1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-((1r,3R)-3-(trifluoromethyl)cyclobutyl)urea Synthetic Scheme
Figure imgf000449_0001
(R)-1-(3-(trifluoromethoxy)phenyl)ethan-1-ol To a solution of BH3-Me2S (279 mg, 3.676 mmol, 1.5 equiv) and (S)-(-)-2-Methyl-CBS- oxazaborolidine (135 mg, 0.490 mmol, 0.2 equiv) in THF (6 mL) was added 1-(3- (trifluoromethoxy)phenyl)ethan-1-one (500 mg, 2.450 mmol, 1 equiv) at 0 °C. The resulting mixture was stirred for 2 h at 0 °C. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford (R)-1-(3-(trifluoromethoxy)phenyl)ethan-1- ol (0.4 g, 80.00%) as a yellow oil.
Figure imgf000450_0001
(R)-7-bromo-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (R)-1-(3-(trifluoromethoxy)phenyl)ethan-1-ol (0.4 g, 1.942 mmol, 1 equiv) in DCM (6 mL) was added (R)-7-bromo-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one (469 mg, 1.942 mmol, 1 equiv), triphenylphosphine (510mg, 1.942 mmol, 1 equiv) and DEAD (338 mg, 1.942 mmol, 1 equiv) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (3:1) to afford (R)-7-bromo-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)- 2H-benzo[b][1,4]oxazin-3(4H)-one (0.3 g, 36.06%) as a yellow solid.
Figure imgf000450_0002
(R)-7-amino-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (R)-7-bromo-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one (0.3 g, 0.699 mmol, 1 equiv) in EtOH (6 mL) & H2O (2 mL) was added sodium ascorbate (28 mg, 0.140 mmol, 0.2 equiv), N1,N2-dimethylcyclohexane-1,2-diamine (20 mg , 0.140 mmol, 0.2 equiv), CuSO4 (23 mg , 0.14 mmol, 0.2 equiv) and NaN3 (136 mg, 2.097 mmol, 3 equiv) at rt. The resulting mixture was stirred 4 h at 80 °C. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (1:1) to afford (R)-7-amino- 2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (166 mg, 65.02%) as a yellow solid.
Figure imgf000451_0001
1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-((1r,3R)-3-(trifluoromethyl)cyclobutyl)urea To a mixture of (R)-7-amino-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (60 mg, 0.163 mmol, 1 equiv) in DCM (4 mL) was stirred under nitrogen atmosphere and added triphosgene (15 mg, 0.149 mmol, 0.3 equiv) at 0 °C. The reaction was stirred at rt for 1 h. TEA (41 mg, 0.407 mmol, 2.5 equiv) was added into the mixture at 0 °C. The reaction was stirred at rt for 1 h. (1r,3r)-3- (trifluoromethyl)cyclobutan-1-amine (23 mg, 0.163 mmol, 1.0 equiv) was added into the mixture at 0 °C. The reaction was stirred at rt for 1 h. The reaction mixture was quenched by 1N hydrochloric acid (5 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-((1r,3R)-3-(trifluoromethyl)cyclobutyl)urea (25.5 mg, 29.44%) as a white solid. MS (ESI): mass calcd. for C24H23F6N3O4: 531.15 m/z, found: 532.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.45 (s, 1H), 7.40 – 7.60 (m, 1H), 7.19 – 7.40 (m, 4H), 6.72 – 6.85 (m, 1H), 6.62 – 6.72 (m, 1H), 6.52 – 6.62 (m, 1H), 5.96 – 6.19 (m, 1H), 4.66 – 4.85 (m, 1H), 4.13 – 4.31 (m,1H), 2.91 – 3.18 (m,1H), 2.31 – 2.43 (m,2H), 2.15 – 2.31 (m, 2H), 1.68 – 1.87 (m, 3H), 1.38 – 1.48 (m, 3H). Example 325: 1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-((1s,3S)-3-(trifluoromethyl)cyclobutyl)urea
Figure imgf000451_0002
1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-((1s,3S)-3-(trifluoromethyl)cyclobutyl)urea To a mixture of (R)-7-amino-2-methyl-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (60 mg, 0.163 mmol, 1 equiv) in DCM (4 mL) was stirred under nitrogen atmosphere and added triphosgene (15 mg, 0.149 mmol, 0.3 equiv) at 0 °C. The reaction was stirred at rt for 1 h. TEA (41 mg, 0.407 mmol, 2.5 equiv) was added into the mixture at 0 °C. The reaction was stirred at rt for 1 h. (1s,3s)-3- (trifluoromethyl)cyclobutan-1-amine (23 mg, 0.163 mmol, 1.0 equiv) was added into the mixture at 0 °C. The reaction was stirred at rt for 1 h. The reaction mixture was quenched by 1N hydrochloric acid (5 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford 1-((R)-2-methyl-3-oxo-4-((S)-1-(3-(trifluoromethoxy)phenyl)ethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-((1s,3S)-3-(trifluoromethyl)cyclobutyl)urea (30.5 mg, 35.20%) as a white solid. MS (ESI): mass calcd. for C24H23F6N3O4: 531.15 m/z, found: 532.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.32 (s, 1H), 7.40 – 7.60 (m, 1H), 7.19 – 7.22 (m, 4H), 6.72 – 6.85 (m, 1H), 6.62 – 6.72 (m, 1H), 6.52 – 6.62 (m, 1H), 5.96 – 6.19 (m, 1H), 4.66 – 4.85 (m, 1H), 4.02 – 4.22 (m,1H), 2.79 – 2.98 (m,1H), 2.35 – 2.49 (m,2H), 1.85 – 2.05 (m, 2H), 1.72 – 1.85 (m, 3H), 1.38 – 1.46 (m, 3H). Example 326: (R)-1-(4-benzyl-6-fluoro-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000452_0001
methyl (R)-2-(5-bromo-4-fluoro-2-nitrophenoxy)propanoate A solution of 5-bromo-4-fluoro-2-nitrophenol (1 g, 4.237 mmol, 1 equiv), methyl (2S)-2- hydroxypropanoate (0.66 g, 6.356 mmol, 1.5 equiv) and PPh3 (1.67 g, 6.356 mmol, 1.5 equiv) in THF (10 mL) was stirred for 1 h at rt under N2 atmosphere. To the above mixture was added DEAD (1.11 g, 6.356 mmol, 1.5 equiv) dropwise over 10 min at 0°C. The resulting mixture was stirred for additional 2h at rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give methyl (2R)-2-(5-bromo-4-fluoro-2-nitrophenoxy)propanoate (1.21 g, 88.66%) as a white solid. MS (ESI): mass calcd. for C10H9BrFNO5: 320.96 m/z, found 322.05, 324.05[M+H, M+H+2]+ .
Figure imgf000453_0001
(R)-7-bromo-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-(5-bromo-4-fluoro-2-nitrophenoxy)propanoate (1.2 g, 3.726 mmol, 1 equiv) in EtOH (12.0 mL)/H2O(1.2 mL) was added Fe (2.1 g, 37.260 mmol, 10 equiv) and NH4Cl (2.0 g, 37.260 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-7-bromo-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (900 mg, 92.89%) as a white solid. MS (ESI): mass calcd. for C9H7BrFNO2: 258.96 m/z, found 260.06, 262.06 [M+H, M+H+2]+ .
Figure imgf000453_0002
(R)-4-benzyl-7-bromo-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (250 mg, 0.961 mmol, 1 equiv) in DMF (3 mL) was added benzyl bromide (493.26 mg, 2.883 mmol, 3 equiv) and K2CO3 (401.48 mg, 2.883 mmol, 3 equiv). The reaction was stirred at rt for 1 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-benzyl-7-bromo- 6-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (250 mg, 74.26%) as a white solid. MS (ESI): mass calcd. for C16H13BrFNO2: 349.01 m/z, found 349.95, 351.95 [M+H, M+H+2]+.
Figure imgf000454_0001
(R)-7-amino-4-benzyl-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-benzyl-7-bromo-6-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (240 mg, 0.578 mmol, 1 equiv) in EtOH (5 mL)/H2O (1 mL) was added N1,N2-dimethylcyclohexane-1,2- diamine (49.33 mg, 0.347 mmol, 0.6 equiv), CuSO4(92.25 mg, 0.578 mmol, 1 equiv), sodium ascorbate (230.19 mg, 1.156 mmol, 2 equiv), and NaN3 (112.74 mg, 1.734 mmol, 3 equiv). The reaction was stirred at 80 °C under N2 for 2 h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-benzyl- 6-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (150 mg, 90.64%) as a white solid. LC/MS (ESI): mass calcd. for C16H15FN2O2: 286.11 m/z, found: 287.20 [M+H]+.
Figure imgf000454_0002
(R)-1-(4-benzyl-6-fluoro-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert- butyl)urea To a solution of (2R)-7-amino-4-benzyl-6-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (130 mg, 0.454 mmol, 1 equiv) and DMAP (5.55 mg, 0.045 mmol, 0.1 equiv) in DCM (3 mL) was added 2-isocyanato-2-methylpropane (135.04 mg, 1.362 mmol, 3 equiv) and TEA (137.84 mg, 1.362 mmol, 3 equiv). The reaction was stirred at rt for 24 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1-[(2R)-4-benzyl-6- fluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea (34.9 mg, 19.74%) as a white solid. LC/MS (ESI): mass calcd. for C21H24FN3O3: 385.18 m/z, found: 408.10 [M+Na]+.19F NMR (282 MHz, DMSO) δ -137.66.1H NMR (300 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.80-7.83 (d, J = 7.7 Hz, 1H), 7.30 – 7.40 (m, 2H), 7.22 – 7.26 (m, 3H), 6.94 – 6.98 (d, J = 12 Hz, 1H), 6.51 (s, 1H), 5.11 (s, 2H), 4.77 – 4.84 (m, 1H), 1.46 – 1.48 (d, J = 6 Hz, 3H), 1.27 (s, 9H). Example 327: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-6-fluoro-2-methyl-3-oxo- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000455_0001
(R)-7-bromo-4-(3-(difluoromethyl)benzyl)-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (2R)-7-bromo-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (250 mg, 0.961 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3-(difluoromethyl)benzene (637.48 mg, 2.883 mmol, 3 equiv) and K2CO3 (401.48 mg, 2.883 mmol, 3 equiv). The reaction was stirred at rt for 1h. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give(2R)-7-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-6-fluoro-2-methyl-2H-1,4- benzoxazin-3-one (350 mg, 90.98%) as a yellow solid. LC/MS: mass calcd. for C17H13BrF3NO2:399.01 m/z, found: 399.90, 401.90 [M+H, M+H+2]+.
Figure imgf000456_0001
(R)-7-amino-4-(3-(difluoromethyl)benzyl)-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (2R)-7-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-6-fluoro-2-methyl-2H-1,4- benzoxazin-3-one (340 mg, 0.850 mmol, 1 equiv) in EtOH (8 mL) and H2O (2 mL) was added (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (72.51 mg, 0.510 mmol, 0.6 equiv), CuSO4 (135.59 mg, 0.850 mmol, 1 equiv), NaN3 (165.70 mg, 2.550 mmol, 3 equiv) and sodium ascorbate (338.33 mg, 1.700 mmol, 2 equiv). The reaction was stirred at 80 °C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-6-fluoro-2-methyl-2H-1,4-benzoxazin-3- one (100 mg, 35.00%) as a yellow solid. LC/MS: mass calcd. for C17H15F3N2O2:336.11 m/z, found:337.10 [M+H]+.
Figure imgf000456_0002
(R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-6-fluoro-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-6-fluoro-2-methyl-2H-1,4- benzoxazin-3-one (100 mg, 0.297 mmol, 1 equiv) in DCM (5 mL) and toluene (2.5 mL) was added 2-isocyanato-2-methylpropane (88.43 mg, 0.891 mmol, 3 equiv). The reaction was stirred at 90°C under N2 for 3h. After cooling down to rt, the reaction was quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 42% to 64% (v/v) CH3CN and Water (10mmol/L NH4HCO3)) to afford the title compound 3-tert-butyl-1-[(2R)-4-{[3- (difluoromethyl)phenyl]methyl}-6-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (22.9 mg, 17.69%) as a white solid. LC/MS: mass calcd. for C22H24F3N3O3:435.18 m/z, found:458.10 [M+Na]+. 1H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 2.6 Hz, 1H), 7.84 (d, J = 7.7 Hz, 1H), 7.37 – 7.52 (m, 4H), 6.84 – 7.21 (m, 2H), 6.51 (s, 1H), 5.10 – 5.25 (m, 2H), 4.83 (q, J = 6.7 Hz, 1H), 1.48 (d, J = 6.8 Hz, 3H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -109.7463, - 137.0823. Example 328: 1-(tert-butyl)-3-((R)-6-fluoro-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea
Figure imgf000457_0001
(R)-7-bromo-6-fluoro-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of (2R)-7-bromo-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.154 mmol, 1 equiv) in DCM (5 mL) was treated with 1-phenyl-ethanol (140.93 mg, 1.154 mmol, 1 equiv) and PPh3 (453.86 mg, 1.731 mmol, 1.5 equiv) for 1h at rt under nitrogen atmosphere followed by the addition of DEAD (200.90 mg, 1.154 mmol, 1 equiv) dropwise at 0°C. The resulting mixture was stirred for 2h at rt under N2 atmosphere. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-6-fluoro-2-methyl-4- [(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (195 mg, 46.41%) as a white solid. MS (ESI): mass calcd. for C17H15BrFNO2: 363.03 m/z, found 364.10, 366.10 [M+H, M+H+2]+ .
Figure imgf000458_0001
(R)-7-amino-6-fluoro-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-6-fluoro-2-methyl-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3- one (175 mg, 0.408 mmol, 1 equiv) in EtOH (4 mL)/H2O (1 mL) was added N1,N2- dimethylcyclohexane-1,2-diamine (34.80 mg, 0.245 mmol, 0.6 equiv), CuSO4 (65.07 mg, 0.408 mmol, 1 equiv), sodium ascorbate (162.36 mg, 0.816 mmol, 2 equiv) and NaN3 (79.52 mg, 1.224 mmol, 3 equiv). The reaction was stirred at 80°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-6- fluoro-2-methyl-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (60 mg, 49.00%)as a white solid. MS (ESI): mass calcd. for C17H17FN2O2: 300.13 m/z, found 301.05 [M+H]+ .
Figure imgf000458_0002
1-(tert-butyl)-3-((R)-6-fluoro-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-6-fluoro-2-methyl-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-3- one (60 mg, 0.200 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (59.41 mg, 0.600 mmol, 3 equiv) in DCM (3 mL) was added TEA (60.65 mg, 0.600 mmol, 3 equiv) and DMAP (2.44 mg, 0.020 mmol, 0.1 equiv). The reaction was stirred at rt for 24 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1- [(2R)-6-fluoro-2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-2H-1,4-benzoxazin-7-yl]urea (14.5 mg, 18.06%) as a white solid. LC/MS (ESI): mass calcd. for C22H26FN3O3:399.20 m/z, found:400.10 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.80-7.82 (d, J = 6 Hz, 1H), 7.33 - 7.44 (m, 2H), 7.23 - 7.33 (m, 3H), 6.48 - 6.59 (m, 2H), 6.04 -6.06 (m, 1H), 4.72 - 4.75 (m, 1H), 1.74 - 1.76 (d, J = 6 Hz, 3H), 1.41 -1.44 (d, J = 9 Hz, 3H), 1.26 (s, 9H).19F NMR (282 MHz, DMSO) δ- 137.16. Example 329: (R)-1-(4-benzyl-6,8-difluoro-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000459_0001
methyl (R)-2-(2,4-difluoro-6-nitrophenoxy)propanoate To a solution of 2,4-difluoro-6-nitrophenol (2 g, 11.423 mmol, 1 equiv) in THF (20 mL) was added methyl (2S)-2-hydroxypropanoate (1.19 g, 11.423 mmol, 1 equiv) and PPh3 (4.49 g, 17.134 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DIAD (2.21 g, 17.134 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at 60°C. The reaction was monitored by TLC. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give tmethyl (2R)-2-(2,4-difluoro- 6-nitrophenoxy)propanoate (2.5 g, 83.80%) as a white oil. LC/MS: mass calcd. for C10H9F2NO5:261.04m/z.
Figure imgf000460_0001
(R)-6,8-difluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-(2,4-difluoro-6-nitrophenoxy)propanoate (2.5 g, 9.572 mmol, 1 equiv) in EtOH (30 mL) and H2O (3 mL) was added Fe (5.35 g, 95.720 mmol, 10 equiv) and NH4Cl (5.12 g, 95.720 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with EtOH (3 x 100 mL). The filtrate was concentrated under reduced pressure. Diluted with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6,8-difluoro-2-methyl-2,4- dihydro-1,4-benzoxazin-3-one (1.74 g, 91.28%) as white oil. LC/MS: mass calcd. for C9H7F2NO2:199.04 m/z, found:197.90 [M-H]-.
Figure imgf000460_0002
(R)-6,8-difluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6,8-difluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (800 mg, 4.017 mmol, 1 equiv) in H2SO4 (10 mL) was added KNO3 (406.12 mg, 4.017 mmol, 1 equiv) at 0°C. The reaction was stirred at rt for 1h. The reaction was quenched with water/Ice at 0°C. Diluted with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6,8-difluoro-2-methyl- 7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (900 mg, 91.77%) as a yellow solid. LC/MS: mass calcd. for C9H6F2N2O4:244.03 m/z, found:242.95 [M-H]-.
Figure imgf000461_0002
(R)-4-benzyl-6,8-difluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6,8-difluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.229 mmol, 1 equiv) in DMF (10 mL) was added K2CO3 (513.17 mg, 3.687 mmol, 3 equiv) and benzyl bromide (315.24 mg, 1.844 mmol, 1.5 equiv) at 0 °C. The reaction was stirred at rt for 1h. The reaction was monitored by TLC. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-4-benzyl-6,8-difluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (390 mg, 94.95%) as a yellow solid.
Figure imgf000461_0001
(R)-7-amino-4-benzyl-6,8-difluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-benzyl-6,8-difluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (380 mg, 1.137 mmol, 1 equiv) in MeOH (10 mL) and H2O (1 mL) was added Fe (634.83 mg, 11.370 mmol, 10 equiv) and NH4Cl (608.06 mg, 11.370 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 50 mL). The filtrate was concentrated under reduced pressure. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-benzyl- 6,8-difluoro-2-methyl-2H-1,4-benzoxazin-3-one (320 mg, 92.51%) as a yellow solid. LC/MS: mass calcd. for C16H14F2N2O2: 304.10 m/z, found:305.15 [M+H]+.
Figure imgf000462_0001
(R)-1-(4-benzyl-6,8-difluoro-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3- (tert-butyl)urea To a solution of (2R)-7-amino-4-benzyl-6,8-difluoro-2-methyl-2H-1,4-benzoxazin-3-one (280 mg, 0.920 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (109.21 mg, 0.368 mmol, 0.4 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (279.34 mg, 2.760 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (201.90 mg, 2.760 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 36% to 66% (v/v) CH3CN and Water (10mmol/L NH4HCO3)) to afford the title compound 1-[(2R)-4-benzyl-6,8-difluoro-2- methyl-3-oxo-2H-1,4-benzoxazin-7-yl]-3-tert-butylurea (65.3 mg, 17.26%) as a white solid. LC/MS: mass calcd. for C21H23F2N3O3:403.17 m/z, found:404.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.50 (s, 1H), 7.31 – 7.39 (m, 2H), 7.26 (td, J = 5.2, 3.0 Hz, 3H), 6.90 (dd, J = 11.4, 1.9 Hz, 1H), 6.15 (s, 1H), 5.09 – 5.23 (m, 2H), 4.98 (q, J = 6.7 Hz, 1H), 1.54 (d, J = 6.7 Hz, 3H), 1.25 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -126.0506, -139.2961. Example 330: (R)-1-(4-benzyl-2-methyl-3-oxo-6-(trifluoromethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000463_0001
methyl (R)-2-(5-bromo-2-nitro-4-(trifluoromethyl)phenoxy)propanoate To a stirred solution of methyl (2S)-2-hydroxypropanoate (181 mg, 1.736 mmol, 1 equiv) in THF was added 60% NaH (83.3 mg, 2.083 mmol, 1.2 equiv) at 0 °C. The final reaction mixture was stirred for 30 min at 0 °C. To the above mixture was added 1-bromo-5-fluoro-4-nitro-2-(trifluoromethyl)benzene (500 mg, 1.736 mmol, 1 equiv) at room temperature. The reaction was stirred for 2 h at rt. The reaction was quenched with water (10 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The residue was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford methyl (R)-2-(5-bromo-2-nitro-4- (trifluoromethyl)phenoxy)propanoate (410 mg, 63.47%) as a white solid. (ESI): mass calcd. for C11H9BrF3NO5: 370.96 m/z, found: 372.09 [M+H]+.
Figure imgf000463_0002
(R)-7-bromo-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a mixture of methyl (R)-2-(5-bromo-2-nitro-4-(trifluoromethyl)phenoxy)propanoate (0.4 g, 1.075 mmol, 1 equiv) in EtOH (25 mL) and water (5 mL). Fe (0.6 g, 10.75 mmol, 10 equiv) and NH4Cl (569 mg, 10.75 mmol, 10 equiv) was added into the mixture. The resulting solution was stirred at 70 °C for 1 h. The reaction was quenched with water (30 mL) at rt. After filtration, the filter cake was washed with EA (3 x 30 mL). The filtrate was added water (30 mL) and extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford (R)-7-bromo-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one as a light yellow oil. (ESI): mass calcd. for C10H7BrF3NO2: 308.96 m/z, found: 310.07 [M+H]+.
Figure imgf000464_0001
(R)-4-benzyl-7-bromo-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of (R)-7-bromo-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (300 mg, 0.968 mmol, 1 equiv) and benzyl bromide (248 mg, 1.452 mmol, 1.5 equiv) and K2CO3 (404 mg, 2.904 mmol, 3 equiv) in DMF was stirred for 3 h at room temperature. The reaction was quenched with water (10 mL) at room temperature. The aqueous layer was extracted with EtOAc (3 x 10 mL). The residue was purified by silica gel column chromatography, eluted with PE/EA (1:1) to afford (R)-4-benzyl-7-bromo-2-methyl-6- (trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (310 mg, 80.06%) as a white solid. (ESI): mass calcd. for C17H13BrF3NO2: 399.01 m/z, found: 400.19 [M+H]+.
Figure imgf000464_0002
(R)-7-amino-4-benzyl-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (R)-4-benzyl-7-bromo-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one (279 mg, 0.699 mmol, 1 equiv) in EtOH (6 mL) & H2O (2 mL) was added sodium ascorbate (28 mg, 0.140 mmol, 0.2 equiv), N1,N2-dimethylcyclohexane-1,2-diamine (20 mg , 0.140 mmol, 0.2 equiv), CuSO4 (23 mg , 0.14 mmol, 0.2 equiv) and NaN3 (136 mg, 2.097 mmol, 3 equiv) at rt. The resulting mixture was stirred 4 h at 80 °C. The reaction mixture was quenched by water (10 mL) and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (30 mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was purified by flash chromatography with PE/EA (1:1) to afford (R)-7-amino-4-benzyl- 2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (177 mg, 75.51%) as a yellow oil. (ESI): mass calcd. for C17H15F3N2O2: 336.10 m/z, found: 337.11 [M+H]+.
Figure imgf000465_0001
(R)-1-(4-benzyl-2-methyl-3-oxo-6-(trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)-3-(tert-butyl)urea To a mixture of 2-(1-(7-amino-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)ethyl)-4- chlorobenzonitrile (34 mg, 0.101 mmol, 1 equiv) and TEA (21 mg, 0.202 mmol, 2 equiv) in DCM (4 mL) was added 2-isocyanato-2-methylpropane (30 mg, 0.303 mmol, 3 equiv) at 0 °C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column with PE/EA (1:1) and Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water (10mmol/L NH4HCO3+0.05%NH3H2O), Mobile Phase B: ACN; Flow rate: 60 mL/min mL/min; Gradient: 50% B to 72% B in 7 min; Wave Length: 254nm/220nm nm; RT1(min): 6.07) to afford (R)-1-(4-benzyl-2-methyl-3-oxo-6-(trifluoromethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (15 mg, 34.09%) as a white solid. (ESI): mass calcd. for C22H24F3N3O3: 435.18 m/z, found: 436.18 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.69 (s, 1H), 7.57 (s, 1H), 7.33 – 7.41 (m, 2H), 7.21 – 7.30 (m, 3H), 7.12 (s, 1H), 6.94 (s, 1H), 5.13 – 5.28 (m, 2H), 4.92 – 5.03 (m, 1H), 1.51 – 1.59 (m, 3H), 1.27 (s, 9H). Example 331: 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000466_0001
(R)-7-bromo-2-methyl-4-((R)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of (2R)-7-bromo-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (600 mg, 2.479 mmol, 1 equiv) in DCM (10 mL) was treated with (S)-1-phenylethanol (454.20 mg, 3.719 mmol, 1.5 equiv) and PPh3 (975.18 mg, 3.719 mmol, 1.5 equiv) for 1h at rt under nitrogen atmosphere followed by the addition of DEAD (647.49 mg, 3.719 mmol, 1.5 equiv) dropwise at 0°C. The resulting mixture was stirred for 2h at rt under N2 atmosphere. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-2-methyl-4-[(1R)-1-phenylethyl]-2H-1,4- benzoxazin-3-one (400 mg, 46.61%) as a white solid. MS (ESI): mass calcd. for C17H16BrNO2: 345.04 m/z, found 346.00, 348.00 [M+H, M+H+2]+.
Figure imgf000466_0002
(R)-7-amino-2-methyl-4-((R)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-7-bromo-2-methyl-4-[(1R)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (390 mg, 0.948 mmol, 1 equiv) in EtOH (4 mL) was added N1,N2-dimethylcyclohexane-1,2-diamine (80.94 mg, 0.569 mmol, 0.6 equiv), CuSO4 (151.36 mg, 0.948 mmol, 1 equiv), sodium ascorbate (377.66 mg, 1.896 mmol, 2 equiv) and NaN3 (184.96 mg, 2.844 mmol, 3 equiv). The reaction was stirred at 80 °C under N2 for 1h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-2-methyl-4-[(1R)-1- phenylethyl]-2H-1,4-benzoxazin-3-one (250 mg, 93.37%) as a white solid. MS (ESI): mass calcd. for C17H18N2O2: 282.14 m/z, found 283.10 [M+H]+.
Figure imgf000467_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((R)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-2-methyl-4-[(1R)-1-phenylethyl]-2H-1,4-benzoxazin-3-one (294 mg, 1.041 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (309.68 mg, 3.123 mmol, 3 equiv) in DCM (10 mL) was added TEA (316.11 mg, 3.123 mmol, 3 equiv) and DMAP (12.72 mg, 0.104 mmol, 0.1 equiv). The reaction was stirred at rt for 24 h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1- [(2R)-2-methyl-3-oxo-4-[(1R)-1-phenylethyl]-2H-1,4-benzoxazin-7-yl]urea (58.6 mg, 14.53%) as a white solid. LC/MS (ESI): mass calcd. for C22H27N3O3:381.21 m/z, found:382.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.25 – 7.39 (m, 6H), 6.62-6.63 (m, 1H), 6.60-6.61 (m, 1H), 6.15-6.17 (m, 1H), 5.94 (s, 1H), 4.69-4.75 (m, 1H), 1.66-1.69 (d, J = 9.0 Hz, 3H), 1.46-1.48 (d, J = 6.0 Hz, 3H), 1.26 (s, 9H). Example 332: (R)-1-(4-benzyl-2,6-dimethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7- yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000468_0001
methyl (2R)-2-(4-methyl-2-nitrophenoxy)propanoate To a solution of 2-nitro-P-cresol (1 g, 6.530 mmol, 1 equiv) in THF (15 mL) was added methyl (2S)-2-hydroxypropanoate (0.82 g, 7.836 mmol, 1.2 equiv) and PPh3 (2.57 g, 9.795 mmol, 1.5 equiv) under N2 at 0°C. The reaction was stirred at 0 degrees C for 0.5 h. Then DEAD (1.71 g, 9.795 mmol, 1.5 equiv) was added at 0°C. The reaction was stirred at 60 degrees C for 2 h. The mixture was allowed to cool down to room temperature. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give methyl (2R)-2-(4-methyl-2-nitrophenoxy)propanoate (1.3 g, 83.22%) as a yellow solid. LC/MS (ESI): mass calcd. for C11H13NO5:239.08 m/z, found:240.05 [M+H]+.
Figure imgf000468_0002
(2R)-2,6-dimethyl-2,4-dihydro-1,4-benzoxazin-3-one To a solution of methyl (2R)-2-(4-methyl-2-nitrophenoxy)propanoate (700 mg, 2.926 mmol, 1 equiv) in EtOH (10 mL) and H2O (2 mL) was added Fe (1.63 g, 29.260 mmol, 10 equiv) and NH4Cl (1.57 g, 29.260 mmol, 10 equiv). The reaction was stirred at 70 degrees C for 2 h. The mixture was allowed to cool down to room temperature. The resulting mixture was filtered, the filter cake was washed with DCM (3 x 50 mL). The filtrate was concentrated under reduced pressure. Quenched with water (100 mL) and extracted with DCM (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-2,6- dimethyl-2,4-dihydro-1,4-benzoxazin-3-one (450 mg, 86.79%) as a yellow solid. LC/MS (ESI): mass calcd. for C10H11NO2:177.08 m/z, found:178.15 [M+H]+.
Figure imgf000469_0001
(2R)-2,6-dimethyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one To a solution of (2R)-2,6-dimethyl-2,4-dihydro-1,4-benzoxazin-3-one (420 mg, 2.370 mmol, 1 equiv) in H2SO4 (5 mL) was added KNO3 (311.52 mg, 3.081 mmol, 1.3 equiv) at 0°C. The reaction was stirred at 0 degrees C for 1 h. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-2,6-dimethyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (160 mg, 30.38%) as a yellow solid. TLC showed the desired product was generated.
Figure imgf000469_0002
(2R)-4-benzyl-2,6-dimethyl-7-nitro-2H-1,4-benzoxazin-3-one To a solution of (2R)-2,6-dimethyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (160 mg, 0.720 mmol, 1 equiv) in DMF (5 mL) was added benzyl bromide (160.11 mg, 0.936 mmol, 1.3 equiv) and K2CO3 (300.73 mg, 2.160 mmol, 3 equiv). The reaction was stirred at 25 degrees C for 16 h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-benzyl-2,6- dimethyl-7-nitro-2H-1,4-benzoxazin-3-one (200 mg, 88.93%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H16N2O4:312.11 m/z, found:310.95 [M-H]-.
