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WO2015010297A1 - Therapeutically active compounds and their methods of use - Google Patents

Therapeutically active compounds and their methods of use Download PDF

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Publication number
WO2015010297A1
WO2015010297A1 PCT/CN2013/080105 CN2013080105W WO2015010297A1 WO 2015010297 A1 WO2015010297 A1 WO 2015010297A1 CN 2013080105 W CN2013080105 W CN 2013080105W WO 2015010297 A1 WO2015010297 A1 WO 2015010297A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
mmol
optionally substituted
mhz
nmr
Prior art date
Application number
PCT/CN2013/080105
Other languages
French (fr)
Inventor
Rene M. Lemieux
Janeta Popovici-Muller
Jeremy M. Travins
Zhenwei CAI
Dawei Cui
Ding Zhou
Original Assignee
Agios Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Agios Pharmaceuticals, Inc. filed Critical Agios Pharmaceuticals, Inc.
Priority to PCT/CN2013/080105 priority Critical patent/WO2015010297A1/en
Priority to JP2016528335A priority patent/JP2016525130A/en
Priority to PCT/CN2014/082869 priority patent/WO2015010626A1/en
Priority to MX2016000696A priority patent/MX2016000696A/en
Priority to CA2918500A priority patent/CA2918500C/en
Priority to EP14828790.7A priority patent/EP3024824A4/en
Priority to US14/341,426 priority patent/US20150031627A1/en
Priority to ARP140102794A priority patent/AR097082A1/en
Priority to TW103125441A priority patent/TWI662023B/en
Publication of WO2015010297A1 publication Critical patent/WO2015010297A1/en
Priority to MX2021001425A priority patent/MX2021001425A/en
Priority to US15/347,407 priority patent/US20170057994A1/en
Priority to US15/915,213 priority patent/US10689414B2/en
Priority to JP2019001587A priority patent/JP6640390B2/en
Priority to US16/871,777 priority patent/US11021515B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/397—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having four-membered rings, e.g. azetidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
    • A—HUMAN NECESSITIES
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
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    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
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    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
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    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C—CHEMISTRY; METALLURGY
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    • C07D—HETEROCYCLIC COMPOUNDS
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    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/18—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member
    • C07D207/22—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member 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
    • C07D207/24—Oxygen or sulfur atoms
    • C07D207/26—2-Pyrrolidones
    • C07D207/273—2-Pyrrolidones 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 other ring carbon atoms
    • C07D207/277—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D207/28—2-Pyrrolidone-5- carboxylic acids; Functional derivatives thereof, e.g. esters, nitriles
    • C—CHEMISTRY; METALLURGY
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    • 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
    • C07D401/04—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 directly linked by a ring-member-to-ring-member bond
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    • 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
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    • C07D—HETEROCYCLIC COMPOUNDS
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    • 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
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Definitions

  • Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate (i. ⁇ ?., a-ketoglutarate). These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is ahomodimer.
  • IDH1 isocitrate dehydrogenase 1 (NADP+), cytosolic
  • IDP isocitrate dehydrogenase 1
  • IDCD ID PC or PICD
  • Theprotein encoded by this gene is the NADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence.
  • This enzyme inperoxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2, 4-dienoyl- CoAs to 3-enoyl-CoAs, aswell as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation ofphytanic acid.
  • the cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production.
  • the human IDH 1 gene encodes a protein of 414 amino acids.
  • the nucleotide and amino acid sequences for human IDH 1 can be found as GenBank entries NM 005896.2 and
  • NP 005887.2 respectively.
  • the nucleotide and amino acid sequences for IDH1 are also described in, e.g., Nekrutenko et al., M ol. Biol. Evol. 15:1674-1684(1998); Geisbrecht et al., J. BioL Chem. 274 :30527-30533(1999); Wiemann et al., Genome Res. 1 1:422-435(2001); The MGC Project Team, Genome Res.
  • Non-mutant e.g., wild type
  • IDH 1 catalyzes the oxidative decarboxylation of isocitrate to a-ketoglutarate thereby reducing NAD + (NADP + ) to NADH (NADPH), e.g., in the forward reaction:
  • Described herein are methods of treating a cancer characterized by the presence of a mutant allele of IDH l or IDH2.
  • the methods comprise the step of administering to a subject in need thereof a compound of formula I, or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate the I
  • R 1 is optionally substituted C4-C6 carbocyclyl
  • each R 2 and R 3 is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
  • R 4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl;
  • ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R 5 groups;
  • each R 5 is independently halo; -CF ; -CN; -OR 6 ;-N(R 6 ) 2 ; -C(0)C,-C 4 alkyl; C 1 -C4 haloalkyl; C ,-C 4 alkyl optionally substituted with -OR 6 or -N(R 6 ) 2 ; -0-Ci-C 4 alkyl optionally substituted with halo, -OR 6 or -N(R 6 ) 2 ; -S0 2 N(R 6 ) 2 ; -S0 2 (C ,-C 4 alkyl); -NR 6 S0 2 R 6 ; C3-C5 carbocyclyl optionally substituted with one or two R 6 groups; -0-(C 3 -C6 carbocyclyl optionally substituted with one or two R 6 groups); 5-6 membered heteroaryl; -Ci-C 4 alkyl-C(0)0-Ci-C4 alkyl; or -C(0)0-
  • each R 6 is independently H or C 1 -C3 alkyl.
  • the compound of formula I inhibits mutant IDH 1/2, particularly mutant IDH 1 having alpha hydroxyl neoactivity. Also described herein are pharmaceutical compositions comprising a compound of formula I.
  • halo or halogen refers to any radical of fluorine, chlorine, bromine or iodine.
  • alky 1 refers to a hydrocarbon chain that may be a straight chain or branched chain, containing the indicated number of carbon atoms.
  • -C 12 alkyl indicates thatthe group may have from 1 to 12 (inclusive) carbon atoms in it.
  • haloalkyl refers to an alkyl in which one or more hydrogen atoms are replaced by halo, and includes alkyl moieties in which all hydrogens have been replaced by halo (e.g., perfiuoroalkyl).
  • arylalkyl or “aralkyl” refer to an alkyl moiety in which an alkyl hydrogen atom is replaced by an ary 1 group .
  • Arylalkyl or aralkyl includes groups in which more than one hydrogen atom has been rep laced by an ary 1 group .
  • Examp les of " arylalky 1" or " aralky 1" include benzy 1, 2- pherylethyl, 3-pherylpropyl, 9-fiuorenyl, benzhydryl, and trityl groups.
  • heteroarylalkyl or “hetero aralkyl” refer to an alkyl moiety in which an alkyl hydrogen atom is replaced by a heteroaryl group .
  • Heteroarylalkyl or heteroaralkyl includes groups in which more than one hydrogen atom has been rep laced by a heteroaryl group.
  • alkylene refers to a divalent akyl, e.g, -CH 2 -, -CH 2 CH 2 -, and -CH 2 CH 2 CH 2 -.
  • alkenyl refers to a straight or branched hydrocarbon chain containing 2- 12 carbon atoms and having one or more double bonds. Examples of alkenyl groups include, but are not limited to, allyl, propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups. One of the double bond carbons may optionally be the point of attachment of the alkenyl substituent.
  • alkynyl refers to a straight or branched hydrocarbon chain containing2-12 carbon atoms and characterized in having one or more triple bonds.
  • alkynyl groups include, but are not limited to, ethynyl, propargyl, and 3-hexynyl.
  • One ofthe triple bond carbons may optionally be the point of attachment of the alkynyl substituent.
  • alkoxy refers to an -O-alkyl radical.
  • t erm halo alkoxy refers to an alkoxy in which one or more hydrogen atoms are replaced by halo, and includes alkoxy moieties in which allhydrogens have been rep laced by halo (e.g., perfiuoroalkoxy).
  • carbocyclyl refers to amonocyclic, bicyclic or tricyclic, hydrocarbon ring system that is not fully aromatic, wherein any ringatom capable of substitution can be substituted by one or more substituents.
  • a carbocyclyl can be fully or partially saturated.
  • a bicyclic or tricylic carbocyclyl may contain one (in the case of abicycle) or up to two (in the case of a tricycle) aromatic rings, as long as at least one ring in the carbocyclylis non-aromatic. Unless otherwise specified, any ring atom capable of substitution in a carbocyclyl can be substituted by one or more substituents.
  • aryl refers to afully aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ringsystem
  • aryl moieties are phenyl, naphthyl, and anthracenyl. Unless otherwise specified, any ring atom in an aryl can be substituted by one or more substituerts.
  • cycloalkyl refers to a saturated cyclic, bicyclic, tricyclic, or poty cyclic hydrocarbon group . Unless otherwise specified, any ring atom can be substituted by one or more substituents.
  • the cycloalkyl groups can contain fused rings. Fused rings are rings that share a common carbon atom. Examples of cycloalkyl moieties include, but are not limited to, cyclop ropy 1, cyclohexyl, methylcyclohexyl, adamantyl, and norbornyl. Unless otherwise specified, any ring atom can be substituted by one or more substituents.
  • heterocyclyl refers to amonocyclic, bicyclic or tricyclic, ringstructure that is not fully aromatic and includes one to four heteroatoms independently selected from N, O, or S in one or more of the rings.
  • a heterocyclyl can be fully or partially saturated.
  • a bicyclic or tricylic heterocyclyl may contain one (in the case of a bicycle) or up to two (inthe case of a tricycle) aromatic rings, as long as at least one ring in the heterocyclyl is non-aromatic. Unless otherwise specified, any ring atom capable of substitution in a heterocyclyl can be substituted by one or more substituents.
  • Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, 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, phenant uridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine,phenothiazine, furazan, phenoxazine, pyrrolidine, ox
  • heteroaryl refers to a monocyclic, bicyclic, or tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms independently selected from O, N, or S, wherein each ringin a heteroaryl is fully aromatic. Unless otherwise specified, any ring atom capable of substitution in a heteroaryl can be substituted by one or more substituents.
  • heteroalkyl and “heter oar alky 1”, as used herein, refers to an alkyl group substituted with aheteroaryl group .
  • the ring heteroatoms of the compounds provided herein include N-O, S(O), and S(0)2.
  • substituted refers to the replacement of a hydrogen atom with another moiety.
  • substituents include alky 1 (e.g., CI , C2, C3, C4, C5, C6, C7, C8, C9, CIO, Cl l, C12 straight or branched chain alkyl), cycloalkyl, haloalkyl (e. g., perfluoroalkyl such as CF 3 ), aryl, heteroaryl, aralkyl, heter oaralkyl, heterocyclyl, akenyl, alkynyl, cycloalkenyl,
  • heterocycloalkenyl, alkoxy, haloakoxy e. g., perfluoroakoxy such as OCF 3
  • haloakoxy e. g., perfluoroakoxy such as OCF 3
  • halo e. g., perfluoroakoxy such as OCF 3
  • halo e. g., perfluoroakoxy such as OCF 3
  • halo e. g., perfluoroakoxy such as OCF 3
  • halo e. g., perfluoroakoxy such as OCF 3
  • tautomer refers to each of two or more isomers of a compound (e.g., a compound described herein) that exist together in equilibrium, and are readily interchangeable by migration of a hydrogen atom or proton, accomp anied by a switch of a single bond and an adjacent double bond.
  • the term “elevated levels of 2HG” means 10%, 20% 30%, 50%, 75%, 100%, 200%, 500% or more 2HG than is present in a subject that does not carry a mutant IDH1 or IDH2 allele.
  • the term “elevated levels of 2HG” may refer to the amount of 2HG within a cell, within a tumor, within an organ comprising a tumor, or within abodily fluid.
  • the term "bodily fluid” includes one or more of amniotic fluid surrounding a fetus, aqueous humour, blood (e.g., blood plasma), serum, Cerebrospinal fluid, cerumen, chyme, Cowper's fluid, female ejaculate, interstitial fluid, lymph, breast milk, mucus (e.g., nasal drainage or phlegm), pleural fluid, pus, saliva, sebum, semen, serum, sweat, tears, urine, vaginal secretion, or vomit.
  • blood e.g., blood plasma
  • serum Cerebrospinal fluid
  • cerumen cerumen
  • chyme chyme
  • Cowper's fluid female ejaculate
  • interstitial fluid lymph
  • breast milk mucus (e.g., nasal drainage or phlegm)
  • mucus e.g., nasal drainage or phlegm
  • pleural fluid pus, saliva, sebum, semen, serum
  • inhibitor or “prevent” include both comp lete and partial inhibition and prevention.
  • An inhibitor may completely or partially inhibit.
  • treat means decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a cancer (e. g., a cancer delineated herein), lessen the severity of the cancer or improve the symptoms associated with the cancer.
  • an amount of a compound effective to treat a disorder or a
  • “therap eutically effective amount” refers to an amount of the comp ound which is effective, upon single or multiple dose administration to a subject, in treating a cell, or in curing alleviating, relieving or improving a subject with a disorder beyond that expected in the absence of such treatment.
  • the term "subject” is intended to include human and non-human animals.
  • exemplary human subjects include a human patient having a disorder, e. g., a disorder described herein or a normal subj ect.
  • non-human animals of the invention includes all vertebrates, e.g., non-mammals (such as chickens, amphibians, reptiles) and mammals, such as non-human primates, domesticated and/or agriculturally useful animals, e.g., sheep, dog, cat, cow, pig, etc.
  • R 1 is optionally substituted C 4 -C 6 carbocyclyl
  • each R and R is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
  • R 4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl;
  • ring A is 4-6 membered non-aromatic ring having 0-1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R 5 groups;
  • each R 5 is independently halo; -CF 3 ; -CN; -OR 6 ;-N(R 6 ) 2 ; -C(0)Ci-C 4 alkyl; Ci-C 4 haloalkyl; C, -C 4 alkyl optionally substituted with -OR 6 or -N(R 6 ) 2 ; -O-C 1 -C4 alkyl optionally substituted with halo, -OR 6 or -N(R 6 ) 2 ; -S0 2 N(R 6 ) 2 ; -S0 2 (C ,-C 4 alkyl); -NR 6 S0 2 R 6 ; C 3 -C 5 carbocyclyl optionally substituted with one or two R 6 groups; -0-(C 3 -C 6 carbocyclyl optionally substituted with one or two R 6 groups); 5-6 membered heteroaryl; -C
  • each R 6 is independently H or C 1 -C3 alkyl.
  • R 1 is optionally substituted C4-C 6 carbocyclyl
  • each R 2 and R 3 is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
  • R 4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyi;
  • ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R 5 groups;
  • each R 5 is independently halo, -CF 3 , -CN, -OR 6 , -N(R 6 ) 2 , -C(0)CH 3 ; C,-C 3 haloalkyl, C,- C 3 alkyl optionally substituted with -OR 6 or -N(R 6 ) 2 ; or
  • each R 6 is independently H or C
  • R 1 is C4-C6 carbocyclyl optionally substituted with one to three R 7 groups;
  • each R and R is independently selected from aryl or heteroaryl, wherein said aryl or heteroaryl is independently optionally substituted with one to three R 7 groups;
  • R 4 is alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyi, wherein said aryl, heteroaryl, aralkyl, and heteroaralkyi are each independently optionally substituted with one to three R 7 groups;
  • ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R 5 groups;
  • each R 5 and R 7 is independently halo; -CF 3 ; -CN; -OR 6 ;-N(R 6 ) 2 ; -C(0)Ci-C 4 alkyl; C 1 -C4 haloalkyl; C 1 -C4 alkyl optionally substituted with -OR 6 or -N(R 6 ) 2 ; -O-C 1 -C4 alkyl optionally substituted with halo, -OR 6 or -N(R 6 ) 2 ; -S0 2 N(R 6 ) 2 ; -S0 2 (C i-C 4 alkyl); -S(0)-Ci.
  • each R 6 is independently H or C 1 -C4 alkyl.
  • R 1 is C4-C6 carbocyclyl optionally substituted with one to three R 7 groups;
  • each R 2 and R 3 is independently selected from aryl or heteroaryl, wherein said aryl or heteroaryl is independently optionally substituted with one to three R groups;
  • R 4 is alkyl, aryl, heteroaryl, aralkyi, or heteroaralkyi, wherein said aryl, heteroaryl, aralkyi, and heteroaralkyi are each independently optionally substituted with one to three R 7 groups;
  • ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R 5 groups;
  • each R 5 and R 7 is independently halo, -CF 3 , -CN, -OR 6 , -N(R 6 ) 2 , -C(0)CH 3 ; C,-C 3 haloalkyl, Ci -C 3 alkyl optionally substituted with -OR 6 or -N(R 6 ) 2 ; or
  • each R 6 is independently H or C i-C 3 alkyl.
  • R 1 is optionally substituted C4-C6 cycloalkyl. In one aspect of this embodiment, R 1 is C4-C cycloalkyl optionally substituted with one to three R 7 groups. In another aspect of this embodiment, R 1 is C4, C 5 , or C 6 cycloalkyl optionally substituted with one to two R 7 groups and R 7 is halo. In another aspect of this embodiment, R 1 is C4 or C 6 cycloalkyl optionally substituted with one to two R 7 groups and R 7 is halo. In yet another aspect of this embodiment, R 1 is
  • R 1 is
  • R is optionally substituted aryl. In one aspect of this specification,
  • R is aryl optionally substituted with oneto three R groups.
  • R is phenyl optionally substituted with one totwo R groups and R is -CI.
  • R is optionally substituted aryl or optionally substituted aryl
  • R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another aspect of this embodiment, R is optionally substituted heteroaryl. In another
  • R is heteroaryl optionally substituted with one to three R group .
  • R 3 is pyridinyl, indazolyl, benzoimidazolyl, indolyl, orN-
  • each R is optionally substituted with one R wherein R is -F.
  • R is optionally substituted aryl.
  • R is optionally substituted aryl.
  • R is aryl optionally substituted with oneto three R groups.
  • R is phenyl optionally substituted with one R wherein R is -F.
  • R 3 is phenyl optionally substituted with one or two R 7 s wherein each R 7 is independently halo; -CN; -N(R 6 ) 2 ; C r C 4 alkyl optionally substituted with -OR 6 ; -0-C C 4 alkyl optionally substituted with halo, or -OR 6 ; -S0 2 N(R 6 ) 2 ; -S0 2 (C 1 -C4 alkyl); -S(0)-C !
  • R is
  • R 4 is optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl.
  • R 4 is aryl, heteroaryl, aralkyl, or heteroaralkyl, wherein said aryl, heteroaryl, aralkyl, and heteroaralkyl are each independently optionally substituted with oneto three R groups.
  • R 4 is aryl or heteroaryl, each aryl or heteroaryl is optionally substituted with one to three R 7 groups.
  • R 4 is 6- membered aryl or 5-6 membered heteroaryl, wherein said aryl or heteroaryl is optionally substituted with one to three R 7 groups.
  • R 4 is 6- membered aryl or 5-6 membered heteroaryl, wherein said aryl or heteroaryl is optionally substituted with one to three R 7 groups.
  • each member of R 4 is optionally substituted with one or two R 7 groups and each R 7 is independently F, CI, methyl, CF 3 , CN, OMe, or N(R 6 ) 2 .
  • each R 7 is independently F, CI, methyl, CF 3 , CN, OMe, or N(R 6 ) 2 .
  • R 4 is:
  • R 100 is independently H, methyl, F, CI, CF 3 , CN, OCH 3 , or N(R 6 ) 2 .
  • R 4 is:
  • R 4 is:
  • ring A is
  • each R 5 is independently halo; -OR 6 ; -C(0)d -C 4 alkyl; C ,-C 4 alkyl optionally substituted with -OR 6 ; -C3-C5 carbocyclyl optionally substituted with one or two R 6 groups; -C 1 -C4 alkyl-C(0)0-Ci-C 4 alkyl; or -C(0)0- C 1 -C4 alkyl.
  • each R 5 is
  • each R 5 is independently methyl or -C(0)CH 3 .
  • ring A is:
  • ring A is:
  • R 1 , R 4 , ring A and R 7 are as defined in formula I or any one of the above embodiments and R 10 is CR 1 1 or N wherein R 1 1 is -F, -S0 2 NH 2 , -S0 2 CH 3 , -CN, methoxy, -OCH 2 OH, -CH 2 OH, -S0 2 N(CH 3 ) 2 , -S0 2 NHCH 3 , -NHS0 2 CH 3 , -CH 2 CH 2 OH, -N(CH 3 ) 2, t-butyl, cyclopropyl, -C(OH)(CH 3 ) 2 , -OCF 3 , -OCHF 2 , -O-cyclopropyl, -1 -methyl-cyclopropyl, or pyrazolyl.
  • R 1 , R 4 , and ring A are as defined in formula I or any one of the above embodiments; R 7 is H or CI; and and R 10 is CR 1 1 or N wherein R 1 1
  • R 1 , R 4 , and ring A are as defined in formula 1 or any one of the above embodiments and R 7 is H or CI.
  • R 4 is:
  • R 4 is H or R 4 and R 1 , R 2 , R 3 , R 4 and ring A as defined in Formula I or in any of the embodiments described herein.
  • R 4 is alkyl.
  • R 4 is optionally substituted aryl or optionally substituted heteroaryl; and R 1 , R 2 , R 3 , R 4 and ring A as defined in Formula I or in any of the embodiments described herein.
  • R 4 is aryl or heteroaryl, each independently substituted with one to three R 7 groups.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and ring A are as defined in any of the embodiments herein.
  • the com ounds ofthis invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, scalemic mixtures, and diastereomeric mixtures, as well as single enantiomers or individual stereoisomers that are substantially free from another possible enantiomer or stereoisomer.
  • substantially free of other stereoisomers means a preparation enriched in acompound having a selected stereochemistry at one or more selected stereocenters by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
  • enriched means that at least the designated percentage of a preparation is the compound having a selected stereochemistry at one or more selected stereocenters.
  • M ethods of obtaining or synthesizing an individual enantiomer or stereoisomer for agiven compound are known in the art and may be applied as practicable to final compounds or to starting material or intermediates.
  • the compound is enriched in a specific stereoisomer by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
  • the compounds of formulal, II, Il-a, II-a-1 , Il-b or II-b-1 may also comprise one or more isotopic substitutions.
  • H may be in any isotopic form, including ' ⁇ , 2 H (D or
  • H T or tritium
  • C may be in any isotopic form, including C, C, and C
  • O may be in any isotopic form, including 16 0 and 18 0; and the like.
  • the compound is enriched in a specific isotopic form of H, C and/or O by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
  • the compounds ofthis invention may also be represented in multiple tautomeric forms, in such instances, the invention expressly includes all tautomeric forms of the compounds described herein, even though only a single tautomeric form may be represented (e.g., alkylation of aring system may result in alkylation at multiple sites, the invention expressly includes all such reaction products). All such isomeric forms of such compounds are expressly included in the present invention.