Figure imgf000470_0002
(2R)-7-amino-4-benzyl-2,6-dimethyl-2H-1,4-benzoxazin-3-one To a solution of (2R)-4-benzyl-2,6-dimethyl-7-nitro-2H-1,4-benzoxazin-3-one (190 mg, 0.608 mmol, 1 equiv) in MeOH (4 mL) and H2O (1 mL) was added Fe (339.73 mg, 6.080 mmol, 10 equiv) and NH4Cl (325.40 mg, 6.080 mmol, 10 equiv). The reaction was stirred at 70 degrees C for 2 h. The mixture was allowed to cool down to room temperature. The resulting mixture was filtered, the filter cake was washed with DCM (3 x 50 mL). The filtrate was concentrated under reduced pressure. Diluted with water (40 mL) and extracted with DCM (3 x 40 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7- amino-4-benzyl-2,6-dimethyl-2H-1,4-benzoxazin-3-one (130 mg, 75.69%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H18N2O2:282.14 m/z, found:283.10 [M+H]+.
Figure imgf000470_0001
(R)-1-(4-benzyl-2,6-dimethyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert- butyl)urea To a solution of (R)-7-amino-4-benzyl-2,6-dimethyl-2H-benzo[b][1,4]oxazin-3(4H)-one (150 mg, 0.531 mmol, 1 equiv) in DCM (5 mL) was added NEt3 (322.61 mg, 3.186 mmol, 6 equiv) , 2- isocyanato-2-methylpropane (210.69 mg, 2.124 mmol, 4 equiv) and DMAP (6.49 mg, 0.053 mmol, 0.1 equiv) at 0°C. The reaction was stirred at 25 °C for 48 hours. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge C18 Column 250 mm x 19 mm x 5 μm column (eluent: 53% to 80% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to give the (R)-1-(4-benzyl-2,6-dimethyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea (63.1 mg, 30.91%) as a white solid. MS (ESI): mass calcd. for C22H27N3O3: 381.2 m/z, found 382.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.65 (s, 1H), 7.40 (s, 1H), 7.30 – 7.36 (m, 2H), 7.21 – 7.27 (m, 3H), 6.82 (s, 1H), 6.48 (s, 1H), 5.10 (s, 2H), 4.76 (q, J = 6.7 Hz, 1H), 2.03 (s, 3H), 1.46 (d, J = 6.7 Hz, 3H), 1.28 (s, 9H). Example 333 and Example 334: (S)-1-(1-benzyl-7-fluoro-3-methyl-2-oxo-1,2,3,4- tetrahydroquinolin-6-yl)-3-(tert-butyl)urea and (R)-1-(1-benzyl-7-fluoro-3-methyl-2-oxo- 1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000471_0001
7-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 7-fluoro-3,4-dihydro-1H-quinolin-2-one (1 g, 6.054 mmol, 1 equiv) in H2SO4 (10 mL) was added HNO3 (0.38 g, 6.054 mmol, 1 equiv) at 0°C. The reaction was stirred at 0°C for 1 h. Quenched with ice water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7- fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (1.1 g, 86.45%) as a white solid. MS (ESI): mass calcd. for C9H7FN2O3: 210.04 m/z, found 209.05 [M-H]- .
Figure imgf000472_0001
1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2(1H)-one To a solution of 7-fluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (1.1 g, 5.234 mmol, 1 equiv) in DMF (10 mL) was added benzyl bromide (2.68 g, 15.702 mmol, 3 equiv) and Cs2CO3(5.13 g, 15.702 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2-one (1.4 g, 89.07%) as a white solid. MS (ESI): mass calcd. for C16H13FN2O3: 300.09 m/z, found 300.95 [M+H]+.
Figure imgf000472_0002
1-benzyl-7-fluoro-3-methyl-6-nitro-3,4-dihydroquinolin-2(1H)-one A solution of 1-benzyl-7-fluoro-6-nitro-3,4-dihydroquinolin-2-one (600 mg, 1.998 mmol, 1 equiv) in tetrahydrofuran (6 mL) was treated with LiHDMS (401.20 mg, 2.398 mmol, 1.2 equiv) for 1 h at -78°C under nitrogen atmosphere followed by the addition of MeI (283.60 mg, 1.998 mmol, 1 equiv) dropwise at -78°C. The resulting mixture was stirred for 1 h at rt under N2 atmosphere. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1- benzyl-7-fluoro-3-methyl-6-nitro-3,4-dihydroquinolin-2-one (275 mg, 43.79%) as a white solid. MS (ESI): mass calcd. for C17H15FN2O3: 314.11 m/z, found 315.10 [M+H]+.
Figure imgf000473_0001
6-amino-1-benzyl-7-fluoro-3-methyl-3,4-dihydroquinolin-2(1H)-one To a solution of 1-benzyl-7-fluoro-3-methyl-6-nitro-3,4-dihydroquinolin-2-one (270 mg, 0.859 mmol, 1 equiv) in EtOH (5 mL)/H2O (0.5 mL) was added Fe (479.71 mg, 8.590 mmol, 10 equiv) and NH4Cl (459.48 mg, 8.590 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 6-amino-1-benzyl-7-fluoro-3-methyl-3,4-dihydroquinolin-2-one (220 mg, 90.07%) as a white solid. MS (ESI): mass calcd. for C17H17FN2O: 284.13 m/z, found 285.10 [M+H]+.
Figure imgf000473_0002
1-(1-benzyl-7-fluoro-3-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 6-amino-1-benzyl-7-fluoro-3-methyl-3,4-dihydroquinolin-2-one (210 mg, 0.739 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato-2-methylpropane (219.65 mg, 2.217 mmol, 3 equiv) and TEA (224.21 mg, 2.217 mmol, 3 equiv). The reaction was stirred at rt for 24 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 1-(1-benzyl-7- fluoro-3-methyl-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (70 mg, 24.72%) as a white solid. MS (ESI): mass calcd. for C22H26FN3O2: 383.20 m/z, found 382.00 [M-H]-.
Figure imgf000474_0001
(S)-1-(1-benzyl-7-fluoro-3-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea and (R)-1-(1-benzyl-7-fluoro-3-methyl-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert- butyl)urea A sample of 1-(1-benzyl-7-fluoro-3-methyl-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea (70 mg, 0.183 mmol, 1 equiv) was separated by CHIRALPAK IG, 5*25 cm, 5 μm (eluent: 30% to 30% (v/v) Hex(0.1% 2M NH3-MeOH)—HPLC and EtOH to yield 1-[(3S)-1-benzyl-7-fluoro-3- methyl-2-oxo-3,4-dihydroquinolin-6-yl]-3-tert-butylurea (29.3 mg, 41.21%) (first enantiomer to elute, labelled *S) MS (ESI): mass calcd. for C22H26FN3O2: 383.20 m/z, found 382.00 [M-H]-. 1H NMR (300 MHz, DMSO-d6) δ 7.92-7.95 (m, 2H), 7.27 – 7.38 (m, 2H), 7.15 – 7.27 (m, 3H), 6.75-6.79 (d, J = 12 Hz, 1H), 6.43 (s, 1H), 5.10 (s, 2H), 2.91-2.95 (d, J = 12 Hz, 1H), 2.59 – 2.74 (m, 2H), 1.27 (s, 9H), 1.15 – 1.19 (m, 3H). 19F NMR (282 MHz, DMSO) δ-131.68 and 1-[(3R)- 1-benzyl-7-fluoro-3-methyl-2-oxo-3,4-dihydroquinolin-6-yl]-3-tert-butylurea (18.6 mg, 26.42%) (second enantiomer to elute, labelled *R) MS (ESI): mass calcd. for C22H26FN3O2: 383.20 m/z, found 382.00 [M-H]-. 1H NMR (300 MHz, DMSO-d6) δ 7.92-7.95 (m, 2H), 7.27 – 7.38 (m, 2H), 7.15 – 7.27 (m, 3H), 6.75-6.79 (d, J = 12 Hz, 1H), 6.43 (s, 1H), 5.10 (s, 2H), 2.91-2.95 (m, 1H), 2.59 – 2.79 (m, 2H), 1.27 (s, 9H), 1.17-1.19 (d, J = 6 Hz, 3H).19F NMR (282 MHz, DMSO) δ - 132.02. Example 335: 3-tert-butyl-1-[(2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2- methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea Synthetic Scheme
Figure imgf000475_0001
(2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-7-nitro-2H-1,4- benzoxazin-3-one To a stirred mixture of (2R)-6,8-difluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (200 mg, 0.819 mmol, 1 equiv) and 1-(bromomethyl)-3-(difluoromethoxy)benzene (388.35 mg, 1.638 mmol, 2 equiv) in DMF (10 mL) was added K2CO3 (342.11 mg, 2.457 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 4 h at 80°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. TLC showed the desired product was generated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to afford (2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-7-nitro- 2H-1,4-benzoxazin-3-one (314 mg, 95.76%) as a white solid.
Figure imgf000476_0001
(2R)-7-amino-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-2H-1,4- benzoxazin-3-one To a stirred mixture of (2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-7- nitro-2H-1,4-benzoxazin-3-one (300 mg, 0.749 mmol, 1 equiv) and NH4Cl (400.89 mg, 7.490 mmol, 10 equiv) in MeOH:H2O=10:1 was added Fe (418.54 mg, 7.490 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 80°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was used in the next step directly without further purification. The residue afforded (2R)-7-amino-4-{[3- (difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-2H-1,4-benzoxazin-3-one (207 mg, 74.59%) as a yellow oil.
Figure imgf000476_0002
3-tert-butyl-1-[(2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-3-oxo- 2H-1,4-benzoxazin-7-yl]urea To a stirred solution of (2R)-7-amino-4-{[3-(difluoromethoxy)phenyl]methyl}-6,8-difluoro-2- methyl-2H-1,4-benzoxazin-3-one (120 mg, 0.324 mmol, 1 equiv) and triphosgene (57.69 mg, 0.194 mmol, 0.6 equiv) in DCM (5 mL) was added TEA (98.38 mg, 0.972 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added 2-methylpropan- 2-amine (23.70 mg, 0.324 mmol, 1 equiv) dropwise at room temperature. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched with water (20 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford 3-tert-butyl-1-[(2R)-4-{[3- (difluoromethoxy)phenyl]methyl}-6,8-difluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (46.4 mg, 30.46%) as a white solid. LC/MS (ESI): mass calcd. for C22H23F4N3O4:469.1 m/z, found:470.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.51 (s, 1H), 7.01-7.47 (m, 5H), 6.80- 7.00 (m, 1H), 6.15 (s, 1H), 5.06-5.27 (m, 2H),4.90-5.06 (m, 1H), 1.45-1.61 (m, 3H), 1.09-1.35 (m, 9H).19F NMR (376 MHz, DMSO) δ -81.89, -126.18 -139.24. Example 336: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-6,8-difluoro-2-methyl-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000477_0001
Figure imgf000478_0001
(2R)-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-7-nitro-2H-1,4- benzoxazin-3-one To a stirred solution of (2R)-6,8-difluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (340 mg, 1.393 mmol, 1 equiv) and 1-(bromomethyl)-3-(difluoromethyl)benzene (338.60 mg, 1.532 mmol, 1.1 equiv) in DMF (10 mL) was added K2CO3 (581.59 mg, 4.179 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at room temperature under air atmosphere. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3x20 mL), dried over anhydrous MgSO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:3) to afford (2R)-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-7-nitro-2H-1,4- benzoxazin-3-one (350 mg, 65.40%) as a yellow solid. TLC showed the desired product was generated.
Figure imgf000478_0002
(2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-2H-1,4- benzoxazin-3-one To a stirred mixture of (2R)-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-7- nitro-2H-1,4-benzoxazin-3-one (350 mg, 0.911 mmol, 1 equiv) and Fe (508.62 mg, 9.110 mmol,10 equiv) in MeOH / water(10:1, 5 mL ) was added NH4Cl (487.17 mg, 9.110 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at 70 °C under air atmosphere. Desired product could be detected by LCMS. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with water (3x10 mL), dried over anhydrous MgSO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (2R)-7-amino-4-{[3- (difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-2H-1,4-benzoxazin-3-one (280 mg, 86.77%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H14F4N2O2:354.10 m/z, found: 354.95 [M+H] +.
Figure imgf000479_0001
3-tert-butyl-1-[(2R)-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-3- oxo-2H-1,4-benzoxazin-7-yl]urea A solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-2H- 1,4-benzoxazin-3-one (60 mg, 0.169 mmol, 1 equiv) in DCM (5 mL) was treated with triphosgene (100.50 mg, 0.338 mmol, 2 equiv) for 1 h at 0 C under nitrogen atmosphere followed by the addition of triethylamine (34.27 mg, 0.338 mmol, 2 equiv) in portions at 0 °C. To the above mixture was added erbumine (24.77 mg, 0.338 mmol, 2 equiv) and triethylamine (34.27 mg, 0.338 mmol, 2 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 1 h at room temperature. Desired product could be detected by LCMS. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with water (3x10 mL), dried over anhydrous MgSO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using a XBridge Prep OBD 250 mm x 19 mm x 5 μm column (eluent: 42% to 72% (v/v) ACN and Water with 10 mmol/L NH4HCO3) to afford 3-tert-butyl-1-[(2R)-4-{[3- (difluoromethyl)phenyl]methyl}-6,8-difluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (3.6 mg, 4.55%) as a white solid. LC/MS (ESI): mass calcd. for C22H23F4N3O3:453.17 m/z, found:452.00 [M-H] -.1H NMR (400 MHz, Acetonitrile-d3) δ 7.45 – 7.60 (m, 4H), 6.60 – 7.00 (m, 2H), 6.10 – 6.30 (s, 1H), 5.20 – 5.40 (s, 1H), 5.10 – 5.20 (m, 2H), 4.80 – 4.95 (m, 1H),1.55 – 1.65 (m, 3H),1.28 – 1.38 (s, 9H). 19F NMR (376 MHz, Acetonitrile-d3) δ -111.38, -128.07, - 141.04. Example 337: 1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(4-(trifluoromethyl)pyridin-2-yl)ethyl)- 1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000480_0001
1-(1-methylcyclobutyl)-3-(2-oxo-1-(1-(4-(trifluoromethyl)pyridin-2-yl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a solution of 6-amino-1-{1-[4-(trifluoromethyl)pyridin-2-yl]ethyl}quinoxalin-2-one (70 mg, 0.209 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (62.13 mg, 0.209 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (63.57 mg, 0.627 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added 1-methylcyclobutan-1-amine (53.49 mg, 0.627 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column30*150 mm 5μm column (eluent: 33% to 47% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-(1- methylcyclobutyl)-1-(2-oxo-1-{1-[4-(trifluoromethyl)pyridin-2-yl]ethyl}quinoxalin-6-yl)urea (43 mg, 46.05%) as a yellow solid. LC/MS: mass calcd. for C21H22F3N5O2:445.17 m/z, found:446.05 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.78 (d, J = 5.1 Hz, 1H), 8.46 (s, 1H), 8.20 (s, 1H), 7.96 (d, J = 2.5 Hz, 1H), 7.75 (s, 1H), 7.69 (d, J = 5.2 Hz, 1H), 7.10 – 7.56 (m, 2H), 6.54 (s, 1H), 6.33 (s, 1H), 2.29 (dt, J = 11.4, 9.1 Hz, 2H), 1.84 – 1.97 (m, 5H), 1.70 – 1.83 (m, 2H), 1.40 (s, 3H).19F NMR (376 MHz, DMSO) δ(ppm): -663.0913. Example 338: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethoxy)benzyl)-6-fluoro-2-methyl-3-oxo- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000481_0001
methyl (R)-2-(4-fluoro-2-nitrophenoxy)propanoate A solution of 4-fluoro-2-nitrophenol (2 g, 12.731 mmol, 1 equiv) in THF (20 mL) was treated with methyl (2S)-2-hydroxypropanoate (1.99 g, 19.096 mmol, 1.5 equiv) and PPh3 (5.01 g, 19.096 mmol, 1.5 equiv) for 1 h at rt under nitrogen atmosphere followed by the addition of DEAD (3.33 g, 19.096 mmol, 1.5 equiv) dropwise at 0°C. The resulting mixture was stirred for 2 h at 60°C under N2 atmosphere. The mixture was allowed to cool down to rt. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give methyl (2R)-2-(4-fluoro-2- nitrophenoxy)propanoate (1 g, 32.30%) as a white solid. MS (ESI): mass calcd. for C10H10FNO5: 243.05 m/z, found 244.15 [M+H]+ .
Figure imgf000482_0001
(R)-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-(4-fluoro-2-nitrophenoxy)propanoate (1 g, 4.112 mmol, 1 equiv) in EtOH (10 mL)/H2O (1 mL) was added Fe (2.30 g, 41.120 mmol, 10 equiv) and NH4Cl (2.20 g, 41.120 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (650 mg, 87.25%) as a white solid. MS (ESI): mass calcd. for C9H8FNO2: 181.05 m/z, found 182.05 [M+H]+.
Figure imgf000482_0002
(R)-6-fluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one A solution of (2R)-6-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (1 g, 5.520 mmol, 1 equiv) and KNO3 (558.06 mg, 5.520 mmol, 1 equiv) in H2SO4 (10 mL) was stirred for 1 h at 0°C. The reaction was stirred at rt for 1 h. Quenched with ice water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-fluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (660 mg, 52.87%) as a white solid. MS (ESI): mass calcd. for C9H7FN2O4: 226.04 m/z, found 224.95 [M-H]-.
Figure imgf000483_0001
(R)-4-(3-(difluoromethoxy)benzyl)-6-fluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (2R)-6-fluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.326 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-3-(difluoromethoxy)benzene (377.32 mg, 1.591 mmol, 1.2 equiv) and K2CO3 (553.99 mg, 3.978 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6-fluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3- one (450 mg, 88.74%) as a white solid. MS (ESI): mass calcd. for C17H13F3N2O5: 382.08 m/z, found 383.10 [M+H]+.
Figure imgf000483_0002
(R)-7-amino-4-(3-(difluoromethoxy)benzyl)-6-fluoro-2-methyl-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (2R)-4-{[3-(difluoromethoxy)phenyl]methyl}-6-fluoro-2-methyl-7-nitro-2H- 1,4-benzoxazin-3-one (514 mg, 1.345 mmol, 1 equiv) in EtOH (10 mL) was added Fe (750.84 mg, 13.450 mmol, 10 equiv) and NH4Cl (719.18 mg, 13.450 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-{[3- (difluoromethoxy)phenyl]methyl}-6-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (450 mg, 95.00%) as a white solid. MS (ESI): mass calcd. for C17H15F3N2O3: 352.10 m/z, found 353.05 [M+H]+.
Figure imgf000484_0001
(R)-1-(tert-butyl)-3-(4-(3-(difluoromethoxy)benzyl)-6-fluoro-2-methyl-3-oxo-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethoxy)phenyl]methyl}-6-fluoro-2-methyl-2H- 1,4-benzoxazin-3-one (200 mg, 0.568 mmol, 1 equiv) in DCM (5 mL) was added 2-isocyanato- 2-methylpropane (168.74 mg, 2.840 mmol, 5 equiv) and TEA (287.22 mg, 2.840 mmol, 5 equiv). The reaction was stirred at rt for 24 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-4-{[3- (difluoromethoxy)phenyl]methyl}-6-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (55.4 mg, 21.60%) as a off-white solid. LC/MS (ESI): mass calcd. for C22H24F3N3O4: 451.17 m/z, found: 452.05 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.82-7.84 (d, J = 6 Hz, 1H), 6.95 – 7.46 (m, 6H), 6.51 (s, 1H), 5.12 (s, 2H), 4.78-4.85 (m, 1H), 1.46-1.48 (d, J = 6.0 Hz, 3H), 1.27 (s, 9H).19F NMR (282 MHz, DMSO) δ-81.91 -137.08. Example 339: (S)-1-(tert-butyl)-3-(7-fluoro-2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000485_0001
7-fluoro-6-nitro-1H-quinoxalin-2-one To a solution of 4-fluoro-5-nitrobenzene-1,2-diamine (2 g, 11.687 mmol, 1 equiv) in ethyl alcohol (10 mL) was added ethyl glyoxylate (1.19 g, 11.687 mmol, 1 equiv). The resulting mixture was stirred for 2 h at 80 °C. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 7-fluoro-6-nitroquinoxalin- 2(1H)-one (1.7 g, 69.55%) as a white solid. MS (ESI): mass calcd. for C8H4FN3O3: 209.17 m/z, found: 208 [M-H]-.
Figure imgf000485_0002
(S)-7-fluoro-6-nitro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 7-fluoro-6-nitroquinoxalin-2(1H)-one (1.6 g, 7.651 mmol, 1.2 equiv) in DCM (40 mL) was added (1R)-1-[3-(trifluoromethoxy)phenyl]ethanol (1.31 g, 6.376 mmol, 1 equiv) , PPh3 (2.51 g, 9.564 mmol, 1.5 equiv) and DEAD (1.39 g, 7.970 mmol, 1.25 equiv). The resulting mixture was stirred for 2 h at 25 °C under N2 and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (S)-7-fluoro-6-nitro-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one (2 g, 78.96%) as a white solid. MS (ESI): mass calcd. for C17H11F4N3O4: 397.17 m/z, found: 398.05 [M+H]+.
Figure imgf000486_0001
(S)-6-amino-7-fluoro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of (S)-7-fluoro-6-nitro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)- one (2 g, 5.034 mmol, 1 equiv) in EtOH (20 mL) and H2O (2 mL) was added Fe (2.81 g, 50.340 mmol, 10 equiv) and NH4Cl (2.69 g, 50.340 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 80 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with DCM (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (S)-6-amino-7-fluoro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin- 2(1H)-one (350 mg, 87.28%) as a white solid. MS (ESI): mass calcd. for C17H13F4N3O2: 367.17 m/z, found: 368.15 [M+H]+.
Figure imgf000486_0002
(S)-1-(tert-butyl)-3-(7-fluoro-2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a solution of (S)-6-amino-7-fluoro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)- one (80 mg, 0.218 mmol, 1 equiv) in DCM (20 mL) was added 2-isocyanato-2-methylpropane (64.77 mg, 0.654 mmol, 3 equiv), DMAP (2.66 mg, 0.022 mmol, 0.1 equiv)and TEA (66.12 mg, 0.654 mmol, 3 equiv). The resulting mixture was stirred for 24 h at rt. The reaction mixture was quenched by water and extracted with DCM (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep- HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 53% B in 9 min, 53% B; Wave Length: 254/220 nm; RT1(min): 8.52; Number Of Runs: 0) to afford (S)-1- (tert-butyl)-3-(7-fluoro-2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6- yl)urea (26 mg, 25.52%)as an white solid. MS (ESI): mass calcd. For C22H22F4N4O3: 466.17 m/z, found: 467.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 8.7 Hz, 1H), 8.23 – 7.27 (m, 2H), 7.44 – 7.50 (m, 1H), 7.30 – 7.35 (m, 3H), 7.13 – 7.17 (m, 1H), 6.58 (s, 1H), 6.46 (br, 1H), 1.88 (d, J = 7.0 Hz, 3H), 1.30 (s, 9H). Example 340: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-5,6-difluoro-2-methyl-3- oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000487_0001
Figure imgf000488_0001
6-bromo-3,4-difluoro-2-nitrophenol To a solution of 2-bromo-4,5-difluorophenol (1.5 g, 7.177 mmol, 1 equiv) and HNO3 (0.45 g, 7.177 mmol, 1 equiv) in H2SO4 (15 mL) was stirred for 1h at 0°C. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-bromo-3,4-difluoro-2-nitrophenol (1.78 g, 83.00%) as a brown solid. LC/MS: mass calcd. for C6H2BrF2NO3: 252.92 m/z found: 253.92, 255.92 [M+H, M+H+2]+.
Figure imgf000488_0002
methyl (R)-2-(6-bromo-3,4-difluoro-2-nitrophenoxy)propanoate To a solution of methyl (2S)-2-hydroxypropanoate (0.73 g, 7.008 mmol, 1 equiv) in DCM (20 mL) was added 6-bromo-3,4-difluoro-2-nitrophenol (1.78 g, 7.008 mmol, 1 equiv) and PPh3 (2.76 g, 10.512 mmol, 1.5 equiv). The resulting mixture was stirred for 1h at rt under N2. DEAD (1.43 g, 8.211 mmol, 1 equiv) was added the mixture at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (1:1) to afford methyl (R)-2-(6-bromo-3,4- difluoro-2-nitrophenoxy) propanoate (2 g, 71.33%) as yellow oil. LC/MS: mass calcd. for C10H8BrF2NO5: 338.96 m/z found: 339.96, 341.96 [M+H, M+H+2]+
Figure imgf000488_0003
(R)-8-bromo-5,6-difluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a mixture of methyl (2R)-2-(6-bromo-3,4-difluoro-2-nitrophenoxy) propanoate (2 g, 5.881 mmol, 1 equiv) in EtOH (10 mL) and H2O (2.5 mL) was added Fe (3.28 g, 58.810 mmol, 10 equiv) and NH4Cl (3.15 g, 58.810 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 70 °C. The resulting mixture was filtered, the filter cake was washed with EtOH. The filtrate was concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (2R)-8-bromo-5,6-difluoro-2-methyl-2,4-dihydro-1,4- benzoxazin-3-one (1.35 g, 82.56%) as a yellow semi-solid. LC/MS: mass calcd. for C9H6BrF2NO2: 276.95 m/z found: 277.95, 279.95 [M+H, M+H+2]+
Figure imgf000489_0001
(R)-8-bromo-5,6-difluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-8-bromo-5,6-difluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (1000 mg, 3.60 mmol, 1 equiv) and KNO3 (727.23 mg, 7.20 mmol, 2 equiv) in H2SO4 (20 mL) was stirred for 12h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford (2R)-8-bromo-5,6-difluoro-2-methyl-2,4-dihydro-1,4- benzoxazin-3-one (1.35 g, 82.56%) as a yellow semi-solid. LC/MS: mass calcd. for C9H5BrF2N2O4: 321.94 m/z found: 322.94, 324.94 [M+H, M+H+2]+
Figure imgf000489_0002
(R)-8-bromo-4-(3-(difluoromethyl)benzyl)-5,6-difluoro-2-methyl-7-nitro-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-8-bromo-5,6-difluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (390 mg, 1.207 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-3-(difluoromethyl) benzene (266.86 mg, 1.207 mmol, 1 equiv) and K2CO3 (333.69 mg, 2.414 mmol, 2 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 50 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (2R)-8-bromo-4-{[3- (difluoromethyl)phenyl]methyl}-5,6-difluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (260 mg, 39.52%) as yellow oil. LC/MS: mass calcd. for C17H11BrF4N2O4: 461.98 m/z found: 462.98, 464.98 [M+H, M+H+2]+.
Figure imgf000490_0001
(R)-7-amino-4-(3-(difluoromethyl)benzyl)-5,6-difluoro-2-methyl-2H-benzo[b][1,4]oxazin- 3(4H)-one A mixture of (2R)-8-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-5,6-difluoro-2-methyl-7- nitro-2H-1,4-benzoxazin-3-one (250 mg, 0.540 mmol, 1 equiv) and Pd/C (574.40 mg, 5.400 mmol, 10 equiv) in MeOH (5 mL) was stirred for 1h at rt under H2 atmosphere. The resulting mixture was filtered, the filter cake was washed with MeOH (20 mL). The filtrate was concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (2R)-7-amino-4-{[3-(difluoromethyl) phenyl]methyl}-5,6-difluoro-2-methyl- 2H-1,4-benzoxazin-3-one (100 mg, 44.45%) as a yellow solid. LC/MS: mass calcd. for C17H14F4N2O2: 354.10 m/z found: 355.10 [M+H]+.
Figure imgf000490_0002
(R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl) benzyl)-5,6-difluoro-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4] oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-5,6-difluoro-2-methyl-2H- 1,4-benzoxazin-3-one (90 mg, 0.254 mmol, 1 equiv) in DCM (5 mL) was added TEA (77.11 mg, 0.762 mmol, 3 equiv) and 2-isocyanato-2-methylpropane (76.2 mg, 0.762 mmol, 3 equiv). The resulting mixture was stirred for 16 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 53% B in 9 min, 53% B; Wave Length: 254/220 nm; RT1(min): 8.52; Number Of Runs: 0) to afford 3-tert-butyl-1-[(2R)-4-{[3- (difluoromethyl)phenyl]methyl}-5,6-difluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (4.7 mg, 4.02%) as a white solid. LC/MS: mass calcd. for C22H23F4N3O3: 453.17 m/z found: 452.10 [M-H]-.1H NMR (300 MHz, Chloroform-d) δ 7.68 (dd, J = 7.0, 2.3 Hz, 1H), 7.34 – 7.44 (m, 2H), 7.31 (s, 1H), 6.62 (s, 1H), 6.26 (s, 1H), 5.35 (d, J = 15.6 Hz, 1H), 5.18 (d, J = 15.5 Hz, 1H), 4.65 (s, 1H), 4.60 (q, J = 6.7 Hz, 2H), 1.59 (d, J = 6.8 Hz, 3H), 1.40 (s, 9H). 19F-NMR (282 MHz, CDCl3) δ -110.63, -147.41, -161.57. Example 341: 1-(4-benzyl-6-cyano-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-3-(tert- butyl)urea Synthetic Scheme
Figure imgf000491_0001
methyl (R)-2-(4-cyano-2-nitrophenoxy) propanoate To a stirred solution of 4-hydroxy-3-nitrobenzonitrile (2.5 g, 15.233 mmol, 1 equiv) and methyl (2S)-2-hydroxypropanoate (1.59 g, 15.233 mmol, 1 equiv) in THF (20 mL) was added PPh3 (5.99 g, 22.849 mmol, 1.5 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 0.5 h at room temperature under nitrogen atmosphere. Then, to the above solution was added DEAD (3.98 g, 22.849 mmol, 1.5 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 2 h at room temperature under nitrogen atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford methyl (R)-2- (4-cyano-2-nitrophenoxy) propanoate (3 g, 74.78%) as a yellow green oil. LC/MS: mass calcd for C11H10N2O5:250.06, found: 251.05 [M+H]+.
Figure imgf000492_0001
(R)-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b] [1,4]oxazine-6-carbonitrile To a stirred solution of methyl (R)-2-(4-cyano-2-nitrophenoxy) propanoate (3 g, 11.990 mmol, 1 equiv) and NH4Cl (6413.41 mg, 119.900 mmol, 10 equiv) in EtOH/H2O (4:1, 40 mL) was added Fe (6695.78 mg, 119.900 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. Desired product could be detected by LCMS. After cooling down to rt, the reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-2-methyl- 3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (1.6 g, 49.64%) as a yellow oil. LC/MS: mass calcd for C10H8N2O2 ; 188.06, found: 189.05 [M+H]+.