  • Synthetic chemistry transformations andprotecting group methodologies useful in synthesizing the applicable compounds are known in the art and include, for example, those described in Larock R, Comprehensive Organic Transformations , VCH Publishers (1989); Greene, TW et al, Protective Groups in Organic
  • a salt may be formed with a suitable cation
  • suitable inorganic cations include, but are not limited to, alkali metal ions such as Na + and K + ,
  • alkaline earth cations such as Ca and Mg
  • other cations such as Al
  • suitable organic cations include, but are not limited to, ammonium ion (i.e., NH 4 + ) and substituted ammonium ions (e.g., NH 3 R , NH 2 R , NHR , NR ).
  • suitable substituted ammonium ions are those derived from ethylamine, diethylamine,
  • dicy clone xylamine triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, andtromethamine, as well as amino acids, such as lysine and argjnine.
  • An example of a common quaternary ammonium ion is N(CH 3 ) 4 + .
  • a salt may be formed with a suitable anion.
  • suitable inorganic anions include, but are not limitedto, those derived from the following inorganic acids:
  • hydrochloric hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous.
  • Suitable organic anions include, but are not limitedto, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, asp artic, benzoic,
  • suitable polymeric organic anions include, but are not limitedto, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.
  • a reference to a particular compound also includes salt forms thereof.
  • compositions may be formulated together with a pharmaceutically acceptable carrier or adjuvant into pharmaceutically acceptable compositions prior to be administered to a subject.
  • pharmaceutically acceptable compositions further comprise additional therapeutic agents in amounts effective for achieving a modulation of disease or disease symptoms, including those described herein.
  • pharmaceutically acceptable carrier or adjuvant refers to a carrier or adjuvant that may be administered to a subject, together with a compound of this invention, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the compound.
  • Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self- emulsifying drug delivery systems (SEDDS) such as d- ⁇ -tocopherol polyethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, poly
  • Cyclodextrins such as ⁇ -, ⁇ -, and ⁇ -cyclodextrin, or chemically modified derivatives such as hydroxy alkylcyclodextrins, including 2- and 3-hydroxypropyl- -cyclodextrins, or other solubilized derivatives may also be advantageously usedto enhance delivery of compounds of the formulae described herein.
  • compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally , nasally, buccally, vaginally or via an implanted reservoir, preferably by oral administration or administration by injection.
  • the pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically -accept able carriers, adjuvants or vehicles.
  • thepH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form.
  • parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
  • compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension.
  • This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents
  • the sterile inject able preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol.
  • a non-toxic parenterally acceptable diluent or solvent for example, as a solution in 1 ,3-butanediol.
  • acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as asolvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or diglycerides.
  • Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions.
  • Other commonly used surfactants such as Tweens or Spans and/or other similar emulsifyingagents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
  • compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions.
  • carriers which are commonly used include lactose and corn starch.
  • Lubricatingagents such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • compositions of this invention may also be administered in the form of suppositories for rectal administration.
  • These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components.
  • suitable non-irritating excipient include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.
  • Topical administration of the pharmaceutical compositions of this invention is useful when the desired treatment involves areas or organs readily accessible by topical application.
  • the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier.
  • Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, poly oxy ethylene
  • the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier with suitable emulsifying agents.
  • suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • the pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically -transdermal patches are also included in this invention.
  • compositions of this invention may be administered by nasal aerosol or inhalation
  • Such compositions are prepared accordingto techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fiuorocarbons, and/or other solubilizing or dispersing agents known in the art.
  • compositions of this invention comprise a combination of a compound of the formulae described herein and one or more additional therapeutic or prophylactic agents
  • both the compound and the additional agent should be present at dosage levels of between about 1 to 100%, and more preferably between about 5 to 95% of the dosage normally administered in a monotherapy regimen.
  • the additional agents may be administered separately, as part of a multiple dose regimen, from the compounds of this invention. Alternatively, those agents may be part of a single dosage form, mixed together with the compounds of this invention in a single composition
  • the compounds described herein can, for example, be administered by injection, intravenously, intraarterially, subdermally, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosahy , topically, in an ophthalmic preparation, or by inhalation, with a dosage ranging from about 0.5 to about 100 mg kg of body weight, alternatively dosages between 1 mg and 1000 mgdose, every 4 to 120 hours, or according to the requirements of theparticular drug
  • the methods herein contemplate administration of an effective amount of compound or compound composition to achieve the desired or stated effect.
  • the pharmaceutical compositions of this invention will be administered from about 1 to about 6 times per day or alternatively, as a continuous infusion.
  • Such administration can be used as a chronic or acute therapy.
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form willvary dep ending up on the host treated and the particular mode of administration.
  • a typical preparation will contain from about 5% to about 95% active compound (w/w).
  • such preparations contain from about 20% to about 80% active compound.
  • a maintenance dose of a compound, composition or combination of this invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level. Subjects may, however, require intermittent treatment on a long- term basis upon any recurrence of disease symptoms.
  • compositions described above comprising a comp ound of formulal, II, Il-a, II-a-1 , Il-b, or II-b-1 or a compound described in any one of the embodiments herein, may further comprise another therapeutic agent useful for treating cancer.
  • a method for inhibiting a mutant IDH1 or IDH2 activity comprising contacting a subject in need thereof with acompound (includingits tautomers and/or
  • the cancer to be treated is characterized by a mutant allele of IDH1 or IDH2 wherein the lDH l or IDH2 mutation results in a new ability of the enzyme to catalyzethe NAPH-dependent reduction of a-ketoglutarate to ?(-)-2-hydroxyglutarate in a subject.
  • the mutant IDH 1 has an R132X mutation.
  • the R132X mutation is selected fromR132H, R132C, R132L, R132V, R132S and R132G. In another aspect, the R132X mutation is R132H or R132C. Inyet another aspect, the R132X mutation is R132H. Also provided are methods of treating a cancer characterized by the presence of a mutant allele of IDH1 comprisingthe step of administering to subject in need thereof (a) a compound of formula I, II, Il-a, ⁇ -a-l , Il-b, or II-b-1, or a compound described in any one of the embodiments herein, or a pharmaceutically acceptable salt thereof, or (b) a pharmaceutical composition comprising(a) and a pharmaceutically acceptable carrier.
  • the cancer to be treated is characterized by a mutant allele of IDH 1 wherein the IDH 1 mutation results in a new ability of the enzyme to catalyze the NAPH- dependent reduction of ⁇ -ketoglutarate to i?(-)-2-hydroxyglutarate in a patient.
  • the IDH1 mutation is an R132X mutation. In another asp ect ofthis
  • the R132X mutation is selected fromR132H, R132C, R132L, R132V, R132S and R132G. In another asp ect, the R132X mutation is R132 H or R132C.
  • a cancer can be analyzed by sequencing cell samples to determine the presence and specific nature of (e.g., the changed amino acid present at) a mutation at amino acid 132 of IDH 1.
  • the comp ounds and methods of this invention are useful to treat ary type of cancer that is characterized by the presence of a mutant allele of IDH1 imparting such acitivity and in particular an IDH 1 R132H or R132C mutation.
  • the efficacy of cancer treatment is monitored by measuring the levels of 2HG in the subject.
  • levels of 2HG are measured prior to treatment, wherein an elevated level is indicated for the use of the compound of formula I, II, ⁇ -a, II-a-1 , Il-b, or II-b-1 or a comp ound described in any one of the embodiments described herein to treat the cancer.
  • the level of 2HG is determined during the course of and/or following termination of treatment to establish efficacy .
  • the level of 2HG is only determined during the course of and/or following termination of treatment. A reduction of 2HG levels during the course of treatment and foUowing treatment is indicative of efficacy.
  • a determination that 2HG levels are not elevated during the course of or foUowingtreatment is also indicative of efficacy.
  • the these 2HG measurements will be utilized together with other well-known determinations of efficacy of cancer treatment, such as reduction in numb er and size of tumors and/or other cancer- associated lesions, improvement in the general health ofthe subject, and alterations in other biomarkers that are associated with cancer treatment efficacy.
  • 2HG can be detected in a sample by LC/MS.
  • the sample is mixed 80:20 with methanol, and centrifuged at 3,000 rpm for 20 minutes at 4 degrees Celsius.
  • the resulting supernatant can be collected and stored at -80 degrees Celsius prior to LC-M S MS to assess 2-bydroxyglutarate levels.
  • LC-M S MS liquid chromatography
  • M etabolites can be separated by reversed phase chromatography using 10 mM tributyl-amine as an ion pairing agent in the aqueous mobile phase, according to a variant of apreviously reported method (Luo et al. J Chromatogr A 1 147, 153-64, 2007).
  • Another method is specific for 2-hydroxyglutarate, running a fast linear gradient from 50% -95% B (buffers as defined above) over 5 minutes.
  • a Synergi Hydro-RP, 100mm x 2 mm, 2.1 ⁇ particle size (Phenomonex) canbe used as the column, as described above.
  • M etabolites can be quantified by comp arison of peak areas with pure metabolite standards at known concentration.
  • M etabolite flux studies from 13 C-glutamine can be performed as described, e.g., in Munger et al. Nat Biotechnol 26, 1 179-86, 2008.
  • 2HG is directly evaluated.
  • a derivative of 2HG formed in process of p erforming the analytic method is evaluated.
  • a derivative can be a derivative formed in M S analysis.
  • Derivatives can include asalt adduct, e.g., a Na adduct, a hydration variant, or a hydration variant which is also a salt adduct, e.g., aNa adduct, e.g., as formed in M S analysis.
  • a metabolic derivative of 2HG is evaluated.
  • examples include species that build up or are elevated, or reduced, as a result of the presence of 2HG, such as glutarate or glutamate that will be correlated to 2HG, e.g., R-2HG.
  • Exemplary 2HG derivatives include dehydrated derivatives such as the compounds rovided below or a salt adduct thereof:
  • the cancer is atumor wherein at least 30, 40, 50, 60, 70, 80 or 90% of the tumor cells carry an IDHl mutation, and in particular an IDHl R132H or R132C mutation, at the time of diagnosis or treatment.
  • IDHl R132X mutations are known to occur in certain types of cancers as indicated in Table 2, below.
  • IDHl R132H mutations have been identified in glioblastoma, acute myelogenous leukemia, sarcoma, melanoma, non-smallcell lung cancer, cho lan gio carcinomas,
  • chondrosarcoma m elodysplastic syndromes (MDS), myeloproliferative neo lasm (MPN), colon cancer, and angio-immunoblastic non-Hodgkin's lymphoma (NHL).
  • MDS m elodysplastic syndromes
  • MPN myeloproliferative neo lasm
  • colon cancer colon cancer
  • NDL angio-immunoblastic non-Hodgkin's lymphoma
  • the methods described herein are used to treat glioma (glioblastoma), acute myelogenous leukemia, sarcoma, melanoma, non-small cell lun g cancer (NSC LC) or cho lan gio carcinomas, chondrosarcoma, my elodys lastic syndromes (MDS), myeloproliferative neoplasm (MPN), colon cancer, or angio-immunoblastic non-Hodgkin's lymphoma (NHL) in a patient.
  • glioma glioblastoma
  • NSC LC non-small cell lun g cancer
  • MDS myelodys lastic syndromes
  • MDN myeloproliferative neoplasm
  • COL angio-immunoblastic non-Hodgkin's lymphoma
  • the cancer is a cancer selected from any one of the cancer types listed in Table 2, and the IDH R132X mutation is one or more of the IDHl R132X mutations listed in Table 2 for that particular cancer type.
  • Treatment methods described herein can additionally comprise various evaluation steps prior to and/or following treatment with acompound of formulal, II, Il-a, II-a-1, Il-b, or II-b-1 or acompound described in any one of the embodiments described herein.
  • the method further comprises the step of evaluating the growth, size, weight, invasiveness, stage and/or other phenotype of the cancer.
  • the method further comprises the step of evaluating the IDHl genotype ofthe cancer. This may be achieved by ordinary methods in the art, such as DNA sequencing, immuno analysis, and/or evaluation of the presence, distribution or level of 2HG.
  • the method further comprises the step of determining the 2HG level in the subject. This may be achieved by spectroscopic analysis, e.g., magnetic resonance-based analysis, e.g., MRI and/or M RS measurement, sample analysis of bodily fluid, such as serum or spinal cord fluid analysis, or by analysis of surgical material, e.g., by mass -sp ectroscopy .
  • spectroscopic analysis e.g., magnetic resonance-based analysis, e.g., MRI and/or M RS measurement
  • sample analysis of bodily fluid such as serum or spinal cord fluid analysis
  • surgical material e.g., by mass -sp ectroscopy
  • the methods described herein comprise the additional step of coadministering to a subject in need thereof a second therapy e.g., an additional cancer therapeutic agent or an additional cancer treatment.
  • additional cancer therapeutic agents include for example, chemotherapy, targeted therapy, antibody therapies, immunotherapy, and hormonal therapy .
  • Additional cancer treatments include, for example: surgery, and radiation therapy . Examples of each of these treatments are provided below.
  • co-administering means that the additional cancer therapeutic agent may be administered together with a compound of this invention as p art of a single dosage form (such as a composition of this invention comprising a compound of the invention and an second therapeutic agent as described above) or as separate, multiple dosage forms.
  • the additional cancer therapeutic agent may be administered prior to, consecutively with, or following the administration of a compound of this invention.
  • both the compounds of this invention and the second thera eutic agent(s) are administered by
  • composition of this invention comprising both a compound of the invention and a second therap eutic agent
  • administration of a composition of this invention, comprising both a compound of the invention and a second therap eutic agent, to a subject does not preclude the separate administration of that same therapeutic agent, any other second therapeutic agent or any compound of this invention to said subject at another time during a course of treatment.
  • co-administering as used herein with respect to an additional cancer treatment means that the additional cancer treatment may occur prior to, consecutively with, concurrently with or following the administration of a comp ound of this invention.
  • the additional cancer therapeutic agent is a chemotherapy agent.
  • chemotherapeutic agents used in cancer therapy include, for example,
  • antimetabolites e.g., folic acid, purine, and pyrimidine derivatives
  • alky lating agents e.g., nitrogen mustards, nitrosoureas, platinum, alkyl sulfonates, hydrazines, triazenes, aziridines, sp indle poison, cytotoxic agents, topoisomerase inhibitors and others
  • hyp omethy lating agents e.g., decitabine (5-aza-deoxycytidine), zebularine, isothiocyanates, azacitidine (5- azacytidine, 5-flouro-2'-deoxycytidine, 5,6-dihydro-5-azacytidine and others).
  • agents include Aclarubicin, Actinomycin, Alitretinoin, Altretamine, Aminopterin, Aminolevulinic acid, Amrubicin, Amsacrine, Anagrelide, Arsenic trioxide, Asparaginase, Atrasentan, Belotecan, Bexarotene, bendamustine, Bleomycin, Bortezomib, Busulfan, Camptothecin, Capecitabine, Carbop latin, Carboquone, Carmofur, Carmustine, Celecoxib, Chlorambucil, Chlormethine, Cisplatin, Cladribine, Clofarabine, Crisantaspase, Cyclophosphamide, Cytarabine, dacarbazine, Dactinomycin, Daunorubicin, Decitabine, Demecolcine, Docetaxel, Doxorubicin, Efaproxiral, Elesclomol, Elsamitrucin, En
  • Floxuridine Fludarabine, Fluorouracil (5FU), Fotemustine, Gemcitabine, Gliadel implants, Hydroxy carbamide, Hydroxyurea, Idarubicin, Ifosfamide, Irinotecan, Irofulven, Ixabepilone, Larotaxel, Leucovorin, Liposomal doxorubicin, Liposomal daunorubicin, Lonidamine,
  • two or more drugs are often given at the same time.
  • two or more chemotherapy agents are used as combination chemotherapy.
  • the additional cancer therapeutic agent is a differentiation agent.
  • Such differentiation agent includes retinoids (such as all-trans-retinoic acid (ATRA), 9-cis retinoic acid, 13-cis-retinoic acid (13-cRA) and 4-hydroxy-phenretinamide (4-HPR)); arsenic trioxide; histone deacetylase inhibitors HDACs (such as azacytidine (Vidaza) and butyrates (e.g., sodium pheny lbutyr ate)); hybrid polar compounds (such as hexamethylene bisacetamide ((HMB A)); vitamin D; and cytokines (such as colony -stimulating factors including G-CSF and GM-CSF, and interferons).
  • retinoids such as all-trans-retinoic acid (ATRA), 9-cis retinoic acid, 13-cis-retinoic acid (13-cRA) and 4-hydroxy-phenretinamide (4-HPR)
  • the additional cancer therapeutic agent is a targeted therapy agent.
  • Targeted therapy constitutes the use of agents specific for the deregulated proteins of cancer cells.
  • Small molecule targeted therapy drugs are generally inhibitors of enzymatic domains on mutated, overexpressed, or otherwise critical proteins within the cancer cell.
  • Prominent examp les are the tyrosine kinase inhibitors such as Axitinib, Bosutinib, Cediranib, dasatinib, erlotinib, imatinib, gefitinib, lapatinib, Lestaurtinib, ilotinib, Semaxanib, Sorafenib, Sunitinib, and Vandetanib, and also cyclin-dependent kinase inhibitors such as Alvocidib and Seliciclib.
  • Monoclonal antibody therapy is another strategy in which the therapeutic agent is an antibody which sp ecifically binds to a protein on the surface of the cancer cells.
  • Examples include the anti- HER2/neu antibody trastuzumab (HERCEPTIN®) typically used in breast cancer, and the anti- CD20 antibody rituximab and Tositumomab typically used in a variety of B-cell malignancies.
  • Other exemplary antibodies include Cetuximab, Panitumumab, Trastuzumab, Alemtuzumab, Bevacizumab, Edreco lomab, and Gemtuzumab.
  • Exemplary fusion proteins include Aflib ercept and Denileukin diftitox.
  • the targeted therapy can be used in combination with a compound described herein, e.g., abiguanide such as metformin or phenformin, preferably phenformin.
  • Targeted therapy can also involve small peptides as "homing devices” which can bind to cell surface receptors or affected extracellular matrix surrounding the tumor. Radionuclides which are attached to these peptides (e.g., RGDs) eventually kill the cancer cell if the nuclide decays in the vicinity of the cell.
  • RGDs Radionuclides which are attached to these peptides
  • An example of such therapy includes BEXXAR®.
  • the additional cancer therapeutic agent is an immunotherapy agent.
  • Cancer immunotherapy refers to a diverse set of therap eutic strategies designed to induce the subject's own immune system to fight the tumor. Contemporary methods for generating an immune resp onse against tumors include intravesicular BCG immunotherapy for superficial bladder cancer, and use of interferons and other cytokines to induce an immune response in renal cell carcinoma and melanoma subjects.
  • Allogeneic hematopoietic stem cell transplantation can be considered a form of immunotherapy, since the donor's immune cells will often attack the tumor in a graft -versus- tumor effect.
  • the immunotherapy agents can be used in combination with a compound or composition described herein.
  • the additional cancer therapeutic agent is a hormonal therapy agent.
  • the growth of some cancers can be inhibited by providing or blocking certain hormones.
  • hormone-sensitive tumors include certain types of breast andprostate cancers. Removingor blo cking estrogen or testosterone is often an important additional treatment.
  • administration of hormone agonists, such as progestogens may be therapeutically beneficial.
  • the hormonal therapy agents can be used in combination with a comp ound or a composition described herein.
  • therap eutic modalities include imatinib, gene therapy, peptide and dendritic cell vaccines, synthetic chlorotoxins, and radiolabeled drugs and antibodies.
  • Step A Tert-butyl 3-oxocyclobutylcarbamate.
  • 3-oxocyclobutanecarboxylic acid 10 g 88 mmol
  • dry DCM 60 mL
  • SOCl 2 20 mL
  • the mixture was heatedto reflux for 1.5 h and then evaporated in vacuo.
  • the resulting mixture was co-evaporated twice with toluene (2 x 8 mL) and the residue was dissolved in acetone (30 mL), followed by adding dropwise to a solution of NaN 3 (12 g 185.0 mmol) in H 2 0 (35 mL) at 0 °C.
  • StepB Tert-butyl 3,3-difluorocyclobutylcarbamate.
  • DAST diethylaminosulfur trifluoride
  • Step C N-(3,3-difhorocyclobutyl)formamide.
  • M eOH 170 mL
  • CH 3 COCl 65 mL
  • tert-butyl 3,3-difluoro -cyclobutylcarbamate 12.1 g, 58.42 mmol
  • the reaction mixture was stirred at 0 °C for 20 min, and then allowed to warm up to r.t and stirred for another 1.5 h.
  • the reaction mixture was concentrated and dissolved in H 2 0 (200 mL).
  • Step D l,l-Difluoro-3-isocyanocyclobutane.
  • N-(3,3-difluorocyclobutyl) - formamide 2.0 g, 14.81 mmol
  • PPh 3 4.27 g, 16.29 mmol
  • DCM 35 mL
  • CCU 1 .43 mL, 14.81 mmol
  • TEA 2.06 mL, 14.81 mmol
  • the reaction mixture was stirred at 45 °C overnight under a N 2 atmosphere.
  • the resulting mixture was evaporated in vacuo at 0 °C.
  • the residue was suspended in Et20 (25 mL) at 0 °C for 30 min and then filtered.
  • the filtrate was evaporated to about 5 mL at 0 °C under reduced pressure.
  • the residue was purified by column chromatography using Et 2 0 as eluent to afford the desired product which was used directly in the next step.
  • Step A Benzyl 3-oxocyclobutanecarboxylate.
  • a mixture of 3-oxocycIobutanecarboxylic acid (5 g, 44 mmol), potassium carbonate ( 12 g, 88 mmol) and benzyl bromide (1 1.2 g, 66 mmol) in acetone (50 mL) was refluxed for 1 h.
  • the solvent was then removed under reduced pressure and the residue was partitioned between ethyl acetate and water. Combined organic layers were dried over anhydrous MgSC , filtered and concentrated.
  • Step B Benzyl 3, 3-difluorocyclobutanecarboxylate.
  • DCM 35 mL
  • DAST 0.8 mL, 6.03 mmol
  • Step C 3,3-Difluorocyclobutanecarboxylic acid Benzyl 3,3-difiuorocyclobutanecarboxylate (0.84 g, 3.72 mo 1) was dissolved m ethanol (40 mL), and approximately 0.02 gpalladium on activated carbon was added. The mixture was stirred at room temperature for 12 h under the atmosphere of H 2 and then filtered through a pad of Celite. The filtrates were concentrated and dried in vacuo to give the desired compound. 1H NMR (400 MHz, CDC1 3 ): ⁇ 3.16— 2.55 (m, 5H). StepD: Tert-butyl 3,3-difluorocyclobutylcarbamate.
  • Step E 3,3-Difluorocyclobutanamine hydrochloride.
  • M eOH 170 mL
  • CH 3 COCl 65 mL
  • iert-butyl 3,3-difiuorocyclobutylcarbamate 12.1 g, 58.4 mmol
  • the mixture was stirred at 0 °C for 20 min and then allowed to warm up to roomtemperature.
  • the reaction mixture was stirred for another 1.5 h and then concentrated to give the crude product which was precipitated in ether to givethe desired product as a white solid.
  • Step F N-(3,3-difhorocyclobutyl)formamide.
  • the mixture of 3,3-difluorocyclobutanamine hydrochloride (6.5 g, 60.7 mmol) and TEA (3 eq) in HCOOEt (90 mL) was stirred at 80 °C overnight in a sealed pressure tube.
  • the solvent was removed in vacuo and the residue was purified by column chromatography with 50% petroleum ether / EtOAc to 25%petroleum ether /
  • Step A Tert-butyl 3-hydroxycyclobutylcarbamate.
  • /er/-butyl 3- oxocyclobutylcarbamate 2 g, 10.8 mmol, 2 eq
  • EtOH 20 mL
  • NaBH 4 204 mg, 1 eq
  • the mixture was then allowed to warm to room temperature and stirred for 30 min.
  • the mixture was concentrated in vacuo and the residue was purified by column chromatography using petroleum ether / EtOAc (V:V, 2: 1 to pure EtOAc) as eluent to afford the desired product as a white solid.
  • Step B Tert-butyl 3-fluorocyclobutylcarbamate.
  • DCM dry DCM (20 mL) at -70 °C
  • DAST dropwise l g, 0.85 mL, 1 .17 eq
  • the mixture was then slowly warmed to room temperature and stirred overnight.
  • the resulting mixture was washed with diluted aq. NaHC0 3 .
  • the organic layer was dried over anhydrous Mg 2 S0 4 and concentrated.
  • Step C 3-Fluorocyclobutanamine.
  • the compound was synthesized as outlined in step E of method A set forth above.
  • Step D N ⁇ (3-fluorocyclobutyl)formamide.
  • the compound was synthesized as outlined in step F of method A set forth above.
  • ⁇ 1MMR 400 MHz, CDC1 3 ): ⁇ 8.10 (s, 1 H), 5.94-5.89 (brs, 1 H), 5.32-5.25 (m, 0.5H), 5.18-5.1 1 (m, 0.5H), 4.63-4.42 (m, 1 H), 2.76-2.62 (m, 2H), 2.44-2.31 (m, 2H).
  • Step E l-Fluoro-3-isocyanocyclobutane.
  • the compound was synthesized via the general procedure as the step G in method A set forth above.
  • Step A Tert-butyl 4-hydroxycyclohexylcarbamate.
  • 4-aminocyclohexanol 23 g, 0.2 mol
  • Et 3 N 60 g, 0.6 mol
  • THF 230 mL
  • EtOAc 3 x 200 mL
  • the combined organic layers were washed with water (2 x 200 mL) and brine (200 mL), dried over anhydrous Na 2 S04 and concentrated.
  • the residue was purified by column chromatography on silica gel using DCM/ MeOH (V:V, 20: 1 ) to afford the desired product as a white solid.
  • Step B Tert-butyl 4-oxocyclohexylcarbamate.
  • DCM DCM
  • Dess-Martin periodinane 39.4 g, 92.9 mmol
  • the resulting solution was stirred at room temperature overnight, quenched with aq. Na 2 S 2 0 3 solution and extracted with DCM (3 x 100 mL).
  • the combined organic layers were washed with water (2 x 100 mL) and brine (100 mL), dried over anhydrous aiSC , and concentrated.
  • the residue was purified by column chromatography on silica gel using petroleum ether / EtOAc (V:V, 10: 1 ) to afford desired product as a white solid.
  • Step C Tert-butyl 4,4-difluorocyclohexylcarbamate.
  • DCM dry DCM
  • DAST 2.58 g, 16 mmol
  • the reaction mixture was stirred at r.t overnight.
  • the reaction mixture was poured into ice water slowly and extracted with DCM ( 3 x 100 mL). The combined organic layers were washed with 2 N aq. NaHC0 3 and brine, dried over anhydrous Na 2 S0 4 , filtered and concentrated in vacuo.
  • Step D 4,4-Difluorocyclohexanamine hydrochloride.
  • a mixture of iert-butyl4,4-difluorocyclo- hexyl carbamate (6.0 g, 25.5 mmol) and 6 N HCl/MeOH (60 mL) was stirred at room temperature for 2 h.
  • the reaction mixture was concentrated to give the crude product which was directly used in next step without further purification.
  • M S 136.1 (M + l) + .
  • Step E N-(4,4-difhorocyclohexyl)formamide.
  • a mixture of 4,4-difluorocyclohexanamine (crude 3.4 g 25.2 mmol), TEA (3 eq) and ethyl formate (35 mL) was stirred at 1 10 °C overnight in a sealed tank. The solvent was removed and the residue was purified by column
  • Step F l,l-Difluoro-4-isocyanocyclohexane.
  • a mixture of N-(4,4-difiuorocyclohexyI) - formamide (2.5 g, 15.3 mmol), PPh 3 (4.4 g, 16.8 mmol), CC1 4 (2.3 g, 15.1 mmol), Et 3 N (1.5 g, 14.9 mmol) and DCM (50 mL) was heatedto 45 °C and stirred overnight. The resulting mixture was evaporated in vacuo and the residue was suspended in Et 2 0 (125 mL) at 0 °C. The filtrate was concentrated and the residue was purified by column chromatography on silica geleluting with Et 2 0 to afford the desired product as a yellow oil which was used directly in the next step.
  • Step A 2-(3-N rophenoxy)ethanol.
  • a suspension of 3-nitrophenol ( 1 g, 7.2 mmol), 2- bromoethanol ( 1 .2 g, 9.6 mmol) and K 2 C0 3 (2 g, 14.4 mmol) in MeCN ( 12 mL) was stirred at 90 °C overnight.
  • the precipitate was collected by filtration to give the first batch of product.
  • the filtrate was concentrated and the residue was purified by column chromatography to afford another batch of the desired product as a yellow solid.
  • Step B 2-(3-Aminophenoxy)ethanol.
  • 2-(3-nitrophenoxy)ethanol 500 mg, 2.7 mmol
  • NH 4 C1 720 mg, 1 3.5 mmol
  • EtOH 10 mL
  • iron powder 900 mg, 16.2 mmol
  • the reaction was then stirred at 90 °C for 2 hr and subsequently cooled.
  • the mixture was filtered and the filtrate was concentrated.
  • the resulting residue was purified by column chromatography to afford the desired product as a yellow solid.
  • Step A Tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole-l-carboxylate.
  • Step B 4-(3-Nitrophenyl)-l H-pyrazole.
  • tert-butyl 4-(4,4,5,5-tetramethyl-l ,3,2- dioxaborolan-2-yl)- l H-pyrazole- l -carboxylate 300 mg, 0.82 mmol
  • l -bromo-3-nitrobenzene 137 mg, 0.68 mmol
  • Na 2 C0 3 216 mg, 2.04 mmol
  • DME/H 2 0 5 mL/ 1 mL
  • N 2 was added Pd(PPh 3 ) 2 Cl2 (24 mg, 0.034 mmol).
  • Step C 3-(lH ⁇ pyrazol-4-yl)aniline.
  • Iron powder (296 mg, 5.30 mmol) was added to a solution of 4-(3-nitrophenyl)- l H-pyrazole (200 mg, 1 .06 mmol) in AcOH/EtOH (2 mL/3 mL).
  • the reaction mixture was stirred at 90 °C for 2 hr and then cooled to room temperature.
  • the reaction mixture was filtered through Celite. The filter cake was washed with H 2 0.
  • the combined organic layers were washed with brine, dried over anhydrous Na 2 S0 4 , and concentrated.
  • the resulting residue was purified by column chromatography to afford the desired product.
  • MS 160.2 (M+ l ) + .
  • Step A Ethyl 3-(dibenzylamino)benzoate 2.
  • Et 3 N 5.26 mL, 0.036 mmol
  • CH 3 CN 30 mL
  • BnBr 4.32 mL, 0.036 mmol
  • the reaction mixture was heated to reflux for 18 hr and then cooled to room temperature.
  • Step B 2-(3-(dibenzylamino)phenyl)propan-2-ol.
  • MeMgBr 3 M sol. in THF, 5.58 mL, 16.7 mmol
  • the reaction was stirred at room temperature overnight and quenched by addition of saturated NH 4 C1.
  • the resulting mixture was extracted with ethyl acetate (3 x 50 mL).
  • Step C 2-(3-aminophenyl)propan-2-ol.
  • 2-(3-(dibenzylamino)phenyl)propan-2- ol (268 mg. 0.81 mmol) in MeOH (5 mL) was added 10% Pd/C (27 mg) in one portion. The reaction mixture was hydrogenated at room temperature overnight under hydrogen atmosphere.
  • Step A Methyl 3-fluoro-5-nltrobenzoate.
  • Thionyl chloride (488 mg, 4.1 mmol) was added dropwise to a solution of 3-fluoro-5-nitrobenzoic acid (500 mg, 2.7 mmol) in dry methanol (10 mL) at 0 °C under nitrogen atmosphere.
  • the reaction was warmed to room temperature and stirred for 6 hr.
  • the reaction mixture was concentrated under reduced pressure to obtain the corresponding methyl ester hydrochloride as a waxy solid which was used directly in the next step.
  • MS 200 (M+ l ) + .
  • Step B Methyl 3-atnino-5-fluorobenzoate.
  • methyl 3-fluoro-5-nitrobenzoate 400 mg, 2 mmol
  • iron powder 560 mg, 10 mmol
  • ammonium chloride 540 mg, 10 mmol
  • the reaction mixture was stirred at 80 °C for 1 hr. After cooling the reaction, the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure to give the desired product.
  • Step C Methyl 3-(dibenzylamino)-5-fluorobenzoate.
  • methyl 3-amino-5- fluorobenzoate 440 mg, 2.6 mmol
  • NaH 1 87 mg, 7.8 mmol
  • benzyl bromide 1 . 1 g, 6.5 mmol
  • Step D 2-(3-(Dibenzylamino)-5-fluorophenyl)propan-2-ol.
  • Methylmagnesium bromide (1 M in THF, 2.4 mL, 2.4 mmol) was dissolved in THF (5 mL) and placed in an ice-water bath.
  • Methyl 3-(dibenzylamino)-5-fluorobenzoate (280 mg, 0.8 mmol) in THF (5 mL) was then slowly added to the reaction mixture. This mixture was stirred for 3 hr while maintaining an internal temperature range between 1 5 to 25 °C.
  • Step A Ethyl 3-(dlbenzylamino)benzoate.
  • Et 3 N 5.26 mL, 0.036 mmol
  • CH 3 CN 30 mL
  • BnBr 4.32 mL, 0.036 mmol
  • Step A (3-Fluoro-5-nitrophenyl)(methyl)sulfane
  • 3-fiuoro-5-nitroaniline 200 mg, 1.28 mmol
  • l ,2-dimethyldisulfane 121 mg, 1.29 mmol
  • CH 3 CN 3 CN
  • Neat isoamyl nitrite 150 mg, 1.28 mmol
  • the reaction mixture was slowly heatedto reflux over 10 min and maintained at a gentle refluxuntilN 2 evolution ceased (30-60 min).
  • the reaction mixture was cooled and the solvent was removed in vacuo to afford a dark oil.
  • the resulting oil was purified by column chromatography to give the desired product as apale yellow solid.
  • StepB 3-Fluoro-5-(methylthio)aniline.
  • M eOH M eOH
  • Pd/C 10% Pd/C
  • Step A (S )-3-Amino-2-(((benzyloxy)carbonyl)amino)propanoic acid.
  • PIAD PI-decanediol
  • Step B (S)-M ethyl 3-amino-2-(((benzyloxy)carbonyl)amino)propanoate hydrochloride.
  • SOCl 2 5 mL
  • the resulting mixture was stirred at 0 °C for 0.5 h before (S)-3-amino-2-(((benzyloxy)carbonyl)amino) propanoic acid (2.6 g, 10 mmol) was added. Then the reaction mixture was stirred at room temperature overnight and concentrated in vacuo to afford the desired product.
  • M S 253.1 (M+l) + .
  • Step C (S)-M ethyl 3-((2-(benzyloxy)-2-oxoethyl)amino)-2-(((benzyloxy)carbonyl)amino)pro- paneate.
  • Step D ( S )-M ethyl 3-((2-(benzyloxy)-2-oxoethyl)( tert-butoxycarbonyl)amino)-2-( ((benzyloxy) carbonyl)amino)pmpanoate.
  • Step E (S)-2-((2-Amino-3-methoxy-3-oxopropyl)(tert-butoxycarbonyl)amino)acetic acid.
  • (S)-methyl 3-((2-(benzyloxy)-2-oxoethyl)(tert-butoxycarbonyl)amino)-2-(((benzyl- oxy)carbonyl)amino)propanoate (2.5 g, 5 mmol) in M eOH (30 mL) was added 10% PdC (250 mg). The mixture was stirred under hydrogen atmosphere at room temperature overnight. The resulting suspension was filtered through Celite, and the filtrate was concentrated in vacuo to afford the desired product.
  • Step E (S )-l-tert-Butyl 3-methyl 5-oxopiperazine-l,3-dicarboxylate.
  • Step F (S)-4-(tert-Butoxycarbonyl)-6-oxopiperazine-2-carboxylic acid
  • MeOH MeOH
  • THF 20 mL
  • a solution ofLiOH * H 2 0 159 mg, 3.8 mmol
  • H 2 0 10 mL
  • EtOAc 25 mL
  • Step A 2-Hydroxy-4-carboxyaIdehyde oxime.
  • Step B 2-Bromopyrimidine-4-carbonitrile.
  • a mixture of 2-hydroxy-4-carboxyaldehyde oxime (9 g, 28.8 mmol), tetrabutyl ammonium bromide ( 10 g, 71 .9 mmol) and phosphorus pentoxide (2 g, 14.4 mmol) in toluene (300 mL) was stirred at 120 °C for 2 h. The resulting mixture was filtered and the filtrate was concentrated. The resulting residue was purified by column
  • n 1 ,2
  • Compound 2 was prepared according to the following scheme, using the following protocol.
  • Step A (S)-Methyl l-methyl-5-oxopyrrolidine-2-carboxylate.
  • DMF 50 mL
  • anhydrous K 2 C0 3 16 g, 1 16 mmol
  • iodomethane 16.4 g, 1 16 mmol
  • Step B (S)-l-Methyl-5-oxopyrrolidine-2-carboxylic acid.
  • MeOH MeOH
  • THF 2 mL
  • H 2 0 2 mL
  • NaOH 0.45 g, 1 1 .4 mmol
  • Step C Compound2. 2-Chlorobenzaldehyde (1 17 mg, 0.83 mmol), 3-fluoroaniline (92.5 mg, 0.83 mmol), crude (S)-l-methyl-5-oxopyrrolidine-2-carboxylic acid (200 mg, -60% purity, 0.83 mmol) and l,l-difluoro-3-isocyanocyclobutane (119 mg, 90% purity, 1.0 mmol) were used in the UGI reaction to give the desired product (diastereomeric mixture). !
  • Step A (S )-N-( l-( 2-Ch loroph enyl)-2-( ( 4,4-difluorocycloh exyl )amino)-2-oxoethyl )-N-( 3-fluoro -phenyl)-5-oxopyrmlidine-2-carboxamide.
  • Compound 44 was prepared according to the following scheme, using the following protocol.
  • Compound 9 was prepared according to the following scheme, using the following protocol.
  • Step A (S)-2-Oxooxazolidine-4-carboxylic acid.
  • (S)-2-(benzyloxycarbonylamino)-3-hydroxypropanoic acid 1 g, 4.2 mmol
  • the mixture was extracted with EtOAc (4 x 10 mL). The combined organic layers were dried over anhydrous Na 2 S0 4 and concentrated in vacuo to give the desired product as a white solid.
  • Step B (4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluoro - phenyl)-2 ⁇ oxooxazolidine-4-carboxamide.
  • 2-Chlorobenzaldehyde 160 mg, 1 .14 mmol
  • 3- fluoroaniline 127 mg, 1 .
  • Step C (S)-N-((R)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3- - fluorophenyl )-2-oxo-3-(pyrimidin-2-yl )oxazolidine-4-carboxamide and(S )-N-( ( S )-l-(2-chloro ⁇ h yl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(pyrimidin- 2-yl)oxazolidine-4-carboxamide.
  • Step A (S)-6-Oxopiperidine-2-carboxylic acid.
  • a solution of (S)-2-aminohexanedioic acid (470 mg, 2.9 mmol) in 20% AcOH (5 mL) was stirred at 1 10 °C overnight.
  • the solvent was removed in vacuo and the residue was dissolved in EtOH ( 10 mL).
  • the unreacted amino acid was precipitated and filtered off. The filtrate was concentrated to give the crude desired product which was used directly in the next step.
  • MS: 142. 1 (M- l ) '
  • Step B (S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-o ⁇
  • Step C (S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxo - fluowphenyl)-6-oxo-l-(pyrimidin-2-yl)piperidine-2-carboxamide and (S)-N-((R)-l-(2-chloro - phenyl)-2-((3,3-difluorocyclobutyl)aniino)-2-oxoethyl)-N-(3-fluorophenyl)-6-oxo-l- (pyrimidin-2-yl)piperidine-2-carboxamide
  • a mixture consisting of ( l /?)-N-( l -(2-chlorophenyl) - 2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoropheny
  • Compound 30 waspr ⁇ ared according to the following scheme, using the following protocol.
  • Step A (S)-3-Hydroxy-2-(4-methoxybenzylamino)propanoic acid.
  • StepB (S)-Benzyl4-(4-methoxybenzyl)-5-oxornorpholine-3-carboxylate.
  • Step C (S Benzyl 5-oxomorpholine-3-carboxylate.
  • CAN eerie ammonium nitrate
  • CAN eerie ammonium nitrate
  • the resulting mixture was stirred at 0 °C for 1 h.
  • DIP EA was added at 0 °C to adjust thepH to 6 ⁇ 7 and the mixture was concentrated in vacuo.
  • the residue was purified by column chromatography to afford the desired product as a white solid M S: 236.1 (M+ l) + .
  • StepD (S )-5-Oxomorpholine-3-carboxylic acid.
  • MS 146.1 (M + l) + .
  • Step E ( S )-N-( ( S )-!-( 2-Chlorophenyl )-2-( ( 3,3-difluorocyclobutyl )amino)-2-oxoethyl )-N-( 3 - fluorophenyl)-5-oxomorpholine-3-carboxamide.
  • Step F Compound 30.
  • a mixture of (5 -N-((5 -l-(2-chlorophenyl)-2-((3,3-difluoro - cyclobutyl)amino)-2-oxoethyl)-N-(3-fiuorophenyl)-5-oxomorpholine-3-carboxamide (100 mg, 0.2 mmol), 2-bromopyrimidine (36 mg, 022 mmol), Pd 2 (dba) 3 (28 mg, 0.03 mmol), XantPhos (16 mg, 0.03 mmol) and Cs 2 C0 3 (160 mg, 0.5 mmol) in 1,4-dioxane (4 mL) was stirred at 100 °C for 3.5 h under N 2 .
  • Step A 5-Nitroisophthaloyl dichloride. To a solution of 5-nitroisophthalic acid (2.3 g, 1 1 mmol) in S0C1 2 (6 mL) was added a drop of DMF and the mixture was stirred at reflux for 3 hr. The resulting reaction mixture was concentrated to give the crude product which was used directly in the next step.
  • Step B 5-Nitroisophthalamide.
  • 5-Nitroisophthaloyl dichloride (2.7 g, 9.7 mmol) was added portionwise to a cold solution of NH 3 ' H 2 0 (40 mL) at 0 °C. The reaction mixture was stirred overnight and a white precipitate formed. The mixture was then filtered, washed with excess of water, and dried at 1 10 °C to give the crude product which was used directly in the next step.
  • Step C 5-Aminoisophthalamide. To a solution of 5-nitroisophthalamide (2 g, 9.6 mmol) in MeOH (200 mL) was added Pd/C (200 mg). The reaction was stirred overnight under a hydrogen atmosphere. The suspension was filtered and the filtrate was concentrated to afford the desired product which was used directly in the next step.
  • StepD 5-((2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)l-(4cyano pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)isophthaIamide.
  • Step E (2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoet yl)l-(4-cyano pyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide.
  • Compound 97 was synthesized via the UGI reaction procedure set forth herein, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring and purified via standard methods.
  • DIPEA (0.055 mL, 0.34 mmol) was added to the mixture followed by Ac 2 0 (0.03 1 mL, 0.34 mmol) at 0°C. Then the mixture was stirred at room temperature for 2 hr. The solution was concentrated and the residue was purified by a standard method to afford the desired product.
  • Step A - Oxazole-5-carboxamide.
  • Ethyl oxazole-5-carboxylate (2 g, 14.2 mmol) was dissolved in NH 3 solution (7 M in MeOH, 25 mL). The solution was stirred at room temperature for 2 hr and filtered. The solid was dried to give the desired product ( 1 .5 g, 92% yield) as a white powder which was used directly in the next step.
  • Step B 2-Iodooxazole-5-carboxamide.
  • Oxazole-5-carboxamide (560 mg, 5.0 mmol) was dissolved in anhydrous THF (7.5 mL) and flushed with N 2 .
  • the solution was cooled to -42 °C and treated with fresh LiHMDS ( 15 mL, 1 M in THF).
  • the solution became dark yellow was stirred for 20 min and fol lowed by the addition of a solution of ZnCl 2 (30 mL, 0.5 M in THF).
  • the reaction was warmed to 0 °C for 1 hr.
  • Step C 2-((S)-2-(((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)a
  • Step D ( S)-N-( ( S)-l-( 2-Chloroph enyl )-2-( ( 3,3-difluomcyclobutyl )amino)-2-oxoethyl )-!-( 5- cyanooxazol-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2- rboxamide.
  • Step A (2S,4R)-l-tert-Butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)pyrrolidine-l,2- dicarboxylate.
  • Imidazole (2.8 g, 40.8 mmol) was added to a solution of (2S,4R)- l -tert-butyl 2- methyl 4-hydroxypyrrolidine- l ,2-dicarboxylate (5.0 g, 20.4 mmol) and TBSC1 (4.6 g, 30.6 mmol) in anhydrous DMF ( 100 mL). The mixture was stirred at room temperature overnight and then partitioned between EtOAc and H 2 0. The organic layer was separated, washed with aq.
  • Step B (2S,4R)-l-tert-Butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-l,2- dicarboxylate.
  • NaI0 4 7.5 g, 35.0 mmol
  • Ru0 2 370 mg, 2.8 mmol
  • Step C ( 2S,4R )-4-(( tert-Butyl dimethyl ' silyl )oxy)-5-oxopyrrolidin e-2-carboxylic acid.
  • TF A (6 mL) was added to a solution of (2S,4R)- l -tert-butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)-5- oxopyrrolidine- l ,2-dicarboxylate (2.5 g, 6.68 mmo l) in DCM (18 mL) at 0 °C.
  • the mixture was stirred at room temperature for 1 hthen concentrated.