Figure imgf000492_0002
(R)-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4]oxazine-6-carbonitrile To a stirred solution of (R)-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carbonitrile (1.6 g, 8.502 mmol, 1 equiv) in H2SO4 (20 mL, 375.242 mmol, 44.13 equiv) was added KNO3 (859.59 mg, 8.502 mmol, 1 equiv) in portions at 0 oC under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. Desired product could be detected by LCMS. Pour the reaction into ice/water. The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (313 mg, 15.00%) as a yellow oil. LC/MS: mass calcd for C10H7N3O4: 233.04, found: 234.05 [M+H]+ .
Figure imgf000493_0001
(R)-4-benzyl-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4]oxazine-6-carbonitrile To a stirred solution of (R)-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6- carbonitrile (150 mg, 0.643 mmol, 1 equiv) and benzyl bromide (165.03 mg, 0.965 mmol, 1.5 equiv) in DMF (5 mL) was added K2CO3 (268.66 mg, 1.929 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)- 4-benzyl-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (124 mg, 56.64%) as a yellow solid. LC/MS: mass calcd for C17H13N3O4: 323.09, found: 324.01 [M+H]+ .
Figure imgf000493_0002
(R)-7-amino-4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b] [1,4]oxazine-6-carbonitrile To a stirred solution of (R)-4-benzyl-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (60 mg, 0.186 mmol, 1 equiv) and NH4Cl (99.27 mg, 1.860 mmol, 10 equiv) in MeOH/H2O (10:1, 5 mL) was added Fe (103.64 mg, 1.860 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. Desired product could be detected by LCMS. After cooling down to rt, the reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-7-amino-4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b] [1,4]oxazine-6-carbonitrile (40 mg, 58.78%) as a yellow oil. LC/MS: mass calcd for C17H15N3O2: 293.12, found:294.05 [M+H]+.
Figure imgf000494_0001
(R)-1-(4-benzyl-6-cyano-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4] oxazin-7-yl)-3-(tert- butyl)urea To a stirred solution of (R)-7-amino-4-benzyl-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (35 mg, 0.119 mmol, 1 equiv) and 2-isocyanato-2-methylpropane (35.49 mg, 0.357 mmol, 3 equiv) in DCM (3 mL) was added TEA (36.22 mg, 0.357 mmol, 3 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 4 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The resulting mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product (30 mg) was purified by Prep-HPLC with the following conditions (Column: XBridge Prep C18 OBD Column, 19*250 mm, 5μm; Mobile Phase A: Water (10mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min mL/min; Gradient: 50% B to 80% B in7min; Wave Length: 254nm nm; RT1(min): 5.9; Number Of Runs: 2) to afford (R)-1-(4-benzyl-6-cyano-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b] [1,4] oxazin-7-yl)-3-(tert-butyl)urea (23.2 mg, 49.12%) as a white solid. LC/MS: mass calcd for C22H24N4O3: 392.18, found: 391.00 [M-H]-.1H NMR (400 MHz, DMSO- d6) δ 8.28 (s, 1H), 7.83 (s, 1H), 7.20 – 7.49 (m, 6H), 6.97 (s, 1H), 5.20(s, 2H), 4.10 – 5.08 (m, 1H), 1.45 – 1.61 (m, 3H), 1.28-1.38 (m, 9H). Example 342: (R)-1-(tert-butyl)-3-(6-cyano-4-(3-(difluoromethyl)benzyl)-2-methyl-3-oxo- 3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000495_0001
(R)-4-(3-(difluoromethyl) benzyl)-2-methyl-7-nitro-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazine-6-carbonitrile To a stirred solution of (2R)-2-methyl-7-nitro-3-oxo-2,4-dihydro-1,4-benzoxazine-6-carbonitrile (91.35 mg, 0.392 mmol, 1 equiv) and 1-(bromomethyl)-3-(difluoromethyl) benzene (129.89 mg, 0.588 mmol, 1.5 equiv) in DMF (4 mL) was added K2CO3 (163.61 mg, 1.176 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-4-(3-(difluoromethyl) benzyl) -2-methyl-7-nitro-3-oxo-3,4-dihydro-2H- benzo[b] [1,4] oxazine-6-carbonitrile (51 mg, 34.87%) as a yellow oil. LC/MS: mass calcd for C18H13F2N3O4: 373.09, found 374.05 [M+H]+.
Figure imgf000495_0002
(R)-7-amino-4-(3-(difluoromethyl) benzyl)-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazine-6-carbonitrile To a stirred mixture of (R)-4-(3-(difluoromethyl) benzyl) -2-methyl-7-nitro-3-oxo-3,4-dihydro- 2H-benzo[b][1,4]oxazine-6-carbonitrile (50 mg, 0.134 mmol, 1 equiv) and Fe (74.80 mg, 1.340 mmol, 10 equiv) in MeOH/H2O (10:1, 3 mL) was added NH4Cl (71.64 mg, 1.340 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. Desired product could be detected by LCMS. After cooling down to rt, the reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-7-amino-4-(3-(difluoromethyl) benzyl)-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b] [1,4] oxazine-6-carbonitrile (32 mg, 69.59%) as a yellow oil. LC/MS: mass calcd. for C18H15F2N3O2: 343.11, found: 344.15 [M+H]+ .
Figure imgf000496_0001
(R)-1-(tert-butyl)-3-(6-cyano-4-(3- (difluoromethyl)benzyl)-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4] oxazin-7-yl)urea To a stirred solution of (R)-7-amino-4-(3-(difluoromethyl) benzyl)-2-methyl-3-oxo-3,4-dihydro- 2H-benzo[b] [1,4] oxazine-6-carbonitrile (25 mg, 0.081 mmol, 1 equiv) and 2-isocyanato-2- methylpropane (24.19 mg, 0.243 mmol, 3 equiv) in DCM (2 mL) was added TEA (24.69 mg, 0.243 mmol, 3 equiv) and in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 4 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The resulting mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product (20 mg ) was purified by Prep-HPLC with the following conditions (Column: XBridge Prep C18 OBD Column, 19*250 mm, 5μm; Mobile Phase A: Water (10mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min mL/min; Gradient: 50% B to 80% B in 7min ; Wave Length: 254 nm ; RT1(min): 5.9; Number of Runs: 2) to afford (R)-1-(tert-butyl)-3- (6-cyano-4-(3-(difluoromethyl) benzyl)-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b] [1,4] oxazin-7- yl) urea (10 mg, 30.04%) as a white solid. LC/MS: mass calcd for C23H24F2N4O3 : 442.18, found: 441.05 [M-H]-.1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 7.84 (s, 1H), 7.45 – 7.61 (m, 3H), 7.38 - 7.46(m, 2H), 6.82 - 7.20(d, J = 10.7 Hz,2H), 5.21 (s, 2H), 4.93 - 5.08 (q, J = 6.7 Hz, 1H), 1.48 - 1.60(d, J = 6.8 Hz, 3H), 1.49 (s, 9H).19F NMR (376 MHz, DMSO) δ -109.72. Example 343: (S)-1-(tert-butyl)-3-(2-oxo-1-(1-phenylethyl)-7-(trifluoromethyl)-1,2,3,4- tetrahydroquinolin-6-yl) urea Synthetic Scheme
Figure imgf000497_0001
7-(trifluoromethyl)-3,4-dihydro-1H-quinolin-2-one To a stirred solution of 2-[2-chloro-4-(trifluoromethyl) phenyl]-4,4,5,5-tetramethyl-1,3,2- dioxaborolane (1 g, 3.262 mmol, 1 equiv) and polyacrylamide (0.23 g, 3.262 mmol, 1 equiv), XPhos (0.62 g, 1.305 mmol, 0.4 equiv) in t-AmOH (10 mL)/MeOH (1 mL) were added K3PO4 (1.52 g, 7.176 mmol, 2.2 equiv), [Rh(COD)Cl]2 (0.16 g, 0.326 mmol, 0.1 equiv) and [PdCl(allyl)]2 (0.12 g, 0.326 mmol, 0.1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 110 °C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 7-(trifluoromethyl)-3,4-dihydro-1H- quinolin-2-one (610 mg, 86.90%) as a yellow oil. MS (ESI): mass calcd. For C10H8F3NO: 215.06 m/z, found 216.20 [M +H]+.
Figure imgf000497_0002
6-nitro-7-(trifluoromethyl)-3,4-dihydro-1H-quinolin-2-one A solution of 7-(trifluoromethyl)-3,4-dihydro-1H-quinolin-2-one (580 mg, 2.695 mmol, 1 equiv) and KNO3 (272.52 mg, 2.695 mmol, 1 equiv) in H2SO4 (5 mL) was stirred for 2 h at room temperature under air atmosphere. The reaction was quenched with water/ice at 0°C. The precipitated solids were collected by filtration and washed with H2O (3x20 mL). The crude product was used in the next step directly without further purification. This resulted in 6-nitro-7- (trifluoromethyl)-3,4-dihydro-1H-quinolin-2-one (570 mg, 81.28%) as a yellow solid.
Figure imgf000498_0001
6-nitro-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-2-one To a stirred solution of 6-nitro-7-(trifluoromethyl)-3,4-dihydro-1H-quinolin-2-one (200 mg, 0.769 mmol, 1 equiv) and 1-phenyl-ethanol (93.91 mg, 0.769 mmol, 1 equiv) in DCM (5 mL) was added PPh3 (302.44 mg, 1.153 mmol, 1.5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (200.82 mg, 1.153 mmol, 1.5 equiv) dropwise over 1 min at 0 °C. The resulting mixture was stirred for additional 1 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 40 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-nitro-1- [(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-2-one (190 mg, 67.84%) as a yellow oil. MS (ESI): mass calcd. for C18H15F3N2O3: 364.10 m/z, found 365.05 [M+H]+.
Figure imgf000498_0002
6-amino-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-2-one To a stirred solution of 6-nitro-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin- 2-one (190 mg, 0.522 mmol, 1 equiv) and Fe (291.24 mg, 5.220 mmol, 10 equiv) in MeOH (5 mL)/H2O (0.5 mL) was added NH4Cl (278.96 mg, 5.220 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at 80 °C under air atmosphere. The mixture was allowed to cool down to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-amino-1-[(1S)-1-phenylethyl]-7- (trifluoromethyl)-3,4-dihydroquinolin-2-one (50 mg, 28.68%) as a yellow oil. MS (ESI): mass calcd. for C18H17F3N2O: 334.13 m/z, found 335.25 [M+H]+.
Figure imgf000499_0001
3-tert-butyl-1-{2-oxo-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-6-yl} urea A solution of 6-amino-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-2-one (30 mg, 0.090 mmol, 1 equiv) in DCM (5 mL) was treated with triphosgene (26.89 mg, 0.090 mmol, 1 equiv) for 1 h at 0°C under nitrogen atmosphere followed by the addition of TEA (45.40 mg, 0.450 mmol, 5 equiv) dropwise at 0°C. The resulting mixture was stirred for 1 h at room temperature under nitrogen atmosphere. To the above mixture was added erbumine (19.69 mg, 0.270 mmol, 3 equiv) dropwise over 1 min at room temperature. The resulting mixture was stirred for additional 1 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm column (eluent: 42% to 54% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.1%NH3.H2O) to afford 3-tert-butyl-1-{2-oxo-1-[(1S)-1- phenylethyl]-7-(trifluoromethyl)-3,4-dihydroquinolin-6-yl} urea (18.8 mg, 48.14%) as a white solid. MS (ESI): mass calcd. for C23H26F3N3O2: 433.20 m/z, found 434.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 7.76 (s, 1H), 7.49 (s, 1H), 7.21 – 7.45 (m, 5H), 7.75 – 7.89 (m, 1H), 6.62 – 7.75 (m, 1H), 6.28 (q, J = 7.0 Hz, 1H), 2.93 (d, J = 6.7 Hz, 2H), 2.62 – 2.75 (m, 2H), 1.62 – 1.85 (m, 3H), 1.15 – 1.35 (m, 9H).19F NMR (282 MHz, DMSO) δ -59.825. Example 344: (S)-1-(tert-butyl)-3-(7-chloro-2-oxo-1-(1-phenylethyl)-1,2-dihydroquinoxalin- 6-yl)urea Synthetic Scheme
Figure imgf000500_0001
7-chloro-6-nitroquinoxalin-2(1H)-one To a solution of 7-chloro-1H-quinoxalin-2-one (1 g, 5.537 mmol, 1 equiv) in H2SO4 (10 mL) was added KNO3 (0.56 g, 5.537 mmol, 1 equiv) at 0°C. The reaction was stirred at 0 °C for 0.5h. Then, the resulting mixture was stirred for 1h at 0°C. The reaction was quenched with water/Ice at 0°C. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-chloro-6-nitro-1H- quinoxalin-2-one (300 mg, 24.02%) as a yellow solid. LC/MS: mass calcd. for C8H4ClN3O3:224.99 m/z, found:226.00 [M+H]+.
Figure imgf000500_0002
(S)-7-chloro-6-nitro-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 7-chloro-6-nitro-1H-quinoxalin-2-one (300 mg, 1.330 mmol, 1 equiv) in DCM (10 mL) was added (R)-1-phenylethan-1-ol (162.46 mg, 1.330 mmol, 1 equiv) and PPh3 (523.21 mg, 1.995 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (347.40 mg, 1.995 mmol, 1.5 equiv) at 0°C under N2. The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give 7-chloro-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (70 mg, 15.96%) as a yellow solid. LC/MS: mass calcd. for C16H12ClN3O3:329.06 m/z, found:330.10 [M+H]+.
Figure imgf000501_0001
(S)-6-amino-7-chloro-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 7-chloro-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (60 mg, 0.182 mmol, 1 equiv) in MeOH (5 mL) and H2O (0.5 mL) was added Fe (101.62 mg, 1.820 mmol, 10 equiv) and NH4Cl (97.33 mg, 1.820 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 30 mL). The filtrate was concentrated under reduced pressure. Diluted with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-chloro-1-[(1S)-1-phenylethyl]quinoxalin-2- one (40 mg, 73.33%) as a yellow solid. LC/MS: mass calcd. for C16H14ClN3O:299.08 m/z, found:300.00 [M+H]+.
Figure imgf000501_0002
(S)-1-(tert-butyl)-3-(7-chloro-2-oxo-1-(1-phenylethyl)-1,2-dihydroquinoxalin-6-yl)urea To a solution of 6-amino-7-chloro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (30 mg, 0.100 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (29.70 mg, 0.100 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (30.38 mg, 0.300 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (31.32 mg, 0.429 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 38% to 68% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.05%NH3.H2O)) to afford the title compound 3-tert-butyl-1-{7-chloro-2-oxo-1-[(1S)- 1-phenylethyl]quinoxalin-6-yl}urea (15.7 mg, 38.63%) as a yellow solid. LC/MS: mass calcd. for C21H23ClN4O2:398.15 m/z, found:399.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.27 (s, 1H), 7.97 (s, 1H), 7.33 – 7.41 (m, 2H), 7.25 – 7.33 (m, 3H), 7.23 (s, 1H), 6.94 (s, 1H), 6.53 (d, J = 7.5 Hz, 1H), 1.87 (d, J = 7.1 Hz, 3H), 1.30 (s, 9H). Example 345: 1-(1-benzyl-5,7-difluoro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl) urea Synthetic Scheme
Figure imgf000502_0001
ethyl (2E)-3-(2,4-difluoro-6-nitrophenyl) prop-2-enoate To a stirred solution of 2-bromo-1,5-difluoro-3-nitrobenzene (2 g, 8.404 mmol, 1 equiv) and ethyl (2E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) prop-2-enoate (2.28 g, 10.085 mmol, 1.2 equiv) in 1,4-dioxane (50 mL)/H2O (5 mL) was added Pd(dppf)Cl2 (0.68 g, 0.840 mmol, 0.1 equiv) and K2CO3 (2.32 g, 16.808 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 90 °C under nitrogen atmosphere. The mixture was allowed to cool down to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford ethyl (2E)-3-(2,4-difluoro-6-nitrophenyl) prop-2-enoate (1.81 g, 83.74%) as a white oil. MS (ESI): mass calcd. For C11H9F2NO4: 257.05 m/z, found 258.15[M +H]+.
Figure imgf000503_0001
ethyl 3-(2-amino-4,6-difluorophenyl) propanoate A mixture of ethyl (2E)-3-(2,4-difluoro-6-nitrophenyl) prop-2-enoate (1.81 g, 7.038 mmol, 1 equiv) and Pd/C (600 mg, 5.638 mmol, 0.80 equiv) in MeOH (10 mL) was stirred for 1 h at room temperature under hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with MeOH (5 x 20 mL). The filtrate was concentrated under reduced pressure. The crude product was used in the next step directly without further purification. This resulted in ethyl 3-(2-amino-4,6-difluorophenyl) propanoate (1.5 g, 92.98%) as a black oil. MS (ESI): mass calcd. For C11H13F2NO2: 229.09 m/z, found 230.20[M +H]+.
Figure imgf000503_0002
5,7-difluoro-3,4-dihydro-1H-quinolin-2-one A solution of ethyl 3-(2-amino-4,6-difluorophenyl) propanoate (1.57 g, 6.849 mmol, 1 equiv) in AcOH (20 mL) was stirred for 1 h at 90 oC under air atmosphere. The resulting mixture was concentrated under reduced pressure. The mixture was neutralized to pH = 7 with saturated NaHCO3 (aq.). The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 5,7-difluoro-3,4-dihydro-1H-quinolin-2-one (1.1 g, 87.69%) as a yellow oil. MS (ESI): mass calcd. for C9H7F2NO: 183.05 m/z, found 184.25 [M+H]+.
Figure imgf000504_0001
5,7-difluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one A solution of 5,7-difluoro-3,4-dihydro-1H-quinolin-2-one (800 mg, 4.368 mmol, 1 equiv) and KNO3 (441.59 mg, 4.368 mmol, 1 equiv) in H2SO4 (10 mL) was stirred for 1 h at room temperature under air atmosphere. The reaction was quenched with water/ice at 0 °C. The precipitated solids were collected by filtration and washed with H2O (3x20 mL). The crude product was used in the next step directly without further purification. This resulted in 5,7- difluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (790 mg, 79.27%) as a yellow oil. MS (ESI): mass calcd. for C9H6F2N2O3: 228.03 m/z, found 229.15 [M+H]+.
Figure imgf000504_0002
1-benzyl-5,7-difluoro-6-nitro-3,4-dihydroquinolin-2-one To a stirred solution of 5,7-difluoro-6-nitro-3,4-dihydro-1H-quinolin-2-one (500 mg, 2.191 mmol, 1 equiv) and BnBr (374.83 mg, 2.191 mmol, 1 equiv) in DMF (5 mL) was added K2CO3 (605.75 mg, 4.382 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at room temperature under air atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 1-benzyl-5,7-difluoro-6-nitro-3,4- dihydroquinolin-2-one (200 mg, 28.67%) as a yellow oil. MS (ESI): mass calcd. for C16H12F2N2O3: 318.08 m/z, found 319.20 [M+H]+.
Figure imgf000505_0001
6-amino-1-benzyl-5,7-difluoro-3,4-dihydroquinolin-2-one To a stirred solution of 1-benzyl-5,7-difluoro-6-nitro-3,4-dihydroquinolin-2-one (150 mg, 0.471 mmol, 1 equiv) and Fe (263.19 mg, 4.710 mmol, 10 equiv) in MeOH (5 mL)/H2O (0.5 mL) was added NH4Cl (252.09 mg, 4.710 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 h at 80 °C under air atmosphere. The mixture was allowed to cool down to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 6-amino-1-benzyl-5,7-difluoro-3,4-dihydroquinolin-2-one (120 mg, 88.32%) as a yellow solid. MS (ESI): mass calcd. for C16H14F2N2O: 288.11 m/z, found 289.25 [M+H]+.
Figure imgf000505_0002
1-(1-benzyl-5,7-difluoro-2-oxo-3,4-dihydroquinolin-6-yl)-3-tert-butylurea A solution of 6-amino-1-benzyl-5,7-difluoro-3,4-dihydroquinolin-2-one (110 mg, 0.382 mmol, 1 equiv) in DCM (5 mL) was treated with triphosgene (113.22 mg, 0.382 mmol, 1 equiv) for 1 h at 0 °C under nitrogen atmosphere followed by the addition of TEA (193.05 mg, 1.910 mmol, 5 equiv) dropwise at 0 °C. The resulting mixture was stirred for 1 h at room temperature under nitrogen atmosphere. To the above mixture was added erbumine (67.66 mg, 1.146 mmol, 3 equiv) dropwise over 1 min at room temperature. The resulting mixture was stirred for additional 1 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm column (eluent: 40% to 50% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.1%NH3.H2O) to afford 1-(1-benzyl-5,7-difluoro-2-oxo-3,4- dihydroquinolin-6-yl)-3-tert-butylurea (52.7 mg, 35.36%) as a white solid. MS (ESI): mass calcd. for C21H23F2N3O2: 387.18 m/z, found 388.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 7.40 (s, 1H), 7.28 – 7.38 (m, 2H), 7.24 (td, J = 4.9, 2.8 Hz, 3H), 6.72 (dd, J = 12.0, 1.8 Hz, 1H), 6.11 (s, 1H), 5.16 (s, 2H), 2.91 (t, J = 7.4 Hz, 2H), 2.73 (dd, J = 8.7, 6.1 Hz, 2H), 1.20 – 1.30 (m, 9H).19F NMR (282 MHz, DMSO) δ -120.687, -123.203. Example 346: 3-tert-butyl-1-{2-oxo-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-6- yl}urea Synthetic Scheme
Figure imgf000506_0001
Synthesis of 7-(trifluoromethyl)-1H-quinoxalin-2-one To a stirred solution of 4-(trifluoromethyl)benzene-1,2-diamine (2 g, 11.354 mmol, 1 equiv) in EtOH(20 mL) was added ethyl glyoxylate (1.16 g, 11.354 mmol, 1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at 60 ℃ under air atmosphere.The reaction was monitored by LCMS. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 7-(trifluoromethyl)-1H-quinoxalin-2- one (1 g, 41.13%) as a yellow solid. LC/MS (ESI): mass calcd. for C9H5F3N2O:214.04 m/z, found:215.0 [M+H]+.
Figure imgf000507_0001
Synthesis of 6-nitro-7-(trifluoromethyl)-1H-quinoxalin-2-one To a stirred solution of 7-(trifluoromethyl)-1H-quinoxalin-2-one (1 g, 4.670 mmol, 1 equiv) in H2SO4 (10 mL) was added KNO3 (1.42 g, 14.010 mmol, 3 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was monitored by LCMS. The reaction was quenched with water/Ice at 0 °C. The precipitated solids were collected by filtration and washed with H2O (3x20 mL). The crude product was used in the next step directly without further purification. This resulted in 6-nitro-7- (trifluoromethyl)-1H-quinoxalin-2-one (600 mg, 49.58%) as a brown solid.LC/MS (ESI): mass calcd. for C9H4F3N3O3:259.02 m/z, found:260.0 [M+H]+.
Figure imgf000507_0002
Synthesis of 6-nitro-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-2-one To a stirred solution of 6-nitro-7-(trifluoromethyl)-1H-quinoxalin-2-one (590 mg, 2.277 mmol, 1 equiv) and 1-phenyl-ethanol (278.14 mg, 2.277 mmol, 1 equiv) in DCM (6 mL) was added PPh3 (895.75 mg, 3.416 mmol, 1.5 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 15 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (594.76 mg, 3.416 mmol, 1.5 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 2 h at room temperature. The reaction was monitored by LCMS. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-nitro-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-2-one (100 mg, 12.09%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H12F3N3O3:363.0 m/z, found:364.0 [M+H]+.
Figure imgf000508_0001
Synthesis of 6-amino-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-2-one To a stirred solution of 6-nitro-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-2-one (90 mg, 0.248 mmol, 1 equiv) in MeOH/H2O(10:1,1.1 mL) was added Fe (15.37 mg, 0.280 mmol, 10 equiv) and NH4Cl (132.51 mg, 2.480 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. The reaction was monitored by LCMS. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-1-[(1S)-1- phenylethyl]-7-(trifluoromethyl)quinoxalin-2-one (30 mg, 36.33%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H14F3N3O:333.1 m/z, found:334.1 [M+H]+.
Figure imgf000508_0002
Synthesis of 3-tert-butyl-1-{2-oxo-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)quinoxalin-6- yl}urea To a stirred solution of 6-amino-7-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (40 mg, 0.143 mmol, 1 equiv) in DCM(0.5 mL) was added TEA (22.77 mg, 0.225 mmol, 3 equiv) and 2- isocyanato-2-methylpropane (22.31 mg, 0.225 mmol, 3 equiv) at room temperature under air atmosphere. The resulting mixture was stirred for 1 days at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (5 mL) and extracted with DCM (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 42% to 72% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.05%NH3H2O) to afford 3-tert-butyl-1-{7-methyl-2-oxo-1- [(1S)-1-phenylethyl]quinoxalin-6-yl}urea (2.7 mg, 4.96%) as a yellow solid. LC/MS (ESI): mass calcd. for C22H23F3N4O2:432.2 m/z, found:433.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.34 (s, 1H), 7.65 (s, 1H), 7.37 – 7.31 (m, 6H), 6.90 (s, 1H), 6.62 (q, J = 7.2 Hz, 1H), 1.86 (d, J = 7.2 Hz, 3H), 1.28 (s, 9H).19F NMR (282 MHz, DMSO-d6) δ -60.29. Example 347: 3-tert-butyl-1-{7-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea Synthetic Scheme
Figure imgf000509_0001
7-methyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one To a stirred solution of 7-bromo-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (200 mg, 0.534 mmol, 1 equiv) and methylboronic acid (63.99 mg, 1.068 mmol, 2 equiv) in toluene/H2O(7:1, 2.4 mL) was added Pd(OAc)2 (24.00 mg, 0.107 mmol, 0.2 equiv), Butyldi-1-adamantylphosphine (1.92 mg, 0.005 mmol, 0.2 equiv) and Na2CO3 (169.95 mg, 1.602 mmol, 3 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at 90 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 7-methyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (100 mg, 60.49%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H15N3O3:309.1 m/z, found:310.0 [M+H]+.
Figure imgf000510_0001
6-amino-7-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-onee To a stirred solution of 7-methyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (90 mg, 0.291 mmol, 1 equiv) in MeOH/H2O(10:1, 1.1 mL) was added Fe (18.05 mg, 0.320 mmol, 10 equiv) and NH4Cl (155.63 mg, 2.910 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. The reaction was monitored by LCMS. After cooling down to rt, quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated.The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-7-methyl-1-[(1S)-1- phenylethyl]quinoxalin-2-one (70 mg, 86.13%) as a yellow solid.LC/MS (ESI): mass calcd. for C17H17N3O:279.14 m/z, found:280.1 [M+H]+.
Figure imgf000510_0002
3-tert-butyl-1-{7-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea To a stirred solution of 6-amino-7-methyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (80 mg, 0.286 mmol, 1 equiv) in DCM (1 mL) was added 2-isocyanato-2-methylpropane (85.17 mg, 0.858 mmol, 3 equiv) and TEA (86.94 mg, 0.858 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 days at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using XSelect CSH Fluoro Pheny 250 mm x 19 mm x 5 μm column (eluent: 38% to 68% (v/v) CH3CN and H2O with 0.1% FA) to afford 3-tert-butyl-1-{7-methyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea (4.1 mg, 3.76%) as a yellow solid.LC/MS (ESI): mass calcd. for C22H26N4O2:378.2 m/z, found:379.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.34 (s, 1H), 8.18 (s, 1H), 7.56 (s, 1H), 7.37 – 7.24 (m, 5H), 7.07 (s, 1H), 6.55 (d, J = 8.4 Hz, 2H), 2.12 (s, 3H), 1.89 (d, J = 6.9 Hz, 3H), 1.29 (s, 9H). Example 348: 3-tert-butyl-1-{7-cyclopropyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6- yl}urea Synthetic Scheme
Figure imgf000511_0001
7-cyclopropyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one To a stirred solution of 7-bromo-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (200 mg, 0.534 mmol, 1 equiv) and cyclopropylboronic acid (229.56 mg, 2.670 mmol, 5 equiv) in toluene/H2O (7:1, 2.4 mL) were added Na2CO3 (169.95 mg, 1.602 mmol, 3 equiv), Pd(OAc)2 (24.00 mg, 0.107 mmol, 0.2 equiv) and Butyldi-1-adamantylphosphine (38.33 mg, 0.107 mmol, 0.2 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 3 h at 90 °C under nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 7-cyclopropyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one as a yellow solid. LC/MS (ESI): mass calcd. for C19H17N3O3:335.13 m/z, found:336.0 [M+H]+.
Figure imgf000512_0001
6-amino-7-cyclopropyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one To a stirred solution of 7-cyclopropyl-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (90 mg, 0.268 mmol, 1 equiv) in MeOH/H2O (10:1, 1.1 mL) was added Fe (16.65 mg, 0.300 mmol, 10 equiv) and NH4Cl (143.55 mg, 2.680 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (8 mL) and extracted with EA (3 x 8 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-7- cyclopropyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (80 mg, 97.62%) as a yellow solid. LC/MS (ESI): mass calcd. for C19H19N3O:305.15 m/z, found:306.05 [M+H]+.
Figure imgf000512_0002
Synthesis of 3-tert-butyl-1-{7-cyclopropyl-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea To a stirred solution of 6-amino-7-cyclopropyl-1-[(1S)-1-phenylethyl]quinoxalin-2-one (80 mg, 0.262 mmol, 1 equiv) and TEA (79.53 mg, 0.786 mmol, 3 equiv) in DCM (1 mL) was added 2- isocyanato-2-methylpropane (77.91 mg, 0.786 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 days at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (8 mL) and extracted with DCM (3 x 8 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep-HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 36% to 46% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.05%NH3H2O) to afford 3-tert-butyl-1-{7-cyclopropyl-2-oxo- 1-[(1S)-1-phenylethyl]quinoxalin-6-yl}urea (16.7 mg, 15.65%) as a yellow solid. LC/MS (ESI): mass calcd. for C24H28N4O2:404.2 m/z, found:405.2 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.32 (s, 1H), 8.21 (s, 1H), 7.71 (s, 1H), 7.40 – 7.36 (m, 2H), 7.35 – 7.26 (m, 3H), 6.64 (s, 2H), 6.52 (s, 1H), 1.84 (d, J = 7.2 Hz, 3H), 1.80 – 1.75 (m, 1H), 1.30 (s, 9H), 0.90 – 0.87 (m, 2H), 0.28 – 0.01 (m, 2H). Example 349: 1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-6- (trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000513_0001
(R)-7-bromo-2-methyl-4-((S)-1-phenylethyl)-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of (2R)-7-bromo-2-methyl-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin-3-one (350 mg, 1.129 mmol, 1 equiv) in DCM (10 mL) was added (R)-1-phenylethan-1-ol (137.90 mg, 1.129 mmol, 1 equiv) and PPh3 (444.10 mg, 1.694 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (294.88 mg, 1.694 mmol, 1.5 equiv) at 0°C under N2. The resulting mixture was stirred for additional 1h at rt. Quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-bromo-2-methyl-4-[(1S)-1-phenylethyl]-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (240 mg, 51.33%) as a yellow solid. LC/MS: mass calcd. for C18H15BrF3NO2:413.02 m/z, found:413.95, 415.95 [M+H, M+H+2]+.