  • the residue was dissolved in M eOH/THF (1 OmL/1 OmL) followed by addition of a solution of LiOH (842 mg, 20.1 mmol) in water (5 mL).
  • Step D The same as general procedure for UGI reaction set forth herein.
  • Step E The same as general procedure for Buchwald reaction set forth herein.
  • Step F (2S,4R )-N-( l-(2-Chlorophenyl )-2-( ( 3,3-difluorocyclobutyl )amino)-2-oxoethyl )-N-(3- cyano-ph enyl )-!-( 4-cyan opyridin-2-yl )-4-hydroxy-5 -oxopyrrolidin e-2-carboxamide.
  • Step A (2S)-1 -tert-Butyl 2-methyl 4-( tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine- 1 ,2-dicarboxylate.
  • LiHM DS (1 M in THF, 22.6 mL, 22.6 mmol) was added into a mixture of (2S,4R)-l -tert -butyl 2-methyl 4-(tert-butyldimethylsi ⁇ loxy)-5-oxopyrrolidine- l,2-dicar-boxylate (6.5 g, 17.4 mmol) in THF (60 mL) at -78°C under N 2 .
  • the mixture was stirred at -78°C for 1 hr.
  • Step B (2S,4S)-M ethyl 4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-2-carboxylate.
  • a solution of (2S)- l -tert-butyl 2-methyl 4-(tert-butyldimetbylsilyloxy)-4-methyl-5-oxopyrroli-dine- 1 ,2-dicarbo xylate (960 mg, 25 mmo l) in TFA/DQVI (V : V 1 : 3) was stirred at room temperature for 1 h. The mixture was then concentrated to give the desired product which was used directly in the next step .
  • Step C (2S)-4-(tert-Butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidin e-2-carboxylic acid.
  • (2S)-methyl 4-(tert -bury ldimethylsiryloxy)-4-methyl-5-oxopyrrolidine-2-carbo-xy late 400 mg, 1.4 mmol
  • LiOH 50 mg 2.1 mmol
  • Step D (2S)-4-(tert-Butyldimeth ⁇ silyloxy)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobut- ylamino)-2-oxoethyl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidin e-2-carboxamide.
  • Step E (2S)-4-(tert-Butyldimethylsilyloxy)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutyl- amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2- carboxamide.
  • Step F ( 2S)-N-( (R)-l-( 2-Ch loroph enyl )-2-( 3,3-dflu orocyclobutyl amino )-2-oxoethyl)-l-( 4- cyanopyridin-2-yl )-N-( 3 -fluorophenyl )-4-hydroxy-4-methyl-5-oxopyrrolidine-2-carboxamide.

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Abstract

Provided are methods of treating a cancer characterized by the presence of a mutant allele of IDH1/2 comprising administering to a subject in need thereof a compound described here.

Description

THERAPEUTIC ALLY ACTIVE COMPOUNDS AND THEIR METHODS OF US E
BACKGROUND OF INVENTION
Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate (i. <?., a-ketoglutarate). These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is ahomodimer.
IDH1 (isocitrate dehydrogenase 1 (NADP+), cytosolic) is also known as IDH; IDP; IDCD; ID PC or PICD. Theprotein encoded by this gene is the NADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence. The presence of this enzyme inperoxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2, 4-dienoyl- CoAs to 3-enoyl-CoAs, aswell as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation ofphytanic acid. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production.
The human IDH 1 gene encodes a protein of 414 amino acids. The nucleotide and amino acid sequences for human IDH 1 can be found as GenBank entries NM 005896.2 and
NP 005887.2 respectively. The nucleotide and amino acid sequences for IDH1 are also described in, e.g., Nekrutenko et al., M ol. Biol. Evol. 15:1674-1684(1998); Geisbrecht et al., J. BioL Chem. 274 :30527-30533(1999); Wiemann et al., Genome Res. 1 1:422-435(2001); The MGC Project Team, Genome Res. 14:2121-2127(2004); Lubec et al, Submitted (DEC-2008) to UniProtKB; Kullmann et al, Submitted (JUN-1996) to the EM BL/GenBank/DDB J databases; and Sjoeblom et al, Science 314:268-274(2006).
Non-mutant, e.g., wild type, IDH 1 catalyzes the oxidative decarboxylation of isocitrate to a-ketoglutarate thereby reducing NAD+ (NADP+) to NADH (NADPH), e.g., in the forward reaction:
Isocitrate + NAD+ (NADP+)→ a-KG + C02 + NADH (NADPH) + H+. It has been discovered that mutations of IDH l present in certain cancer cells result in a new ability of the enzyme to catalyze the NAPH-dependent reduction of a-ketoglutarate to R(-)- 2-hydroxyglutarate (2HG). The production of 2HG is believed to contribute to the formation and progression of cancer (Dang, L et al, Nature 2009, 462:739-44).
The inhibition of mutant IDH l and its neoactivity is therefore a potential therapeutic treatment for cancer. Accordingly, there is an ongoing need for inhibitors of IDH l mutants having alpha hydroxyl neoactivity.
SUMMARY OF INVENTION
Described herein are methods of treating a cancer characterized by the presence of a mutant allele of IDH l or IDH2. The methods comprise the step of administering to a subject in need thereof a compound of formula I, or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate the
Figure imgf000003_0001
I
R1 is optionally substituted C4-C6 carbocyclyl;
each R2 and R3 is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
R4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl;
ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 is independently halo; -CF ; -CN; -OR6;-N(R6)2; -C(0)C,-C4 alkyl; C1 -C4 haloalkyl; C ,-C4 alkyl optionally substituted with -OR6 or -N(R6)2; -0-Ci-C4 alkyl optionally substituted with halo, -OR6 or -N(R6)2; -S02N(R6)2; -S02(C ,-C4 alkyl); -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -0-(C3-C6 carbocyclyl optionally substituted with one or two R6 groups); 5-6 membered heteroaryl; -Ci-C4 alkyl-C(0)0-Ci-C4 alkyl; or -C(0)0-C,-C4 alkyl;or
each R6 is independently H or C 1 -C3 alkyl. The compound of formula I inhibits mutant IDH 1/2, particularly mutant IDH 1 having alpha hydroxyl neoactivity. Also described herein are pharmaceutical compositions comprising a compound of formula I.
DETAILED DES CRIPTION OF THE INVENTION
The details of construction and the arrangement of components set forth in the following description or illustrated in the drawings are not meant to be limiting. Other embodiments and different ways to practice the invention are expressly included. Also, the phraseology and terminology used herein is for thepurpose of description and should not be regarded as limiting. The use of "including," "comprising," or "having" "containing", "involving", and variations thereof herein, is meant to encompass the items listed thereafter and equivalents thereof as well as additional items.
Definitions:
Theterm "halo" or "halogen" refers to any radical of fluorine, chlorine, bromine or iodine.
Theterm "alky 1" refers to a hydrocarbon chain that may be a straight chain or branched chain, containing the indicated number of carbon atoms. For example, -C12 alkyl indicates thatthe group may have from 1 to 12 (inclusive) carbon atoms in it. The term "haloalkyl" refers to an alkyl in which one or more hydrogen atoms are replaced by halo, and includes alkyl moieties in which all hydrogens have been replaced by halo (e.g., perfiuoroalkyl). The terms "arylalkyl" or "aralkyl" refer to an alkyl moiety in which an alkyl hydrogen atom is replaced by an ary 1 group . Arylalkyl or aralkyl includes groups in which more than one hydrogen atom has been rep laced by an ary 1 group . Examp les of " arylalky 1" or " aralky 1" include benzy 1, 2- pherylethyl, 3-pherylpropyl, 9-fiuorenyl, benzhydryl, and trityl groups. The terms
"heteroarylalkyl" or "hetero aralkyl" refer to an alkyl moiety in which an alkyl hydrogen atom is replaced by a heteroaryl group . Heteroarylalkyl or heteroaralkyl includes groups in which more than one hydrogen atom has been rep laced by a heteroaryl group.
Theterm "alkylene" refers to a divalent akyl, e.g, -CH2-, -CH2CH2-, and -CH2CH2CH2-. Theterm "alkenyl" refers to a straight or branched hydrocarbon chain containing 2- 12 carbon atoms and having one or more double bonds. Examples of alkenyl groups include, but are not limited to, allyl, propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups. One of the double bond carbons may optionally be the point of attachment of the alkenyl substituent. Theterm "alkynyl" refers to a straight or branched hydrocarbon chain containing2-12 carbon atoms and characterized in having one or more triple bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, propargyl, and 3-hexynyl. One ofthe triple bond carbons may optionally be the point of attachment of the alkynyl substituent.
Theterm "alkoxy" refers to an -O-alkyl radical. The t erm" halo alkoxy" refers to an alkoxy in which one or more hydrogen atoms are replaced by halo, and includes alkoxy moieties in which allhydrogens have been rep laced by halo (e.g., perfiuoroalkoxy).
Theterm "carbocyclyl" refers to amonocyclic, bicyclic or tricyclic, hydrocarbon ring system that is not fully aromatic, wherein any ringatom capable of substitution can be substituted by one or more substituents. A carbocyclyl can be fully or partially saturated. A bicyclic or tricylic carbocyclyl may contain one (in the case of abicycle) or up to two (in the case of a tricycle) aromatic rings, as long as at least one ring in the carbocyclylis non-aromatic. Unless otherwise specified, any ring atom capable of substitution in a carbocyclyl can be substituted by one or more substituents.
Theterm "aryl" refers to afully aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ringsystem Examples of aryl moieties are phenyl, naphthyl, and anthracenyl. Unless otherwise specified, any ring atom in an aryl can be substituted by one or more substituerts.
Theterm "cycloalkyl" as employed herein refers to a saturated cyclic, bicyclic, tricyclic, or poty cyclic hydrocarbon group . Unless otherwise specified, any ring atom can be substituted by one or more substituents. The cycloalkyl groups can contain fused rings. Fused rings are rings that share a common carbon atom. Examples of cycloalkyl moieties include, but are not limited to, cyclop ropy 1, cyclohexyl, methylcyclohexyl, adamantyl, and norbornyl. Unless otherwise specified, any ring atom can be substituted by one or more substituents.
Theterm "heterocyclyl" refers to amonocyclic, bicyclic or tricyclic, ringstructure that is not fully aromatic and includes one to four heteroatoms independently selected from N, O, or S in one or more of the rings. A heterocyclyl can be fully or partially saturated. A bicyclic or tricylic heterocyclyl may contain one (in the case of a bicycle) or up to two (inthe case of a tricycle) aromatic rings, as long as at least one ring in the heterocyclyl is non-aromatic. Unless otherwise specified, any ring atom capable of substitution in a heterocyclyl can be substituted by one or more substituents. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, 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, phenant uridine, 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.
Theterm "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms independently selected from O, N, or S, wherein each ringin a heteroaryl is fully aromatic. Unless otherwise specified, any ring atom capable of substitution in a heteroaryl can be substituted by one or more substituents. The terms "hetaralkyl" and "heter oar alky 1", as used herein, refers to an alkyl group substituted with aheteroaryl group . The ring heteroatoms of the compounds provided herein include N-O, S(O), and S(0)2.
Theterm "substituted" refers to the replacement of a hydrogen atom with another moiety. Typical substituents include alky 1 (e.g., CI , C2, C3, C4, C5, C6, C7, C8, C9, CIO, Cl l, C12 straight or branched chain alkyl), cycloalkyl, haloalkyl (e. g., perfluoroalkyl such as CF3), aryl, heteroaryl, aralkyl, heter oaralkyl, heterocyclyl, akenyl, alkynyl, cycloalkenyl,
heterocycloalkenyl, alkoxy, haloakoxy (e. g., perfluoroakoxy such as OCF3), halo, hydroxy, carboxy, carboxylate, cyano, nitro, amino, alkyl amino, S03H, sulfate, phosphate,
methylenedioxy (-0-CH2-0- wherein oxygens are attached to vicinal atoms), ethylenedioxy, oxo (not a substituent on heteroaryl), thioxo (e. g., C=S) (not a substituent onheteroaryl), imino (akyl, aryl, aralkyl), S^^lkyl (where n is 0-2), S(0)n aryl (where n is 0-2), S(0)n heteroaryl (where n is 0-2), S(0)n heterocyclyl (where n is 0-2), amine (mono-, di-, alkyl, cycloalkyl, aralkyl, hetero aralkyl, aryl, heteroaryl, and combinations thereof), ester (alkyl, aralkyl, hetero aralkyl, aryl, heteroaryl), amide (mono-, di-, alkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and combinations thereof), sulfonamide (mono-, di-, alkyl, aralkyl, heteroaralkyl, and combinations thereof). In one asp ect, the substituents on a group are independently any one single, or any subset of the aforementioned substituents. In another aspect, a substituent may itselfbe substituted with any one of the above substituents.
The term "tautomer" refers to each of two or more isomers of a compound (e.g., a compound described herein) that exist together in equilibrium, and are readily interchangeable by migration of a hydrogen atom or proton, accomp anied by a switch of a single bond and an adjacent double bond.
As used herein, the term "elevated levels of 2HG" means 10%, 20% 30%, 50%, 75%, 100%, 200%, 500% or more 2HG than is present in a subject that does not carry a mutant IDH1 or IDH2 allele. The term "elevated levels of 2HG" may refer to the amount of 2HG within a cell, within a tumor, within an organ comprising a tumor, or within abodily fluid.
The term "bodily fluid" includes one or more of amniotic fluid surrounding a fetus, aqueous humour, blood (e.g., blood plasma), serum, Cerebrospinal fluid, cerumen, chyme, Cowper's fluid, female ejaculate, interstitial fluid, lymph, breast milk, mucus (e.g., nasal drainage or phlegm), pleural fluid, pus, saliva, sebum, semen, serum, sweat, tears, urine, vaginal secretion, or vomit.
As used herein, the terms "inhibit" or "prevent" include both comp lete and partial inhibition and prevention. An inhibitor may completely or partially inhibit.
The term "treat" means decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a cancer (e. g., a cancer delineated herein), lessen the severity of the cancer or improve the symptoms associated with the cancer.
As used herein, an amount of a compound effective to treat a disorder, or a
"therap eutically effective amount" refers to an amount of the comp ound which is effective, upon single or multiple dose administration to a subject, in treating a cell, or in curing alleviating, relieving or improving a subject with a disorder beyond that expected in the absence of such treatment.
As used herein, the term "subject" is intended to include human and non-human animals. Exemplary human subjects include a human patient having a disorder, e. g., a disorder described herein or a normal subj ect. The term "non-human animals" of the invention includes all vertebrates, e.g., non-mammals (such as chickens, amphibians, reptiles) and mammals, such as non-human primates, domesticated and/or agriculturally useful animals, e.g., sheep, dog, cat, cow, pig, etc.
Compounds
Provided is a compound having formula 1 or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate the
Figure imgf000008_0001
I
R1 is optionally substituted C4-C6 carbocyclyl;
each R and R is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
R4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl;
ring A is 4-6 membered non-aromatic ring having 0-1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 is independently halo; -CF3; -CN; -OR6;-N(R6)2; -C(0)Ci-C4 alkyl; Ci-C4 haloalkyl; C, -C4 alkyl optionally substituted with -OR6 or -N(R6)2; -O-C 1 -C4 alkyl optionally substituted with halo, -OR6 or -N(R6)2; -S02N(R6)2; -S02(C ,-C4 alkyl); -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -0-(C3-C6 carbocyclyl optionally substituted with one or two R6 groups); 5-6 membered heteroaryl; -C|-C4 alkyl-C(0)0-Ci -C alkyl; or -C(0)0-C,-C4 alkyl; or
each R6 is independently H or C 1 -C3 alkyl.
Provided is also a compound having formula I or a pharmaceutically acceptable salt or hydrate thereof, wherein:
Figure imgf000009_0001
R1 is optionally substituted C4-C6 carbocyclyl;
each R2 and R3 is independently selected from optionally substituted aryl or optionally substituted heteroaryl;
R4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyi;
ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 is independently halo, -CF3, -CN, -OR6, -N(R6)2, -C(0)CH3; C,-C3 haloalkyl, C,- C3 alkyl optionally substituted with -OR6 or -N(R6)2; or
each R6 is independently H or C |-C3 alkyl.
Provided is also a compound having formula 1 or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate thereof, wherein:
Figure imgf000009_0002
I
R1 is C4-C6 carbocyclyl optionally substituted with one to three R7 groups;
each R and R is independently selected from aryl or heteroaryl, wherein said aryl or heteroaryl is independently optionally substituted with one to three R7 groups;
R4 is alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyi, wherein said aryl, heteroaryl, aralkyl, and heteroaralkyi are each independently optionally substituted with one to three R7 groups;
ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 and R7 is independently halo; -CF3; -CN; -OR6;-N(R6)2; -C(0)Ci-C4 alkyl; C 1 -C4 haloalkyl; C 1 -C4 alkyl optionally substituted with -OR6 or -N(R6)2; -O-C 1 -C4 alkyl optionally substituted with halo, -OR6 or -N(R6)2; -S02N(R6)2; -S02(C i-C4 alkyl); -S(0)-Ci.4 alky 1, -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -0-(C3- C6 carbocyclyl optionally substituted with one or two R6 groups); 5-6 membered heteroaryl; -Ci- C4 aIkyl-C(0)0-Ci-C4 alky!; or -C(0)0- -C alkyl; or
each R6 is independently H or C 1 -C4 alkyl.
Provided is also a compound having formula I or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate thereof, wherein:
Figure imgf000010_0001
I
R1 is C4-C6 carbocyclyl optionally substituted with one to three R7 groups;
each R2 and R3 is independently selected from aryl or heteroaryl, wherein said aryl or heteroaryl is independently optionally substituted with one to three R groups;
R4 is alkyl, aryl, heteroaryl, aralkyi, or heteroaralkyi, wherein said aryl, heteroaryl, aralkyi, and heteroaralkyi are each independently optionally substituted with one to three R7 groups;
ring A is 4-6 membered non-aromatic ring having 0- 1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 and R7 is independently halo, -CF3, -CN, -OR6, -N(R6)2, -C(0)CH3; C,-C3 haloalkyl, Ci -C3 alkyl optionally substituted with -OR6 or -N(R6)2; or
each R6 is independently H or C i-C3 alkyl.
In one embodiment, R1 is optionally substituted C4-C6 cycloalkyl. In one aspect of this embodiment, R1 is C4-C cycloalkyl optionally substituted with one to three R7 groups. In another aspect of this embodiment, R1 is C4, C5, or C6 cycloalkyl optionally substituted with one to two R7 groups and R7 is halo. In another aspect of this embodiment, R1 is C4 or C6 cycloalkyl optionally substituted with one to two R7 groups and R7 is halo. In yet another aspect of this embodiment, R1 is
Figure imgf000010_0002
n yet another as ect of this embodiment, R1 is
Figure imgf000011_0001
In another embodiment, R is optionally substituted aryl. In one aspect of this
2 · · 7
embodiment, R is aryl optionally substituted with oneto three R groups. In another aspect of
2 7 7 this embodiment, R is phenyl optionally substituted with one totwo R groups and R is -CI.
3
In another embodiment, R is optionally substituted aryl or optionally substituted aryl
3
heteroaryl. In one aspect of this embodiment, R is optionally substituted heteroaryl. In another
3 · · 7
aspect of this embodiment, R is heteroaryl optionally substituted with one to three R group . In yet another asp ect of this embodiment, R3 is pyridinyl, indazolyl, benzoimidazolyl, indolyl, orN-
3 · · 7 7
methylindolyl, wherein each R is optionally substituted with one R wherein R is -F. In
3
another aspect of this embodiment, R is optionally substituted aryl. In another asp ect of this
3 7
embodiment, R is aryl optionally substituted with oneto three R groups. In yet another aspect
3 · · · 7 · 7
of this embodiment, R is phenyl optionally substituted with one R wherein R is -F. In yet another aspect of this embodiment, R3 is phenyl optionally substituted with one or two R7s wherein each R7 is independently halo; -CN; -N(R6)2; CrC4 alkyl optionally substituted with -OR6; -0-C C4 alkyl optionally substituted with halo, or -OR6; -S02N(R6)2; -S02(C1 -C4 alkyl); -S(0)-C!_4 alkyl, -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one R6; -O- (C3-C6 carbocyclyl); 5-memberedheteroaryl. In yet another aspect of this embodiment, R is
7 7
phenyl optionally substituted with one ortwo R swherein each R is
independently -F, -S02NH2, -S02CH3, -S(0)CH3, -CN,
methoxy, -OCH2OH, -CH2OH, -S02N(CH3)2, -S02NHCH3, -NHS02CH3, -CH2CH2OH, -N(CH3) 2, t -butyl, cyclopropyl, -C(OH)(CH3)2, -OCF3, -OCHF2, -cyclop ropy \ -1-methyl-cyclopropyl, or pyrazolyl.
In another embodiment, R4 is optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl. In one aspect of this embodiment, R4 is aryl, heteroaryl, aralkyl, or heteroaralkyl, wherein said aryl, heteroaryl, aralkyl, and heteroaralkyl are each independently optionally substituted with oneto three R groups. In another aspect of this embodiment, R4 is aryl or heteroaryl, each aryl or heteroaryl is optionally substituted with one to three R7 groups. In another aspect of this embodiment, R4 is 6- membered aryl or 5-6 membered heteroaryl, wherein said aryl or heteroaryl is optionally substituted with one to three R7 groups. In yet another aspect of this embodiment, R4
Figure imgf000012_0001
each member of R4 is optionally substituted with one or two R7 groups and each R7 is independently F, CI, methyl, CF3, CN, OMe, or N(R6)2. In yet another aspect of this
embodiment, R4 is:
Figure imgf000012_0002
R100 is independently H, methyl, F, CI, CF3, CN, OCH3, or N(R6)2. In yet another aspect of this embodiment, R4 is:
Figure imgf000013_0001
CI, CF3, CN, OCH3, or N(R6)2. In yet another embodiment, R4 is:
Figure imgf000013_0002
In another embodiment, ring A is
Figure imgf000014_0001
Figure imgf000014_0002
wherein denotes ring A's attachment to the amide moiety of formula and *> denotes ring 's attachment to R4; and each member of ring A is optional e or two R5
Figure imgf000014_0003
Figure imgf000014_0004
wherein ^ denotes ring A's attachment to the amide moiety of formula and denotes ring A's attachment to R ; and each member of ring A is optionally substituted with one or two R5 groups. In one aspect of this embodiment, each R5 is independently halo; -OR6; -C(0)d -C4 alkyl; C ,-C4 alkyl optionally substituted with -OR6; -C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -C 1 -C4 alkyl-C(0)0-Ci-C4 alkyl; or -C(0)0- C 1 -C4 alkyl. In one aspect of this embodiment, each R5 is
independently -OH, -F, -CH2CH2OH, -CH2C(0)OCH2CH3, -C(0)0-t-butyl, cyclopropyl, methyl or -C(0)CH3. In one aspect of this embodiment, each R5 is independently methyl or -C(0)CH3. In another aspect of this embodiment, ring A is:
Figure imgf000015_0001
In another aspect of this embodiment, ring A is:
Figure imgf000015_0002
Provided is also a compound having formula II or a pharmaceutically acceptable salt or hydrate thereof, wherein R1 , R2, R3, ring A and R4 are as defined in formula 1 or any one of the above embodiments.
Figure imgf000015_0003
formula II
Provided is also a compound having formula Il-a or a pharmaceutically acceptable salt or hydrate thereof, wherein R 1 , R4, ring A and R7 are as defined in formula I or any one of the above embodiments.
Figure imgf000016_0001
formula II-a
Provided is also a compound having formula II-a- 1 or a pharmaceutically acceptable salt or hydrate thereof, wherein R1 , R4, ring A and R7 are as defined in formula I or any one of the above embodiments and R10 is CR1 1 or N wherein R1 1 is -F, -S02NH2, -S02CH3, -CN, methoxy, -OCH2OH, -CH2OH, -S02N(CH3)2, -S02NHCH3, -NHS02CH3, -CH2CH2OH, -N(CH3) 2, t-butyl, cyclopropyl, -C(OH)(CH3)2, -OCF3, -OCHF2, -O-cyclopropyl, -1 -methyl-cyclopropyl, or pyrazolyl.
Figure imgf000016_0002
formula Il-a- 1
Provided is also a compound having formula Il-b or a pharmaceutically acceptable salt or hydrate thereof, wherein R1 , R4, and ring A are as defined in formula I or any one of the above embodiments; R7 is H or CI; and and R10 is CR1 1 or N wherein R1 1
is -F, -S02NH2, -S02CH3, -CN,
methoxy. -OCH2OH, -CH2OH, -S02N(CH3)2, -S02NHCH3, -NHS02CH3, -CH2CH2OH, -N(CH3) ?, t-butyl, cyclopropyl, -C(OH)(CH3)2, -OCF3, -OCHF2, -O-cyclopropyl, -1-methyl-cyclopropyl, or pyrazolyl.
Figure imgf000017_0001
Il-b
Provided is also a compound having formula ll-b-l or a pharmaceutically acceptable salt or hydrate thereof, wherein R1, R4, and ring A are as defined in formula 1 or any one of the above embodiments and R7 is H or CI.
Figure imgf000017_0002
formula II-b-1
In another embodiment of formula II, Il-a, II-a-1, Il-b, or II-b-1,
1 is:
Figure imgf000017_0003
R4is:
Figure imgf000018_0001
Further embodiments provided herein include combinations of one or more of the icular embodiments set forth above.
In another embodiment, exemplary compounds of formula I are depicted below in
Figure imgf000018_0002
18
Figure imgf000019_0001
19
Figure imgf000020_0001
20
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
23
Figure imgf000024_0001
Figure imgf000025_0001
25
Figure imgf000026_0001
26
Figure imgf000027_0001
27
Figure imgf000028_0001
28
Figure imgf000029_0001
29
Figure imgf000030_0001
30
Figure imgf000031_0001
31
Figure imgf000032_0001
32
Figure imgf000033_0001
33
Figure imgf000034_0001
34
Figure imgf000035_0001
35
Figure imgf000036_0001
36
Figure imgf000037_0001
37
Figure imgf000038_0001
38
Figure imgf000039_0001
39
Figure imgf000040_0001
40
Figure imgf000041_0001
41
Figure imgf000042_0001
42
Figure imgf000043_0001
43
Figure imgf000044_0001
Figure imgf000045_0001
Included herein are also methods for making compounds of Formula I or a compound of any one of the embodiments described herein comprising reacting R'NC with R2CHO, R3NH2
and
Figure imgf000045_0002
, wherein R4 is H or R4 and R1 , R2, R3, R4 and ring A as defined in Formula I or in any of the embodiments described herein. In one aspect of the preceding methods, R4 is alkyl.
Also included herein are methods for making compounds of Formula I or a compound of any one of the embodiments described herein comprising ( 1 ) reacting R'NC with R2CHO,
Figure imgf000045_0003
wherein R4 is optionally substituted aryl or optionally substituted heteroaryl; and R1 , R2, R3, R4 and ring A as defined in Formula I or in any of the embodiments described herein. In one aspect of the preceding methods, R4 is aryl or heteroaryl, each independently substituted with one to three R7 groups. In another aspect of the preceding method, R1 , R2, R3, R4, R5, R6, R7 and ring A are as defined in any of the embodiments herein. The com ounds ofthis invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, scalemic mixtures, and diastereomeric mixtures, as well as single enantiomers or individual stereoisomers that are substantially free from another possible enantiomer or stereoisomer. The term "substantially free of other stereoisomers" as used herein means a preparation enriched in acompound having a selected stereochemistry at one or more selected stereocenters by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. The term "enriched" means that at least the designated percentage of a preparation is the compound having a selected stereochemistry at one or more selected stereocenters. M ethods of obtaining or synthesizing an individual enantiomer or stereoisomer for agiven compound are known in the art and may be applied as practicable to final compounds or to starting material or intermediates.
In one embodiment, the compound is enriched in a specific stereoisomer by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
The compounds of formulal, II, Il-a, II-a-1 , Il-b or II-b-1 may also comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including 'Η, 2H (D or
3 12 13 14 deuterium), and H (T or tritium); C may be in any isotopic form, including C, C, and C; O may be in any isotopic form, including 160 and 180; and the like. For example, the compound is enriched in a specific isotopic form of H, C and/or O by at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.
Unless otherwise indicated when a disclosed compound is named or depicted by a structure wihout specifying the stereochemistry and has one or more chiral centers, it is understood to rep resent allpossible stereoisomers of the compound.
The compounds ofthis invention may also be represented in multiple tautomeric forms, in such instances, the invention expressly includes all tautomeric forms of the compounds described herein, even though only a single tautomeric form may be represented (e.g., alkylation of aring system may result in alkylation at multiple sites, the invention expressly includes all such reaction products). All such isomeric forms of such compounds are expressly included in the present invention.
Compounds described herein may be prepared following procedures detailed in the examples and other analo gous methods known to one skilled in the art. Compounds produced by any of the schemes set forth below may be further modified (e. g, through the addition of substituentsto rings, etc.) to produce additional compounds. The specific approaches and compounds shown herein are not intendedto be limiting. The suitability of a chemical group in a comp ound structure for use in the synthesis of another comp ound is within the knowledge of one of ordinary skill in the art. Synthetic chemistry transformations andprotecting group methodologies (protection and deprotection) useful in synthesizing the applicable compounds are known in the art and include, for example, those described in Larock R, Comprehensive Organic Transformations , VCH Publishers (1989); Greene, TW et al, Protective Groups in Organic
rd
Synthesis, 3 Ed., John Wiley and Sons (1999); Fieser, L et al., Fieser and Fieser 's Reagents for Organic Synthesis, John Wiley and Sons (1994); and Paquette, L, ed., Encyclopedia ofReagentsor Organic Synthesis , John Wiley and Sons (1995) and subsequent editions thereof.
Combinations of substituerts and variables envisioned by this invention are only those that result inthe formation of stable compounds.
It may be convenient or desirable to prepare, purify, and/or handle a corresponding salt of the active compound, for example, a pharmaceutically acceptable salt. Examples of
pharmaceutically acceptable salts are discussed in Berge et ah, 1977, "Pharmaceutically
Acceptable Salts." J. Pharm Sci. Vol. 66, pp . 1-19.
For example, if the compound is anionic, or has afunctional group which may be anionic {e.g., -COOH may be -COO"), then a salt may be formed with a suitable cation Examples of suitable inorganic cations include, but are not limited to, alkali metal ions such as Na+ and K+,
2+ 2+ · 3+
alkaline earth cations such as Ca and Mg , and other cations such as Al . Examples of suitable organic cations include, but are not limited to, ammonium ion (i.e., NH4 +) and substituted ammonium ions (e.g., NH3R , NH2R , NHR , NR ). Examples of some suitable substituted ammonium ions are those derived from ethylamine, diethylamine,
dicy clone xylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, andtromethamine, as well as amino acids, such as lysine and argjnine. An example of a common quaternary ammonium ion is N(CH3)4 +.
If the compound is cationic, or has a functional group that may be cationic (e.g., -NH2 may be -NH3 ), then a salt may be formed with a suitable anion. Examples of suitable inorganic anions include, but are not limitedto, those derived from the following inorganic acids:
hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous.
Examples of suitable organic anions include, but are not limitedto, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, asp artic, benzoic,
camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric,
glucohep tonic, gluconic, glutamic, gly colic, hydroxy maleic, hydroxy naphthalene carboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, prop ionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examples of suitable polymeric organic anions include, but are not limitedto, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.
Unless otherwise specified, a reference to a particular compound also includes salt forms thereof.
Compositicns androutes of administration
The compounds utilized in the methods described here in may be formulated together with a pharmaceutically acceptable carrier or adjuvant into pharmaceutically acceptable compositions prior to be administered to a subject. In another embodiment, such pharmaceutically acceptable compositions further comprise additional therapeutic agents in amounts effective for achieving a modulation of disease or disease symptoms, including those described herein.
Theterm "pharmaceutically acceptable carrier or adjuvant" refers to a carrier or adjuvant that may be administered to a subject, together with a compound of this invention, and which does not destroy the pharmacological activity thereof and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the compound.
Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self- emulsifying drug delivery systems (SEDDS) such as d- α -tocopherol polyethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, poly ethylene glycol and wool fat.
Cyclodextrins such as α-, β-, and γ-cyclodextrin, or chemically modified derivatives such as hydroxy alkylcyclodextrins, including 2- and 3-hydroxypropyl- -cyclodextrins, or other solubilized derivatives may also be advantageously usedto enhance delivery of compounds of the formulae described herein.
The pharmaceutical compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally , nasally, buccally, vaginally or via an implanted reservoir, preferably by oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically -accept able carriers, adjuvants or vehicles. In some cases, thepH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents
(such as, for example, Tween 80) and suspending agents. The sterile inject able preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as asolvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens or Spans and/or other similar emulsifyingagents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions and/or emulsions are administered orally, the active ingredient may be suspended or dissolved in an oily phase is combined with emulsifyingand/or suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
The pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.
Topical administration of the pharmaceutical compositions of this invention is useful when the desired treatment involves areas or organs readily accessible by topical application. For application top ically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, poly oxy ethylene
polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier with suitable emulsifying agents. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically -transdermal patches are also included in this invention.
The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation Such compositions are prepared accordingto techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fiuorocarbons, and/or other solubilizing or dispersing agents known in the art.
When the compositions of this invention comprise a combination of a compound of the formulae described herein and one or more additional therapeutic or prophylactic agents, both the compound and the additional agent should be present at dosage levels of between about 1 to 100%, and more preferably between about 5 to 95% of the dosage normally administered in a monotherapy regimen. The additional agents may be administered separately, as part of a multiple dose regimen, from the compounds of this invention. Alternatively, those agents may be part of a single dosage form, mixed together with the compounds of this invention in a single composition
The compounds described herein can, for example, be administered by injection, intravenously, intraarterially, subdermally, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosahy , topically, in an ophthalmic preparation, or by inhalation, with a dosage ranging from about 0.5 to about 100 mg kg of body weight, alternatively dosages between 1 mg and 1000 mgdose, every 4 to 120 hours, or according to the requirements of theparticular drug The methods herein contemplate administration of an effective amount of compound or compound composition to achieve the desired or stated effect. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 6 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form willvary dep ending up on the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w/w). Alternatively, such preparations contain from about 20% to about 80% active compound.
Lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular subject will depend up on avariety of factors, including the activity ofthe specific comp ound emp loyed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptoms, the subject's disposition to the disease, condition or symptoms, and the judgment ofthe treating physician.
Upon improvement of a subject's condition, a maintenance dose of a compound, composition or combination of this invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level. Subjects may, however, require intermittent treatment on a long- term basis upon any recurrence of disease symptoms.
The pharmaceutical compositions described above comprising a comp ound of formulal, II, Il-a, II-a-1 , Il-b, or II-b-1 or a compound described in any one of the embodiments herein, may further comprise another therapeutic agent useful for treating cancer.
Methods of Use
Provided is a method for inhibiting a mutant IDH1 or IDH2 activity comprising contacting a subject in need thereof with acompound (includingits tautomers and/or
isotopologues) of structural formula I, II, Π-a, II-a-1 , Il-b, or II-b-1 or acompound described in any one of the embodiments herein, or a pharmaceutically acceptable salt thereof. In one embodiment, the cancer to be treated is characterized by a mutant allele of IDH1 or IDH2 wherein the lDH l or IDH2 mutation results in a new ability of the enzyme to catalyzethe NAPH-dependent reduction of a-ketoglutarate to ?(-)-2-hydroxyglutarate in a subject. In one aspect of this embodiment, the mutant IDH 1 has an R132X mutation. In one aspect ofthis embodiment, the R132X mutation is selected fromR132H, R132C, R132L, R132V, R132S and R132G. In another aspect, the R132X mutation is R132H or R132C. Inyet another aspect, the R132X mutation is R132H. Also provided are methods of treating a cancer characterized by the presence of a mutant allele of IDH1 comprisingthe step of administering to subject in need thereof (a) a compound of formula I, II, Il-a, Π-a-l , Il-b, or II-b-1, or a compound described in any one of the embodiments herein, or a pharmaceutically acceptable salt thereof, or (b) a pharmaceutical composition comprising(a) and a pharmaceutically acceptable carrier.
In one embodiment, the cancer to be treated is characterized by a mutant allele of IDH 1 wherein the IDH 1 mutation results in a new ability of the enzyme to catalyze the NAPH- dependent reduction of α-ketoglutarate to i?(-)-2-hydroxyglutarate in a patient. In one aspect of this embodiment, the IDH1 mutation is an R132X mutation. In another asp ect ofthis
embodiment, the R132X mutation is selected fromR132H, R132C, R132L, R132V, R132S and R132G. In another asp ect, the R132X mutation is R132 H or R132C. A cancer can be analyzed by sequencing cell samples to determine the presence and specific nature of (e.g., the changed amino acid present at) a mutation at amino acid 132 of IDH 1.
Without being bound by theory, applicants believe that mutant alleles of IDH 1 wherein the IDH1 mutation results in a new ability of the enzyme to catalyze the NAPH-dependent reduction of a-ketoglutarate to i?(-)-2-hydroxyglutarate, and in particular R132H mutations of IDH1 , characterize a subset of all types of cancers, without regard to their cellular nature or location in the body. Thus, the comp ounds and methods of this invention are useful to treat ary type of cancer that is characterized by the presence of a mutant allele of IDH1 imparting such acitivity and in particular an IDH 1 R132H or R132C mutation.
In one aspect of this embodiment, the efficacy of cancer treatment is monitored by measuring the levels of 2HG in the subject. Typically levels of 2HG are measured prior to treatment, wherein an elevated level is indicated for the use of the compound of formula I, II, Π-a, II-a-1 , Il-b, or II-b-1 or a comp ound described in any one of the embodiments described herein to treat the cancer. Once the elevated levels are established, the level of 2HG is determined during the course of and/or following termination of treatment to establish efficacy . In certain embodiments, the level of 2HG is only determined during the course of and/or following termination of treatment. A reduction of 2HG levels during the course of treatment and foUowing treatment is indicative of efficacy. Similarly, a determination that 2HG levels are not elevated during the course of or foUowingtreatment is also indicative of efficacy. Typically, the these 2HG measurements will be utilized together with other well-known determinations of efficacy of cancer treatment, such as reduction in numb er and size of tumors and/or other cancer- associated lesions, improvement in the general health ofthe subject, and alterations in other biomarkers that are associated with cancer treatment efficacy.
2HG can be detected in a sample by LC/MS. The sample is mixed 80:20 with methanol, and centrifuged at 3,000 rpm for 20 minutes at 4 degrees Celsius. The resulting supernatant can be collected and stored at -80 degrees Celsius prior to LC-M S MS to assess 2-bydroxyglutarate levels. A variety of different liquid chromatography (LC) separation methods can be used. Each method can be coupled by negative electrospray ionization (ESI, -3.0 kV) to triple-quadrupole mass spectrometers operating in multiple reaction monitoring (MRM) mode, with M S parameters optimized on infused metabolite standard solutions. M etabolites can be separated by reversed phase chromatography using 10 mM tributyl-amine as an ion pairing agent in the aqueous mobile phase, according to a variant of apreviously reported method (Luo et al. J Chromatogr A 1 147, 153-64, 2007). One method allows resolution of TCA metabolites: t = 0, 50% B; t = 5, 95% B; t= 7, 95% B; t= 8, 0% B, where B refers to an organic mobile phase of 100% methanol. Another method is specific for 2-hydroxyglutarate, running a fast linear gradient from 50% -95% B (buffers as defined above) over 5 minutes. A Synergi Hydro-RP, 100mm x 2 mm, 2.1 μηι particle size (Phenomonex) canbe used as the column, as described above. M etabolites can be quantified by comp arison of peak areas with pure metabolite standards at known concentration. M etabolite flux studies from 13C-glutamine can be performed as described, e.g., in Munger et al. Nat Biotechnol 26, 1 179-86, 2008.
In one embodiment 2HG is directly evaluated.
In another embodiment a derivative of 2HG formed in process of p erforming the analytic method is evaluated. By way of example such a derivative can be a derivative formed in M S analysis. Derivatives can include asalt adduct, e.g., a Na adduct, a hydration variant, or a hydration variant which is also a salt adduct, e.g., aNa adduct, e.g., as formed in M S analysis.
In another embodiment a metabolic derivative of 2HG is evaluated. Examples include species that build up or are elevated, or reduced, as a result of the presence of 2HG, such as glutarate or glutamate that will be correlated to 2HG, e.g., R-2HG. Exemplary 2HG derivatives include dehydrated derivatives such as the compounds rovided below or a salt adduct thereof:
Figure imgf000055_0001
In one embodiment the cancer is atumor wherein at least 30, 40, 50, 60, 70, 80 or 90% of the tumor cells carry an IDHl mutation, and in particular an IDHl R132H or R132C mutation, at the time of diagnosis or treatment.
IDHl R132X mutations are known to occur in certain types of cancers as indicated in Table 2, below.
Table 2. IDH mutations associated with certain cancers
Cancer Tvpe IDH1 R132X Tumor Tvpe
Mutati on
brain tumors R132H primary tumor
R132C primary tumor
R132S primary tumor
R132G primary tumor
R132L primary tumor
R132V primary tumor
fibrosarcoma R132C HT 1080 fibrosarcoma cell
line
Acute Myeloid Leukemia R132H primary tumor
(AML)
R132G primary tumor
R132C primary tumor
Prostate cancer R132H primary tumor
R132C primary tumor
Acute lymphoblastic leukemia R132C primary tumor
(ALL) aragangliomas R132C primary tumor
IDHl R132H mutations have been identified in glioblastoma, acute myelogenous leukemia, sarcoma, melanoma, non-smallcell lung cancer, cho lan gio carcinomas,
chondrosarcoma, m elodysplastic syndromes (MDS), myeloproliferative neo lasm (MPN), colon cancer, and angio-immunoblastic non-Hodgkin's lymphoma (NHL). Accordingly, in one embodiment, the methods described herein are used to treat glioma (glioblastoma), acute myelogenous leukemia, sarcoma, melanoma, non-small cell lun g cancer (NSC LC) or cho lan gio carcinomas, chondrosarcoma, my elodys lastic syndromes (MDS), myeloproliferative neoplasm (MPN), colon cancer, or angio-immunoblastic non-Hodgkin's lymphoma (NHL) in a patient.
Accordingly in one embodiment, the cancer is a cancer selected from any one of the cancer types listed in Table 2, and the IDH R132X mutation is one or more of the IDHl R132X mutations listed in Table 2 for that particular cancer type.
Treatment methods described herein can additionally comprise various evaluation steps prior to and/or following treatment with acompound of formulal, II, Il-a, II-a-1, Il-b, or II-b-1 or acompound described in any one of the embodiments described herein.
In one embodiment, prior to and/or after treatment with acompound of Structural formula I, II, Il-a, Π- a- 1 , Il-b, or Il-b- 1 or a co mpound described in any one of the embodiments described herein, the method further comprises the step of evaluating the growth, size, weight, invasiveness, stage and/or other phenotype of the cancer.
In one embodiment, prior to and/or after treatment with acompound of formulal, II, Il-a,
II-a-1 , Il-b, or II-b-1 or a comp ound described in any one of the embodiments described herein, the method further comprises the step of evaluating the IDHl genotype ofthe cancer. This may be achieved by ordinary methods in the art, such as DNA sequencing, immuno analysis, and/or evaluation of the presence, distribution or level of 2HG.
In one embodiment, prior to and/or after treatment with acompound of formulal, II, Il-a, II-a-1 , Il-b, or II-b-1 or a comp ound described in any one of the embodiments described herein, the method further comprises the step of determining the 2HG level in the subject. This may be achieved by spectroscopic analysis, e.g., magnetic resonance-based analysis, e.g., MRI and/or M RS measurement, sample analysis of bodily fluid, such as serum or spinal cord fluid analysis, or by analysis of surgical material, e.g., by mass -sp ectroscopy .
Combination therapies
In some embodiments, the methods described herein comprise the additional step of coadministering to a subject in need thereof a second therapy e.g., an additional cancer therapeutic agent or an additional cancer treatment. Exemplary additional cancer therapeutic agents include for example, chemotherapy, targeted therapy, antibody therapies, immunotherapy, and hormonal therapy . Additional cancer treatments include, for example: surgery, and radiation therapy . Examples of each of these treatments are provided below.
The term "co-administering" as used herein with respectto an additional cancer therapeutic agents means that the additional cancer therapeutic agent may be administered together with a compound of this invention as p art of a single dosage form (such as a composition of this invention comprising a compound of the invention and an second therapeutic agent as described above) or as separate, multiple dosage forms. Alternatively, the additional cancer therapeutic agent may be administered prior to, consecutively with, or following the administration of a compound of this invention. In such combination therapy treatment, both the compounds of this invention and the second thera eutic agent(s) are administered by
conventional methods. The administration of a composition of this invention, comprising both a compound of the invention and a second therap eutic agent, to a subject does not preclude the separate administration of that same therapeutic agent, any other second therapeutic agent or any compound of this invention to said subject at another time during a course of treatment. The term "co-administering" as used herein with respect to an additional cancer treatment means that the additional cancer treatment may occur prior to, consecutively with, concurrently with or following the administration of a comp ound of this invention.