Figure imgf000514_0001
tert-butyl ((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-6-(trifluoromethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)carbamate To a solution of (2R)-7-bromo-2-methyl-4-[(1S)-1-phenylethyl]-6-(trifluoromethyl)-2H-1,4- benzoxazin-3-one (260 mg, 0.628 mmol, 1 equiv) in dioxane (10 mL) was added tert-butyl carbamate (110.30 mg, 0.942 mmol, 1.5 equiv), Ruphos (29.29 mg, 0.063 mmol, 0.1 equiv), RuPhos Palladacycle Gen.3 (52.50 mg, 0.063 mmol, 0.1 equiv) and Cs2CO3 (613.53 mg, 1.884 mmol, 3 equiv). The reaction was stirred at 90°C under N2 for 1h. The mixture was allowed to cool down to rt. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give tert-butyl N- [(2R)-2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-6-(trifluoromethyl)-2H-1,4-benzoxazin-7- yl]carbamate (250 mg, 88.42%) as a yellow solid. LC/MS: mass calcd. for C23H25F3N2O4:450.18 m/z, found:451.15 [M+H]+.
Figure imgf000514_0002
(R)-7-amino-2-methyl-4-((S)-1-phenylethyl)-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin- 3(4H)-one To a solution of tert-butyl N-[(2R)-2-methyl-3-oxo-4-[(1S)-1-phenylethyl]-6-(trifluoromethyl)- 2H-1,4-benzoxazin-7-yl]carbamate (270 mg, 0.599 mmol, 1 equiv) in DCM (4 mL) was added TFA (1 mL) at 0 °C. The reaction was stirred at rt for 1h. Quenched with NaHCO3 solution (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-2-methyl-4-[(1S)-1-phenylethyl]-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (100 mg, 47.62%) as a yellow solid. LC/MS: mass calcd. for C18H17F3N2O2:350.12 m/z, found:351.05 [M+H]+.
Figure imgf000515_0001
1-(tert-butyl)-3-((R)-2-methyl-3-oxo-4-((S)-1-phenylethyl)-6-(trifluoromethyl)-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-2-methyl-4-[(1S)-1-phenylethyl]-6-(trifluoromethyl)-2H-1,4- benzoxazin-3-one (50 mg, 0.143 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (42.35 mg, 0.143 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (43.33 mg, 0.429 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (31.31 mg, 0.429 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XSelect CSH C18 Column, 19*250 mm, 5μm column (eluent: 58% to 88% (v/v) CH3CN and Water (0.1% FA)) to afford the title compound 3-tert-butyl-1-[(2R)-2-methyl-3-oxo- 4-[(1S)-1-phenylethyl]-6-(trifluoromethyl)-2H-1,4-benzoxazin-7-yl]urea (26.1 mg, 40.55%) as a white solid. LC/MS: mass calcd. for C23H26F3N3O3:449.19 m/z, found:450.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.69 (s, 1H), 7.54 (s, 1H), 7.35 – 7.43 (m, 2H), 7.29 (d, J = 7.3 Hz, 3H), 6.93 (s, 1H), 6.82 (s, 1H), 6.19 (q, J = 7.1 Hz, 1H), 4.92 (q, J = 6.8 Hz, 1H), 1.75 (d, J = 7.2 Hz, 3H), 1.50 (d, J = 6.8 Hz, 3H), 1.27 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -58.9959. Example 350: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-6- (trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000516_0001
(R)-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin- 3-one (100 mg, 0.340 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3- (difluoromethyl)benzene (112.72 mg, 0.510 mmol, 1.5 equiv) and K2CO3 (141.98 mg, 1.020 mmol, 3 equiv). The reaction was stirred at rt for 1h. The reaction was monitored by TLC. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-{[3- (difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H-1,4- benzoxazin-3-one (140 mg, 94.83%) as a yellow solid.
Figure imgf000516_0002
(R)-7-amino-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-6-(trifluoromethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-7-nitro-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (130 mg, 0.299 mmol, 1 equiv) in MeOH (5 mL) and H2O (0.5 mL) was added Fe (167.16 mg, 2.990 mmol, 10 equiv) and NH4Cl (160.12 mg, 2.990 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 30 mL). The filtrate was concentrated under reduced pressure. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (100 mg, 82.63%) as a yellow solid. LC/MS: mass calcd. for C18H14F6N2O2:404.10 m/z, found:405.05 [M+H]+.
Figure imgf000517_0001
(R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-6- (trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (150 mg, 0.371 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (110.09 mg, 0.371 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (112.63 mg, 1.113 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (81.40 mg, 1.113 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep C18 OBD Column, 19*250 mm, 5μm column (eluent: 49% to 79% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-tert-butyl-1-[(2R)-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2- methyl-3-oxo-6-(trifluoromethyl)-2H-1,4-benzoxazin-7-yl]urea (20.3 mg, 10.79%) as a white solid. LC/MS: mass calcd. for C23H23F6N3O3:503.16 m/z, found:504.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 7.31 – 7.40 (m, 2H), 7.23 – 7.31 (m, 3H), 7.06 (d, J = 1.6 Hz, 1H), 6.27 (s, 1H), 5.28 (d, J = 16.4 Hz, 1H), 5.21 (d, J = 16.4 Hz, 1H), 5.14 (q, J = 6.7 Hz, 1H), 1.59 (d, J = 6.7 Hz, 3H), 1.25 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -59.0147, -136.7360. Example 351: (S)-1-(tert-butyl)-3-(5-(hydroxymethyl)-2-oxo-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000518_0001
5-chloro-6-nitroquinoxalin-2(1H)-one To a solution of 5-chloro-1H-quinoxalin-2-one (1 g, 5.537 mmol, 1 equiv) in H2SO4 (20 mL) was added KNO3 (0.56 g, 5.537 mmol, 1 equiv) at 0 °C. The reaction was stirred at rt for 3h. The reaction was quenched by the addition of water/Ice at 0°C. Diluted with water (200 mL) and extracted with EA (3 x 200 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-chloro-6-nitro-1H-quinoxalin-2-one (610 mg, 48.83%) as a yellow solid. LC/MS: mass calcd. for C8H4ClN3O3:224.99 m/z, found:226.15 [M+H]+.
Figure imgf000519_0001
(S)-5-chloro-6-nitro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 5-chloro-6-nitro-1H-quinoxalin-2-one (700 mg, 3.103 mmol, 1 equiv) in DCM (10 mL) was added (1R)-1-[3-(trifluoromethoxy)phenyl]ethanol (639.72 mg, 3.103 mmol, 1 equiv) and PPh3 (1220.83 mg, 4.655 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (810.60 mg, 4.655 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Quenched with water (80 mL) and extracted with DCM (3 x 80 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-chloro-6-nitro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (240 mg, 18.69%) as a yellow solid. LC/MS: mass calcd. for C17H11ClF3N3O4:413.04 m/z, found:414.05 [M+H]+.
Figure imgf000519_0002
(S)-5-(hydroxymethyl)-6-nitro-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)- one To a solution of 5-chloro-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (160 mg, 0.387 mmol, 1 equiv) in dioxane (5 mL) was added X-phos Pd G2 (30.43 mg, 0.039 mmol, 0.1 equiv) and (tributylstannyl)methanol (248.34 mg, 0.774 mmol, 2 equiv). The reaction was stirred at 90°C under N2 for 1h. After cooling down to rt, the reaction was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-(hydroxymethyl)-6-nitro-1-[(1S)-1- [3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (70 mg, 44.22%) as a yellow solid. LC/MS: mass calcd. for C18H14F3N3O5:409.09 m/z, found:410.00 [M+H]+.
Figure imgf000520_0001
(S)-5-(((tert-butyldimethylsilyl)oxy)methyl)-6-nitro-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 5-(hydroxymethyl)-6-nitro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (45 mg, 0.110 mmol, 1 equiv) in DCM (2 mL) was added Imidazole (22.45 mg, 0.330 mmol, 3 equiv) and TBSCl (24.85 mg, 0.165 mmol, 1.5 equiv) at 0 °C. The reaction was stirred at rt for 3h. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 5-{[(tert-butyldimethylsilyl)oxy]methyl}-6-nitro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (45 mg, 78.18%) as a yellow solid. LC/MS: mass calcd. for C24H28F3N3O5Si:523.18 m/z, found:524.15 [M+H]+.
Figure imgf000520_0002
(S)-6-amino-5-(((tert-butyldimethylsilyl)oxy)methyl)-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)quinoxalin-2(1H)-one To a solution of 5-{[(tert-butyldimethylsilyl)oxy]methyl}-6-nitro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (30 mg, 0.057 mmol, 1 equiv) in MeOH (4 mL) and H2O (0.4 mL) was added Fe (32.00 mg, 0.570 mmol, 10 equiv) and NH4Cl (30.65 mg, 0.570 mmol, 10 equiv). The reaction was stirred at 80°C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 30 mL). The filtrate was concentrated under reduced pressure. Diluted with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-5-{[(tert-butyldimethylsilyl)oxy]methyl}-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (25 mg, 88.40%) as a yellow solid. LC/MS: mass calcd. for C24H30F3N3O3Si:493.20 m/z, found:494.15 [M+H]+.
Figure imgf000521_0001
(S)-1-(tert-butyl)-3-(5-(((tert-butyldimethylsilyl)oxy)methyl)-2-oxo-1-(1-(3- (trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6-yl)urea To a solution of 6-amino-5-{[(tert-butyldimethylsilyl)oxy]methyl}-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (20 mg, 0.041 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (12.02 mg, 0.041 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (12.30 mg, 0.123 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (8.89 mg, 0.123 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-tert-butyl- 1-(5-{[(tert-butyldimethylsilyl)oxy]methyl}-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl)urea (20 mg, 83.28%) as a yellow solid. LC/MS: mass calcd. for C29H39F3N4O4Si:592.27 m/z, found:615.10 [M+Na]+.
Figure imgf000521_0002
(S)-1-(tert-butyl)-3-(5-(hydroxymethyl)-2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)urea To a solution of 3-tert-butyl-1-(5-{[(tert-butyldimethylsilyl)oxy]methyl}-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl)urea (20 mg, 0.034 mmol, 1 equiv) in THF (1 mL) was added Et3N.3HF (8.16 mg, 0.051 mmol, 1.5 equiv) at 0 °C. The reaction was stirred at rt for 1h. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC Triart C18 ExRs Column, 20*250mm, 5μm column (eluent: 45% to 75% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-tert-butyl-1-[5-(hydroxymethyl)-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl]urea (9.3 mg, 57.33%) as a white solid. LC/MS: mass calcd. for C23H25F3N4O4:478.18 m/z, found:477.15 [M-H]-.1H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.90 – 8.10 (m, 2H), 7.47 (dd, J = 9.2, 7.2 Hz, 1H), 7.24 – 7.32 (m, 3H), 6.90 (s, 1H), 6.62 (s, 1H), 5.17 (s, 1H), 5.04 (dd, J = 11.7, 4.9 Hz, 1H), 4.96 (dd, J = 11.8, 5.0 Hz, 1H), 1.90 (d, J = 7.1 Hz, 3H), 1.28 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -56.6537. Example 352: 3-tert-butyl-1-{7-cyano-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl}urea
Figure imgf000522_0001
7-bromo-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one To a solution of 7-bromo-6-nitro-1H-quinoxalin-2-one (1 g, 3.703 mmol, 1 equiv) in DCM (15 mL) was added (2R,4E)-3-methylidene-5-(trifluoromethoxy)hex-4-en-2-ol (0.73 g, 3.703 mmol, 1 equiv) and PPh3 (1.46 g, 5.554 mmol, 1.5 equiv). After the mixture was stirred at rt for 1h, to the above mixture was added DEAD (0.64 g, 3.703 mmol, 1 equiv) dropwise at 0 ℃. The resulting mixture was stirred for additional for 2 h at 25 ℃. Quenched with water (100 mL) and extracted with DCM (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-bromo-6-nitro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (130 mg, 7.66%) as a yellow oil. MS (ESI): mass calcd. for C17H11BrF3N3O4:456.99 m/z, found:457.99, 459.99 [ M+H, M+H+2]+.
Figure imgf000523_0001
6-amino-7-bromo-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one To a solution of 7-bromo-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (110 mg, 0.240 mmol, 1 equiv) in MeOH (1 mL) and H2O (10 mL) was added Fe (134.07 mg, 2.400 mmol, 10 equiv) and NH4Cl (128.42 mg, 2.400 mmol, 10 equiv). The reaction was stirred at 70 ℃ for 2 h. The mixture was allowed to cool down to room temperature and quenched with water (50 mL). The resulting mixture was filtered; the filter cake was washed with EA (50 mL). The filtrate was extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-bromo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (50 mg, 48.64%) as a yellow solid. MS (ESI): mass calcd. for C17H13BrF3N3O2:427.01 m/z, found:428.01, 430.01 [M+H, M+H+2]+.
Figure imgf000523_0002
7-amino-3-oxo-4-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxaline-6-carbonitrile To a solution of 6-amino-7-bromo-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (60 mg, 0.140 mmol, 1 equiv) in dimethylformamide (4 mL) was added Zn(CN)2 (19.74 mg, 0.168 mmol, 1.2 equiv) and Ruphos (6.54 mg, 0.014 mmol, 0.1 equiv) RuPhos Pd G3 (11.72 mg, 0.014 mmol, 0.1 equiv). The reaction was stirred at 100 °C for 2 h under N2 atmosphere. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-3-oxo-4-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxaline-6- carbonitrile (15 mg, 28.60%) as a yellow oil. MS (ESI): mass calcd. for C18H13F3N4O2:374.10 m/z, found:375.10 [M+H]+.
Figure imgf000524_0001
3-tert-butyl-1-{7-cyano-2-oxo-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-6- yl}urea To a solution of 7-amino-3-oxo-4-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxaline-6- carbonitrile (15 mg, 0.040 mmol, 1 equiv) in DCM (2 mL) was added triphosgene (5.95 mg, 0.020 mmol, 0.5 equiv) at 0 °C under N2 atmosphere. The resulting mixture was stirred 2 h at 25 °C. Then TEA (16.22 mg, 0.159 mmol, 3 equiv) was added into above system at 0 °C. The resulting mixture was stirred 1 h at 25 °C. To the above mixture were added erbumine (4.40 mg, 0.060 mmol, 1.5 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (30 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD 19*250 mm, 5μm column (eluent: 45% to 75% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{7-cyano-2-oxo-1-[(1S)-1- [3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl}urea (4.1 mg, 21.08%) as a yellow solid. LC/MS (ESI): mass calcd. for C23H22F3N5O3:473.17 m/z, found:474.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.39 (s, 1H), 8.33 (s, 1H), 7.44 – 7.72 (m, 2H), 7.27 – 7.38 (m, 3H), 6.95 (s, 1H), 6.43 (m, 1H), 1.88 (d, J = 7.0 Hz, 3H), 1.32 (s, 9H). 19F NMR (376 MHz, DMSO) δ -56.71. Example 353: 3-tert-butyl-1-{5-fluoro-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl}urea Synthetic Scheme
Figure imgf000525_0001
5-fluoro-1H-quinoxalin-2-one To a stirred solution of 3-fluorobenzene-1,2-diamine (2 g, 15.856 mmol, 1 equiv) in EtOH (20 mL) was added ethyl glyoxylate (1.62 g, 15.856 mmol, 1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 5-fluoro-1H-quinoxalin-2-one (1 g, 38.4%) as a brown solid. LC/MS (ESI): mass calcd. for C8H5FN2O:164.04 m/z, found:165.0 [M+H]+.
Figure imgf000525_0002
5-fluoro-6-nitro-1H-quinoxalin-2-one To a stirred solution of 5-fluoro-1H-quinoxalin-2-one (1 g, 6.092 mmol, 1 equiv) in H2SO4 (10 mL) was added KNO3 (615.95 mg, 6.092 mmol, 1 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was monitored by LCMS. The reaction was quenched with water/Ice (50 mL) at 0 °C, And extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 5-fluoro-6-nitro-1H-quinoxalin-2-one (400 mg, 31.39%) as a brown solid. LC/MS (ESI): mass calcd. for C8H4FN3O3:209.0 m/z, found:210.0 [M+H]+.
Figure imgf000526_0001
5-fluoro-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one To a stirred solution of 5-fluoro-6-nitro-1H-quinoxalin-2-one (400 mg, 1.913 mmol, 1 equiv) and (1R)-1-[3-(trifluoromethoxy)phenyl]ethanol (394.32 mg, 1.913 mmol, 1 equiv) in DCM (4 mL) was added PPh3 (752.50 mg, 2.869 mmol, 1.5 equiv) in portions at rt under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (333.10 mg, 1.913 mmol, 1 equiv) in portions at 0 °C. The resulting mixture was stirred for 2 h at room temperature under nitrogen atmosphere. The reaction was monitored by LCMS. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 5-fluoro-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin- 2-one (80 mg, 10.53%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H11F4N3O4:397.1 m/z, found:398.0 [M+H]+.
Figure imgf000526_0002
6-amino-5-fluoro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one To a stirred solution of 5-fluoro-6-nitro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin- 2-one (80 mg, 0.201 mmol, 1 equiv) in MeOH/H2O(10:1, 1.1 mL) were added NH4Cl (107.71 mg, 2.010 mmol, 10 equiv) and Fe (14.06 mg, 0.250 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-5-fluoro-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (30 mg, 40.56%) as a yellow solid.LC/MS (ESI): mass calcd. for C17H13F4N3O2:367.1 m/z, found:367.95 [M+H]+.
Figure imgf000527_0001
3-tert-butyl-1-{5-fluoro-2-oxo-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-6- yl}urea To a stirred solution of 6-amino-5-fluoro-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (20 mg, 0.054 mmol, 1 equiv) in DCM (1 mL) were added 2-isocyanato-2-methylpropane (16.19 mg, 0.162 mmol, 3 equiv) and TEA (16.53 mg, 0.162 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 days at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (5 mL) and extracted with DCM (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18250 mm x 19 mm x 5 μm column (eluent: 45% to 75% (v/v) CH3CN and H2O with 10mmol/L NH4HCO30.1%NH3.H2O) to afford 3-tert-butyl-1-{5-fluoro-2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl}urea (2.3 mg, 9.03%) as a yellow solid. LC/MS (ESI): mass calcd. for C22H22F4N4O3:466.2 m/z, found:467.2 [M+H]+. 1H NMR (300 MHz, Chloroform-d) δ 8.17 (s, 1H), 7.65 (d, J = 15.3 Hz, 1H), 7.26 – 7.23 (m, 2H), 7.03 – 6.98 (m, 3H), 6.50 (s, 1H), 6.19 (s, 1H), 1.88 – 1.85 (m, 3H), 1.32 (s, 9H).19F NMR (282 MHz, Chloroform-d) δ -57.75. Example 354: (R)-1-(4-benzyl-8-fluoro-2-methyl-3-oxo-6-(trifluoromethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000528_0001
2-fluoro-6-nitro-4-(trifluoromethyl)phenol To a solution of 2-fluoro-4-(trifluoromethyl)phenol (2 g, 11.105 mmol, 1 equiv) in DCE (20 mL) was added H2SO4 (20 mL) at -10°C. And to the above mixture was added KNO3 (1.12 g, 11.105 mmol, 1 equiv) at -10°C. The resulting mixture was stirred for additional 30 min at -10 °C. The reaction was quenched by the addition of water/Ice at 0 °C. Quenched with water (400 mL) and extracted with EA (3 x 400 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. This resulted in 2-fluoro-6-nitro-4-(trifluoromethyl)phenol (2.14 g, 85.61%) as a yellow oil. LC/MS: mass calcd. for C7H3F4NO3:225.00 m/z, found:223.95 [M-H]-.
Figure imgf000528_0002
methyl (R)-2-(2-fluoro-6-nitro-4-(trifluoromethyl)phenoxy)propanoate To a solution of 2-fluoro-6-nitro-4-(trifluoromethyl)phenol (1.4 g, 6.219 mmol, 1 equiv) in THF (10 mL) was added methyl (2S)-2-hydroxypropanoate (647.48 mg, 6.219 mmol, 1 equiv) and PPh3 (2446.98 mg, 9.329 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DIAD (1886.46 mg, 9.329 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at 60°C. After cooling down to rt, the reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give methyl (2R)-2-[2-fluoro-6- nitro-4-(trifluoromethyl)phenoxy]propanoate (1.14 g, 58.90%) as a yellow oil. LC/MS: mass calcd. for C11H9F4NO5:311.04 m/z, found:311.90 [M+H]+.
Figure imgf000529_0001
(R)-8-fluoro-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-[2-fluoro-6-nitro-4-(trifluoromethyl)phenoxy]propanoate (1.14 g, 3.663 mmol, 1 equiv) in EtOH (8 mL) and H2O (2 mL) was added Fe (2.05 g, 36.630 mmol, 10 equiv) and NH4Cl (1.96 g, 36.630 mmol, 10 equiv). The reaction was stirred at 70 °C for 1h. The mixture was allowed to cool down to rt. The filtrate was concentrated under reduced pressure. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-8-fluoro-2-methyl-6- (trifluoromethyl)-2,4-dihydro-1,4-benzoxazin-3-one (720 mg, 78.88%) as a yellow solid. LC/MS: mass calcd. for C10H7F4NO2:249.04 m/z, found:249.95 [M+H]+.
Figure imgf000529_0002
(R)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-8-fluoro-2-methyl-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin-3-one (700 mg, 2.809 mmol, 1 equiv) in H2SO4 (10 mL) was added KNO3 (284.03 mg, 2.809 mmol, 1 equiv) at 0 °C. The reaction was stirred at rt for 2h. The reaction was quenched with water/Ice at 0°C. Diluted with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-8-fluoro-2- methyl-7-nitro-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin-3-one (400 mg, 48.40%) as a yellow solid. LC/MS: mass calcd. for C10H8F4N2O4:294.03 m/z, found:292.95 [M-H]-.
Figure imgf000530_0001
(R)-4-benzyl-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)- one To a solution of (2R)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin- 3-one (420 mg, 1.428 mmol, 1 equiv) in DMF (10 mL) was added benzyl bromide (366.31 mg, 2.142 mmol, 1.5 equiv) and K2CO3 (596.30 mg, 4.284 mmol, 3 equiv) at 0 °C. The reaction was stirred at rt for 1h. The reaction was monitored by TLC. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-benzyl-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)- 2H-1,4-benzoxazin-3-one (400 mg, 72.90%) as a yellow solid.
Figure imgf000530_0002
(R)-7-amino-4-benzyl-8-fluoro-2-methyl-6-(trifluoromethyl)-2H-benzo[b][1,4]oxazin-3(4H)- one To a solution of (2R)-4-benzyl-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H-1,4-benzoxazin- 3-one (400 mg, 1.041 mmol, 1 equiv) in MeOH (10 mL) and H2O (1 mL) was added Fe (581.28 mg, 10.410 mmol, 10 equiv) and NH4Cl (556.77 mg, 10.410 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 50 mL). The filtrate was concentrated under reduced pressure.Diluted with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino- 4-benzyl-8-fluoro-2-methyl-6-(trifluoromethyl)-2H-1,4-benzoxazin-3-one (300 mg, 81.35%) as a yellow solid. LC/MS: mass calcd. for C17H14F4N2O2:354.10 m/z, found:355.10 [M+H]+.
Figure imgf000531_0001
(R)-1-(4-benzyl-8-fluoro-2-methyl-3-oxo-6-(trifluoromethyl)-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea To a solution of (2R)-7-amino-4-benzyl-8-fluoro-2-methyl-6-(trifluoromethyl)-2H-1,4- benzoxazin-3-one (70 mg, 0.198 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (58.62 mg, 0.198 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (19.99 mg, 0.198 mmol, 1 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (14.45 mg, 0.198 mmol, 1 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP 18 OBD Column, 19*250 mm, 5μm column (eluent: 52% to 82% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 1-[(2R)-4-benzyl-8-fluoro-2-methyl-3-oxo-6-(trifluoromethyl)-2H-1,4-benzoxazin-7-yl]-3-tert- butylurea (27.2 mg, 30.36%) as a white solid. LC/MS: mass calcd. for C22H23F4N3O3:453.17 m/z, found:454.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 7.31 – 7.40 (m, 2H), 7.23 – 7.31 (m, 3H), 7.06 (d, J = 1.6 Hz, 1H), 6.27 (s, 1H), 5.28 (d, J = 16.4 Hz, 1H), 5.21 (d, J = 16.4 Hz, 1H), 5.14 (q, J = 6.7 Hz, 1H), 1.59 (d, J = 6.7 Hz, 3H), 1.25 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -59.0147, -136.7360. Example 355: 1-[(3R)-1-benzyl-7-chloro-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6-yl]- 3-tert-butylurea Synthetic Scheme
Figure imgf000532_0001
methyl (2R)-2-[(5-chloro-3-nitropyridin-2-yl)oxy]propanoate To a stirred solution of methyl (2R)-2-hydroxypropanoate (1.08 g, 10.364 mmol, 1 equiv) in THF (30 mL) was added NaH (0.75 g, 31.092 mmol, 3 equiv) in portions at 0°C under air atmosphere. The resulting mixture was stirred for 30 min at room temperature under air atmosphere. To the above mixture was added 2,5-dichloro-3-nitropyridine (2 g, 10.364 mmol, 1 equiv) in portions at 0°C. The resulting mixture was stirred for additional 1h at room temperature. The reaction was quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%)to afford methyl (2R)-2-[(5-chloro-3-nitropyridin-2-yl)oxy]propanoate (1 g, 37.02%) as a white solid. LC/MS (ESI): mass calcd. for C9H9ClN2O5:260.0 m/z, found:261.10 [M+H]+.
Figure imgf000532_0002
(3R)-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one To a stirred mixture of methyl (2R)-2-[(5-chloro-3-nitropyridin-2-yl)oxy]propanoate (800 mg, 3.069 mmol, 1 equiv) and NH4Cl (1641.87 mg, 30.690 mmol, 10 equiv) in MeOH:H2O=10:1 (15 mL) was added Fe (1714.15 mg, 30.690 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 80°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (3R)-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (300 mg, 49.21%) as a yellow oil. LC/MS (ESI): mass calcd. for C8H7ClN2O2:198.0 m/z, found:199.15 [M+H]+.
Figure imgf000533_0001
(3R)-6-bromo-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one To a stirred solution of (3R)-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (210 mg, 1.057 mmol, 1 equiv) in DMF (5 mL) were added NBS (1129.15 mg, 6.342 mmol, 6 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 4 h at 100°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to afford (3R)-6-bromo-7-chloro- 3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (170 mg, 57.94%) as a yellow solid. LC/MS (ESI): mass calcd. for C8H6BrClN2O2: 275.9 m/z, found:274.80., 272.80 [M-H, M-H+2]-.
Figure imgf000533_0002
(3R)-1-benzyl-6-bromo-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one To a stirred mixture of (3R)-6-bromo-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (160 mg, 0.577 mmol, 1 equiv) and benzyl bromide (197.23 mg, 1.154 mmol, 2 equiv) in DMF (5 mL) was added K2CO3 (240.80 mg, 1.731 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 4h at rt under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. Desired product could be detected by LCMS. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (3R)-1-benzyl-6-bromo- 7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one (127 mg, 59.92%) as a yellow solid. LC/MS (ESI): mass calcd. for C15H12BrClN2O2:366.0 m/z, found: 367.0, 369.0 [M+H, M+H+2]+.
Figure imgf000534_0001
(3R)-6-amino-1-benzyl-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one To a stirred mixture of (3R)-1-benzyl-6-bromo-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin- 2-one (85 mg, 0.231 mmol, 1 equiv) and NaN3 (45.08 mg, 0.693 mmol, 3 equiv) in EtOH:H2O = 10:1 (5 mL) were added N1,N2-dimethylcyclohexane-1,2-diamine (19.73 mg, 0.139 mmol, 0.6 equiv) and CuSO4 (36.89 mg, 0.231 mmol, 1 equiv) and sodium ascorbate (92.05 mg, 0.462 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 80°C under nitrogen atmosphere. After cooling down to room temperature, the reaction quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (3R)-6-amino-1-benzyl-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one (56 mg, 80.0%) as a yellow solid. LC/MS (ESI): mass calcd. for C15H14ClN3O2:303.1 m/z, found:304.25 [M+H]+.
Figure imgf000535_0001
1-[(3R)-1-benzyl-7-chloro-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6-yl]-3-tert- butylurea To a stirred solution of (3R)-6-amino-1-benzyl-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin- 2-one (50 mg, 0.165 mmol, 1 equiv) and triphosgene (61.79 mg, 0.209 mmol, 1.2 equiv) in DCM (5 mL) was added TEA (49.97 mg, 0.495 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added 2-methylpropan-2-amine (12.04 mg, 0.165 mmol, 1 equiv) dropwise at room temperature. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford 1-[(3R)-1-benzyl-7-chloro-3-methyl-2-oxo-3H-pyrido[2,3- b][1,4]oxazin-6-yl]-3-tert-butylurea (8.3 mg, 12.37%) as a white solid. LC/MS (ESI): mass calcd. for C20H23ClN4O3:402.1 m/z, found:403.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.89 (d, J = 13.4 Hz, 2H), 7.62 (s, 1H), 7.32-7.40 (m, 2H), 7.19-7.32 (m, 3H), 5.16-5.23 (m, 3H), 1.56 (d, J = 6.8 Hz, 3H), 1.32 (s, 9H). Example 356: (R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-6- (trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000536_0001
(R)-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2,4-dihydro-1,4-benzoxazin- 3-one (100 mg, 0.340 mmol, 1 equiv) in DMF (10 mL) was added 1-(bromomethyl)-3- (difluoromethyl)benzene (112.72 mg, 0.510 mmol, 1.5 equiv) and K2CO3 (141.98 mg, 1.020 mmol, 3 equiv). The reaction was stirred at rt for 1h. The reaction was monitored by TLC. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-{[3- (difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-7-nitro-6-(trifluoromethyl)-2H-1,4- benzoxazin-3-one (140 mg, 94.83%) as a yellow solid.