In some embodiments, the additional cancer therapeutic agent is a chemotherapy agent. Examples of chemotherapeutic agents used in cancer therapy include, for example,
antimetabolites (e.g., folic acid, purine, and pyrimidine derivatives), alky lating agents (e.g., nitrogen mustards, nitrosoureas, platinum, alkyl sulfonates, hydrazines, triazenes, aziridines, sp indle poison, cytotoxic agents, topoisomerase inhibitors and others) and hyp omethy lating agents (e.g., decitabine (5-aza-deoxycytidine), zebularine, isothiocyanates, azacitidine (5- azacytidine, 5-flouro-2'-deoxycytidine, 5,6-dihydro-5-azacytidine and others). Exemplary agents include Aclarubicin, Actinomycin, Alitretinoin, Altretamine, Aminopterin, Aminolevulinic acid, Amrubicin, Amsacrine, Anagrelide, Arsenic trioxide, Asparaginase, Atrasentan, Belotecan, Bexarotene, bendamustine, Bleomycin, Bortezomib, Busulfan, Camptothecin, Capecitabine, Carbop latin, Carboquone, Carmofur, Carmustine, Celecoxib, Chlorambucil, Chlormethine, Cisplatin, Cladribine, Clofarabine, Crisantaspase, Cyclophosphamide, Cytarabine, Dacarbazine, Dactinomycin, Daunorubicin, Decitabine, Demecolcine, Docetaxel, Doxorubicin, Efaproxiral, Elesclomol, Elsamitrucin, Enocitabine, Epirubicin, Estramustine, Etoglucid, Etoposide,
Floxuridine, Fludarabine, Fluorouracil (5FU), Fotemustine, Gemcitabine, Gliadel implants, Hydroxy carbamide, Hydroxyurea, Idarubicin, Ifosfamide, Irinotecan, Irofulven, Ixabepilone, Larotaxel, Leucovorin, Liposomal doxorubicin, Liposomal daunorubicin, Lonidamine,
Lomustine, Lucanthone, Mannosulfan, Masoprocol, M elphalan, M ercaptop urine, M esna, M ethotrexate, M ethyl aminolevulinate, Mitobronitol, M itoguazone, M itotane, Mitomycin, M itoxantrone, Nedap latin, Nimustine, Oblimersen, Omacetaxine, Ortataxel, O xalip latin, Paclitaxel, Pegaspargase, Pemetrexed, Pentostatin, Pirarubicin, Pixantrone, Plicamycin, Porfimer sodium, Prednimustine, Procarbazine, Raltitrexed, Ranimustine, Rubitecan, Sapacitabine, Semustine, Sitimagene ceradenovec, Stratap latin, Streptozocin, Talaporfin, Tegafur-uracil, Temoporfin, Temozolomide, Teniposide, Tesetaxel, Testolactone, Tetranitrate, Thiotφa, Tiazofurine, Tioguanine, Tipifarnib, Topotecan, Trabectedin, Triaziquone, Triethylenemelamine, Triplatin, Tretinoin, Treosulfan, Trofosfamide, Uramustine, Valrubicin, Verteporfin, Vinblastine, Vincristine, Vindesine, Vinflunine, Vinorelbine, Vorinostat, Zorubicin, and other cytostatic or cytotoxic agents described herein.
Because some drugs work better togetherthan alone, two or more drugs are often given at the same time. Often, two or more chemotherapy agents are used as combination chemotherapy.
In some embodiments, the additional cancer therapeutic agent is a differentiation agent. Such differentiation agent includes retinoids (such as all-trans-retinoic acid (ATRA), 9-cis retinoic acid, 13-cis-retinoic acid (13-cRA) and 4-hydroxy-phenretinamide (4-HPR)); arsenic trioxide; histone deacetylase inhibitors HDACs (such as azacytidine (Vidaza) and butyrates (e.g., sodium pheny lbutyr ate)); hybrid polar compounds (such as hexamethylene bisacetamide ((HMB A)); vitamin D; and cytokines (such as colony -stimulating factors including G-CSF and GM-CSF, and interferons).
In some embodiments the additional cancer therapeutic agent is a targeted therapy agent. Targeted therapy constitutes the use of agents specific for the deregulated proteins of cancer cells. Small molecule targeted therapy drugs are generally inhibitors of enzymatic domains on mutated, overexpressed, or otherwise critical proteins within the cancer cell. Prominent examp les are the tyrosine kinase inhibitors such as Axitinib, Bosutinib, Cediranib, dasatinib, erlotinib, imatinib, gefitinib, lapatinib, Lestaurtinib, ilotinib, Semaxanib, Sorafenib, Sunitinib, and Vandetanib, and also cyclin-dependent kinase inhibitors such as Alvocidib and Seliciclib. Monoclonal antibody therapy is another strategy in which the therapeutic agent is an antibody which sp ecifically binds to a protein on the surface of the cancer cells. Examples include the anti- HER2/neu antibody trastuzumab (HERCEPTIN®) typically used in breast cancer, and the anti- CD20 antibody rituximab and Tositumomab typically used in a variety of B-cell malignancies. Other exemplary antibodies include Cetuximab, Panitumumab, Trastuzumab, Alemtuzumab, Bevacizumab, Edreco lomab, and Gemtuzumab. Exemplary fusion proteins include Aflib ercept and Denileukin diftitox. In some embodiments, the targeted therapy can be used in combination with a compound described herein, e.g., abiguanide such as metformin or phenformin, preferably phenformin.
Targeted therapy can also involve small peptides as "homing devices" which can bind to cell surface receptors or affected extracellular matrix surrounding the tumor. Radionuclides which are attached to these peptides (e.g., RGDs) eventually kill the cancer cell if the nuclide decays in the vicinity of the cell. An example of such therapy includes BEXXAR®.
In some embodiments, the additional cancer therapeutic agent is an immunotherapy agent. Cancer immunotherapy refers to a diverse set of therap eutic strategies designed to induce the subject's own immune system to fight the tumor. Contemporary methods for generating an immune resp onse against tumors include intravesicular BCG immunotherapy for superficial bladder cancer, and use of interferons and other cytokines to induce an immune response in renal cell carcinoma and melanoma subjects.
Allogeneic hematopoietic stem cell transplantation can be considered a form of immunotherapy, since the donor's immune cells will often attack the tumor in a graft -versus- tumor effect. In some embodiments, the immunotherapy agents can be used in combination with a compound or composition described herein.
In some embodiments, the additional cancer therapeutic agent is a hormonal therapy agent. The growth of some cancers can be inhibited by providing or blocking certain hormones. Common examples of hormone-sensitive tumors include certain types of breast andprostate cancers. Removingor blo cking estrogen or testosterone is often an important additional treatment. In certain cancers, administration of hormone agonists, such as progestogens may be therapeutically beneficial. In some embodiments, the hormonal therapy agents can be used in combination with a comp ound or a composition described herein.
Other possible additional therap eutic modalities include imatinib, gene therapy, peptide and dendritic cell vaccines, synthetic chlorotoxins, and radiolabeled drugs and antibodies.
EXAMPLES
The chemical name of each compound described below is generated by ChemBioOffice software.
DCM = dichloromethane TEA = triethylamine
DPPA = dipheny Iphosphoryl azide TFA = trifiuoroacetic acid
DIPEA = AyV- Dii sop ropy let hy lamine TFAA = trifiuoroacetic anhy dride
General procedures for the preparation of l ,l-difluoro-3-isocyanocyclobutane
Method A :
Figure imgf000060_0001
HCJ e O H F 1 I
Step A : Tert-butyl 3-oxocyclobutylcarbamate. To a solution of 3-oxocyclobutanecarboxylic acid (10 g 88 mmol) in dry DCM (60 mL) at 0 °C, SOCl2 (20 mL) was added dropwise. The mixture was heatedto reflux for 1.5 h and then evaporated in vacuo. The resulting mixture was co-evaporated twice with toluene (2 x 8 mL) and the residue was dissolved in acetone (30 mL), followed by adding dropwise to a solution of NaN3 (12 g 185.0 mmol) in H20 (35 mL) at 0 °C. After addition, the mixture was stirred for another 1 h and then quenched with ice (1 10 g). The resulting mixture was extracted with Et20 (2 xlOO mL). Combined organic layers were washed with brine, dried over anhydrous Mg2S04 and concentrated to about 15 mL solution. Toluene (2 x 30 mL) was added into the residue and the mixture was co-evaporated twice to remove Et20 (about 30 mL solution left each time to avoid explosion). The resulting toluene solution was heated to 90°C until the evolution of N2 ceased. Next, 40 mL of t-BuOH was added into the reaction mixture and the resultingmixture was stirred overnight at 90 °C. The mixture was cooled and concentrated. The residue was purified by column chromatography using petroleum ether / EtOAc (V:V, 7 :1 to 5:1) as eluent to afford the desired product as a white solid.M S: 186.1 (M+l)+.
StepB: Tert-butyl 3,3-difluorocyclobutylcarbamate. To a solution of ieri-butyl-3-oxocyclo - butylcarbamate (2.56 g, 1 11.07 mmol) in dry DCM (190 mL), DAST (diethylaminosulfur trifluoride) (41.0 mL, 222.14 mmol) was added dropwise at 0 °C underthe atmosphere of N2. The mixture was then allowed to warm up to r.t and stirred overnight. The resulting mixture was slowly added into apre-cooled saturated aq. NaHC03 solution and extracted with DCM (3 x 200 mL). Combined organic layers were washed with brine, dried over anhydrous MgS04, and concentrated in vacuo. The residuewas purified by column chromato graphy using petroleum ether / EtOAc (V:V, 15 :1) as eluent to afford the desired product. !H NMR (400 MHz, DM SO- d6): δ 4.79 (s, 1H), 4.07 (s, 1H), 2.98 (s, 2H), 2.58 - 2.29 (m, 2H), 1.46 (s, 9H).M S: 208.1 (M+l)+.
Step C: N-(3,3-difhorocyclobutyl)formamide. To a solution of M eOH (170 mL) and CH3COCl (65 mL), tert-butyl 3,3-difluoro -cyclobutylcarbamate (12.1 g, 58.42 mmol) was added in one portion dropwise at 0 °C. The reaction mixture was stirred at 0 °C for 20 min, and then allowed to warm up to r.t and stirred for another 1.5 h. The reaction mixture was concentrated and dissolved in H20 (200 mL). The resulting mixture was extracted by Et20 (150 mL) and the aqueous layer was adjustedto pH=l 1 with solid Na2C03 and extracted by DCM (2 x 150 mL). The combined organic layers were dried over anhydrous M gS04, filtered and concentrated in vacuo usinga co ld-water bath (<20 °C). The residue was dissolved in HCOOEt (90 mL), and transferred into a sealed pressure tube. This reaction mixture was heatedto 80 °C and stirred overnight. The solvent was removed, and the residue was purified by column chromatography using petroleum ether / EtOAc (V:V, 1 : 1 to 1 :3) as eluent to afford the desired product. MS: 136. 1 (M+ l )+.
Step D: l,l-Difluoro-3-isocyanocyclobutane. To a solution of N-(3,3-difluorocyclobutyl) - formamide (2.0 g, 14.81 mmol) and PPh3 (4.27 g, 16.29 mmol) in DCM (35 mL) were added CCU ( 1 .43 mL, 14.81 mmol) and TEA (2.06 mL, 14.81 mmol). The reaction mixture was stirred at 45 °C overnight under a N2 atmosphere. The resulting mixture was evaporated in vacuo at 0 °C. The residue was suspended in Et20 (25 mL) at 0 °C for 30 min and then filtered. The filtrate was evaporated to about 5 mL at 0 °C under reduced pressure. The residue was purified by column chromatography using Et20 as eluent to afford the desired product which was used directly in the next step.
Method B:
ι DPPA, Et3N, Toluene p
COOH NHBoc
Figure imgf000062_0001
Step A: Benzyl 3-oxocyclobutanecarboxylate. A mixture of 3-oxocycIobutanecarboxylic acid (5 g, 44 mmol), potassium carbonate ( 12 g, 88 mmol) and benzyl bromide (1 1.2 g, 66 mmol) in acetone (50 mL) was refluxed for 1 h. The solvent was then removed under reduced pressure and the residue was partitioned between ethyl acetate and water. Combined organic layers were dried over anhydrous MgSC , filtered and concentrated. The residue was purified with silica gel chromatography eluting with a gradient of 100% hexane to 96% hexane / EtOAc to give the desired compound. Ή NMR (400 MHz, CDC13): δ 7.45 - 7.27 (m, 5H), 5.19 (s, 2H), 3.55 - 3.36 (m, 2H), 3.33 - 3.1 1 (m, 3H).
Step B: Benzyl 3, 3-difluorocyclobutanecarboxylate. To a solution of benzyl 3- oxocyclobutanecarboxylate (1 .23g, 6.03 mmol) in DCM (35 mL) was added DAST (0.8 mL, 6.03 mmol) dropwise under nitrogen. The mixture was stirred at room temperature for 16 h and then diluted with DCM . After successive washes with saturated sodium bicarbonate, IN aq.
hydrochloride acid, and brine, the organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product waspurifiedby silica gel chromatography with 93% hexane / EtOAc as eluent to give the desired compound as an oiL H NMR (400MHz, CDC13): δ 7.47 - 7.27 (m, 5H), 5.16 (s, 2H), 3.09 - 2.95 (m, 1H), 2.90 - 2.60 (m, 4H).
Step C: 3,3-Difluorocyclobutanecarboxylic acid Benzyl 3,3-difiuorocyclobutanecarboxylate (0.84 g, 3.72 mo 1) was dissolved m ethanol (40 mL), and approximately 0.02 gpalladium on activated carbon was added. The mixture was stirred at room temperature for 12 h under the atmosphere of H2 and then filtered through a pad of Celite. The filtrates were concentrated and dried in vacuo to give the desired compound. 1H NMR (400 MHz, CDC13): δ 3.16— 2.55 (m, 5H). StepD: Tert-butyl 3,3-difluorocyclobutylcarbamate. B enzyl 3,3-difiuorocyclobutanecarboxy lie acid (3.7 g, 27.3 mmol), DPPA (7.87 g, 27 mmol) and TEA (2.87 g, 28.4 mmol) were dissolved in i-BuOH (25 mL). The mixture was refluxed for 5 h and then diluted with ethyl acetate (about 200 mL). The organic phase was washed twice with 5% citric acid and saturated sodium hydrogen carbonate respectively, dried over anhydrous Mg2S04 and evaporated under reduced pressure. The residue was purified by silica gel chromatography with 50% hexane / EtOAc to give the desired product. MS: 208.1 (M + l)+.
Step E: 3,3-Difluorocyclobutanamine hydrochloride. To a cold solution of M eOH (170 mL) and CH3COCl (65 mL) was added iert-butyl 3,3-difiuorocyclobutylcarbamate (12.1 g, 58.4 mmol) dropwise at 0 °C. After completion of the addition, the mixture was stirred at 0 °C for 20 min and then allowed to warm up to roomtemperature. The reaction mixture was stirred for another 1.5 h and then concentrated to give the crude product which was precipitated in ether to givethe desired product as a white solid. MS: 108.1 (M+l)+.
Step F: N-(3,3-difhorocyclobutyl)formamide. The mixture of 3,3-difluorocyclobutanamine hydrochloride (6.5 g, 60.7 mmol) and TEA (3 eq) in HCOOEt (90 mL) was stirred at 80 °C overnight in a sealed pressure tube. The solvent was removed in vacuo and the residue was purified by column chromatography with 50% petroleum ether / EtOAc to 25%petroleum ether /
EtOAc to give the desired product. 1H NMR (400MHz, DM SO-d6): δ 8.54 (s, 1H), 8.01 - 7.89 (m, 1H), 4.16 - 3.84 (m, 1H), 3.06 - 2.73 (m, 2H), 2.72 -2.33 (m, 2H). M S: 136.1 (M+l)+. Step G: l,l-Difluoro-3-isocyanocyclob tane. The compound was synthesized as outlined in step D of method A set forth above.
General procedures for the preparation of l-fluoro-3-isocyanocyclobutane
°Υ-ι NaBH4 — j PAST F\ MeOH/ HCI F\— , HCOOEt^ *Ν— | PP 3; CCI ; TEA^
^HBoc NHBoc NaC°3 ΝΗ2 ^ ^NHCHO ∞M NC
Step A: Tert-butyl 3-hydroxycyclobutylcarbamate. To a solution of /er/-butyl 3- oxocyclobutylcarbamate (2 g, 10.8 mmol, 2 eq) in EtOH (20 mL) was added NaBH4 (204 mg, 1 eq) at 0 °C. The mixture was then allowed to warm to room temperature and stirred for 30 min. The mixture was concentrated in vacuo and the residue was purified by column chromatography using petroleum ether / EtOAc (V:V, 2: 1 to pure EtOAc) as eluent to afford the desired product as a white solid. MS: 1 88.1 (M+ l )+.
Step B: Tert-butyl 3-fluorocyclobutylcarbamate. To a solution tert-butyl 3-hydroxycyclobutyl - carbamate ( 1 g, 5.35 mmol) in dry DCM (20 mL) at -70 °C was added DAST dropwise (l g, 0.85 mL, 1 .17 eq) under the atmosphere of N2. The mixture was then slowly warmed to room temperature and stirred overnight. The resulting mixture was washed with diluted aq. NaHC03. The organic layer was dried over anhydrous Mg2S04 and concentrated. The residue was purified by flash chromatography using petroleum ether / EtOAc (V:V, 20: 1 to 2: 1 ) as eluent to afford a white solid as the desired product. MS : 190.1 (M+ l )+.
Step C: 3-Fluorocyclobutanamine. The compound was synthesized as outlined in step E of method A set forth above.
Step D: N~(3-fluorocyclobutyl)formamide. The compound was synthesized as outlined in step F of method A set forth above. Ή 1MMR (400 MHz, CDC13): δ 8.10 (s, 1 H), 5.94-5.89 (brs, 1 H), 5.32-5.25 (m, 0.5H), 5.18-5.1 1 (m, 0.5H), 4.63-4.42 (m, 1 H), 2.76-2.62 (m, 2H), 2.44-2.31 (m, 2H).
Step E: l-Fluoro-3-isocyanocyclobutane. The compound was synthesized via the general procedure as the step G in method A set forth above.
General procedures for the preparation of l,l-difluoro-4-isocyanocyclohexane
Figure imgf000065_0001
Step A: Tert-butyl 4-hydroxycyclohexylcarbamate. To a solution of 4-aminocyclohexanol (23 g, 0.2 mol) and Et3N (60 g, 0.6 mol) in THF (230 mL) was added (Boc)20 (87 g, 0.4 mol). The resulting solution was stirred at room temperature overnight. The solvent was removed under reduced pressure and the residue was extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with water (2 x 200 mL) and brine (200 mL), dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography on silica gel using DCM/ MeOH (V:V, 20: 1 ) to afford the desired product as a white solid. MS: 216.2 (M+l )+.
Step B: Tert-butyl 4-oxocyclohexylcarbamate. To a solution of tert-buty\ 4-hydroxycyclohexylcarbamate ( 1 0.0 g, 46.5 mmol) in DCM ( 100 mL) was added Dess-Martin periodinane (39.4 g, 92.9 mmol) portionwise. The resulting solution was stirred at room temperature overnight, quenched with aq. Na2S203 solution and extracted with DCM (3 x 100 mL). The combined organic layers were washed with water (2 x 100 mL) and brine (100 mL), dried over anhydrous aiSC , and concentrated. The residue was purified by column chromatography on silica gel using petroleum ether / EtOAc (V:V, 10: 1 ) to afford desired product as a white solid.
Step C: Tert-butyl 4,4-difluorocyclohexylcarbamate. To a solution of ter/-butyl 4-oxocyclohexylcarbamate (2.13 g, 1 0 mmol) in dry DCM (25 mL) was added DAST (2.58 g, 16 mmol) dropwise at -5 °C under nitrogen. After addition, the reaction mixture was stirred at r.t overnight. The reaction mixture was poured into ice water slowly and extracted with DCM ( 3 x 100 mL). The combined organic layers were washed with 2 N aq. NaHC03 and brine, dried over anhydrous Na2S04, filtered and concentrated in vacuo. The residue was purified by column chromatography using petroleum ether / EtOAc (V:V, 5 : 1 ) as eluent to afford a mixture of the title compound (-70%) and the byproduct tert-butyl 4-fluorocyclohex-' 3-enylcarbamate (-30%) as a light-yellow solid.
To the above mixtures (2.52 g, 10.7 mmol) in DCM (25 mL) was added m-CPBA (2.20 g, 12.9 mmol) portionwise at 0 °C while keeping the internal temperature below 5 °C. After addition, the reaction mixture was stirred at room tem erature overnight. To the reaction mixture was added saturated aq. a2S203 (8.0 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 40 min, and then extracted by DCM (3 x 5.0 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04, and evaporated in vacuo. The residue was used directly in the next step without further purification.
To the above residue in M eOH (15 mL) was added NaBH4 (0.202 g, 5.35 mmol) at 0 °C. The reaction mixture was stirred at room temperature overnight. Water (0.38 g) was added dropwiseto quench the reaction at 0 °C. The resulting mixture was stirred at 0 °C for 30 min, and concentrated in vacuo. The residue was purified by column chromatography using DCM as eluent to afford the pure comp ound as a white solid. 1H MR (400 MHz, CDC13): δ 4.46 (s, lH), 3.59 (s, 1H), 2.25 - 1.69 (m, 6H), 1.61 - 1.20 (m, 1 1H).M S: 236.2 (M+l)+.
Step D: 4,4-Difluorocyclohexanamine hydrochloride. A mixture of iert-butyl4,4-difluorocyclo- hexyl carbamate (6.0 g, 25.5 mmol) and 6 N HCl/MeOH (60 mL) was stirred at room temperature for 2 h. The reaction mixture was concentrated to give the crude product which was directly used in next step without further purification. ¾ NMR (400 MHz, CD3OD): δ 4.89 (s, 2H), 3.32-3.26 (m, 1H), 2.14-2.01 (m, 4H), 2.02-1.85 (m, 2H), 1.74-1.65 (m, 2H). M S: 136.1 (M + l)+.
Step E: N-(4,4-difhorocyclohexyl)formamide. A mixture of 4,4-difluorocyclohexanamine (crude 3.4 g 25.2 mmol), TEA (3 eq) and ethyl formate (35 mL) was stirred at 1 10 °C overnight in a sealed tank. The solvent was removed and the residue was purified by column
chromatography using DCM /M eOH (V:V, 10: l) as eluent to afford the desired pro duct. !H
MR (400 MHz, CDC13): δ 8.14 (s, 1H), 5.98 (s, 1H), 3.93 (m, 1H), 2.54 - 2.19 (m, lH), 2.15 - 1.39 (m, 7H). M S: 164.1 (M+l)+.
Step F: l,l-Difluoro-4-isocyanocyclohexane. A mixture of N-(4,4-difiuorocyclohexyI) - formamide (2.5 g, 15.3 mmol), PPh3 (4.4 g, 16.8 mmol), CC14 (2.3 g, 15.1 mmol), Et3N (1.5 g, 14.9 mmol) and DCM (50 mL) was heatedto 45 °C and stirred overnight. The resulting mixture was evaporated in vacuo and the residue was suspended in Et20 (125 mL) at 0 °C. The filtrate was concentrated and the residue was purified by column chromatography on silica geleluting with Et20 to afford the desired product as a yellow oil which was used directly in the next step.
General procedures for the preparation of 2-(3-aminophenoxy)ethanol
Figure imgf000067_0001
Step A: 2-(3-N rophenoxy)ethanol. A suspension of 3-nitrophenol ( 1 g, 7.2 mmol), 2- bromoethanol ( 1 .2 g, 9.6 mmol) and K2C03 (2 g, 14.4 mmol) in MeCN ( 12 mL) was stirred at 90 °C overnight. The precipitate was collected by filtration to give the first batch of product. The filtrate was concentrated and the residue was purified by column chromatography to afford another batch of the desired product as a yellow solid.
Step B: 2-(3-Aminophenoxy)ethanol. To a solution of 2-(3-nitrophenoxy)ethanol (500 mg, 2.7 mmol) and NH4C1 (720 mg, 1 3.5 mmol) in EtOH (10 mL) was added iron powder (900 mg, 16.2 mmol) at room temperature. The reaction was then stirred at 90 °C for 2 hr and subsequently cooled. The mixture was filtered and the filtrate was concentrated. The resulting residue was purified by column chromatography to afford the desired product as a yellow solid. MS: 154.1 (M+ l )+.
General procedures for the preparation of 3-(lH-pyrazoI-4-yl)aniline
Figure imgf000067_0002
Step A: Tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole-l-carboxylate.
To a solution of 4-(4,4,5,5-tetramethyl- l ,3,2-dioxaborolan-2-yl)- l H-pyrazole (500 mg, 2.57 mmol) and (Boc)20 (672 mg, 3.08 mmol) in DMF (1 .0 mL) was added DMAP (63 mg, 0.52 mmol) in one portion. The mixture was stirred at room temperature overnight, and then partitioned between EtOAc and saturated aq. NH4CI. The organic layer was separated, washed with brine, dried over anhydrous a2S04, and concentrated to afford the crude product.
Step B: 4-(3-Nitrophenyl)-l H-pyrazole. To a solution of tert-butyl 4-(4,4,5,5-tetramethyl-l ,3,2- dioxaborolan-2-yl)- l H-pyrazole- l -carboxylate (300 mg, 0.82 mmol), l -bromo-3-nitrobenzene ( 137 mg, 0.68 mmol) and Na2C03 (216 mg, 2.04 mmol) in DME/H20 (5 mL/ 1 mL) under N2, was added Pd(PPh3)2Cl2 (24 mg, 0.034 mmol). The mixture was stirred at 85 °C overnight, and then quenched with HbO. The resulting mixture was extracted with EtOAc (3 x 25 mL). The organic layer was separated, washed with brine, dried over anhydrous Na2S04, and concentrated. The resulting residue was purified by column chromatography to afford the desired product. MS: 190.2 (M+ l )+.
Step C: 3-(lH~pyrazol-4-yl)aniline. Iron powder (296 mg, 5.30 mmol) was added to a solution of 4-(3-nitrophenyl)- l H-pyrazole (200 mg, 1 .06 mmol) in AcOH/EtOH (2 mL/3 mL). The reaction mixture was stirred at 90 °C for 2 hr and then cooled to room temperature. The reaction mixture was filtered through Celite. The filter cake was washed with H20. The filtrate was neutralized with 1 N NaOH to pH = 8 and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04, and concentrated. The resulting residue was purified by column chromatography to afford the desired product. MS: 160.2 (M+ l )+.
General procedures for the preparation of 2-(3-aminophenyl)propan-2-ol
Figure imgf000068_0001
Step A: Ethyl 3-(dibenzylamino)benzoate 2. To a solution of ethyl 3-aminobenzoate (2 g, 0.012 mmol) and Et3N (5.26 mL, 0.036 mmol) in CH3CN (30 mL), was added BnBr (4.32 mL, 0.036 mmol) in one portion. The reaction mixture was heated to reflux for 18 hr and then cooled to room temperature. The mixture was concentrated to dryness in vacuo and the residue was purified by column chromatography (PE : EtOAc = 10 : 1 as eluent) to afford the desired product as a white solid. MS: 346.1 (M+l )+.
Step B: 2-(3-(dibenzylamino)phenyl)propan-2-ol. To a solution of ethyl 3- (dibenzylamino)benzoate ( 1 .85 g, 5.58 mmol) in anhydrous THF (1 5 mL) at 0 °C under nitrogen atmosphere was added MeMgBr (3 M sol. in THF, 5.58 mL, 16.7 mmol) dropwise over 30 min. The reaction was stirred at room temperature overnight and quenched by addition of saturated NH4C1. The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with NaHC03, water and brine, dried over anhydrous Na2S04, filtered and then concentrated to dryness. The residue was purified by column chromatography (PE : EtOAc = 2 : 1 as eluent) to afford the desired product as a colorless oil. MS: 332.1 (M+l )+. Step C: 2-(3-aminophenyl)propan-2-ol. To a solution of 2-(3-(dibenzylamino)phenyl)propan-2- ol (268 mg. 0.81 mmol) in MeOH (5 mL) was added 10% Pd/C (27 mg) in one portion. The reaction mixture was hydrogenated at room temperature overnight under hydrogen atmosphere. The catalyst was filtered off through Celite and the filtrate was concentrated to dryness. The residue was purified by column chromatography (PE : EtOAc = 1 : 2 as eluent) to afford the desired product as a yellow solid. MS: 1 52.1 (M+ l )+.
General procedures for the preparation of 2-(3-amino-5-fluorophenyl)propan-2-ol
Figure imgf000069_0001
Step A. Methyl 3-fluoro-5-nltrobenzoate. Thionyl chloride (488 mg, 4.1 mmol) was added dropwise to a solution of 3-fluoro-5-nitrobenzoic acid (500 mg, 2.7 mmol) in dry methanol (10 mL) at 0 °C under nitrogen atmosphere. The reaction was warmed to room temperature and stirred for 6 hr. The reaction mixture was concentrated under reduced pressure to obtain the corresponding methyl ester hydrochloride as a waxy solid which was used directly in the next step. MS: 200 (M+ l )+.
Step B. Methyl 3-atnino-5-fluorobenzoate. To a solution of methyl 3-fluoro-5-nitrobenzoate (400 mg, 2 mmol) in ethanol ( 10 mL) was added iron powder (560 mg, 10 mmol) and ammonium chloride (540 mg, 10 mmol) in one portion. The reaction mixture was stirred at 80 °C for 1 hr. After cooling the reaction, the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure to give the desired product. MS: 170 (M+ l ) +.
Step C. Methyl 3-(dibenzylamino)-5-fluorobenzoate. To a solution of methyl 3-amino-5- fluorobenzoate (440 mg, 2.6 mmol) in dry DMF ( 10 mL) was added NaH ( 1 87 mg, 7.8 mmol) portionwise, followed by addition of benzyl bromide (1 . 1 g, 6.5 mmol). The reaction mixture was stirred at 40 °C for 16 hr and concentrated. The resulting residue was purified by column chromatography to give the desired product. MS: 350 (M+ l )+.
Step D. 2-(3-(Dibenzylamino)-5-fluorophenyl)propan-2-ol. Methylmagnesium bromide (1 M in THF, 2.4 mL, 2.4 mmol) was dissolved in THF (5 mL) and placed in an ice-water bath. Methyl 3-(dibenzylamino)-5-fluorobenzoate (280 mg, 0.8 mmol) in THF (5 mL) was then slowly added to the reaction mixture. This mixture was stirred for 3 hr while maintaining an internal temperature range between 1 5 to 25 °C. Then the mixture was cooled to 0 °C and treated with ammonium chloride solution, then extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous a^SC , and concentrated in vacuo. The residue was purified by column chromatography on silica gel to give the desired product. MS: 350 (M+l )+. Step E. 2-(3-Amino-5-fluorophenyl)propan-2-ol. To a solution of 2-(3-(dibenzylamino)-5- fluorophenyl)propan-2-ol ( 150 mg, 0.43 mmol) in ethanol (5 mL) was added 10 % Pd/C (15 mg) under a hydrogen atmosphere. The reaction mixture was stirred at room temperature for 16 hr. The suspension was then filtered through Celite and the filtrate was concentrated in vacuo. The residue was purified by column chromatography to give the desired product. MS : 170 (M+l )+.
General procedures for the preparation of ethyl l-(3-aminophenyl)cyclopropanol
Figure imgf000070_0001
Step A. Ethyl 3-(dlbenzylamino)benzoate. To a solution of ethyl 3-aminobenzoate (2 g, 0.012 mmol) and Et3N (5.26 mL, 0.036 mmol) in CH3CN (30 mL) was added BnBr (4.32 mL, 0.036 mmol) in one portion. The reaction mixture was heated to reflux for 18 h and cooled down to room temperature. The mixture was concentrated in vacuo and the resulting residue was purified by column chromatography to afford the desired product as a white solid. MS: 346.1 (M+l )+. Step B. l-(3-(Dibenzylamino)phenyl)cyclopropanol. To a solution of ethyl 3- (dibenzylamino)benzoate ( 1 .85 g, 5.58 mmol) in anhydrous THF (20 mL) at room temperature under N2 was added titanium tetraisopropoxide (0.25 mL, 0.84 mmol) dropwise over 10 min. After one hour of stirring, EtM gBr (THF solution, 4.1 mL, 12.3 mmol) was added dropwise over 30 min. The reaction mixture was stirred at roomtemperature for 3 h. The resulting mixture was quenched by addition of saturated aq. NH4CI, and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with NaHC03, water and brine, dried over anhydrous Na2S04, and concentrated in vacuo. The residue was purified by column chromatography (PE :
EtOAc = 5 : 1 as eluent) to afford the desired product as a colorless oil. !H NMR (400MHz, CDC13) : δ 7.33 - 7.28 (m, 5H), 7.25 - 7.18 (m, 5H), 7.1 1 (t, J= 8.0 Hz, 1H), 6.80 - 6.75 (m, lH), 6.61 - 6.56 (m, 2H), 4.65 (s, 4H), 1.17 - 1.13 (m, 2H), 0.93 - 0.90 (m, 2H). M S: 330.1 (M+ l)+. Step C. Ethyl l-(3-aminophenyl)cyclopropanol. To a solution of l-(3- (dibenzylamino)phenyl)cyclopropanol (1.8 g, 5.45 mmol) in M eOH (lO mL) at room
temperature was added 10% Pd/C (200 mg) in one portion. The reaction mixture was stirred at roomtemperature under ahydrogen atmosphere overnight. The suspensino was filtered through Celite, and the filtrate was concentrated in vacuo. The residue was purified by column
chromatography (PE : EtOAc = 2 : 1 as eluent) to afford the desired product as a yellow solid. 1H NMR (400 MHz, CDC13) : δ 7.10 (t, J= 7.8 Hz, 1H), 6.69 (t, J= 2.0 Hz, 1H), 6.63 - 6.60 (m, 1H), 6.56 - 6.53 (m, 1H), 1.22 - 1.19 (m, 2H), 1.01 - 0.98 (m, 2H). M S: 150.1 (M + l)+.
General procedures for the preparation of 3-fluoro-5-(meth)lthio)aniline
Figure imgf000071_0001
Step A. (3-Fluoro-5-nitrophenyl)(methyl)sulfane A solution of 3-fiuoro-5-nitroaniline (200 mg, 1.28 mmol), l ,2-dimethyldisulfane (121 mg, 1.29 mmol) and CH3CN (3 mL) was stirred at 30 °C. Neat isoamyl nitrite (150 mg, 1.28 mmol) was slowly added via syringe over 5 min. The reaction mixture was slowly heatedto reflux over 10 min and maintained at a gentle refluxuntilN2 evolution ceased (30-60 min). The reaction mixture was cooled and the solvent was removed in vacuo to afford a dark oil. The resulting oil was purified by column chromatography to give the desired product as apale yellow solid.
StepB: 3-Fluoro-5-(methylthio)aniline. To a solution of (3-fiuoro-5- nitrophenyl)(methyl)sulfane (90 mg, 0.48 mmol) in M eOH (10 mL) was added 10% Pd/C (9 mg) in one ortion. The resulting mixture was purged with H2 three times and stirred at room temperature for 1 h. The suspension was filtered through Celite, and the filter cake was washed withM eOH (5 mL). The filtrate was concentrated in vacuo to afford the desired pro duct which was used directly in next step . M S: 158.0(M + 1)+.
General procedure for the preparation of (S )-2-oxo-l,3-oxazinane- -carboxylic acid
Figure imgf000072_0001
To amixture of (S)-2-amino-4-hydroxybutanoic acid (10 g, 84.0 mmol) and 250 mL of aq. NaOH (2 mol/L, 20.4 g, 510 mmol) at 0 °C was added a solution of triphosgene in dioxane (25.3 g in 125 mL dioxane) dropwise over 1 h.The internal temperature was kept below 5 °C during the addition. The mixture was then stirred at room temperature for 2 days. The reaction mixture was then concentrated in vacuo, followed by addition of 200 mL of CH3CN. The resulting mixture was then heated to 60 °C and stirred vigorously for 0.5 h. The hot mixture was filtered immediately. The filtrate was then concentrated to 100 mL and the desired product was precipitated out. The crudeproduct was collected by filtration and used directly in the next step without further purification. M S: 146.0(M+ 1)+.
General procedure for the preparation of (S )-4-(tert-butoxycarbonyl)-6-oxopiperazine-2- carboxylic acid
Figure imgf000073_0001
Figure imgf000073_0002
B oc Boc
I I
DCC , iHLMiOtL
U0H-¾0 HOOCAAO
Step A : (S )-3-Amino-2-(((benzyloxy)carbonyl)amino)propanoic acid. To a mixture of(S)-4- amino-2-(((benzyloxy)carbonyl)amino)-4-o xobutanoic acid (3 g, 1 1.3 mmol) in M eCN (20 mL), EtOAc (20 mL ) and H20 (10 mL), was added PIAD (4.38 g, 13.5 mmol) in one portion. The reaction mixture was stirred at room temperature overnight. The resulting mixture was filtered, and the filtrate was concentrated in vacuo to afford the desired product. M S: 239.1 (M +l)+. Step B:(S)-M ethyl 3-amino-2-(((benzyloxy)carbonyl)amino)propanoate hydrochloride. To a stirred solution ofMeOH (50 mL) was added SOCl2 (5 mL) dropwise at 0 °C. The resulting mixture was stirred at 0 °C for 0.5 h before (S)-3-amino-2-(((benzyloxy)carbonyl)amino) propanoic acid (2.6 g, 10 mmol) was added. Then the reaction mixture was stirred at room temperature overnight and concentrated in vacuo to afford the desired product. M S: 253.1 (M+l)+.
Step C: (S)-M ethyl 3-((2-(benzyloxy)-2-oxoethyl)amino)-2-(((benzyloxy)carbonyl)amino)pro- paneate. To a solution of (S)-methyl 3-amino-2-(((benzyloxy)carbonyl)amino)propanoate hydrochloride (2.6 g, 0.01 mol) in THF (40 mL) was added DIPEA (4.0 g, 0.03 mol) at 0 °C. The mixture was stirred at 0 °C for 5 min, followed by addition of benzyl 2-bromoacetate (4.7 g, 0.02 mol). Then the mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was quenched by addition of H2Oand then extracted with EtOAc (3 x40 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04, and concentrated The resulting residue was purified by column chromato graphy to afford the desired product. M S: 401.2 (M + l)+.
Step D: ( S )-M ethyl 3-((2-(benzyloxy)-2-oxoethyl)( tert-butoxycarbonyl)amino)-2-( ((benzyloxy) carbonyl)amino)pmpanoate. To a solution of (S)-methyl 3-((2-(benzyloxy)-2-oxoethyl)amino)- 2-(((benzyloxy)carbonyl)amino)propanoate (3.0 g, 7.5 mmol) in THF (40 mL) was added DIPEA (2.9 g, 22.5 mmol) at 0 C. The mixture was stirred at 0 C for 5 min followed by addition of di-tert-butyl dicarbonate (3.27 g, 15 mmol). Then the mixture was allowed to warm to room temperature and stirred overnight. After quenching with a saturated. NaHC03 solution, the resulting mixture was extracted with EtOAc (3 x 60 mL) and concentrated. The resulting residue was purified by column chromato graphy to afford the desired product. MS: 501.2 (M+l)+.
Step E: (S)-2-((2-Amino-3-methoxy-3-oxopropyl)(tert-butoxycarbonyl)amino)acetic acid. To a solution of (S)-methyl 3-((2-(benzyloxy)-2-oxoethyl)(tert-butoxycarbonyl)amino)-2-(((benzyl- oxy)carbonyl)amino)propanoate (2.5 g, 5 mmol) in M eOH (30 mL) was added 10% PdC (250 mg). The mixture was stirred under hydrogen atmosphere at room temperature overnight. The resulting suspension was filtered through Celite, and the filtrate was concentrated in vacuo to afford the desired product. M S: 277.1 (M+ l)+.
Step E :(S )-l-tert-Butyl 3-methyl 5-oxopiperazine-l,3-dicarboxylate. To a solution of(S)-2-((2- amino-3-methoxy-3-oxopropyl)(tert-butoxycarbonyl)amino)acetic acid (1.2 g, 4 mmo l) in DCM (100 mL) was added DCC (1.34 g, 6 mmol) at 5 °C. The mixture was stirred at 10 °C for 4 h followed by addition of Et3N (0.88 g, 8 mmol). The resulting mixture was stirred at room temperature for 18 h and then concentrated. The residue was added to EtOAc (20 mL) and the precipitate was filtered. The filtrate was concentrated and the residue was purified by column chromatography to afford the desired product. MS: 259.1 (M+ l)+.
Step F :(S)-4-(tert-Butoxycarbonyl)-6-oxopiperazine-2-carboxylic acid To a mixture of (S)- l- tert4outyl 3-methyl 5-oxopiperazine- l ,3-dicarboxylate (500 mg, 1.9 mmo l) in MeOH (20 mL) and THF (20 mL) was added a solution ofLiOH*H20 (159 mg, 3.8 mmol) in H20 (10 mL) at 0 °C. The mixture was stirred at room temperature for 2h and then partitioned between EtOAc (25 mL) and H20. The aqueous layer was acidified with 2N HC1 to pH 3-4 and then extracted with EtOAc (3 x20 mL). The comb ined organic layers were washed with brine, dried over anhydrous a2SC>4 and concentrated to afford the desired product which was used directly in the next reaction. MS: 245.1 (M+l )+.
General procedure for the preparation of 2-bromopyrimidine-4-carbonitrile
OH OH Br
Ν Ν HCI NaNQ2 TBAB,P2Q5 ,
AcOH L> . , N toluene
OH ^
Step A: 2-Hydroxy-4-carboxyaIdehyde oxime. 2-Hydroxy-4-methyl pyrimidine hydrochloride
(25.0 g 171 mmol) and sodium nitrate ( 17.7 mg, 260 mmol) were slowly added to 200 mL of
50% acetic acid at 0 °C. The reaction mixture was stirred at room temperature for 3 h. The resulting suspension and the solids were filtered, washed with water and dried to afford the desired product. Ή NMR (400 MHz, DMSO-d6): δ 12.42 (s, 1 H), 1 1.89 (s, 1 H), 7.92 (d, J= 6.4
Hz, 1 H), 7.75 (s, 1 H), 6.43 (d, J = 6.4 Hz, 1 H). MS: 140.0 (M+ l )+.
Step B: 2-Bromopyrimidine-4-carbonitrile. A mixture of 2-hydroxy-4-carboxyaldehyde oxime (9 g, 28.8 mmol), tetrabutyl ammonium bromide ( 10 g, 71 .9 mmol) and phosphorus pentoxide (2 g, 14.4 mmol) in toluene (300 mL) was stirred at 120 °C for 2 h. The resulting mixture was filtered and the filtrate was concentrated. The resulting residue was purified by column
chromatography to give the desired compound as a yellow solid. Ή NMR (400 MHz, CDC13): δ 8.82 (d, J = 4.8 Hz, 1 H), 7.66 (d, J = 4.8 Hz, 1 H). MS: 185.0 (M+l )+.
General synthetic procedures for making compounds of formula I:
Figure imgf000075_0001
n=1 ,2
X=CH2,0,NH,CH(OH)(S and ),F
General procedures for the UGI reaction:
A mixture of aldehyde (3.5 mmol) and aniline (3.5 mmol) in MeOH (8 mL) was stirred at room temperature for 30 min. Then the acid (3.5 mmol) was added and the reaction mixture was stirred for another 30 min, followed by addition of the isocyanide (3.5 mmol). The resulting mixture was then stirred at room temperature overnight and quenched with H20. The resulting mixture was partitioned between EtOAc and H20. The organic layer was washed with brine, dried over anhydrous Na2S04, and then concentrated. The resulting residue was purified by a standard method to afford the desired product.
General procedures for the Buchwald reaction:
A mixture of amine (0.30 mmol), aryl bromide (0.30 mmol), CS2CO3 (129 mg, 0.39 mmol), Pd2(dba)3 ( 1 8 mg, 0.02 mmol) and Xant-Phos (9.4 mg, 0.02 mmol) in 1 ,4-dioxane (10 mL) was stirred under N2 at 80 °C overnight. After filtration, the filtrate was concentrated in vacuo and the residue was purified by a standard method to give the desired products.
Example 1. Preparation of (S)-methyl l-methyl-5-oxopyrrolidine-2-carboxylate.
Compound 2 was prepared according to the following scheme, using the following protocol.
Figure imgf000076_0001
Step A: (S)-Methyl l-methyl-5-oxopyrrolidine-2-carboxylate. To a mixture of (5)-5-oxopyro- lidine-2-carboxylic acid (5.0 g, 38.8 mmol) in DMF (50 mL) were added anhydrous K2C03 (16 g, 1 16 mmol) and iodomethane ( 16.4 g, 1 16 mmol) at room temperature The resulting mixture was warmed to 40°C,stirred for 24 h and concentrated in vacuo. The residue was precipitated with EtOAc (80 mL) and filtered. The filter cake was washed with EtOAc (2 10 mL). The combined filtrates were concentrated and the residue was purified by column chromatography on silica gel to give the desired product. Ή-NMR (400 MHz, CDC13): δ 4. 1 8 - 4.1 1 (m, 1 H), 3.70 (s, 3H), 2.87 (s, 3H), 2.56 - 2.29 (m, 3H), 2.16 - 2.04 (m, 1 H). MS: 158.1 (M+ l )+.
Step B: (S)-l-Methyl-5-oxopyrrolidine-2-carboxylic acid. To a solution of (5)-methyl 1 -methyl- 5-oxopyrrolidine-2-carboxyIate (0.6 g, 3.8 mmol) in MeOH (6 mL) were added THF (2 mL), H20 (2 mL) and NaOH (0.45 g, 1 1 .4 mmol) at room temperature The resulting mixture was stirred at room temperature for 18 h and then acidified with 2 "N HC1 to pH=3-4 at 0°C. The mixture was extracted with EtOAc (3 x 30 mL),the combined organic layers were dried over anhydrous Na2S04and concentrated to give the crude product as a yellow solid (0.8 g) which was used directly in the next step . M S: 142.1 (M -l)~.