Figure imgf000537_0001
(R)-7-amino-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-6-(trifluoromethyl)-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-7-nitro-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (130 mg, 0.299 mmol, 1 equiv) in MeOH (5 mL) and H2O (0.5 mL) was added Fe (167.16 mg, 2.990 mmol, 10 equiv) and NH4Cl (160.12 mg, 2.990 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 30 mL). The filtrate was concentrated under reduced pressure. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (100 mg, 82.63%) as a yellow solid. LC/MS: mass calcd. for C18H14F6N2O2:404.10 m/z, found:405.05 [M+H]+.
Figure imgf000537_0002
(R)-1-(tert-butyl)-3-(4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-6- (trifluoromethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-6- (trifluoromethyl)-2H-1,4-benzoxazin-3-one (150 mg, 0.371 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (110.09 mg, 0.371 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (112.63 mg, 1.113 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (81.40 mg, 1.113 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep C18 OBD Column, 19*250 mm, 5μm column (eluent: 49% to 79% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-tert-butyl-1-[(2R)-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2- methyl-3-oxo-6-(trifluoromethyl)-2H-1,4-benzoxazin-7-yl]urea (20.3 mg, 10.79%) as a white solid. LC/MS: mass calcd. for C23H23F6N3O3:503.16 m/z, found:504.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 7.31 – 7.40 (m, 2H), 7.23 – 7.31 (m, 3H), 7.06 (d, J = 1.6 Hz, 1H), 6.27 (s, 1H), 5.28 (d, J = 16.4 Hz, 1H), 5.21 (d, J = 16.4 Hz, 1H), 5.14 (q, J = 6.7 Hz, 1H), 1.59 (d, J = 6.7 Hz, 3H), 1.25 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -59.0147, -136.7360. Example 357: (S)-1-(tert-butyl)-3-(1-(1-(3-chlorophenyl)ethyl)-7-fluoro-2-oxo-1,2- dihydroquinoxalin-6-yl) Urea Synthetic Scheme
Figure imgf000538_0001
7-fluoro-6-nitroquinoxalin-2(1H)-one To a solution of 4-fluoro-5-nitrobenzene-1,2-diamine (2 g, 11.687 mmol, 1 equiv) in ethyl alcohol (10 mL) was added ethyl glyoxylate (1.19 g, 11.687 mmol, 1 equiv). The resulting mixture was stirred for 2 h at 80 °C. The reaction mixture was quenched by water and extracted with DCM (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 7-fluoro-6-nitroquinoxalin-2(1H)-one (1.7 g, 69.55%) as a white solid. MS (ESI): mass calcd. for C8H4FN3O3:209.17 m/z, found:208 [M-H]-.
Figure imgf000539_0001
(S)-1-(1-(3-chlorophenyl)ethyl)-7-fluoro-6-nitroquinoxalin-2(1H)-one To a solution of 7-fluoro-6-nitroquinoxalin-2(1H)-one (1 g, 4.782 mmol, 1 equiv) in DCM (40 mL) was added (1R)-1-(3-chlorophenyl)ethanol (0.75 g, 4.782 mmol, 1 equiv) PPh3 (1.88 g, 7.173 mmol, 1.5 equiv) and DEAD (1.25 g, 7.173 mmol, 1.5 equiv). The resulting mixture was stirred for 2 h at 25 °C under N2 and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with DCM (3 x 30 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (S)-1-(1-(3-chlorophenyl)ethyl)-7-fluoro-6- nitroquinoxalin-2(1H)-one (400 mg, 24.06%) as a white solid. MS (ESI): mass calcd. for C16H11ClFN3O3: 347.17 m/z, found: 348.20 [M+H]+ .
Figure imgf000539_0002
(S)-6-amino-1-(1-(3-chlorophenyl)ethyl)-7-fluoroquinoxalin-2(1H)-one To a solution of (S)-1-(1-(3-chlorophenyl)ethyl)-7-fluoro-6-nitroquinoxalin-2(1H)-one (300 mg, 0.863 mmol, 1 equiv) in EtOH (1 mL) and H2O (0.1 mL) was added Fe (481 mg, 8.630 mmol, 10 equiv) and NH4Cl (461 mg, 8.630 mmol, 10 equiv). The resulting mixture was stirred for 2 h at 80 °C and stirred until the starting material was totally consumed by TLC. The reaction mixture was quenched by water and extracted with DCM (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford (S)-6-amino-1-(1-(3-chlorophenyl)ethyl)-7-fluoroquinoxalin-2(1H)-one (120 mg, 43.77%) as a white solid. MS (ESI): mass calcd. for C16H13ClFN3O: 317.17 m/z, found: 318.20 [M+H]+ .
Figure imgf000540_0001
(S)-1-(tert-butyl)-3-(1-(1-(3-chlorophenyl)ethyl)-7-fluoro-2-oxo-1,2-dihydroquinoxalin-6- yl)urea To a mixture of (S)-6-amino-1-(1-(3-chlorophenyl)ethyl)-7-fluoroquinoxalin-2(1H)-one (60 mg, 0.189 mmol, 1 equiv) and TEA (57 mg, 0.567 mmol, 3 equiv) in DCM (4 mL) was added 2- isocyanato-2-methylpropane (56 mg, 0.567 mmol, 3 equiv) at 0 °C. The reaction was stirred for overnight at rt. The reaction mixture was quenched by water (10 mL) and extracted with DCM (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column with PE/EA (1:1) & Prep-HPLC (Column: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; Mobile Phase A: Water(10 mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL/min; Gradient: 28% B to 53% B in 9 min, 53% B; Wave Length: 254/220 nm; RT1(min): 8.52; Number Of Runs: 0) to afford (S)-1-(tert-butyl)-3- (1-(1-(3-chlorophenyl)ethyl)-7-fluoro-2-oxo-1,2-dihydroquinoxalin-6-yl)urea (12.3 mg, 15.09%) as a white solid. MS (ESI): mass calcd. For C21H22ClFN4O2: 416.17 m/z, found: 417.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.57 (d, J = 8.7 Hz, 1H), 8.19 (s, 1H), 8.25 (s, 1H), 7.32 – 7.42 (m, 4H), 7.20 – 7.29 (m, 1H), 6.58 (s, 1H), 6.44 (s, 1H), 1.87 (d, J = 7.0 Hz, 3H), 1.30 (s, 9H). Example 358: 3-tert-butyl-1-{1-[(1S)-1-(3-chlorophenyl)ethyl]-2-oxo-7- (trifluoromethyl)quinoxalin-6-yl}urea Synthetic Scheme
Figure imgf000541_0001
7-(trifluoromethyl)-1H-quinoxalin-2-one To a stirred solution of 4-(trifluoromethyl)benzene-1,2-diamine (2 g, 11.354 mmol, 1 equiv) in EtOH(20 mL) was added ethyl glyoxylate (1.16 g, 11.354 mmol, 1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 3 h at 60 ℃ under air atmosphere.The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 7-(trifluoromethyl)-1H-quinoxalin-2-one (1.2 g, 49.35%) as a yellow solid. LC/MS (ESI): mass calcd. for C9H5F3N2O:214.04 m/z, found:215.0 [M+H]+.
Figure imgf000541_0002
6-nitro-7-(trifluoromethyl)-1H-quinoxalin-2-one To a stirred solution of 7-(trifluoromethyl)-1H-quinoxalin-2-one (1.1 g, 5.137 mmol, 1 equiv) in H2SO4 (11 mL) was added KNO3 (1.56 g, 15.411 mmol, 3 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was monitored by LCMS. The reaction was quenched with water/Ice (100 mL) at 0 °C, and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-nitro-7-(trifluoromethyl)-1H- quinoxalin-2-one (600 mg, 45.07%) as a brown solid.LC/MS (ESI): mass calcd. for C9H4F3N3O3:259.02 m/z, found: 260.0 [M+H]+.
Figure imgf000542_0001
1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitro-7-(trifluoromethyl)quinoxalin-2-one To a stirred solution of 6-nitro-7-(trifluoromethyl)-1H-quinoxalin-2-one (600 mg, 2.315 mmol, 1 equiv) and (1R)-1-(3-chlorophenyl)ethanol (362.60 mg, 2.315 mmol, 1 equiv) in DCM (10 mL) was added PPh3 (910.93 mg, 3.473 mmol, 1.5 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 15 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (403.23 mg, 2.315 mmol, 1 equiv) in portions at 0 °C. The resulting mixture was stirred for additional 2 h at room temperature. The reaction was monitored by LCMS. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitro-7-(trifluoromethyl)quinoxalin-2- one (110 mg, 11.94%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H11ClF3N3O3:397.04 m/z, found:398.0 [M+H]+.
Figure imgf000542_0002
6-amino-1-[(1S)-1-(3-chlorophenyl)ethyl]-7-(trifluoromethyl)quinoxalin-2-one To a stirred solution of 1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitro-7-(trifluoromethyl)quinoxalin- 2-one (100 mg, 0.251 mmol, 1 equiv) in MeOH/H2O(10:1, 5 mL) was added Fe (14.04 mg, 0.250 mmol, 10 equiv) and NH4Cl (134.48 mg, 2.510 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. The reaction was monitored by LCMS. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (0-100%) to afford 6-amino-1-[(1S)-1-(3-chlorophenyl)ethyl]-7-(trifluoromethyl)quinoxalin-2-one (55 mg, 59.48%) as a yellow solid. LC/MS (ESI): mass calcd. for C17H14F3N3O:367.1 m/z, found:368.05 [M+H]+.
Figure imgf000543_0001
3-tert-butyl-1-{1-[(1S)-1-(3-chlorophenyl)ethyl]-2-oxo-7-(trifluoromethyl)quinoxalin-6- yl}urea To a stirred solution of 6-amino-1-[(1S)-1-(3-chlorophenyl)ethyl]-7-(trifluoromethyl)quinoxalin- 2-one (50 mg, 0.136 mmol, 1 equiv) in DCM (4 mL) was added TEA (41.27 mg, 0.408 mmol, 3 equiv) and 2-isocyanato-2-methylpropane (40.43 mg, 0.408 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1 days at room temperature under air atmosphere. The reaction was monitored by LCMS. Quenched with water (5 mL) and extracted with DCM (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by Prep- HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5 μm column (eluent: 43% to 59% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.05%NH3H2O) to afford 3-tert-butyl-1-{1- [(1S)-1-(3-chlorophenyl)ethyl]-2-oxo-7-(trifluoromethyl)quinoxalin-6-yl}urea (2.7 mg, 4.23%) as a yellow solid. LC/MS (ESI): mass calcd. for C22H22ClF3N4O2:466.1 m/z, found:467.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.37 (d, J = 3.0 Hz, 2H), 7.68 (s, 1H), 7.44 – 7.37(m, 3H), 7.29 (s, 2H), 6.92 (s, 1H), 6.55 (d, J = 7.2 Hz, 1H), 1.85 (d, J = 7.2 Hz, 3H), 1.29 (s, 9H). 19F NMR (282 MHz, DMSO-d6) δ -60.17. Example 359: cyclopentyl (S)-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2- dihydroquinoxalin-6-yl)carbamate Synthetic Scheme
Figure imgf000544_0001
cyclopentyl (S)-(2-oxo-1-(1-(3-(trifluoromethoxy)phenyl)ethyl)-1,2-dihydroquinoxalin-6- yl)carbamate To a solution of 6-amino-1-[(1S)-1-[3-(trifluoromethoxy)phenyl]ethyl]quinoxalin-2-one (80 mg, 0.229 mmol, 1 equiv) in DCM (5 mL) was added cyclopentyl carbonochloridate (51.05 mg, 0.344 mmol, 1.5 equiv) and pyridine (54.35 mg, 0.687 mmol, 3 equiv) at 0°C. The reaction was stirred at rt for 1h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 49% to 63% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.05%NH3.H2O)) to afford the title compound cyclopentyl N-{2-oxo-1-[(1S)-1-[3- (trifluoromethoxy)phenyl]ethyl]quinoxalin-6-yl}carbamate (32.2 mg, 30.22%) as a yellow solid.LC/MS: mass calcd. for C23H22F3N3O4:461.16 m/z, found:462.10 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 8.26 (s, 1H), 8.00 (s, 1H), 7.43 – 7.51 (m, 1H), 7.46 (s, 1H), 7.29 (q, J = 5.7 Hz, 3H), 7.17 (s, 1H), 6.57 (s, 1H), 5.00 – 5.15 (m, 1H), 1.85 – 1.91 (m, 5H), 1.65 – 1.72 (m, 4H), 1.60 (d, J = 9.4 Hz, 2H).19F NMR (376 MHz, DMSO) δ(ppm): -56.6795. Example 360: (S)-1-(tert-butyl)-3-(7-(hydroxymethyl)-2-oxo-1-(1-phenylethyl)-1,2- dihydroquinoxalin-6-yl)urea
Figure imgf000545_0002
(S)-7-(hydroxymethyl)-6-nitro-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 7-bromo-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (300 mg, 0.802 mmol, 1 equiv) in dioxane (3 mL) was added (tributylstannyl)methanol (386.14 mg, 1.203 mmol, 1.5 equiv) and XPhos Pd G2 (63.08 mg, 0.080 mmol, 0.1 equiv). The reaction was stirred at 90°C for 12 h under N2. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-(hydroxymethyl)-6-nitro-1-[(1S)-1- phenylethyl]quinoxalin-2-one (40 mg, 15.34%) as a white solid. MS (ESI): mass calcd. for C17H15N3O4: 325.11 m/z, found 323.95 [M-H]-.
Figure imgf000545_0001
(S)-6-amino-7-(hydroxymethyl)-1-(1-phenylethyl)quinoxalin-2(1H)-one To a solution of 7-(hydroxymethyl)-6-nitro-1-[(1S)-1-phenylethyl]quinoxalin-2-one (40 mg, 0.123 mmol, 1 equiv) in MeOH (2 mL)/H2O (0.2 mL) was added Fe (68.66 mg, 1.230 mmol, 10 equiv) and NH4Cl (65.77 mg, 1.230 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to 6-amino-7-(hydroxymethyl)-1- [(1S)-1-phenylethyl]quinoxalin-2-one (25 mg, 68.84%) as a white solid. MS (ESI): mass calcd. for C17H17N3O2: 295.13 m/z, found 296.10 [M+H]+.
Figure imgf000546_0001
(S)-6-amino-7-(((tert-butyldimethylsilyl)oxy)methyl)-1-(1-phenylethyl)quinoxalin-2(1H)- one To a solution of 6-amino-7-(hydroxymethyl)-1-[(1S)-1-phenylethyl]quinoxalin-2-one (70 mg, 0.237 mmol, 1 equiv) in DCM (3 mL) was added TBSCl (53.58 mg, 0.355 mmol, 1.5 equiv) and imidazole (48.41 mg, 0.711 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-{[(tert- butyldimethylsilyl)oxy]methyl}-1-[(1S)-1-phenylethyl]quinoxalin-2-one (75 mg, 77.25%) as a white solid. MS (ESI): mass calcd. for C23H31N3O2Si: 409.22 m/z, found 410.25 [M+H]+.
Figure imgf000546_0002
(S)-1-(tert-butyl)-3-(7-(((tert-butyldimethylsilyl)oxy)methyl)-2-oxo-1-(1-phenylethyl)-1,2- dihydroquinoxalin-6-yl)urea To a solution of 6-amino-7-{[(tert-butyldimethylsilyl)oxy]methyl}-1-[(1S)-1- phenylethyl]quinoxalin-2-one (50 mg, 0.122 mmol, 1 equiv) in DCM (1 mL) was added triphosgene (36.22 mg, 0.122 mmol, 1 equiv). The reaction was stirred at 0°C under N2 for 1 h. The reaction was added TEA (74.12 mg, 0.732 mmol, 6 equiv) and stirred at rt under N2 for 1 h. The reaction was added 2-methylpropan-2-amine (26.78 mg, 0.366 mmol, 3 equiv) and stirred at rt under N2 for 1 h. Quenched with water (5 mL) and extracted with DCM (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 3-tert-butyl-1-(7-{[(tert-butyldimethylsilyl)oxy]methyl}-2-oxo-1-[(1S)-1- phenylethyl]quinoxalin-6-yl)urea (20 mg, 32.21%) as a white solid. LC/MS (ESI): mass calcd. for C28H40N4O3Si: 508.29 m/z, found: 509.35 [M+H]+.
Figure imgf000547_0001
1-(1-benzyl-5-chloro-2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-(tert-butyl)urea To a solution of 3-tert-butyl-1-(7-{[(tert-butyldimethylsilyl)oxy]methyl}-2-oxo-1-[(1S)-1- phenylethyl]quinoxalin-6-yl)urea (20 mg, 0.039 mmol, 1 equiv) in THF (1 mL) was added Triethylamine trihydrofluoride (9.51 mg, 0.058 mmol, 1.5 equiv). The reaction was stirred at rt for 1 h. Quenched with water (5 mL) and extracted with EA (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm (eluent: 22% to 38% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[7-(hydroxymethyl)-2-oxo-1-[(1S)-1-phenylethyl]quinoxalin-6-yl]urea (6.2 mg, 39.70%) as a white solid. LC/MS (ESI): mass calcd. for C22H26N4O3: 394.20 m/z, found: 395.15 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 8.21-8.27 (d, J = 18 Hz, 2H), 7.58 (s, 1H), 7.21 – 7.38 (m, 6H), 6.60 (s, 2H), 5.29-5.33 (t, J = 12 Hz, 1H), 4.26-4.42 (m, 2H), 1.87- 1.89 (d, J = 6 Hz, 3H), 1.27 (s, 9H). Example 361: 3-tert-butyl-1-{1-[(1S)-1-(3-chlorophenyl)ethyl]-7-cyano-2-oxoquinoxalin-6- yl}urea Synthetic Scheme
Figure imgf000548_0001
7-bromo-6-nitro-1H-quinoxalin-2-one To a solution of 7-bromo-1H-quinoxalin-2-one (5 g, 22.218 mmol, 1 equiv) in H2SO4 (50 mL) was added KNO3 (2.92 g, 28.883 mmol, 1.3 equiv) at 0 °C. The reaction was stirred at 0 °C for 1 h. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-bromo-6-nitro-1H- quinoxalin-2-one (4.5 g, 75.00%) as a yellow solid. MS (ESI): mass calcd. for C8H4BrN3O3:268.94 m/z, found:269.94, 271.96 [M+H, M+H+2]+.
Figure imgf000548_0002
7-bromo-1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitroquinoxalin-2-one To a solution of 7-bromo-6-nitro-1H-quinoxalin-2-one (1.5 g, 5.555 mmol, 1 equiv) in DCM (20 mL) was added (1R)-1-(3-chlorophenyl)ethanol (0.87 g, 5.555 mmol, 1 equiv) and PPh3 (2.19 g, 8.332 mmol, 1.5 equiv). After the mixture was stirred at rt for 1h, to the above mixture was added DEAD (0.97 g, 5.555 mmol, 1 equiv) dropwise at 0 ℃. The resulting mixture was stirred for 2 h at 25 ℃. Quenched with water (100 mL) and extracted with DCM (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7- bromo-1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitroquinoxalin-2-one (210 mg, 9.25%) as a yellow oil. MS (ESI): mass calcd. for C16H11BrClN3O3:406.97 m/z, found:407.97, 409.97 [M+H, M+H+2]+.
Figure imgf000549_0001
6-amino-7-bromo-1-[(1S)-1-(3-chlorophenyl)ethyl]quinoxalin-2-one To a solution of 7-bromo-1-[(1S)-1-(3-chlorophenyl)ethyl]-6-nitroquinoxalin-2-one (280 mg, 0.685 mmol, 1 equiv) in MeOH (10 mL) and H2O (1 mL) was added Fe (382.65 mg, 6.850 mmol, 10 equiv) and NH4Cl (366.51 mg, 6.850 mmol, 10 equiv). The reaction was stirred at 70 ℃ for 2 h. The mixture was allowed to cool down to room temperature and quenched with water (50 mL). The resulting mixture was filtered; the filter cake was washed with EA (50 mL). The filtrate was extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-bromo-1-[(1S)-1-(3- chlorophenyl)ethyl]quinoxalin-2-one) as a yellow oil. MS (ESI): mass calcd. for C16H13BrClN3O:376.99 m/z, found:377.99, 379.99 [M+H, M+H+2]+.
Figure imgf000549_0002
7-amino-4-[(1S)-1-(3-chlorophenyl)ethyl]-3-oxoquinoxaline-6-carbonitrile To a solution of 6-amino-7-bromo-1-[(1S)-1-(3-chlorophenyl)ethyl]quinoxalin-2-one (140 mg, 0.370 mmol, 1 equiv) in dimethylformamide (8 mL) was added Zn(CN)2 (52.10 mg, 0.444 mmol, 1.2 equiv) and Pd(PPh3)4 (42.73 mg, 0.037 mmol, 0.1 equiv). The reaction was stirred at 100 °C for 2 h under N2 atmosphere. The mixture was allowed to cool down to room temperature. Quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4-[(1S)-1-(3- chlorophenyl)ethyl]-3-oxoquinoxaline-6-carbonitrile (60 mg, 49.97%) as a red solid. MS (ESI): mass calcd. for C17H13ClN4O:324.08 m/z, found:325.08 [M+H]+.
Figure imgf000550_0001
3-tert-butyl-1-{1-[(1S)-1-(3-chlorophenyl)ethyl]-7-cyano-2-oxoquinoxalin-6-yl}urea To a solution of 7-amino-4-[(1S)-1-(3-chlorophenyl)ethyl]-3-oxoquinoxaline-6-carbonitrile (50 mg, 0.154 mmol, 1 equiv) in DCM (8 mL) was added triphosgene (22.84 mg, 0.077 mmol, 0.5 equiv) at 0 °C under N2 atmosphere. The resulting mixture was stirred for 2 h at 25 °C. Then TEA (46.74 mg, 0.462 mmol, 3 equiv) was added into above system at 0 °C. The resulting mixture was stirred for 1 h at 25 °C. To the above mixture was added erbumine (16.89 mg, 0.231 mmol, 1.5 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (30 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using aXselect CSH C18 OBD 30*150 mm, 5μm column (eluent: 42% to 82% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(1S)-1-(3- chlorophenyl)ethyl]-7-cyano-2-oxoquinoxalin-6-yl}urea (14.7 mg, 21.84%) as a yellow solid. LC/MS (ESI): mass calcd. for C22H22ClN5O2:423.15 m/z, found:424.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.49 (s, 1H), 8.40 (s, 1H), 8.32 (s, 1H), 7.53 – 7.90 (m, 1H), 7.40 – 7.45 (m, 1H), 7.32 – 7.40 (m, 2H), 7.23 – 7.40 (m, 1H), 6.96 (s, 1H), 6.38 (s, 1H), 1.87 (d, J = 7.0 Hz, 3H), 1.31 (s, 9H). Example 362: 1-[(3R)-1-benzyl-7-cyano-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6-yl]- 3-tert-butylurea Synthetic Scheme
Figure imgf000551_0001
(3R)-6-amino-1-benzyl-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazine-7-carbonitrile To a stirred mixture of (3R)-6-amino-1-benzyl-7-chloro-3-methyl-3H-pyrido[2,3-b][1,4]oxazin- 2-one (50 mg, 0.165 mmol, 1 equiv) and Zn(CN)2 (38.66 mg, 0.330 mmol, 2 equiv) in DMF (5 mL) was added BrettPhos Pd G3 (298.44 mg, 0.330 mmol, 02 equiv) and BrettPhos (17.67 mg, 0.033 mmol, 0.2 equiv) and Zn (3.23 mg, 0.050 mmol, 0.3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 120°C under nitrogen atmosphere. After cooling down to room temperature, the reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (3R)-6-amino-1-benzyl-3-methyl-2- oxo-3H-pyrido[2,3-b][1,4]oxazine-7-carbonitrile (44 mg, 90.7%) as a white solid. LC/MS (ESI): mass calcd. for C16H14N4O2:294.1 m/z, found:295.2 [M+H]+.
Figure imgf000551_0002
1-[(3R)-1-benzyl-7-cyano-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6-yl]-3-tert- butylurea To a stirred solution of (3R)-6-amino-1-benzyl-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazine-7- carbonitrile (50 mg, 0.170 mmol, 1 equiv) and triphosgene (60.49 mg, 0.204 mmol, 1.2 equiv) in DCM (5 mL) was added TEA (51.57 mg, 0.510 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at 50 oC under nitrogen atmosphere. To the above mixture was added 2-methylpropan-2-amine (24.85 mg, 0.340 mmol, 2 equiv) dropwise at room temperature. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford 1-[(3R)-1-benzyl-7-cyano-3-methyl-2-oxo-3H-pyrido[2,3- b][1,4]oxazin-6-yl]-3-tert-butylurea (2.4 mg, 3.55%) as a white solid. LC/MS (ESI): mass calcd. for C21H23N5O3:393.2 m/z, found:394.20 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1H), 7.82 (s, 1H), 7.32-7.46 (m, 2H), 7.23-7.32 (m, 3H), 6.89 (s, 1H), 5.22-5.35 (m, 1H), 5.15 (s, 2H), 1.58 (d, J = 6.8 Hz, 3H), 1.29 (s, 9H). Example 363: (S)-1-(tert-butyl)-3-(7-cyano-1-(1-(2,5-dichlorophenyl) ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl) urea Synthetic Scheme
Figure imgf000552_0001
(1R)-1-(2,5-dichlorophenyl) ethanol To a stirred solution of CBS (0.56 g, 2.116 mmol, 0.2 equiv) in THF (10 mL) was added BH3- Me2S (0.53 g, 6.983 mmol, 0.66 equiv) and 1-(2,5-dichlorophenyl) ethanone (2 g, 10.580 mmol, 1 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 1 h at 0 °C under air atmosphere. The reaction was quenched with water at 0 °C. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (1R)-1-(2,5- dichlorophenyl) ethanol (1.83 g, 90.54%) as a yellow oil. MS (ESI): mass calcd. For C8H8Cl2O: 190.00 m/z, found 191.15[M +H]+.
Figure imgf000553_0001
7-bromo-1-[(1S)-1-(2,5-dichlorophenyl) ethyl]-6-nitroquinoxalin-2-one To a stirred solution of (1R)-1-(2,5-dichlorophenyl) ethanol (300 mg, 1.570 mmol, 1 equiv) and 7-bromo-6-nitro-1H-quinoxalin-2-one (508.85 mg, 1.884 mmol, 1.2 equiv) in DCM (5 mL) was added PPh3 (617.80 mg, 2.355 mmol, 1.5 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added DEAD (410.21 mg, 2.355 mmol, 1.5 equiv) dropwise over 1 min at 0 °C. The resulting mixture was stirred for additional 1 h at room temperature. The resulting mixture was stirred for 1 h at room temperature under nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford 7-bromo-1- [(1S)-1-(2,5-dichlorophenyl) ethyl]-6-nitroquinoxalin-2-one (50 mg, 7.19%) as a yellow oil. MS (ESI): mass calcd. For C16H10BrCl2N3O3: 440.93 m/z, found 443.85 [M+H, M+H+2]+.
Figure imgf000553_0002
6-amino-7-bromo-1-[(1S)-1-(2,5-dichlorophenyl) ethyl] quinoxalin-2-one A mixture of 7-bromo-1-[(1S)-1-(2,5-dichlorophenyl) ethyl]-6-nitroquinoxalin-2-one (220 mg, 0.497 mmol, 1 equiv), Fe (277.28 mg, 4.970 mmol, 10 equiv) and NH4Cl (265.59 mg, 4.970 mmol, 10 equiv) was added EtOH (10 mL) and H2O (1 mL) and stirred at 70 °C for 1 h. The mixture was allowed to cool down to room temperature. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-bromo-1-[(1S)-1-(2,5-dichlorophenyl) ethyl] quinoxalin-2-one (200 mg, 97.51%) as a yellow solid. MS (ESI): mass calcd. for C16H12BrCl2N3O: 410.95 m/z, found 413.95 [M+H, M+H+2]+.
Figure imgf000554_0001
7-amino-4-[(1S)-1-(2,5-dichlorophenyl) ethyl]-3-oxoquinoxaline-6-carbonitrile A mixture of 6-amino-7-bromo-1-[(1S)-1-(2,5-dichlorophenyl) ethyl] quinoxalin-2-one (200 mg, 0.484 mmol, 1 equiv), Zn(CN)2 (68.22 mg, 0.581 mmol, 1.2 equiv) and Pd(PPh3)4 (55.95 mg, 0.048 mmol, 0.1 equiv) was treated with dimethylformamide (10 mL) and stirred at 110 °C under N2 for 1 h. The mixture was allowed to cool down to room temperature. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4-[(1S)-1-(2,5- dichlorophenyl) ethyl]-3-oxoquinoxaline-6-carbonitrile (120 mg, 69.00%) as a white solid. MS (ESI): mass calcd. for C17H12Cl2N4O: 358.04 m/z, found 359.10 [M+H]+.
Figure imgf000554_0002
3-tert-butyl-1-{7-cyano-1-[(1S)-1-(2,5-dichlorophenyl) ethyl]-2-oxoquinoxalin-6-yl} urea A solution of 7-amino-4-[(1S)-1-(2,5-dichlorophenyl) ethyl]-3-oxoquinoxaline-6-carbonitrile (20 mg, 0.056 mmol, 1 equiv) in DCM (5 mL) was treated with Triphosgene (16.52 mg, 0.056 mmol, 1 equiv) for 1 h at 0 °C under nitrogen atmosphere followed by the addition of TEA (28.17 mg, 0.280 mmol, 5 equiv) dropwise at 0 °C. The resulting mixture was stirred for 1 h at room temperature under nitrogen atmosphere. To the above mixture was added erbumine (12.22 mg, 0.168 mmol, 3 equiv) dropwise over 1 min at room temperature. The resulting mixture was stirred for additional 1 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column 150 mm x 30 mm x 5 μm column (eluent: 44% to 74% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3+0.05%NH3H2O) to afford 3-tert-butyl-1-{7-cyano-1-[(1S)-1-(2,5- dichlorophenyl) ethyl]-2-oxoquinoxalin-6-yl} urea (1 mg, 3.89%) as a yellow solid. MS (ESI): mass calcd. for C22H21Cl2N5O2: 457.11 m/z, found 458.15 [M+H]+.1H NMR (300 MHz, Acetonitrile-d3) δ 8.45 (s, 1H), 8.12 (s, 1H), 7.83 – 7.98 (m, 2H), 7. 28 – 7.45 (m, 2H), 7.15 (s, 1H), 5.92 – 6.20 (m, 1H), 5.75 (s, 1H), 1.90 – 1.95 (m, 3H), 1.38 – 1.41 (m, 9H). Example 364: 3-tert-butyl-1-{1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-7-cyano-2- oxoquinoxalin-6-yl}urea Synthetic Schemes
Figure imgf000555_0001
(1R)-1-(5-chloro-2-fluorophenyl)ethanol To a solution of (S)-(-)-2-Methyl-CBS-oxazaborolidine (1.22 g, 4.636 mmol, 0.4 equiv) in THF (24 mL) was added BH3-Me2S (1.16 g, 15.297 mmol, 1.32 equiv) and 1-(5-chloro-2- fluorophenyl)ethanone (2 g, 11.589 mmol, 1 equiv) at 0°C. The reaction was stirred at 0°C for 2 h. TLC showed the desired product was generated. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (1R)-1-(5-chloro-2-fluorophenyl)ethanol (1.4 g, 69.19%) as a white solid.
Figure imgf000556_0001
7-bromo-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-6-nitroquinoxalin-2-one To a solution of (1R)-1-(5-chloro-2-fluorophenyl)ethanol (970 mg, 5.556 mmol, 1 equiv) in DCM (12 mL) was added 7-bromo-6-nitro-1H-quinoxalin-2-one (1.50 g, 5.556 mmol, 1 equiv) and PPh3 (2.19 g, 8.334 mmol, 1.5 equiv). After the mixture was stirred at rt for 1 h, to the above mixture was added DEAD (0.97 g, 5.556 mmol, 1 equiv) dropwise at 0 ℃. The resulting mixture was stirred for 2 h at 25 ℃. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7- bromo-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-6-nitroquinoxalin-2-one (140 mg, 5.91%) as a yellow oil. MS (ESI): mass calcd. for C16H10BrClFN3O3:424.96 m/z, found:425.96, 427.96 [M+H, M+H+2]+.
Figure imgf000556_0002
6-amino-7-bromo-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]quinoxalin-2-one To a solution of 7-bromo-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-6-nitroquinoxalin-2-one (190 mg, 0.445 mmol, 1 equiv) in MeOH (4 mL) and H2O (0.4 mL) was added Fe (248.71 mg, 4.450 mmol, 10 equiv) and NH4Cl (238.22 mg, 4.450 mmol, 10 equiv). The reaction was stirred at 70 ℃ for 2 h. The mixture was allowed to cool down to room temperature and quenched with water (50 mL). The resulting mixture was filtered, the filter cake was washed with EA (50 mL). The filtrate was extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-bromo-1-[(1S)-1-(5-chloro-2- fluorophenyl)ethyl]quinoxalin-2-one (50 mg, 28.31%) as a yellow oil. MS (ESI): mass calcd. for C16H12BrClFN3O:394.98 m/z, found:395.98, 397.98 [M+H, M+H+2]+.
Figure imgf000557_0001
7-amino-4-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-3-oxoquinoxaline-6-carbonitrile To a solution of 6-amino-7-bromo-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]quinoxalin-2-one (60 mg, 0.151 mmol, 1 equiv) in dimethylformamide (3 mL) was added Zn(CN)2 (21.31 mg, 0.181 mmol, 1.2 equiv) and Pd(PPh3)4 (17.48 mg, 0.015 mmol, 0.1 equiv). The reaction was stirred at 100 °C for 2 h under N2 atmosphere. The mixture was allowed to cool down to room temperature. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-amino-4-[(1S)-1-(5- chloro-2-fluorophenyl)ethyl]-3-oxoquinoxaline-6-carbonitrile (30 mg, 57.86%) as a red solid. MS (ESI): mass calcd. for C17H12ClFN4O:342.07 m/z, found:343.07 [M+H]+.
Figure imgf000557_0002
3-tert-butyl-1-{1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-7-cyano-2-oxoquinoxalin-6- yl}urea To a solution of 7-amino-4-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-3-oxoquinoxaline-6- carbonitrile (30 mg, 0.088 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (12.99 mg, 0.044 mmol, 0.5 equiv) at 0 °C under N2 atmosphere. The resulting mixture was stirred for 1 h at 25 °C. Then TEA (26.57 mg, 0.264 mmol, 3 equiv) was added into above system at 0 °C. The resulting mixture was stirred 1 h at 25 °C. To the above mixture was added erbumine (9.60 mg, 0.132 mmol, 1.5 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using XBridge Prep OBD C1830*150 mm, 5μm column (eluent: 40% to 62% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-{1-[(1S)-1-(5- chloro-2-fluorophenyl)ethyl]-7-cyano-2-oxoquinoxalin-6-yl}urea (12.6 mg, 30.13%) as a yellow solid. MS (ESI): mass calcd. for C22H21ClFN5O2:441.14 m/z, found: 440.10 [M-H]-.1H NMR (300 MHz, DMSO-d6) δ 8.47 – 8.38 (m, 2H), 8.25 – 8.38 (s, 2H), 7.67 – 7.76 (m, 1H), 7.35 – 7.46 (m, 1H), 7.07 – 7.19 (m, 1H), 6.95 (s, 1H), 6.29 – 6.37 (m, 1H), 1.83 (d, J = 6.9 Hz, 3H), 1.36 (s, 9H). 19F NMR (282 MHz, DMSO) δ -118.07. Example 365: (S)-1-(tert-butyl)-3-(7-chloro-1-(1-(2,5-dichlorophenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000558_0001
(S)-7-chloro-1-(1-(2,5-dichlorophenyl)ethyl)-6-nitroquinoxalin-2(1H)-one To a solution of 7-chloro-6-nitro-1H-quinoxalin-2-one (800 mg, 3.546 mmol, 1 equiv) in DCM (10 mL) was added (1R)-1-(2,5-dichlorophenyl)ethanol (1016.27 mg, 5.319 mmol, 1.5 equiv) and PPh3 (1395.23 mg, 5.319 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (926.40 mg, 5.319 mmol, 1.5 equiv) under N2 at 0°C. The resulting mixture was stirred for additional 1h at 0°C. The reaction was monitored by TLC. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-chloro-1-[(1S)-1-(2,5- dichlorophenyl)ethyl]-6-nitroquinoxalin-2-one) as a yellow solid.
Figure imgf000559_0001
(S)-6-amino-7-chloro-1-(1-(2,5-dichlorophenyl)ethyl)quinoxalin-2(1H)-one To a solution of 7-chloro-1-[(1S)-1-(2,5-dichlorophenyl)ethyl]-6-nitroquinoxalin-2-one (60 mg, 0.151 mmol, 1 equiv) in MeOH (5 mL) and H2O (0.5 mL) was added Fe (84.06 mg, 1.510 mmol, 10 equiv) and NH4Cl (80.51 mg, 1.510 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3x50 mL). The filtrate was concentrated under reduced pressure. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-chloro-1-[(1S)-1-(2,5- dichlorophenyl)ethyl]quinoxalin-2-one (30 mg, 54.07%) as a yellow solid. LC/MS: mass calcd. for C16H12Cl3N3O:367.00 m/z, found:367.95 [M+H]+.
Figure imgf000559_0002
(S)-1-(tert-butyl)-3-(7-chloro-1-(1-(2,5-dichlorophenyl)ethyl)-2-oxo-1,2-dihydroquinoxalin- 6-yl)urea To a solution of 6-amino-7-chloro-1-[(1S)-1-(2,5-dichlorophenyl)ethyl]quinoxalin-2-one (30 mg, 0.081 mmol, 1 equiv) in DCM (3 mL) was added triphosgene (24.15 mg, 0.081 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (24.71 mg, 0.243 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (17.86 mg, 0.243 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column30*150 mm column (eluent: 51% to 67% (v/v) CH3CN and Water(10 mmol/L NH4HCO3+0.1%NH3.H2O)) to afford the title compound 3-tert-butyl-1-{7-chloro-1-[(1S)-1-(2,5- dichlorophenyl)ethyl]-2-oxoquinoxalin-6-yl}urea (22 mg, 57.79%) as a yellow solid. LC/MS: mass calcd. for C21H21Cl3N4O2:466.07 m/z, found:467.10 [M+H]+. 1H NMR (300 MHz, DMSO- d6) δ 8.58 (s, 1H), 8.06 (d, J = 9.7 Hz, 2H), 7.92 (s, 1H), 7.84 (d, J = 2.2 Hz, 1H), 7.35 – 7.48 (m, 2H), 6.98 (s, 1H), 6.09 (d, J = 7.0 Hz, 1H), 1.84 (d, J = 6.8 Hz, 3H), 1.32 (s, 9H). Example 366: 1-(tert-butyl)-3-((R)-6-cyano-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro- 2H-benzo[b][1,4]oxazin-7-yl)urea Synthetic Scheme
Figure imgf000560_0001
(R)-6-bromo-2-methyl-7-nitro-4-((S)-1-phenylethyl)-2H-benzo[b][1,4] oxazin-3(4H)-one To a stirred solution of (2R)-6-bromo-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.045 mmol, 1 equiv) and (R)-1-phenylethan-1-ol (127.67 mg, 1.045 mmol, 1 equiv) in DCM (4 mL) were added PPh3 (411.16 mg, 1.567 mmol, 1.5 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 0.5 h at room temperature under nitrogen atmosphere. Then, to the above solution was added DEAD (546.00 mg, 3.135 mmol, 3 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 2 h at room temperature under nitrogen atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (15 mL). The resulting mixture was extracted with ethyl acetate (3 x 15 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-6-bromo-2-methyl-7-nitro-4-((S)-1-phenylethyl)-2H-benzo[b] [1,4] oxazin-3(4H)-one (225 mg, 55%) as a yellow oil. LC/MS: mass calcd for C17H15BrN2O4: 390.02, found: 391.05, 393.05 [M+H, M+H+2]+ .
Figure imgf000561_0001
(R)-7-amino-6-bromo-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one To a stirred solution of (R)-6-bromo-2-methyl-7-nitro-4-((S)-1-phenylethyl)-2H-benzo[b][1,4] oxazin-3(4H)-one (220 mg, 0.562 mmol, 1 equiv) and NH4Cl (300.80 mg, 5.620 mmol, 10 equiv) in MeOH/H2O (10:1, 4 mL) was added Fe (314.04 mg, 5.620 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70 °C under air atmosphere. Desired product could be detected by LCMS. After cooling down to rt, the reaction was quenched with water (15 mL). The resulting mixture was extracted with ethyl acetate (3 x 15 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-7-amino-6-bromo-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b] [1,4] oxazin-3(4H)-one (190 mg, 70.15%) as a yellow solid. LC/MS: mass calcd for C17H17BrN2O2: 360.05, found: 361.02, 363.02 [M+H, M+H+2]+ .
Figure imgf000562_0001
(R)-7-amino-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H-benzo[b][1,4]oxazine-6- carbonitrile To a stirred solution of (R)-7-amino-6-bromo-2-methyl-4-((S)-1-phenylethyl)-2H-benzo[b] [1,4] oxazin-3(4H)-one (190 mg, 0.526 mmol, 1 equiv) and BrettPhos Pd G3 (95.36 mg, 0.105 mmol, 0.2 equiv) in DMF (5 mL) were added BrettPhos (56.47 mg, 0.105 mmol, 0.2 equiv), Zn(CN)2 (12.35 mg, 0.105 mmol, 0.2 equiv), and Zn (68.78 mg, 1.052 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 120 °C under nitrogen atmosphere. Desired product could be detected by LCMS. After cooling down to rt, the reaction was quenched with water (15 mL). The resulting mixture was extracted with ethyl acetate (3 x 15 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (R)-7-amino-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b] [1,4] oxazine-6-carbonitrile (118 mg, 51.10%) as a yellow oil. LC/MS: mass for C18H17N3O2:307.13, found: 308.02 [M+H]+.
Figure imgf000562_0002
1-(tert-butyl)-3-((R)-6-cyano-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4] oxazin-7-yl)urea To a stirred solution of (R)-7-amino-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b] [1,4] oxazine-6-carbonitrile (108 mg, 0.351 mmol, 1 equiv) and 2-isocyanato-2- methylpropane (34.83 mg, 0.351 mmol, 1 equiv) in DCM (4 mL) was added TEA (106.67 mg, 1.053 mmol, 3 equiv) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. Desired product could be detected by LCMS. The reaction was quenched with water (15 mL). The resulting mixture was extracted with ethyl acetate (3 x 15 mL). The crude product (60mg) was purified by Prep-HPLC with the following conditions (Column: XBridge Prep C18 OBD Column, 19*250 mm, 5μm; Mobile Phase A: Water (10mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min mL/min; Gradient: 50% B to 80% B in 7 min; Wave Length: 254 nm; RT1(min): 5.9 ; Number Of Runs: 2) to afford 1-(tert-butyl)-3-((R)-6-cyano-2-methyl-3-oxo-4-((S)-1-phenylethyl)-3,4-dihydro-2H- benzo[b][1,4] oxazin-7-yl)urea(37.7 mg, 26.39%) as a white solid. LC/MS: mass calcd for C23H26N4O3:406.20, found: 407.25 [M+H]+ .1H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.84 (s, 1H), 7.43 – 7.33 (m, 3H), 7.20-7.33 (m, 3H), 6.97 (d, J = 9.2 Hz,2H), 5.95-6.14 (m, 1H),1.78(d, J = 7.1 Hz,3H),1.48(d, J = 6.8 Hz,3H), 1.28 (s, 9H). Example 367: 3-tert-butyl-1-[(2R)-6-cyano-4-[(2,5-dichlorophenyl)methyl]-2-methyl-3-oxo- 2H-1,4-benzoxazin-7-yl]urea Synthetic Scheme
Figure imgf000563_0001
(2R)-6-bromo-4-[(2,5-dichlorophenyl)methyl]-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one To a solution of (2R)-6-bromo-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 1.045 mmol, 1 equiv) in DMF (5 mL) was added 2-(bromomethyl)-1,4-dichlorobenzene (300.87 mg, 1.254 mmol, 1.2 equiv) and K2CO3 (436.45 mg, 3.135 mmol, 3 equiv). The reaction was stirred at 80 °C for 1 h. The mixture was allowed to cool down to room temperature. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-bromo-4-[(2,5- dichlorophenyl)methyl]-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (370 mg, 79.37%) as a white solid. MS (ESI): mass calcd. for C16H11BrCl2N2O4:443.93 m/z, found: 444.92, 446.92 [M+H, M+H+2]+.
Figure imgf000564_0001
(2R)-7-amino-6-bromo-4-[(2,5-dichlorophenyl)methyl]-2-methyl-2H-1,4-benzoxazin-3-one To a solution of (2R)-6-bromo-4-[(2,5-dichlorophenyl)methyl]-2-methyl-7-nitro-2H-1,4- benzoxazin-3-one (370 mg, 0.829 mmol, 1 equiv) in MeOH (0.5 mL) and H2O (5 mL) was added Fe (463.21 mg, 8.290 mmol, 10 equiv) and NH4Cl (443.67 mg, 8.290 mmol, 10 equiv). The reaction was stirred at 70 ℃ for 2 h. The mixture was allowed to cool down to room temperature and quenched with water (50 mL). The resulting mixture was filtered, the filter cake was washed with EA (50 mL). The filtrate was extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-6-bromo-4- [(2,5-dichlorophenyl)methyl]-2-methyl-2H-1,4-benzoxazin-3-one (270 mg, 78.23%) as a yellow oil. MS (ESI): mass calcd. for C16H13BrCl2N2O2:413.95 m/z, found:414.95, 416.95 [M+H, M+H+2]+.
Figure imgf000564_0002
(2R)-7-amino-4-[(2,5-dichlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4-benzoxazine-6- carbonitrile To a solution of (2R)-7-amino-6-bromo-4-[(2,5-dichlorophenyl)methyl]-2-methyl-2H-1,4- benzoxazin-3-one (260 mg, 0.625 mmol, 1 equiv) in DMF (5 mL) was added Zn(CN)2 (88.04 mg, 0.750 mmol, 1.2 equiv) and Pd(PPh3)4 (72.21 mg, 0.063 mmol, 0.1 equiv). The reaction was stirred at 90 °C for 2 h under N2 atmosphere. The mixture was allowed to cool down to room temperature. Quenched with water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-[(2,5- dichlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4-benzoxazine-6-carbonitrile (120 mg, 53.02%) as a white solid. MS (ESI): mass calcd. for C17H13Cl2N3O2:361.04 m/z, found:362.04 [M+H]+.
Figure imgf000565_0001
3-tert-butyl-1-[(2R)-6-cyano-4-[(2,5-dichlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4- benzoxazin-7-yl]urea To a solution of (2R)-7-amino-4-[(2,5-dichlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4- benzoxazine-6-carbonitrile (110 mg, 0.304 mmol, 1 equiv) in DCM (10 mL) was added triphosgene (45.06 mg, 0.152 mmol, 0.5 equiv) at 0°C under N2 atmosphere. The resulting mixture was stirred for 1 h at 25 °C. Then, TEA (92.19 mg, 0.912 mmol, 3 equiv) was added into above system at 0 °C. The resulting mixture was stirred for 1 h at 25 °C. To the above mixture was added erbumine (33.32 mg, 0.456 mmol, 1.5 equiv) dropwise at 0 °C. The reaction was stirred at 25 °C for 2 h. Quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using XBridge Prep OBD C1830*150 mm, 5μm column (eluent: 53% to 67% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-6-cyano-4-[(2,5-dichlorophenyl)methyl]-2-methyl-3-oxo-2H-1,4- benzoxazin-7-yl]urea (68.4 mg, 48.33%) as a white solid. MS (ESI): mass calcd. for C22H22Cl2N4O3:460.11 m/z, found:461.05 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.88 (s, 1H), 7.53 – 7.62 (m, 1H), 7.37 – 7.47 (m, 1H), 7.32 (s, 1H), 7.03 – 7.11 (m, 1H), 6.98 (s, 1H), 4.96 – 5.25 (m, 3H), 1.52 (d, J = 6.8 Hz, 3H), 1.30 (s, 9H). Example 368: (R)-1-(1-benzyl-3-methyl-2-oxo-7-(trifluoromethyl)-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-6-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000566_0001
methyl (R)-2-((3-nitro-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate To a solution of methyl (R)-2-hydroxypropanoate (2.30 g, 22.071 mmol, 1 equiv) in THF (60 mL) was added NaH (794.50 mg, 33.107 mmol, 1.5 equiv) at 0°C under air atmosphere. After 30 min, the 2-chloro-3-nitro-5-(trifluoromethyl)pyridine (5 g, 22.071 mmol, 1 equiv) was added to the mixture at 0°C. The resulting mixture was stirred for 2h at room temperature. Desired product could be detected by LCMS. The mixture was diluted with water (150 mL). The resulting mixture was extracted with EA (3 x 150 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0-20% ethyl acetate/petroleum ether) to afford the methyl (R)-2-((3-nitro-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate (3.5 g, 53.90%) as a white solid. MS (ESI): mass calcd. for C10H9F3N2O5: 294.05 m/z, found 295.05 [M+H] +.
Figure imgf000567_0001
(R)-2-((3-amino-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate To a solution of methyl (R)-2-((3-nitro-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate (3.5 g, 11.897 mmol, 1 equiv) in MeOH (40 mL, 24.699 mmol) was added Pd/C (3.49 g) at 0°C under N2 atmosphere. The resulting mixture was maintained under hydrogen and stirred at 25 °C for 3 h. The resulting mixture was filtered, and the filter cake was washed with MeOH (4 x 40 mL). The filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel chromatography (0 - 50% ethyl acetate/petroleum ether) to afford the methyl (R)-2-((3-amino-5- (trifluoromethyl)pyridin-2-yl)oxy)propanoate (2.8 g, 89.08%) as a yellow oil. LC/MS: mass calcd. for C10H11F3N2O3: 264.07, found: 265.05, 267.05 [M+H, M+H+2]+.
Figure imgf000567_0002
methyl (R)-2-((3-amino-6-bromo-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate To a stirred mixture of methyl (R)-2-((3-amino-5-(trifluoromethyl)pyridin-2-yl)oxy)propanoate (1.5 g, 5.677 mmol, 1 equiv) in DMF (15 mL) was added NBS (1.5 g, 8.515 mmol, 1.5 equiv) at -40°C under air atmosphere. The resulting mixture was maintained under air and stirred at -40 °C for 3 h. The reaction was quenched with water (50 mL). The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-30% ethyl acetate/petroleum ether) to afford the methyl (R)-2-((3-amino-6-bromo-5- (trifluoromethyl)pyridin-2-yl)oxy)propanoate (530 mg, 27.21%) as a yellow oil. LC/MS: mass calcd. for C10H10BrF3N2O3: 341.98, found: 342.90, 344.90 [M+H, M+H+2] +.
Figure imgf000567_0003
(R)-6-bromo-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one A stirred mixture of methyl (R)-2-((3-amino-6-bromo-5-(trifluoromethyl)pyridin-2- yl)oxy)propanoate (700 mg, 2.040 mmol, 1 equiv) in HOAc (10 mL) was stirred at 80 °C for 3 h. After cooling down to room temperature, the reaction was quenched with NaHCO3 solution (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the (R)-6- bromo-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (370 mg, 58.30%) as a yellow oil. LC/MS: mass calcd. for C9H6BrF3N2O2: 309.96, found: 310.85, 312.85 [M+H, M+H+2] +.
Figure imgf000568_0001
(R)-6-azido-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one To a stirred mixture of (R)-6-bromo-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin- 2(3H)-one (300 mg, 0.964 mmol, 1 equiv) in DMF (5 mL) was added NaN3 (94.05 mg, 1.446 mmol, 1.5 equiv) at 25°C under air atmosphere. The resulting mixture was maintained under nitrogen and stirred at 110 °C for 3 h. After cooling down to room temperature, the reaction was quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the (R)-6-azido-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)- one as a yellow oil. LC/MS: mass calcd. for C9H6F3N5O2: 273.05, found: 274.10 [M+H] +.
Figure imgf000568_0002
(R)-6-azido-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one To a stirred mixture of (R)-6-azido-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin- 2(3H)-one (147 mg, 0.538 mmol, 1 equiv) in DMF (5 mL) was added (bromomethyl)benzene (138.06 mg, 0.807 mmol, 1.5 equiv) , K2CO3 (224.74 mg, 1.614 mmol, 3 equiv) at 25°C under air atmosphere. The resulting mixture was maintained under air and stirred at 25 °C for 3 h. The reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the (R)-6-azido-1-benzyl-3-methyl-7-(trifluoromethyl)-1H- pyrido[2,3-b][1,4]oxazin-2(3H)-one (140 mg, 71.61%) as a yellow oil. LC/MS: mass calcd. for C16H12F3N5O2: 363.09, found: 364.20 [M+H] +.
Figure imgf000569_0001
(R)-6-amino-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one To a stirred mixture of (R)-6-azido-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3- b][1,4]oxazin-2(3H)-one (130 mg, 0.358 mmol, 1 equiv) in MeOH (3 mL) was added Pd/C (129.85 mg) at 0°C under air atmosphere. The resulting mixture was maintained under hydrogen and stirred at 25 °C for 3 h. The resulting mixture was filtered, and the filter cake was washed with MeOH (4 x 40 mL). The filtrate was concentrated under reduced pressure. This afforded the (R)- 6-amino-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one (110 mg, 91.14%) as a yellow oil. LC/MS: mass calcd. for C16H14F3N3O2: 337.10, found: 338.15 [M+H]+ .
Figure imgf000569_0002
(R)-1-(1-benzyl-3-methyl-2-oxo-7-(trifluoromethyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-6-yl)-3-(tert-butyl)urea To a stirred mixture of (R)-6-amino-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3- b][1,4]oxazin-2(3H)-one (100 mg, 0.296 mmol, 1 equiv) in DCM (3 mL) was added triphosgene (87.97 mg, 0.296 mmol, 1 equiv) at 0 degrees C under nitrogen atmosphere. After stirring at 0 degrees C for 1h, the TEA (150.00 mg, 1.480 mmol, 5 equiv) was added to the mixture at room temperature. After 30 min, erbumine (65.05 mg, 0.888 mmol, 3 equiv) and TEA (150.00 mg, 1.480 mmol, 5 equiv) was added to the reaction system. The resulting mixture was stirred for 1h at 25 degrees C. Desired product could be detected by LCMS. The mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a X Bridge Prep OBD C18150 mm x 30 mm x 5μm column (eluent: 49% to 71% (v/v) CH3CN and H2O with 10 mmol /L NH4HCO3) to afford the (R)-1-(1-benzyl-3-methyl-2-oxo- 7-(trifluoromethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)-3-(tert-butyl)urea (12.8 mg, 9.52%) as a white solid. MS (ESI): mass calcd. for C21H23F3N4O3: 436.17 m/z, found 437.15 [M +H] +.1H-NMR (400 MHz, DMSO-d6) δ 7.72 (s, 1H), 7.57 (s, 1H), 7.31 – 7.39 (m, 2H), 7.23 – 7.31 (m, 4H), 5.24 – 5.33 (m, 1H), 5.21 (d, J = 7.5 Hz, 2H), 1.60 (d, J = 6.8 Hz, 3H), 1.29 (s, 9H).19F-NMR (376 MHz, DMSO) δ -58.98. Example 369: (R)-1-(4-benzyl-6-cyano-8-fluoro-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazin-7-yl)-3-(tert-butyl)urea Synthetic Scheme
Figure imgf000570_0001
methyl (R)-2-(4-bromo-2-fluoro-6-nitrophenoxy)propanoate A solution of 4-bromo-2-fluoro-6-nitrophenol (5 g, 21.187 mmol, 1 equiv) in tetrahydrofuran (50 mL) was treated with methyl (2S)-2-hydroxypropanoate (2.21 g, 21.187 mmol, 1 equiv) and PPh3 (8.34 g, 31.781 mmol, 1.5 equiv) for 1 h at rt under nitrogen atmosphere followed by the addition of DIAD (6.43 g, 31.781 mmol, 1.5 equiv) dropwise at 0°C. The resulting mixture was stirred for 2 h at rt under N2 atmosphere. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give methyl (2R)-2-(4-bromo-2-fluoro-6-nitrophenoxy)propanoate (5.8 g, 84.99%) as a white solid. Desired product could be detected by TLC.
Figure imgf000571_0001
(R)-6-bromo-8-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of methyl (2R)-2-(4-bromo-2-fluoro-6-nitrophenoxy)propanoate (5.8 g, 18.008 mmol, 1 equiv) in EtOH (60 mL)/H2O (6 mL) was added Fe (10.06 g, 180.080 mmol, 10 equiv) and NH4Cl (9.63 g, 180.080 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (100 mL) and extracted with EA (3 x 100 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-bromo-8-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (4.5 g, 96.09%) as a white solid. MS (ESI): mass calcd. forC9H7BrFNO2: 258.96 m/z, found 259.95, 261.95 [M+H, M+H+2]+.
Figure imgf000571_0002
(R)-6-bromo-8-fluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6-bromo-8-fluoro-2-methyl-2,4-dihydro-1,4-benzoxazin-3-one (1.5 g, 5.768 mmol, 1 equiv) in sulfuric acid (10 mL) was added KNO3 (0.58 g, 5.768 mmol, 1 equiv) at 0°C. The reaction was stirred at 0°C for 1 h. Quenched with ice water (50 mL) and extracted with EA (3 x 50 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-bromo-8-fluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3- one (1.46 g, 82.98%) as a white solid. MS (ESI): mass calcd. for C9H6BrFN2O4: 303.95 m/z, found: 302.95, 304.95 [M-H, M-H+2]-.
Figure imgf000572_0001
(R)-4-benzyl-6-bromo-8-fluoro-2-methyl-7-nitro-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6-bromo-8-fluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (350 mg, 1.147 mmol, 1 equiv) in DMF (3 mL) was added benzyl bromide (294.35 mg, 1.720 mmol, 1.5 equiv) and K2CO3 (479.17 mg, 3.441 mmol, 3 equiv). The reaction was stirred at rt for 1 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-4-benzyl-6- bromo-8-fluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (416 mg, 91.75%) as a white solid. MS (ESI): mass calcd. for C16H12BrFN2O4: 394.00 m/z, found: 395.18, 397.18 [M+H, M+H+2]+.
Figure imgf000572_0002
(R)-7-amino-4-benzyl-6-bromo-8-fluoro-2-methyl-2H-benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-4-benzyl-6-bromo-8-fluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (400 mg, 1.012 mmol, 1 equiv) in EtOH (8 mL)/H2O (1 mL) was added Fe (565.26 mg, 10.120 mmol, 10 equiv) and NH4Cl (541.42 mg, 10.120 mmol, 10 equiv). The reaction was stirred at 70°C for 3 h. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-benzyl-6-bromo-8-fluoro-2-methyl-2H- 1,4-benzoxazin-3-one (250 mg, 67.63%) as a white solid. MS (ESI): mass calcd. for C16H14BrFN2O2: 364.02 m/z, found 365.05, 367.05 [M+H, M+H+2]+.
Figure imgf000573_0001
(R)-7-amino-4-benzyl-8-fluoro-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6- carbonitrile To a solution of (2R)-7-amino-4-benzyl-6-bromo-8-fluoro-2-methyl-2H-1,4-benzoxazin-3-one (240 mg, 0.657 mmol, 1 equiv) in DMF (5 mL) was added Zn(CN)2 (154.33 mg, 1.314 mmol, 2 equiv), Zn (12.89 mg, 0.197 mmol, 0.3 equiv), BrettPhos (70.55 mg, 0.131 mmol, 0.2 equiv) and BrettPhos Pd G3 (119.15 mg, 0.131 mmol, 0.2 equiv). The reaction was stirred at 120°C for 3 h under N2 atmosphere. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-benzyl-8-fluoro-2- methyl-3-oxo-2H-1,4-benzoxazine-6-carbonitrile (160 mg, 78.21%) as a white solid. MS (ESI): mass calcd. for C17H14FN3O2: 311.11 m/z, found: 312.20 [M+H]+.
Figure imgf000573_0002
(R)-1-(4-benzyl-6-cyano-8-fluoro-2-methyl-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)- 3-(tert-butyl)urea To a solution of (2R)-7-amino-4-benzyl-8-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazine-6- carbonitrile (100 mg, 0.321 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (95.31 mg, 0.321 mmol, 1 equiv). The reaction was stirred at 0 °C under N2 for 1 h. The reaction was added TEA (162.52 mg, 1.605 mmol, 5 equiv) and stirred at 0 °C under N2 for 1 h. The reaction was added erbumine (70.48 mg, 0.963 mmol, 3 equiv) and stirred at rt under N2 for 1 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a Aeris PEPTIDE 5um XB-C18 Axia 21.2 mm X 250 mm, 5 μm column (eluent: 43% to 57% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 1-[(2R)-4-benzyl-6-cyano-8-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]-3- tert-butylurea (42.1 mg, 31.66%) as a white solid. LC/MS (ESI): mass calcd. for C22H23FN4O3: 410.18 m/z, found: 411.15 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.31 – 7.42 (m, 3H), 7.21 – 7.31 (m, 3H), 6.32 (s, 1H), 5.21 (s, 2H), 5.10-5.17 (m, 1H), 1.55-1.58 (d, J = 9 Hz, 3H), 1.27 (s, 9H).19F NMR (282 MHz, DMSO) δ -140.79. Example 370: (R)-1-(tert-butyl)-3-(6-cyano-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl- 3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea
Figure imgf000574_0001
(R)-6-bromo-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-7-nitro-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6-bromo-8-fluoro-2-methyl-7-nitro-2,4-dihydro-1,4-benzoxazin-3-one (300 mg, 0.983 mmol, 1 equiv) in DMF (5 mL) was added 1-(bromomethyl)-3- (difluoromethyl)benzene (326.07 mg, 1.474 mmol, 1.5 equiv) and K2CO3 (410.71 mg, 2.949 mmol, 3 equiv). The reaction was stirred at rt for 3 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-6-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-8- fluoro-2-methyl-7-nitro-2H-1,4-benzoxazin-3-one (400 mg, 91.36%) as a white solid. MS (ESI): mass calcd. for C17H12BrF3N2O4: 443.99 m/z, found: 443.00, 445.00 [M-H, M-H+2]-.
Figure imgf000575_0001
(R)-7-amino-6-bromo-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-2H- benzo[b][1,4]oxazin-3(4H)-one To a solution of (2R)-6-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-7-nitro- 2H-1,4-benzoxazin-3-one (600 mg, 1.348 mmol, 1 equiv) in EtOH (6 mL)/H2O (0.6 mL) was added Fe (752.64 mg, 13.480 mmol, 10 equiv) and NH4Cl (720.90 mg, 13.480 mmol, 10 equiv) . The reaction was stirred at 70°C for 3 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to give (2R)-7-amino-6-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl- 2H-1,4-benzoxazin-3-one (376 mg, 67.19%) as a white solid. MS (ESI): mass calcd. forC17H14BrF3N2O2: 414.02 m/z, found 415.05, 417.05 [M+H, M+H+2]+.
Figure imgf000575_0002
(R)-7-amino-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-3,4-dihydro-2H- benzo[b][1,4]oxazine-6-carbonitrile To a solution of (2R)-7-amino-6-bromo-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2- methyl-2H-1,4-benzoxazin-3-one (370 mg, 0.891 mmol, 1 equiv) in DMF (5 mL) was added Zn(CN)2 (313.90 mg, 2.673 mmol, 3 equiv), Brettphos (95.67 mg, 0.178 mmol, 0.2 equiv), BrettPhos Pd G3 (140.23 mg, 0.178 mmol, 0.2 equiv), Zn (17.48 mg, 0.267 mmol, 0.3 equiv). The reaction was stirred at 120°C for 3 h under N2. The mixture was allowed to cool down to rt. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (2R)-7-amino-4-{[3- (difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazine-6-carbonitrile (190 mg, 59.01%) as a white solid. MS (ESI): mass calcd. for C18H14F3N3O2: 361.10 m/z, found: 360.00 [M-H]-.
Figure imgf000576_0001
(R)-1-(tert-butyl)-3-(6-cyano-4-(3-(difluoromethyl)benzyl)-8-fluoro-2-methyl-3-oxo-3,4- dihydro-2H-benzo[b][1,4]oxazin-7-yl)urea To a solution of (2R)-7-amino-4-{[3-(difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-3-oxo- 2H-1,4-benzoxazine-6-carbonitrile (100 mg, 0.277 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (82.12 mg, 0.277 mmol, 1 equiv). The reaction was stirred at 0 °C under N2 for 1 h. The reaction was added TEA (140.03 mg, 1.385 mmol, 5 equiv) and stirred at 0 °C under N2 for 1 h. The reaction was added erbumine (60.73 mg, 0.831 mmol, 3 equiv) and stirred at rt under N2 for 1 h. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a Aeris PEPTIDE 5um XB-C18 Axia 21.2 mm X 250 mm, 5 μm column (eluent: 43% to 57% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(2R)-6-cyano-4-{[3- (difluoromethyl)phenyl]methyl}-8-fluoro-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (32.8 mg, 25.64%) as a white solid. LC/MS (ESI): mass calcd. for C23H23F3N4O3: 460.17 m/z, found: 461.10 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.10 (s, 1H), 7.35 – 7.57 (m, 5H), 7.03 (s, 1H), 6.32 (s, 1H), 5.27 (s, 2H), 5.12-5.18 (m, 1H), 1.56-1.58 (d, J = 6 Hz, 3H), 1.27 (s, 9H).19F NMR (282 MHz, DMSO) δ -109.47, -109.77, -140.68. Example 371: 3-tert-butyl-1-[(3R)-7-cyano-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl- 2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6-yl]urea Synthetic Scheme
Figure imgf000577_0001
(3R)-6-bromo-7-chloro-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-3H-pyrido[2,3- b][1,4]oxazin-2-one To a stirred mixture of (3R)-6-bromo-7-chloro-3-methyl-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (150 mg, 0.541 mmol, 1 equiv) and 1-(bromomethyl)-3-(difluoromethyl)benzene (238.97 mg, 1.082 mmol, 2 equiv) in DMF (10 mL) was added K2CO3 (225.75 mg, 1.623 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 4 h at 80°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (3R)-6-bromo-7-chloro-1- {[3-(difluoromethyl)phenyl]methyl}-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one (150 mg, 66.45%) as a white solid. LC/MS (ESI): mass calcd. for C16H12BrClF2N2O2:416.0 m/z., found:417.0, 419.0 [M+H, M+H+2]+.
Figure imgf000578_0001
(3R)-6-amino-7-chloro-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-3H-pyrido[2,3- b][1,4]oxazin-2-one To a stirred mixture of (3R)-6-bromo-7-chloro-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl- 3H-pyrido[2,3-b][1,4]oxazin-2-one (150 mg, 0.359 mmol, 1 equiv) and NaN3 (30.65 mg, 0.471 mmol, 3 equiv) in EtOH:H2O=10:1 (5 mL) was added N1,N2-dimethylcyclohexane-1,2-diamine (30.65 mg, 0.215 mmol, 0.6 equiv) and CuSO4 (25.09 mg, 0.157 mmol, 1 equiv) and sodium ascorbate (143.03 mg, 0.718 mmol, 2 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 80°C under nitrogen atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3)to afford (3R)-6-amino-7-chloro-1-{[3- (difluoromethyl)phenyl]methyl}-3-methyl-3H-pyrido[2,3-b][1,4]oxazin-2-one (100 mg, 78.71%) as a white solid. LC/MS (ESI): mass calcd. for C16H14ClF2N3O2:353.1 m/z, found:354.1 [M+H]+.
Figure imgf000578_0002
(3R)-6-amino-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-2-oxo-3H-pyrido[2,3- b][1,4]oxazine-7-carbonitrile To a stirred mixture of (3R)-6-amino-7-chloro-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl- 3H-pyrido[2,3-b][1,4]oxazin-2-one (100 mg, 0.283 mmol, 1 equiv) and Zn(CN)2 (66.39 mg, 0.566 mmol, 2 equiv) in DMF (5 mL) was added BrettPhos Pd G3 (51.25 mg, 0.057 mmol, 0.2 equiv) and BrettPhos (17.67 mg, 0.033 mmol, 0.2 equiv) and Zn (5.54 mg, 0.085 mmol, 0.3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 1h at 120°C under nitrogen atmosphere. After cooling down to room temperature, the reaction was quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to afford (3R)-6-amino-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-2-oxo-3H-pyrido[2,3- b][1,4]oxazine-7-carbonitrile (60 mg, 61.5% ) as a white solid. LC/MS (ESI): mass calcd. for C17H14F2N4O2:344.1 m/z, found: 345.1 [M+H]+.
Figure imgf000579_0001
3-tert-butyl-1-[(3R)-7-cyano-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-2-oxo-3H- pyrido[2,3-b][1,4]oxazin-6-yl]urea To a stirred solution of (3R)-6-amino-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-2-oxo- 3H-pyrido[2,3-b][1,4]oxazine-7-carbonitrile (60 mg, 0.174 mmol, 1 equiv) and triphosgene (62.05 mg, 0.209 mmol, 1.2 equiv) in DCM (5 mL) was added TEA (52.90 mg, 0.522 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added 2- methylpropan-2-amine (25.49 mg, 0.348 mmol, 2 equiv) dropwise at room temperature. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford 3-tert-butyl-1-[(3R)-7- cyano-1-{[3-(difluoromethyl)phenyl]methyl}-3-methyl-2-oxo-3H-pyrido[2,3-b][1,4]oxazin-6- yl]urea (2.8 mg, 3.51%) as a white solid. LC/MS (ESI): mass calcd. for C22H23F2N5O3:443.2 m/z, found:444.2 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H), 7.85 (s, 1H), 7.32-7.45 (m, 4H), 6.85-7.20 (m, 2H), 5.25-5.34 (m, 1H), 5.20 (s, 2H), 1.58 (d, J = 6.9 Hz, 3H), 1.29 (s, 9H). 19F NMR (376 MHz, DMSO) δ -109.69. Example 372: (S)-1-(tert-butyl)-3-(7-chloro-1-(1-(5-chloro-2-fluorophenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea Synthetic Scheme
Figure imgf000580_0001
(S)-7-chloro-1-(1-(5-chloro-2-fluorophenyl)ethyl)-6-nitroquinoxalin-2(1H)-one To a solution of 7-chloro-6-nitro-1H-quinoxalin-2-one (500 mg, 2.216 mmol, 1 equiv) in DCM (10 mL) was added (1R)-1-(5-chloro-2-fluorophenyl)ethanol (386.99 mg, 2.216 mmol, 1 equiv) and PPh3 (872.02 mg, 3.324 mmol, 1.5 equiv). The reaction was stirred at rt under N2 for 0.5h. To the above mixture was added DEAD (579.00 mg, 3.324 mmol, 1.5 equiv) at 0°C. The resulting mixture was stirred for additional 1h at 0°C. Quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 7-chloro-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-6-nitroquinoxalin-2- one (120 mg, 14.17%) as a yellow solid. LC/MS: mass calcd. for C16H10Cl2FN3O3:381.01 m/z, found:382.10 [M+H]+.
Figure imgf000580_0002
(S)-6-amino-7-chloro-1-(1-(5-chloro-2-fluorophenyl)ethyl)quinoxalin-2(1H)-one To a solution of 7-chloro-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-6-nitroquinoxalin-2-one (140 mg, 0.366 mmol, 1 equiv) in MeOH (10 mL) and H2O (1 mL) was added Fe (204.58 mg, 3.660 mmol, 10 equiv) and NH4Cl (195.95 mg, 3.660 mmol, 10 equiv). The reaction was stirred at 80 °C for 1h. The mixture was allowed to cool down to rt. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 50 mL). The filtrate was concentrated under reduced pressure. Quenched with water (20 mL) and extracted with EA (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-7-chloro-1-[(1S)- 1-(5-chloro-2-fluorophenyl)ethyl]quinoxalin-2-one (120 mg, 93.01%) as a yellow solid. LC/MS: mass calcd. for C16H12Cl2FN3O:351.03 m/z, found:352.00 [M+H]+.
Figure imgf000581_0001
(S)-1-(tert-butyl)-3-(7-chloro-1-(1-(5-chloro-2-fluorophenyl)ethyl)-2-oxo-1,2- dihydroquinoxalin-6-yl)urea To a solution of 6-amino-7-chloro-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]quinoxalin-2-one (100 mg, 0.284 mmol, 1 equiv) in DCM (5 mL) was added triphosgene (84.25 mg, 0.284 mmol, 1 equiv) under N2 at 0 °C. The reaction was stirred at 0 °C under N2 for 1h. To the above mixture was added TEA (86.20 mg, 0.852 mmol, 3 equiv) at 0°C. The resulting mixture was stirred for additional 1h at rt. Then, to the above mixture was added erbumine (62.30 mg, 0.852 mmol, 3 equiv). The resulting mixture was stirred for additional 1h at rt. Quenched with water (20 mL) and extracted with DCM (3 x 20 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Prep OBD C18 Column, 30*150 mm, 5μm column (eluent: 43% to 73% (v/v) CH3CN and Water (10mmol/L NH4HCO3+0.05%NH3H2O)) to afford the title compound 3-tert-butyl-1- {7-chloro-1-[(1S)-1-(5-chloro-2-fluorophenyl)ethyl]-2-oxoquinoxalin-6-yl}urea (48.8 mg, 37.94%) as a yellow solid. LC/MS: mass calcd. for C21H21Cl2FN4O2:450.10 m/z, found:451.15 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.11 (s, 1H), 8.05 (s, 1H), 7.90 (s, 1H), 7.73 (dd, J = 6.8, 2.7 Hz, 1H), 7.43 (ddd, J = 8.7, 4.2, 2.6 Hz, 1H), 7.15 (dd, J = 10.5, 8.8 Hz, 1H), 6.99 (s, 1H), 6.36 (s, 1H), 1.84 (d, J = 6.8 Hz, 3H), 1.32 (s, 9H).19F NMR (376 MHz, DMSO) δ(ppm): -118.1424. Example 373: 3-tert-butyl-1-[(2R)-4-[(5-chloro-2-fluorophenyl)methyl]-6-cyano-2-methyl- 3-oxo-2H-1,4-benzoxazin-7-yl]urea Synthetic Scheme
Figure imgf000582_0001
(2R)-7-amino-6-bromo-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl-2H-1,4-benzoxazin-3- one To a stirred mixture of (2R)-6-bromo-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl-7-nitro-2H- 1,4-benzoxazin-3-one (300 mg, 0.698 mmol, 1 equiv) and NH4Cl (373.51 mg, 6.980 mmol, 10 equiv) in MeOH:H2O=10:1 (10 mL) was added Fe (389.95 mg, 6.980 mmol, 10 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at 70°C under air atmosphere. After cooling down to room temperature, the reaction was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%)to afford (2R)-7-amino-6-bromo-4- [(5-chloro-2-fluorophenyl)methyl]-2-methyl-2H-1,4-benzoxazin-3-one (236 mg, 84.57%) as a white solid. LC/MS (ESI): mass calcd. for C16H13BrClFN2O2:398.0 m/z, found: 399.0, 401.0 [M+H, M+H+2]+.
Figure imgf000582_0002
(2R)-7-amino-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl-3-oxo-2H-1,4-benzoxazine-6- carbonitrile To a stirred mixture of (2R)-7-amino-6-bromo-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl- 2H-1,4-benzoxazin-3-one (230 mg, 0.576 mmol, 1 equiv) and Zn(CN)2 (81.09 mg, 0.691 mmol, 1.2 equiv) in DMF (10 mL) was added Pd(PPh3)4 (66.51 mg, 0.058 mmol, 0.1 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2h at 90°C under nitrogen atmosphere. After cooling down to room temperature, the reaction was quenched with water (30 mL) and extracted with EA (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. TLC showed the desired product was generated. The residue was purified by column chromatography on silica gel with EA/PE (0- 100%) to afford (2R)-7-amino-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl-3-oxo-2H-1,4- benzoxazine-6-carbonitrile (111 mg, 55.78%) as a white solid.
Figure imgf000583_0001
3-tert-butyl-1-[(2R)-4-[(5-chloro-2-fluorophenyl)methyl]-6-cyano-2-methyl-3-oxo-2H-1,4- benzoxazin-7-yl]urea To a stirred solution of (2R)-7-amino-4-[(5-chloro-2-fluorophenyl)methyl]-2-methyl-3-oxo-2H- 1,4-benzoxazine-6-carbonitrile (60 mg, 0.174 mmol, 1 equiv) and triphosgene (61.79 mg, 0.209 mmol, 1.2 equiv) in DCM (5 mL) was added TEA (52.68 mg, 0.522 mmol, 3 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 30 min at room temperature under nitrogen atmosphere. To the above mixture was added 2-methylpropan-2- amine (25.38 mg, 0.348 mmol, 2 equiv) dropwise at room temperature. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched with water (30 mL) and extracted with DCM (3 x 30 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 17% to 42% (v/v) CH3CN and H2O with 10mmol/L NH4HCO3) to afford 3-tert-butyl-1-[(2R)-4-[(5- chloro-2-fluorophenyl)methyl]-6-cyano-2-methyl-3-oxo-2H-1,4-benzoxazin-7-yl]urea (26.1 mg, 33.81%) as a white solid. LC/MS (ESI): mass calcd. for C22H22ClFN4O3:444.1 m/z, found:445.15 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.86 (s, 1H), 7.38-7.50 (m, 2H), 7.26-7.38 (m, 1H), 7.10-7.26 (m, 1H), 6.99 (s, 1H), 5.09-5.41 (m, 2H), 4.89-5.08 (m, 1H), 1.49 (d, J = 6.8 Hz, 3H), 1.29 (s, 9H).19F NMR (376 MHz, DMSO) δ -119.29. Example 374: (R)-1-(tert-butyl)-3-(1-(3-(difluoromethyl) benzyl)-3-methyl-2-oxo-7- (trifluoromethyl)-2,3-dihydro-1H-pyrido[2,3-b] [1,4] oxazin-6-yl) urea Synthetic Scheme
Figure imgf000584_0001
(3R)-6-azido-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7-(trifluoromethyl)-3H- pyrido[2,3-b] [1,4] oxazin-2-one To a stirred solution of (3R)-6-azido-3-methyl-7-(trifluoromethyl)-1H,3H-pyrido[2,3-b] [1,4] oxazin-2-one (80 mg, 0.293 mmol, 1 equiv) and 1-(bromomethyl)-3-(difluoromethyl) benzene (77.68 mg, 0.352 mmol, 1.2 equiv) in DMF (5 mL) was added K2CO3 (122.31 mg, 0.879 mmol, 3 equiv) in portions at room temperature under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (3R)-6-azido-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7- (trifluoromethyl)-3H-pyrido[2,3-b] [1,4] oxazin-2-one (100 mg, 82.62%) as a yellow oil. MS (ESI): mass calcd. for C17H12F5N5O2: 413.09 m/z, found 414.05 [M+H]+.
Figure imgf000585_0001
(3R)-6-amino-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7-(trifluoromethyl)-3H- pyrido[2,3-b] [1,4] oxazin-2-one A mixture of (3R)-6-azido-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7- (trifluoromethyl)-3H-pyrido[2,3-b] [1,4] oxazin-2-one (100 mg, 0.242 mmol, 1 equiv) and Pd/C (30 mg, 0.282 mmol, 1.17 equiv) in MeOH (5 mL) was stirred for 1 h at room temperature under hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with MeOH (4 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1:1) to afford (3R)-6-amino- 1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7-(trifluoromethyl)-3H-pyrido[2,3-b] [1,4] oxazin-2-one (80 mg, 85.37%) as a yellow oil. MS (ESI): mass calcd. for C17H14F5N3O2: 387.10 m/z, found 388.05 [M+H]+.
Figure imgf000585_0002
(R)-1-(tert-butyl)-3-(1-(3-(difluoromethyl) benzyl)-3-methyl-2-oxo-7-(trifluoromethyl)-2,3- dihydro-1H-pyrido[2,3-b] [1,4] oxazin-6-yl) urea A solution of (3R)-6-amino-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-7- (trifluoromethyl)-3H-pyrido[2,3-b] [1,4] oxazin-2-one (80 mg, 0.207 mmol, 1 equiv) in toluene (5 mL) was treated with Triphosgene (122.58 mg, 0.414 mmol, 2 equiv) for 30 min at room temperature under nitrogen atmosphere followed by the addition of triethylamine (104.51 mg, 1.035 mmol, 5 equiv) in portions at 0 °C. The resulting mixture was stirred for 2 h at 90 °C under nitrogen atmosphere. To the above mixture was added erbumine (45.32 mg, 0.621 mmol, 3 equiv) dropwise over 1 min at room temperature. The resulting mixture was stirred for additional 1 h at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using a XSelect CSH Fluoro Pheny 250 mm x 19 mm x 5 μm column (eluent: 45% to 75% (v/v) CH3CN and H2O with 10 mmol/L 0.05% TFA) to afford 3-tert-butyl-1-[(3R)-1-{[3-(difluoromethyl) phenyl] methyl}-3-methyl-2- oxo-7-(trifluoromethyl)-3H-pyrido[2,3-b] [1,4] oxazin-6-yl] urea (30 mg, 29.38%) as a yellow solid. MS (ESI): mass calcd. for C22H23F5N4O3: 486.17 m/z, found 487.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 7.72 (s, 1H), 7.61 (s, 1H), 7.38 – 7.58 (m, 4H), 7.30 (s, 1H), 7.02 (s, 1H), 5.18 – 5.40 (m, 3H), 1.60 (d, J = 6.8 Hz, 3H), 1.15 – 1.42 (m, 9H).19F NMR (282 MHz, DMSO) δ -59.022, -109.803 Example 375: 3-tert-butyl-1-{1-[(1S)-1-[2-fluoro-5-(2,2,2-trifluoro-1- hydroxyethyl)phenyl]ethyl]-2-oxoquinoxalin-6-yl}urea Synthetic Scheme
Figure imgf000586_0001
2-(3-bromo-4-fluorophenyl)-1,3-dioxolane To a solution of 3-bromo-4-fluorobenzaldehyde (10 g, 49.259 mmol, 1 equiv) in Toluene (50 mL) was added ethylene glycol (9477.90 mg, 152.703 mmol, 3.1 equiv) and para-toluene sulfonate (1696.47 mg, 9.852 mmol, 0.2 equiv). The resulting mixture was stirred for 3 h at 120°C. The mixture was diluted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 2-(3-bromo-4-fluorophenyl)-1,3-dioxolane (10 g, 82.17%) as a white solid. MS (ESI): mass calcd. for C9H8BrFO2: 245.97 m/z, found 247.00 [M+H, M+H+2]+.
Figure imgf000587_0001
1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethanone To a solution of 2-(3-bromo-4-fluorophenyl)-1,3-dioxolane (2 g, 8.095 mmol, 1 equiv) in THF (20 mL) was added butyllithium (601.54 mg, 9.390 mmol, 1.16 equiv) dropwise at -78°C under nitrogen atmosphere. The resulting mixture was stirred for 1 h at -78 °C. N-methoxy-N- methylacetamide (868.17 mg, 8.419 mmol, 1.04 equiv) was added to the reaction at -78 °C under nitrogen atmosphere. The resulting mixture was stirred for 1 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 1-[5- (1,3-dioxolan-2-yl)-2-fluorophenyl]ethanone (1.2 g, 70.52%) as a yellow solid. MS (ESI): mass calcd. for C11H11FO3: 210.07 m/z, found 211.00 [M+H]+.
Figure imgf000587_0002
(1R)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethanol To a solution of CBS (1001.50 mg, 3.806 mmol, 0.4 equiv) in THF (20 mL) was added BH3-Me2S (1084.09 mg, 14.273 mmol, 1.5 equiv) and 1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethanone (2 g, 9.515 mmol, 1 equiv) dropwise at 0°C. The resulting mixture was stirred for 1 h at 0°C. The mixture was diluted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford (1R)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethanol (1.5 g, 74.29%) as a white solid. MS (ESI): mass calcd. for C11H13FO3: 212.08 m/z, found 213.30 [M+H]+.
Figure imgf000588_0001
1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-6-nitroquinoxalin-2-one To a solution of (1R)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethanol (1.6 g, 7.539 mmol, 1 equiv) in DCM (20 mL) was added 6-nitro-1H-quinoxalin-2-one (1.44 g, 7.539 mmol, 1 equiv) and PPh3 (2.97 g, 11.308 mmol, 1.5 equiv) dropwise under nitrogen atmosphere. The resulting mixture was stirred for 0.5 h at rt. DEAD (1.97 g, 11.308 mmol, 1.5 equiv) was added to the reaction under nitrogen atmosphere. The resulting mixture was stirred for 1 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-6-nitroquinoxalin-2-one (1.2 g, 41.30%) as a yellow solid. MS (ESI): mass calcd. for C19H16FN3O5: 385.10 m/z, found 386.00 [M+H]+.
Figure imgf000588_0002
6-amino-1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]quinoxalin-2-one To a solution of 1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-6-nitroquinoxalin-2-one (1.2 g, 3.114 mmol, 1 equiv) in MeOH (15 mL) and H2O (1.5 mL) was added Fe (1.74 g, 31.140 mmol, 10 equiv) and NH4Cl (1.67 g, 31.140 mmol, 10 equiv). The resulting mixture was stirred for 3 h at 70°C. After cooling down to rt, the mixture was diluted with EtOAc (100 mL), washed with water (3 x 50 mL), brine (2 x 50 mL) and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (3:1) to afford 6-amino-1- [(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]quinoxalin-2-one (200 mg, 18.07%) as a white solid. MS (ESI): mass calcd. For C19H18FN3O3: 355.13 m/z, found 356.20 [M+H]+.
Figure imgf000589_0001
3-tert-butyl-1-{1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-2-oxoquinoxalin-6- yl}urea To a solution of 6-amino-1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]quinoxalin-2-one (200 mg, 0.563 mmol, 1 equiv) in DCM (10 mL) was added triphosgene (167.01 mg, 0.563 mmol, 1 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred for 10 min at rt. TEA (113.90 mg, 1.126 mmol, 2 equiv) was added to the reaction under nitrogen atmosphere. The resulting mixture was stirred for 15 min at rt. Then erbumine (82.32 mg, 1.126 mmol, 2 equiv) and TEA (113.90 mg, 1.126 mmol, 2 equiv) was added to the reaction under nitrogen atmosphere. The resulting mixture was stirred for 1h at rt. The reaction mixture was quenched by water and extracted with DCM (3 x 30 mL).The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column, eluted with PE / EA (2:1) to afford 3-tert-butyl-1-{1-[(1S)- 1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-2-oxoquinoxalin-6-yl}urea (150 mg, 58.64%) as a yellow solid. MS (ESI): mass calcd. for C24H27FN4O4: 454.20 m/z, found 455.30 [M+H]+.
Figure imgf000589_0002
3-tert-butyl-1-{1-[(1S)-1-(2-fluoro-5-formylphenyl)ethyl]-2-oxoquinoxalin-6-yl}urea To a solution of 3-tert-butyl-1-{1-[(1S)-1-[5-(1,3-dioxolan-2-yl)-2-fluorophenyl]ethyl]-2- oxoquinoxalin-6-yl}urea (130 mg, 0.286 mmol, 1 equiv) in THF (7 mL) was added 3N HCl (3.5 mL) dropwise at 0°C. The resulting mixture was stirred for 1 h at rt. The mixture was diluted with EtOAc (20 mL), washed with water (3 x 10 mL), brine (2 x 10 mL) and concentrated under vacuum to afford 3-tert-butyl-1-{1-[(1S)-1-(2-fluoro-5-formylphenyl)ethyl]-2-oxoquinoxalin-6-yl}urea (85 mg, 72.4%) as a white solid. MS (ESI): mass calcd. for C22H23FN4O3: 410.17 m/z, found 411.30 [M+H]+.
Figure imgf000590_0001
3-tert-butyl-1-{1-[(1S)-1-[2-fluoro-5-(2,2,2-trifluoro-1-hydroxyethyl)phenyl]ethyl]-2- oxoquinoxalin-6-yl}urea To a solution of 3-tert-butyl-1-{1-[(1S)-1-(2-fluoro-5-formylphenyl)ethyl]-2-oxoquinoxalin- 6-yl}urea (150 mg, 0.365 mmol, 1 equiv) in dimethylformamide (10 mL) was added trimethyl(trifluoromethyl)silane (207.86 mg, 1.460 mmol, 4 equiv) and N,N- dimethylmethanamine oxide (109.80 mg, 1.460 mmol, 4 equiv) dropwise at 0°C. The resulting mixture was stirred for 8 h at rt. The reaction mixture was quenched by water and extracted with EA (3 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to yield a crude product which was directly purified by silica gel column eluted with PE / EA (2:1) & Prep-HPLC (Column: YMC Triart C18 ExRs Column, 20*250mm, 5μm; Mobile Phase A: Water(10mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min mL/min; Gradient: 41%B to 71%B in 7 min; Wave Length: 254nm/220nm nm; RT1(min): 6.42 Column: YMC Triart C18 ExRs Column, 20*250mm, 5μm; Mobile Phase A: Water(10mmol/L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL/min mL/min; Gradient: 41%B to 71%B in 7 min; Wave Length: 254nm/220nm nm; RT1(min): 6.42) to afford 3-tert-butyl-1-{1-[(1S)-1-[2-fluoro-5-(2,2,2-trifluoro-1-hydroxyethyl)phenyl]ethyl]-2- oxoquinoxalin-6-yl}urea (2.8 mg, 1.57%) as an light yellow. MS (ESI): mass calcd. For C23H24F4N4O3, 480.18 m/z, found 481.25 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.17 (s, 1H), 7.79 – 8.00 (m, 2H), 7.29 – 7.55 (m, 3H), 7.13 (dd, J = 10.8, 8.5 Hz, 1H), 6.97 (s, 1H), 6.57 (s, 1H), 6.01 (s, 1H), 5.28 (d, J = 7.3 Hz, 1H), 1.88 (d, J = 7.0 Hz, 3H), 1.29 (s, 9H).19F NMR (282 MHz, DMSO) δ -76.88, -116.10. Example 376: 1-(tert-butyl)-3-((R)-3-methyl-2-oxo-1-((S)-1-phenylethyl)-7- (trifluoromethyl)-2,3-dihydro-1H-pyrido[2,3-b] [1,4] oxazin-6-yl)urea Synthetic Scheme
Figure imgf000591_0001
(R)-6-bromo-3-methyl-1-((S)-1-phenylethyl)-7-(trifluoromethyl)-1H-pyrido[2,3- b][1,4]oxazin-2(3H)-one A solution of (3R)-6-bromo-3-methyl-7-(trifluoromethyl)-1H,3H-pyrido[2,3-b][1,4]oxazin-2-one (50 mg, 0.161 mmol, 1 equiv) in DCM (5 mL) was treated with (R)-1-phenylethan-1-ol (29.46 mg, 0.241 mmol, 1.5 equiv) and PPh3 (63.24 mg, 0.241 mmol, 1.5 equiv) for 1 h at rt under nitrogen atmosphere followed by the addition of DEAD (41.99 mg, 0.241 mmol, 1.5 equiv) dropwise at 0°C. The resulting mixture was stirred for 2 h at rt under N2 atmosphere. Quenched with water (10 mL) and extracted with DCM (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (3R)-6-bromo-3-methyl-1- [(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H-pyrido[2,3-b][1,4]oxazin-2-one (40 mg, 59.93%) as a yellow solid. LC/MS: mass calcd. for C17H14BrF3N2O2: 414.02 m/z, found: 415.10, 417.10 [M+H, M+H+2]+.
Figure imgf000592_0001
(R)-6-azido-3-methyl-1-((S)-1-phenylethyl)-7-(trifluoromethyl)-1H-pyrido[2,3- b][1,4]oxazin-2(3H)-one To a solution of (3R)-6-bromo-3-methyl-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H- pyrido[2,3-b][1,4]oxazin-2-one (100 mg, 0.241 mmol, 1 equiv) in DMSO (3 mL) was added NaN3 (31.31 mg, 0.482 mmol, 2 equiv). The reaction was stirred at 100 °C under N2 for 0.5 h. After cooling down to rt, the reaction was quenched with water (10 mL) and extracted with EA (3 x 10 mL). The mixture was allowed to cool down to rt. The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give (3R)-6-azido-3-methyl-1- [(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H-pyrido[2,3-b][1,4]oxazin-2-one (31 mg, 30.70%) as a white solid. MS (ESI): mass calcd. for C17H14F3N5O2: 377.11 m/z, found: 378.05 [M+H]+.
Figure imgf000592_0002
(R)-6-amino-3-methyl-1-((S)-1-phenylethyl)-7-(trifluoromethyl)-1H-pyrido[2,3- b][1,4]oxazin-2(3H)-one To a solution of 6-azido-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H-pyrido[2,3- b][1,4]oxazin-2-one (60 mg, 0.165 mmol, 1 equiv) in THF (2.5 mL)/H2O (0.5 mL) was added trimethylphosphine (125.65 mg, 1.650 mmol, 10 equiv). The reaction was stirred at rt for 1 h. Quenched with water (5 mL) and extracted with EA (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel with EA/PE (0-100%) to give 6-amino-1-[(1S)-1- phenylethyl]-7-(trifluoromethyl)-3H-pyrido[2,3-b][1,4]oxazin-2-one (50 mg, 80.78%) as a white solid. MS (ESI): mass calcd. for C17H16F3N3O2: 351.12 m/z, found: 352.05 [M+H]+.
Figure imgf000593_0001
1-(tert-butyl)-3-((R)-3-methyl-2-oxo-1-((S)-1-phenylethyl)-7-(trifluoromethyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazin-6-yl)urea To a solution of (3R)-6-amino-3-methyl-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H- pyrido[2,3-b][1,4]oxazin-2-one (100 mg, 0.285 mmol, 1 equiv) in dry-DMF(3 mL) was added NaH (6.83 mg, 0.285 mmol, 1 equiv). The reaction was stirred at rt for 0.5 h. The reaction was added 2-isocyanato-2-methylpropane (84.65 mg, 0.855 mmol, 3 equiv) and stirred at rt for 1 h. Quenched with water (10 mL) and extracted with EA (3 x 10 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a YMC-Actus Triart C18 ExRS 150 mm x 30 mm x 5 μm column (eluent: 49% to 73% (v/v) CH3CN and H2O with 10 mmol/L NH4HCO3) to afford the title compound 3-tert-butyl-1-[(3R)-3-methyl-2-oxo-1-[(1S)-1-phenylethyl]-7-(trifluoromethyl)-3H- pyrido[2,3-b][1,4]oxazin-6-yl]urea (38.2 mg, 29.42%) as a white solid. LC/MS (ESI): mass calcd. for C22H25F3N4O3: 450.19 m/z, found: 451.20 [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 7.69 (s, 1H), 7.19 – 7.47 (m, 7H), 6.07 – 6.26 (m, 1H), 5.13-5.24 (m, 1H), 1.74-1.76 (m, 3H), 1.55-1.57 (m, 3H), 1.28 (s, 9H).19F NMR (282 MHz, DMSO) δ -59.29, -59.33, -59.41. Example 377: (S)-1-(tert-butyl)-3-(6-oxo-5-(1-phenylethyl)-3-(trifluoromethyl)-5,6,7,8- tetrahydro-1,5-naphthyridin-2-yl)urea Synthetic Scheme
Figure imgf000594_0001
ethyl (E)-3-(3-amino-5-(trifluoromethyl)pyridin-2-yl)acrylate To a stirred mixture of 2-chloro-5-(trifluoromethyl) pyridin-3-amine (3 g, 15.263 mmol, 1 equiv) in dioxane (40 mL) and H2O (10 mL) was added ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)acrylate (5.18 g, 22.895 mmol, 1.5 equiv) , Pd(dppf)Cl2 (1.12 g, 1.526 mmol, 0.1 equiv) , Na2CO3 (4.85 g, 45.789 mmol, 3 equiv) and X-phos (727.61 mg, 1.526 mmol, 0.1 equiv) at room temperature under air atmosphere. The resulting mixture was maintained under nitrogen and stirred at 90 °C for 3 h. After cooling down to room temperature, the reaction was quenched with water (60 mL). The resulting mixture was extracted with ethyl acetate (3 x 60 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the ethyl (E)-3-(3-amino-5-(trifluoromethyl) pyridin-2-yl)acrylate (2.5 g, 62.95%) as a yellow oil. LC/MS: mass calcd. for C11H11F3N2O2: 260.08, found: 261.15 [M+H] +.
Figure imgf000594_0002
ethyl 3-(3-amino-5-(trifluoromethyl)pyridin-2-yl)propanoate To a stirred mixture of ethyl (E)-3-(3-amino-5-(trifluoromethyl) pyridin-2-yl)acrylate (1 g, 3.843 mmol, 1 equiv) in THF (15 mL) was added NaBH4 (726.90 mg, 19.215 mmol, 5 equiv) and CoCl2 (249.47 mg, 1.921 mmol, 0.5 equiv) at 0 degrees C under air atmosphere. The resulting mixture was maintained under air and stirred at 25 °C for 3 h. The reaction was quenched with water (30 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the ethyl 3- (3-amino-5-(trifluoromethyl) pyridin-2-yl)propanoate (750 mg, 74.42%) as a yellow oil. LC/MS: mass calcd. for C11H13F3N2O2: 262.09, found: 263.15 [M+H] +.
Figure imgf000595_0001
ethyl 3-(3-amino-6-bromo-5-(trifluoromethyl)pyridin-2-yl)propanoate To a stirred mixture of ethyl 3-(3-amino-5-(trifluoromethyl)pyridin-2-yl)propanoate (700 mg, 2.669 mmol, 1 equiv) in ACN (5 mL) was added NBS (570.13 mg, 3.203 mmol, 1.2 equiv) at 0 degrees C under air atmosphere. The resulting mixture was maintained under air and stirred at 0 °C for 3 h. The reaction was quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-50% ethyl acetate/petroleum ether) to afford the ethyl 3-(3-amino-6-bromo-5-(trifluoromethyl)pyridin- 2-yl)propanoate (350 mg, 38.44%) as a yellow oil. LC/MS: mass calcd. for C11H12BrF3N2O2: 340.00, found: 341.05, 343.05 [M+H, M+H+2] +.
Figure imgf000595_0002
6-bromo-7-(trifluoromethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one The ethyl 3-(3-amino-6-bromo-5-(trifluoromethyl)pyridin-2-yl)propanoate (300 mg, 0.879 mmol, 1 equiv) was dissolved in HOAc (3 mL) at 0 degrees C under air atmosphere. The resulting mixture was maintained under air and stirred at 80 °C for 3 h. After cooling down to room temperature, the reaction was quenched with NaHCO3 solution (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-60% ethyl acetate/petroleum ether) to afford the 6-bromo-7-(trifluoromethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (250 mg, 96.34%) as a yellow oil. LC/MS: mass calcd. for C9H6BrF3N2O: 293.96, found: 295.05, 297.05 [M+H, M+H+2] +.
Figure imgf000596_0001
(S)-6-bromo-1-(1-phenylethyl)-7-(trifluoromethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred mixture of 6-bromo-7-(trifluoromethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (240 mg, 0.813 mmol, 1 equiv) in DCM (5 mL) was added (R)-1-phenylethan-1-ol (149.06 mg, 1.220 mmol, 1.5 equiv) and PPh3 (320.02 mg, 1.220 mmol, 1.5 equiv) at 0 degrees C under air atmosphere. After stirring at 25 °C for 30 min under nitrogen, DEAD (212.49 mg, 1.220 mmol, 1.5 equiv) was added to the reaction system at 0 degrees C. The resulting mixture was maintained under nitrogen and stirred at 25 °C for 3 h. The reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the (S)-6-bromo-1-(1- phenylethyl)-7-(trifluoromethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one (140 mg, 43.11%) as a yellow oil. LC/MS: mass calcd. for C17H14BrF3N2O: 398.02, found: 399.10, 401.10 [M+H, M+H+2] +.
Figure imgf000596_0002
(S)-6-amino-1-(1-phenylethyl)-7-(trifluoromethyl)-3,4-dihydro-1,5-naphthyridin-2(1H)-one To a stirred mixture of (S)-6-bromo-1-(1-phenylethyl)-7-(trifluoromethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (130 mg, 0.326 mmol, 1 equiv) in EtOH (2 mL) and H2O (1 mL) was added NaN3 (63.51 mg, 0.978 mmol, 3 equiv), N1, N2-dimethylcyclohexane-1,2-diamine (27.79 mg, 0.196 mmol, 0.6 equiv), CuSO4 (51.97 mg, 0.326 mmol, 1 equiv) and sodium ascorbate (129.68 mg, 0.652 mmol, 2 equiv) at room temperature under air atmosphere. The resulting mixture was maintained under nitrogen and stirred at 80 °C for 3 h. After cooling down to room temperature, the reaction was quenched with water (10 mL). The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. The residue obtained was purified by silica gel chromatography (0-100% ethyl acetate/petroleum ether) to afford the (S)-6-amino-1-(1-phenylethyl)-7-(trifluoromethyl)- 3,4-dihydro-1,5-naphthyridin-2(1H)-one (50 mg, 45.79%) as a yellow oil. LC/MS: mass calcd. for C17H16F3N3O: 335.12, found: 336.00 [M+H] +.
Figure imgf000597_0001
(R)-6-amino-1-benzyl-3-methyl-7-(trifluoromethyl)-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one To a stirred mixture of (S)-6-amino-1-(1-phenylethyl)-7-(trifluoromethyl)-3,4-dihydro-1,5- naphthyridin-2(1H)-one (45 mg, 0.134 mmol, 1 equiv) in DMF (2 mL) was added NaH (4.83 mg, 0.201 mmol, 1.5 equiv) at 0 degrees C under air atmosphere. After stirring at 0 degrees C for 30 min, 2-isocyanato-2-methylpropane (39.91 mg, 0.402 mmol, 3 equiv) was added to the reaction system. The resulting mixture was stirred at 25 °C for 3h. Quenched with water (5 mL) and extracted with EA (3 x 5 mL). The combined extracts were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC using a XBridge Shield RP18 OBD Column 150 mm x 30 mm x 5 μm column (eluent: 70% to 85% (v/v) Me and Water with 0.1%FA) to afford the title compound (S)-1-(tert-butyl)-3-(6-oxo-5-(1-phenylethyl)- 3-(trifluoromethyl)-5,6,7,8-tetrahydro-1,5-naphthyridin-2-yl)urea (12.9 mg, 21.43%) as a white solid.LC/MS (ESI): mass calcd. for C22H25F3N4O2:434.19 m/z, found:435.25 [M+H] +.1H-NMR (300 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.51 (s, 1H), 7.23 – 7.44 (m, 5H), 7.06 (s, 1H), 6.16 – 6.32 (m, 1H), 2.95 – 3.15 (m, 2H), 2.87 – 2.96(m, 1H), 2.76 – 2.82(m, 1H), 1.68 (d, J = 7.2 Hz, 3H), 1.31 (s, 9H).19F-NMR (282 MHz, DMSO) δ -60.51. Example 378: 1-[(3S)-1-benzyl-3-methyl-2-oxo-7-(trifluoromethyl)-3H-pyrido[2,3- b][1,4]oxazin-6-yl]-3-tert-butylurea Synthetic Scheme
Figure imgf000598_0001
1-[(3S)-1-benzyl-3-methyl-2-oxo-7-(trifluoromethyl)-3H-pyrido[2,3-b][1,4]oxazin-6-yl]-3- tert-butylurea The crude product (130 mg) was separated by chiral-HPLC using a CHIRALPAK IA, 5*25 cm, 5 μm (eluent: 35% to 35%(v/v) : EtOH--HPLC and Hex(0.5% 2M NH3-MeOH) to yield second enantiomer example 1-[(3S)-1-benzyl-3-methyl-2-oxo-7-(trifluoromethyl)-3H-pyrido[2,3- b][1,4]oxazin-6-yl]-3-tert-butylurea (20 mg, 15.09%) as a white solid. LC/MS: mass calcd. for C21H23F3N4O3: 436.2, found: 437.20 [M+H]+.1H NMR (400 MHz, DMSO-d6) δ 7.72 (s, 1H), 7.58 (s, 1H), 7.30 – 7.40 (m, 2H), 7.21 – 7.30 (m, 4H), 5.13 – 5.34 (m, 3H), 1.60 (d, J = 6.8 Hz, 3H), 1.29 (s, 6H).19F NMR (376 MHz, DMSO) δ -58.74. hTSHR TR-FRET cAMP Assay Assays were performed using Expi293F Inducible cells (Invitrogen) stably expressing hTSHR via a pcZeo TetO DNA plasmid. Cell lines were maintained in suspension in Expi293 Expression Medium (ThermoFisher Scientific) supplemented with 10 μg/mL Blasticidin and 10 μg/mL Zeocin and incubated at 37°C, 8% CO2, with shaking. hTSHR cells were incubated overnight at 32°C, 5% CO2, with shaking, but without Doxycycline. Assay-ready aliquots were prepared by harvesting each cell type separately 24-hours post-induction. Cells were pelleted at 4°C, resuspended in Expi293 Expression media + 10% DMSO, aliquoted, and kept frozen at - 80°C until ready for use. For the assay, concentration-response curves of test and reference compounds were added to 384-well plates using an Echo 650 liquid handler (Beckman Coulter) and backfilled with DMSO to a final concentration of 0.5%. cAMP was measured using the cisbio cAMP Gs dynamic HTRF kit (PerkinElmer) according to manufacturer instructions. Aliquots of frozen hTSHR cells were quickly thawed and washed with phosphate-buffered saline (Sigma-Aldrich) to remove media and DMSO. The cells were resuspended in kit-supplied Stimulation Buffer at 0.8 x 106 cells/mL.7.5 μL of the hTSHR cell dilution were added to each well of the assay plate and incubated with the compounds for 15 minutes in a 37°C, 0% CO2 incubator.2.5 μL of Vehicle control or an EC80 concentration of recombinant human anti-TSHR antibody (M22, Creative Biolabs) was added to stimulate cAMP production. Cells were incubated with for 30 minutes at 37°C, 0% CO2. Following this incubation, the cells were lysed and accumulated cAMP was detected through the addition of kit-supplied lysis buffer containing d2-reagent and Eu-cryptate antibody. The HTRF signal was quantified using a BMG Clariostar plate reader optimized for HTRF assays. The HTRF ratio was determined by dividing the signal output at 655 nm by that at 620 nm. Data were normalized to the signal produced by the EC80 concentration of either M22 or FSH (0% inhibition) and vehicle (no agonist, 100% inhibition). Results of the assay are provided in Table 17. Table 17.
Figure imgf000599_0001
Figure imgf000600_0001
Figure imgf000601_0001
Figure imgf000602_0001
Figure imgf000603_0001
Figure imgf000604_0001
Figure imgf000605_0001
Figure imgf000606_0001
Figure imgf000607_0001
Figure imgf000608_0001
Figure imgf000609_0001
Figure imgf000610_0001
Figure imgf000611_0001
Figure imgf000612_0001
Figure imgf000613_0001
Figure imgf000614_0001
Figure imgf000615_0001
Figure imgf000616_0001
Figure imgf000617_0001
* IC50 bins: + >1 µM; ++ = 1 µM-100 nM; and +++ <100 nM INCORPORATION BY REFERENCE All of the U.S. patents and U.S. and PCT patent application publications cited herein are hereby incorporated by reference. EQUIVALENTS Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

We claim: 1. A compound of Formula (I):
Figure imgf000618_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. 2. A compound of Formula (I):
Figure imgf000619_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, (C1-C6)alkoxy, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy. 3. A compound of Formula (II):
Figure imgf000621_0001
or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1, 2, 3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring, 5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1- C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; W represents: –C(R6)2–C(R7)2–, –C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, –N=C(R10) –, or –O–; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy. 4. A compound of Formula (II):
Figure imgf000623_0001
or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, , 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C1-C6)alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; Wa–Wb represents: –C(R6)2–C(R7)2–, –O–C(R8)2–, –C(R8)2–O–, –C(R8)2–NR9–, or –N=C(R10) –; and R6, R7, R8, R9 and R10 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy. 5. A compound of Formula (III):
Figure imgf000624_0001
or a pharmaceutically acceptable salt thereof, wherein: A is (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C8)cycloalkyl, or 4- to 7-membered heterocycloalkyl; n is 0, 1,
2,
3, 4, or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3- C8)cycloalkyl, (C3-C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, 4- to 7-membered heterocycloalkyl, (C1-C6)alkoxy, (C1-C6)alkoxy(C1-C6)alkyl, halo, cyano, amino, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C2-C6)alkynyl, (C2-C6)alkenyl, (C3-C8)cycloalkyl, C3- C8)cycloalkoxy, (C6-C10)aryl, (C6-C10)aryloxy, (C6-C10)aryl(C1-C6)alkyl, (C1-C6)alkoxy, (C1- C6)alkoxy(C1-C6)alkyl, and 4- to 7-membered heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl,
4- to 7-membered heterocycloalkyl, 5- to 6- membered heteroaryl, (C1-C6)alkoxy, (C1-C6)haloalkoxy, (C1-C6)fluoroalkoxy, 5- to 6-membered heteroaryloxy, or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl, 5- to 7-membered heterocycloalkyl ring,
5- to 6- membered heteroaryl ring or phenyl ring, any of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C1-C6)haloalkyl (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, or (C3- C8)cycloalkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl(C1-C6)alkyl), (C6-C10)aryl, (C6- C10)aryl(C1-C6)alkyl), or 4- to 7-membered heterocycloalkyl; each of which is optionally substituted with one or more substituents independently selected from halo, cyano, (C1-C6)alkyl, (C1-C6)haloalkyl, hydroxy, (C1-C6)alkoxy(C1-C6)alkyl, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy, or when Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo, or when Z is C(R5)2, R3 and one R5 taken together with the carbon to which they are attached form a 4, (C3-C8)cycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; R10 is hydrogen, (C1-C6)alkyl,or NH2, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, cyano, hydroxy, (C1-C6)alkoxy(C1- C6)alkyl, and (C1-C6)alkoxy.
6. A compound of Formula (III):
Figure imgf000626_0001
or a pharmaceutically acceptable salt thereof, wherein A is (C6-C10)aryl or 5- to 10-membered heteroaryl; n is 0, 1, 2, 3, 4 or 5; R1 is independently for each occurrence (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7- membered heterocycloalkyl, (C1-C6)alkoxy, halo, cyano, formyl, carboxy, or alkoxycarbonyl, wherein (C1-C6)alkyl, (C3-C8)cycloalkyl, 4- to 7-membered heterocycloalkyl, and (C1-C6)alkoxy are each optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; or two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1-C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy; Q is N or CR2c; R2a, R2b, and R2c, are each independently hydrogen, halo, cyano, (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; R3 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; X and Z are independently NR4, C(R5)2, or O; R4 and R5 are independently for each occurrence hydrogen or (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy; L is (C1-C6)alkylene optionally substituted with one or more substituents independently selected from halo, hydroxy, C1-C6)alkoxy, and cyano; and R10 is hydrogen or (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy.
7. The compound of any one of claims 1 to 6, wherein A is phenyl.
8. The compound of any one of claims 1 to 6, wherein A is a 5- to 7-membered heteroaryl comprising 1 or 2 nitrogen atoms.
9. The compound of claim 8, wherein A is pyridyl or diazolyl. thiazolyl, pyrazinyl, pyrimidinyl,
10. The compound of claim 9, wherein A is pyridyl or diazolyl.
11. The compound of any one of claims 1 to 6, wherein A is isoquinolinyl or indazolyl.
12. The compound of any one of calims 1 to 6, wherein A is cyclopentyl, cyclohexyl, or tetrahydropyranyl.
13. The compound of any one of claims 1 to 12, wherein R1 is independently for each occurrence (C1-C6)alkyl, (C1-C6)alkoxy, fluoro, chloro, or cyano, wherein (C1-C6)alkyl and (C1- C6)alkoxy are optionally substituted with one or more substituents independently selected from fluoro and hydroxyl.
14. The compound of any one of claims 1 to 12, wherein at least one R1 is fluoro, chloro, or cyano.
15. The compound of any one of claims 1 to 14, wherein at least one R1 is (C1-C6)alkyl optionally substituted with one or more substituents independently selected from hydroxyl and fluoro,
16. The compound claim 15, wherein at least one R1 is 2,2,2-trifluoroethyl-1-hydroxyethyl.
17. The compound claim 15, wherein at least one R1 wherein at least one R1 is difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluropropyl, or 4,4,4-trifluorobutyl.
18. The compound of any one of claims 1 to 14, wherein at least one R1 is (C1-C6)alkoxy optionally substituted with at least one fluoro.
19. The compound of claim 18, wherein at least one R1 is difluoromethoxy or trifluoromethoxy.
20. The compound of claim 18, wherein at least one R1 is methoxy or isopropoxy.
21. The compound of any one of claims 1 to 14, wherein at least one R1 is methyl optionally substituted with hydroxy or one or more fluoro.
22. The compound of any one of claims 1 to 14, wherein at least R1 is (C1-C6)alkyl substituted with an amido having the structure:
Figure imgf000628_0001
.
23. The compound of any one of claims 1 to 14, wherein at least one R1 is (C3-C8)cycloalkyl.
24. The compound of claim 23, wherein the (C3-C8)cycloalkyl is cyclopropyl or cyclobutyl.
25. The compound of any one of claims 1 to 14, wherein at least R1 is (C3-C8)cycloalkoxy.
26. The compound of claim 25, wherein the (C3-C8)cycloalkoxy is cyclopentyloxy or cyclohexyloxy.
27. The compound of any one of claims 1 to 14, wherein R1 is (C2-C6)alkynyl optionally substituted with hydroxy.
28. The compound of any one of claims 1 to 12, wherein two vicinal occurrences of R1 taken together with the atoms to which they are attached form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring, either of which is optionally substituted with one or more substituents independently selected from halo, hydroxy, cyano, amido, sulfonamido, (C1- C6)alkyl, (C3-C8)cycloalkyl, and (C1-C6)alkoxy.
29. The compound of claim 28, wherein A is phenyl comprising two vicinal occurrences of R1, wherein the two vicinal occurrences of R1, taken together with the atoms to which they are attached, form a fused 5- to 7-membered cycloalkyl or 5- to 7-membered heterocycloalkyl ring.
30. The compound of any one of claims 1 to 27, wherein n is 2.
31. The compound of any one of claims 1 to 27, wherein n is 1.
32. The compound of any one of claims 1 to 12, wherein n is 0.
33. The compound of any one of claims 1 to 32, wherein Q is CR2c.
34. The compound of any one of claims 1 to 32, wherein Q is N.
35. The compound of any one of claims 1 to 33, wherein R2a, R2b, and R2c are each hydrogen.
36. The compound of any one of claims 1 to 33, wherein at least one of R2a, R2b, and R2c is fluoro, chloro, or cyano.
37. The compound of any one of claims 1 to 36, wherein R3 is (C1-C6)alkyl optionally substituted with hydroxy.
38. The compound of claim 37, wherein R3 is t-butyl.
39. The compound of any one of claims 1 to 36, wherein R3 is (C3-C8)cycloalkyl optionally substituted with one or more (C1-C6)alkyl, (C1-C6)fluoroalkyl, fluoro, or (C1-C6)alkoxy(C1- C6)alkyl.
40. The compound of claim 39, wherein the (C3-C8)cycloalkyl is cyclobutyl, cyclopentyl, cylohexyl, bicyclo[1.1.1]pentanyl, bicyclo[2.2.1]heptanyl,bicycle[2.1.1]hexanyl, or bicyclo[3.1.0]hexanyl.
41. The compound of claim 40 or 41, wherein the (C3-C8)cycloalkyl is optionally substituted with one or more substituents selected from fluoro, trifluoromethyl, methoxymethyl, methyl, and hydroxyl.
42. The compound of any one of claims 1 to 36, wherein R3 is 4- to 7-membered heterocycloalkyl optionally substituted with one or more substituents selected from (C1-C6)alkyl, (C1-C6)fluoroalkyl, or fluoro.
43. The compound of claim 42, wherein the 4- to 7-membered heterocycloalkyl is etrahydropyranyl or oxetanyl.
44. The compound of any one of claims 1 to 36, wherein Z is NR4, R3 and R4 taken together with the nitrogen to which they are attached form a 4- to 7-member heterocycloalkyl optionally substituted with one or more (C1-C6)alkyl or halo.
45. The compound of claim 44, wherein the 4- to 7 membered heterocycloalkyl is azetidine or pyrrolidine,
46. The compound of any one of claims 1 to 45, wherein X is NR4 or O.
47. The compound of any one of claims 1 to 45 wherein X is NH.
48. The compound of any one of claims 1 to 43 and 46 to 47, wherein Z is NR4, or O.
49. The compound of any one of claims 1 to 43 and 46 to 47, wherein Z is NH.
50. The compound of any one of claims 1 to 49, wherein one and only one of X and Z is CH2.
51. The compound of any one of claims 1 to 43 and 46 to 47, wherein X and Z are each NH.
52. The compound of any one of claims 1 to 51, wherein L is unsubstituted (C1-C6)alkylene.
53. The compound of any one of claims 1 to 51, wherein L is (C1-C6)alkylene substituted with hydroxy.
54. The compound of any one of claims 1 to 51, wherein L is (C1-C6)alkylene substituted with 1 to 3 halo atoms.
55. The compound of any one of claims 1 to 51, wherein L is –CH2–, –-CH(CH3)–, –CH(CH2CH3)–, or –CH(CH2OH)–.
56. The compound of any one of claims 2 to 55, wherein Wa–Wb represents: –C(R6)2–C(R7)2–.
57. The compound of claim 56, wherein at least one R6 is (C1-C6)alkyl, wherein (C1-C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, and (C1-C6)alkoxy.
58. The compound of claim 56 or 57, wherein at least one R7 is (C1-C6)alkyl, wherein (C1- C6)alkyl is optionally substituted with one or more substituents independently selected from halo, hydroxy, (C1-C6)alkoxy and cyano.
59. The compound of claim 56, wherein each R6 and each R7 is hydrogen.
60. The compound of any one of claims 2 to 55, wherein: Wa–Wb represents: –O–C(R8)2–.
61. The compound of claim 60, wherein each R8 is hydrogen.
62. The compound of claim 60, wherein one R8 is (C1-C6)alkyl; and one R8 is hydrogen.
63. The compound of claim 62, wherein one R8 is methyl.
64. The compound of any one of claims 2 to 55, wherein: Wa–Wb represents: –C(R8)2–O–.
65. The compound of claim 64, wherein each R8 is hydrogen.
66. The compound of any one of claims 2 to 30, wherein: Wa–Wb represents: –C(R8)2–NR9–.
67. The compound of claim 66, wherein each R8 is hydrogen.
68. The compound of claim 66 or 67, wherein R9 is (C1-C6)alkyl.
69. The compound of claim 68, wherein R9 is methyl.
70. The compound of any one of claims 2 to 55, wherein: Wa–Wb represents: –N=C(R10) –.
71. The compound of claim 70, wherein R10 is hydrogen.
72. The compound of claim 70, wherein R10 is (C1-C6)alkyl.
73. The compound of claim 70, wherein R10 is methyl.
74. A compound selected from the following table:
Figure imgf000631_0001
Figure imgf000632_0001
Figure imgf000633_0001
Figure imgf000634_0001
Figure imgf000635_0001
Figure imgf000636_0001
Figure imgf000637_0001
Figure imgf000638_0001
Figure imgf000639_0001
Figure imgf000640_0001
Figure imgf000641_0001
Figure imgf000642_0001
Figure imgf000643_0001
Figure imgf000644_0001
Figure imgf000645_0001
Figure imgf000646_0001
Figure imgf000647_0001
Figure imgf000648_0001
Figure imgf000649_0001
Figure imgf000650_0001
Figure imgf000651_0001
Figure imgf000652_0001
Figure imgf000653_0001
Figure imgf000654_0001
Figure imgf000655_0001
Figure imgf000656_0001
Figure imgf000657_0001
Figure imgf000658_0001
Figure imgf000659_0001
Figure imgf000660_0001
Figure imgf000661_0001
Figure imgf000662_0001
Figure imgf000663_0001
Figure imgf000664_0001
Figure imgf000665_0001
Figure imgf000666_0001
Figure imgf000667_0001
Figure imgf000668_0001
Figure imgf000669_0001
Figure imgf000670_0001
Figure imgf000671_0001
Figure imgf000672_0001
Figure imgf000673_0001
Figure imgf000674_0001
Figure imgf000675_0001
Figure imgf000676_0001
Figure imgf000677_0001
Figure imgf000678_0001
Figure imgf000679_0001
Figure imgf000680_0001
Figure imgf000681_0001
Figure imgf000682_0001
Figure imgf000683_0001
Figure imgf000684_0001
Figure imgf000685_0001
Figure imgf000686_0001
Figure imgf000687_0001
Figure imgf000688_0001
Figure imgf000689_0001
Figure imgf000690_0002
or a pharmaceutically acceptable salt thereof.
75. A compound selected from the following table:
Figure imgf000690_0001
Figure imgf000691_0001
Figure imgf000692_0001
Figure imgf000693_0001
Figure imgf000694_0001
Figure imgf000695_0001
or a pharmaceutically acceptable salt thereof.
76. A pharmaceutical composition, comprising a compound of any one of claims 1 to 75; and at least one pharmaceutically acceptable excipient.
77. A method of inhibiting a thyroid stimulating hormone receptor, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1 to 75.
78. A method of treating hyperthyroidism, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1 to 75.
79. The method of claim 78, wherein the subject has Graves' disease.
80. The method of claim 79 wherein the subject has Graves' ophthalmopathy.
81. The method of claim 79 or 80, wherein the subject has Graves' dermopathy.
82. The method of claim 78, wherein the subject has thyroid cancer.
PCT/US2023/028950 2022-07-29 2023-07-28 Substituted 3,4-dihydroquinolinone inhibitors of tshr WO2024026076A2 (en)

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