Step C: Compound2. 2-Chlorobenzaldehyde (1 17 mg, 0.83 mmol), 3-fluoroaniline (92.5 mg, 0.83 mmol), crude (S)-l-methyl-5-oxopyrrolidine-2-carboxylic acid (200 mg, -60% purity, 0.83 mmol) and l,l-difluoro-3-isocyanocyclobutane (119 mg, 90% purity, 1.0 mmol) were used in the UGI reaction to give the desired product (diastereomeric mixture). !H MR (400 MHz, CDCb): δ 8.52 (d, J= 4.9 Hz, 0.2H), 8.16 (m, 0.3H), 7.87 - 7.47 (m, 2H), 7.42 - 7.31 (m, 1H), 7.25 - 7.1 1 (m, 2H), 7.08 - 6.89 (m, 3.3H), 6.74 (d, J= 6.0 Hz, 0.7H), 6.57 (m, 2H), 4.42 - 4.26 (m, 1.3H), 4.20 - 4.08 (m, 0.5H), 4.00 (m, lH), 3.00 (m, 2H), 2.74 (m, 3H), 2.63 - 1.82 (m, 6H). MS: 494.1 (M+l)+.
Example 2. Preparation of (S )-N-(l-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2 - oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide.
Compounds 3 and 4 were prepared according to the following scheme, using the following protocol.
Figure imgf000077_0001
Comp ound 4
Step A. (S )-N-( l-( 2-Ch loroph enyl)-2-( ( 4,4-difluorocycloh exyl )amino)-2-oxoethyl )-N-( 3-fluoro -phenyl)-5-oxopyrmlidine-2-carboxamide. 3-Fluoroaniline (86 mg, 0.78 mmol), 2- chlorobenz aldehyde (109 mg, 0.78 mmol), (5 -5-oxopyrrolidine-2-carboxylic acid (100 mg, 0.78 mmol) and l,l-difluoro-4-isocyanocyclohexane (135 mg 0.91 mmol) were used in theUGI reaction to give the desiredproduct. M S: 508.1 (M+l)+. Step B. (S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohex l)amino^ - fluorophenyl)-5-oxo-l-(pyrimidin-2-yl)pyrrolidine-2-carboxamide and (S)-N-((R)-l-(2 - chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-l- (pyrimidin-2-yl)pyrrolidine-2-carboxamide. A mixture of (S)-N-(l-(2-chlorophenyl) -2-((4,4- difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (100 mg, 0.20 mmol), 2-bromopyrimidine (47 mg, 0.30 mmol), CS2CO3 (129 mg, 0.39 mmol), Pd2(dba)3 (18 mg, 0.02 mmol) and Xant-Phos (9.4 mg, 0.02 mmol) in 1,4-dioxane (10 mL) was stirred under N2 at 80 °C overnight. After filtration, the filtrate was concentrated in vacuo and the residue was purified by a standard method to give the desired products.
(S)-N-((S)-l-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(^ - phenyl)-5-oxo-l-(pyrimidin-2-yl)pyrrolidine-2-carboxamide. Compound 4 Ή NMR (400 MHz, CDCI3): δ 8.71 (d, J= 4.8 Hz, 2H), 7.75 (m, 1 H), 7.33 (m, 2H), 7.18 (m, 1 H), 7.09 - 6.87 (m, 511), 6.47 (s, 1H), 5.61 (d,J=7.6 Hz, 1H), 4.86 (d,J=6.6 Hz, 1H), 3.98 (m, 1H), 3.01 -2.84 (m, 2H),2.58(m, 1H), 2.30 - 2.20 (m, 1H), 1.93 (m, 7H), 1.47 (m, 2H); MS: 586.2 (M+l)+.
(S)-N-((R)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethy^
fluorophenyl)-5-oxo-l-(pyrimidin-2-yl)pyrrolidine-2-carboxamide. Compound 3.Ή NMR (400 MHz, CDCI3): δ 8.75 (dd, J= 4.8, 2.0 Hz, 2H), 7.40 (d, J= 7.8 Hz, 1H), 7.23 (s, 3H), 7.08 (dt,J= 11.3,6.3 Hz, 3H), 6.99 (d,J=3.7 Hz, 1H), 6.27 (s, 1H), 6.13-5.92 (m, 1H), 5.02 (m, 1 H), 4.76 (m, 1 H), 3.92 (m, 1 H), 2.88 (m, 1 H), 2.67 - 2.46 (m, 1 H), 2.44 -2.19 (m, 2H), 2.00 (m, 8H). MS: 586.1 (M+l)+.
The following analogs were synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted-aromatic ring or heteroaromatic ring using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
Compound 6
Figure imgf000078_0001
Ή NMR (400 MHz, CDC13): δ 8.75 (d, J= 4.8 Hz, 2H), 7.35 (m, 3H), 7.25 - 6.81 (m, 5H), 6.28 (s, 1H),5.84 (d,J=7.5 Hz, IH), 4.76 (m, IH), 3.98 - 3.59 (m, IH), 2.92 (m, lH),2.58(m, IH), 2.35 -2.20 (m, IH), 2.07 (m, IH), 1.83 (m, 2H), 1.57 (m,4H), 1.46- 1.17 (m, 4H). MS: 550.2
(M+])+.
Compound 7
Figure imgf000079_0001
Ή NMR (400 MHz, CDC13): δ 8.73 (m, 2H), 7.80 (s, IH), 7.35 (s, IH), 7.23 - 6.72 (m, 6H), 6.47 (s, IH), 5.49 (d,J=7.7 Hz, IH), 4.87 (d,J=6.6 Hz, IH), 4.74 -4.42 (m, IH), 3.86 (d,J = 8.0 Hz, 1 H), 3.19 - 2.77 (m, 1 H), 2.56 (m, 1 H), 2.44 - 2.21 (m, 1 H), 2.13 - 1.73 (m, 4H), 1.60 (s, 2H), 1.26 (m, 4H). MS: 550.2 (M+l)+.
Compound 49
Figure imgf000079_0002
Ή NMR (400 MHz, CDC13): δ 8.69 (s, 2H), 7.76 (s, IH), 7.49 - 6.68 (m, 7H), 6.44 (s, IH), 6.19 (s, IH), 4.93 (m, 3H), 2.23 (m, 8H). MS: 540.1 (M+])+.
Compound 51
Figure imgf000079_0003
Ή NMR (400 MHz, CDC13): δ 8.81 (d, J= 4.9 Hz, IH), 8.66 (d, J= 2.7 Hz, IH), 8.04-7.79 (m, IH), 7.49-7.31 (m, IH), 7.13-6.92 (m, 6H), 6.60 (m, IH), 6.25 - 5.95 (m, IH), 5.68 (m, IH), 4.73 (dd,J= 16.0,6.9 Hz, lH),4.39(m, IH), 2.98 (m, 3H), 2.53 (m, 4 ), 2.14 - 1.93 (m, IH). MS: 592.1 (M+l)+.
Compound 5
Figure imgf000080_0001
Ή NMR (400 MHz, CDCI3): 58.46 -8.32 (m, 1.7H), 7.78 - 7.61 (m, 1.5H), 7.39 (m, 1.5H), 7.23 (m, 1.6H), 7.13 - 6.88 (m, 4H), 6.40 (m, IH), 6.11 (m, IH), 5.01 -4.77 (m, 1H),4.26 (m, IH), 3.51 (d,J = 5.5 Hz, 0.3H), 3.13-2.75 (m, 3H), 2.61 -2.22 (m, 3H), 2.17-1.90 (m, IH). MS: 557.1 (M+l)+.
Compound 10
Figure imgf000080_0002
Ή NMR (400 MHz, CDCI3): δ 8.56 (m, 2H), 8.16 (s, 1.3H), 7.74 (s, IH), 7.36 (s, 2.6H), 7.19 (s, IH), 7.12-6.82 (m, 3H), 6.52 (m, 2H), 6.19 (m, IH), 4.65 -4.48 (m, IH), 4.26 (m, 1.3H), 3.90 -3.82 (m, 0.3H), 2.87 (m, 3H), 2.64-1.98 (m, 6H). MS: 557.1 (M+l)+.
Compound 41
Figure imgf000081_0001
Ή NMR (400 MHz, CDC13): δ 7.98 (m, IH), 7.65 (m, 2Η), 7.44 - 7.30 (m, 2H), 7.03 (m, ( 6.51 (m, IH), 6.36 (s, IH), 5.12 (d, J= 6.3 Hz, IH), 4.33 (s, IH), 3.97 (s, 3H), 3.10-2.63 3H), 2.60 -2.00 (m, 5H). MS: 587.1 (M+l)+.
Compound 26
Figure imgf000081_0002
Ή NMR (400 MHz, CDC13): δ 8.32 (m, IH), 8.05 (t,J= 8.6 Hz, IH), 7.69 (s, IH), 7.45 - 7.30 (m, IH), 7.25 - 6.78 (m, 6H), 6.38 (m, 2H), 4.88 (m, IH), 4.33 (s, IH), 3.89 (s, 3H), 3.11 -2.72 (m, 3H), 2.66-2.29 (m, 3H), 2.23 - 1.86 (m, 2H). MS: 587.1 (M+l)+.
Compound 17
Figure imgf000081_0003
Ή NMR (400 MHz, CDC13): δ 7.93 (m, IH), 7.56 (m, 2H), 7.21 (m, 3H), 7.10 - 6.87 (m, 3H), 6.42 (m, 3H), 5.04 (m, IH), 4.25 (m, IH), 3.97 (d, J= 6.1 Hz, 3H), 3.10 - 2.69 (m, 3H), 2.60 - 2.15 (m, 4H), 2.12 - 1.87 (m, IH). MS: 587.2 (M+l)+.
Compound 28
Figure imgf000082_0001
Ή NMR (400 MHz, CDC13): δ 8.19 (m, IH), 7.79 - 7.33 (m, 3H), 7.28 - 7.06 (m, 4H), 7.06 - 6.83 (m, 4H), 6.47 - 6.32 (m, 2H), 5.09 - 4.91 (m, 1 H), 4.25 (m, 1 H), 3.09 - 2.60 (m, 4H), 2.57 (s, 3H), 2.53 - 1.99 (m, 5H). MS: 571.0 (M+l)+.
Compound 21
Figure imgf000082_0002
Ή NMR (400 MHz, CDC13): δ 8.26 (d,J=8.5 Hz, IH), 8.15 (s, IH), 7.64 (s, IH), 7.48 (m, IH), 7.32 (d, J= 7.5 Hz, IH), 7.14 (m, 2H), 7.04-6.83 (m, 3H), 6.40 (s, IH), 6.04 (s, IH), 4.89 (m, 1 H), 4.31 (s, 1 H), 2.89 (m, 3H), 2.48 (m, 2H), 2.40 - 2.27 (m, 3H), 2.26 - 1.84 (m, 3H). MS: 571.2 (M+l)+.
Compound 27
Figure imgf000082_0003
Ή NMR (400 MHz, CDCI3): δ 8.30 - 8.15 (m, 2H), 7.68 (s, IH), 7.38 (m, IH), 7.24 - 6.85 (m, 6H), 6.46 -6.16 (m, 2H), 4.94 (d, J= 6.0 Hz, 1 H), 4.32 (s, 1 H), 3.10 - 2.74 (m, 3H), 2.60 - 2.43 (m, 2H), 2.36 (m, 4H), 2.23 - 1.91 (m, 2H). MS: 571.2 (M+l)+.
Compound 15
Figure imgf000083_0001
Ή NMR (400 MHz, CDC13): δ 8.17 (d, J= 8.3 Hz, IH), 7.56 (m, 2H), 7.25 -6.96 (m, 5H), 6.89 (m, 2H), 6.42 (s, IH), 6.21 (s, IH), 5.12 -4.96 (m, IH), 4.31 (m, IH), 3.14 - 2.74 (m, 3H), 2.55 (s, 3H), 2.51 -2.28 (m, 3H), 2.20 (m, IH), 2.05 - 1.87 (m, IH). MS: 571.2 (M+l)+.
Figure imgf000083_0002
Ή NMR (400 MHz, CDC13): δ 8.72 (m, IH), 7.88 (m, IH), 7.65 (s, IH), 7.57 - 7.30 (m, 2H), 7.23 - 7.09 (m, 2H), 7.02 (s, 2H), 6.96 - 6.83 (m, 1 H), 6.44 (s, 1 H), 6.05 (d, J= 6.5 Hz, 1 H), 5.31 -4.93 (m, IH), 4.33 (s, IH), 3.02 (m, 2H), 2.86 (m, IH), 2.63 - 2.45 (m, 2H), 2.44-2.23 (m, 2H), 2.01 (m, IH). MS: 625.1(M+1)+.
Compound 31
Figure imgf000083_0003
Ή NMR (400 MHz, CDC13): δ 8.91 - 8.34 (m, 2H), 8.03 (s, IH), 7.79 - 7.34 (m, 3H), 7.22 6.75 (m, 5H), 6.46 (s, IH), 6.02 (d,J=6.5 Hz, IH), 4.95 (dd, J= 9.4, 3.1 Hz, 1H),4.35 (s, 3.13-2.76 (m, 3H), 2.68 - 1.83 (m, 5H). MS: 625.1(M+1)+.
Compound 39
Figure imgf000084_0001
Ή NMR (400 MHz, CDC13): δ 8.65 (d, J= 23.6 Hz, 2H), 7.87 (s, 1 H), 7.59 - 7.29 (m, 3H), 7.26 -6.71 (m, 5H), 6.59 (s, IH), 6.28 (s, 1H),4.83 (d,J=8.2 Hz, IH), 4.12 (s, IH), 3.10 - 2.62 (m, 3H), 2.56 (m, IH), 2.36 - 1.84 (m, 4H). MS : 625.1(M+1)+.
Figure imgf000084_0002
Ή NMR (400 MHz, CDC13): δ 8.74 (s, IH), 8.53 (s, IH), 7.71 (s, IH), 7.31 (d,J=8.3 Hz, IH), 7.25 -6.80 (m, 6H), 6.44 (s, IH), 6.08 (s, IH), 4.95 (m, IH), 4.35 (s, 1 H), 3.15 - 2.76 (m, 3H), 2.66-2.17 (m, 4H), 2.03 (s, IH). MS : 625.1(M+1)+.
Compound 11
Figure imgf000084_0003
Ή NMR (400 MHz, CDC13): δ 8.29 (dd, J= 8.1, 2.0 Hz, IH), 7.74 (m, 2H), 7.31 (m, 2H), 7.22 - 7.12 (m, 2H), 7.00 (s, 2H), 6.93 (m, IH), 6.67 (dd,J= 7.9, 2.4 Hz, IH), 6.46 (m, IH), 6.06 (m, lH),4.86(m, 1 H), 4.35 (m, 1H),2.93 (m, 3H), 2.59 - 2.39 (m, 2H), 2.23 (m, lH),2.02(m, IH). MS: 575.1 (M+l)+.
Compound 29
Figure imgf000085_0001
Ή NMR (400 MHz, CDC13): δ 8.40 (m, IH), 8.24 (m, IH), 7.71 (d,J=7.7Hz, IH), 7.49 - 7.30 (m, 2H), 7.28 - 7.21 (m, 1 H), 7.12 (m, 2H), 7.04 - 6.88 (m, 3H), 6.67 (m, IH), 6.42 (s, 2H), 4.90 (m, 1 H), 4.27 (m, 1 H), 3.07 - 2.76 (m, 3H), 2.58 - 2.29 (m, 3H). MS: 575.0 (M+l )+.
Compound 12
Figure imgf000085_0002
Ή NMR (400 MHz, CDC13): δ 8.27 (m, IH), 7.64 - 7.30 (m, 3H), 7.27 - 6.62 (m, 7H), 6.47 - 6.30 (m, IH), 6.28-6.07 (m, IH), 5.00 -4.55 (m, IH), 4.26 (m, IH), 3.12 - 2.67 (m, 3H), 2.65 -2.36 (m, 3H), 2.22 (m, 2H). MS: 575.1(M+1)+.
Compound 34
Figure imgf000085_0003
Ή NMR (400 MHz, CDC13): δ 8.37 (t, J= 8.9 Hz, IH), 7.63 (m, 2H), 7.49 - 6.84 (m, 8H), 6.44 (s, 1H),5.94 (m, 1 H), 5.07 - 4.74 (m, 1H),4.25 (d,J=51.6 Hz, IH), 3.10 - 2.67 (m, 3H), 2.63 - 1.85 (m, 5H), 1.25 (s, IH). MS: 591.1(M+1)+.
Compound 35
Figure imgf000086_0001
Ή NMR (400 MHz, DMSO-d6) : δ 8.38 (d, J= 8.2 Hz, 1 H), 7.86 - 7.34 (m, 4H), 7.25 - 6.79 (m, 6H), 6.46 (s, IH), 5.99 (s, IH), 4.95 (d,J=9.2 Hz, IH), 4.34 (s, IH), 3.12 - 2.70 (m, 3H), 2.63 - 1.87 (m, 6H). MS: 591.1(M+1)+.
Compound 48
Figure imgf000086_0002
Ή NMR (400 MHz, CDC13): δ 8.59 - 8.19 (m, 2H), 7.82 - 7.57 (m, 2H), 7.45 - 7.34 (m, 2H), 7.01 (m, 4H), 6.45 (s, IH), 5.94 (s, IH), 4.89 (dd, J= 9.3, 3.1 Hz, lH),4.30(m, IH), 3.21 -2.69 (m, 3H), 2.61 - 1.88 (m, 5H). MS: 591.1(M+1)+.
Compound 33
Figure imgf000086_0003
Ή NMR (400 MHz, CDC13): δ 8.63 - 8.03 (m, 2H), 7.67 (s, IH), 7.23 - 6.65 (m, 8H), 6.45 - 5.93 (m, 2H), 4.84 (m, 1 H), 4.23 (m, 1 H), 3.04 - 2.65 (m, 4H), 2.65 - 1.83 (m, 5H). MS:
591.1(M+1)+.
Compound 36
Figure imgf000087_0001
Ή NM (400 MHz, CDC13): δ 8.79 - 8.51 (m, 2H), 7.88 (s, IH), 7.51 - 7.29 (m, 2H), 7.22 (m, 2H), 7.08 (t,J= 7.3 Hz, IH), 6.99 (t, J= 7.2 Hz, IH), 6.78 (s, IH), 6.51 (d,J=5.8Hz, IH), 6.28 (s, IH), 4.79 (m, IH), 4.14 (s, IH), 3.02 -2.66 (m, 3H), 2.55 (m, IH), 2.33 - 1.99 (m, 4H). MS: 582.1(M+1)+.
Compound 37
Figure imgf000087_0002
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1 H), 8.52 (s, 1 H), 7.85 - 7.30 (m, 3H), 7.24 - 6.79 (m, 5H), 6.43 (s, IH), 6.12 (s, 1H),4.92 (d,J=6.8Hz, lH),4.34(s, 1H),2.90 (m, 3H), 2.64 - 2.46 (m, IH), 2.46-2.11 (m, 3H), 1.97 (m, IH). MS : 582.1(M+1)+.
Compound 47
Figure imgf000087_0003
Ή NMR (400 MHz, CDC13): δ 8.66 - 8.38 (m, 2H), 7.90 (d, J= 7.0 Hz, IH), 7.68 (s, IH), 7.37 (m, IH), 7.25 -6.80 (m, 6H), 6.44 (s, 1H),5.97 (d,J=6.6 Hz, IH), 4.91 (d,J=6.7Hz, IH), 4.32 (s, IH), 3.30-2.78 (m, 4H), 2.41 (m, 4H), 2.02 (s, IH). MS : 582.1(M+1)+.
Compound 16
Figure imgf000088_0001
Ή NMR (400 MHz, CDC13): δ 8.58 (d, J= 9.3 Hz, IH), 8.11 (d, J= 8.7 Hz, IH), 7.97 (d, J= 8.5 Hz, 1 H), 7.86 - 7.59 (m, 3H), 7.48 (m, 2H), 7.18 (m, 3H), 6.97 (m, 3H), 6.38 (s, IH), 6.11 (s, 1 H), 5.20 (s, 1 H), 4.30 (s, 1 H), 3.09 - 2.77 (m, 3H), 2.67 - 2.44 (m, 2H), 2.36 - 2.21 (m, 2H), 2.10- 1.92 (m, IH). MS: 607.2 (M+l)+.
Compound 1
Figure imgf000088_0002
Ή NMR (400 MHz, CDC13): δ 8.69 (d, J= 4.8 Hz, 2H), 7.71 (s, IH), 7.31 (m, IH), 7.18 (m, IH), 7.13 -6.77 (m, 6H), 6.46 (s, IH), 6.22 (s, 1 H), 5.00 - 4.62 (m, IH), 4.35 (s, IH), 3.19 -2.71 (m, 3H), 2.69 - 1.83 (m, 5H). MS: 451.2 (M+l)+.
Compound 22
Figure imgf000088_0003
Ή NMR (400 MHz, DMSO-d6): δ 8.15-8.01 (m, IH), 7.62-7.52 (m, IH), 7.31-6.69 (m, 9H), 6.24 (s, IH), 5.65-4.66 (m, IH), 2.60 (m, IH), 2.20-2.05 (m, 3H), 1.76-0.83 (m, 4H). MS: 451.2 (M+lf.
Compound 18
Figure imgf000089_0001
1 H NMR (400 MHz, DMSO-d6): 69.70 (s, 1 H), 8.48 - 8.26 (m, 2H), 7.72 (s, 1 H), 7.46 - 7.31 (m, I H), 7.28 -7.15 (m, 2H), 7.13- 6.89 (m, 3H), 6.55 - 6.14 (m, 2H), 4.82 (m, 1 H), 4.26 (m, 1 H), 2.90 (m, 3H), 2.64-2.40 (m, 2H), 2.34 - 1.99 (m, 3H). MS: 558.1 (M+l)+.
Figure imgf000089_0002
Ή NMR (400 MHz, CDC13): δ 7.54 (d, J= 3.5 Hz, 1 H), 7.45 - 7.29 (m, 3H), 7.28 - 6.95 (m, 6H),6.44 (d,J=6.0 Hz, lH),6.24(s, 1H),4.92 (m, 1H),4.25 (s, 1 H), 3.11 -2.79 (m, 3H), 2.61 (m, 1H),2.43 (m, IH), 2.39-2.27 (m, 2H), 2.27-2.11 (m, IH). MS: 563.1 (M+l)+.
Compound 14
Figure imgf000089_0003
Ή NMR (400 MHz, CDCI3): δ 7.66 (s, IH), 7.48 (s, IH), 7.35 (s, IH), 7.26-6.82 (m, 8H), 6.43 (s, IH), 6.09 (d,J= 6.3 Hz, IH), 4.98 (d,J=8.7Hz, IH), 4.34 (s, IH), 3.08 - 2.84 (m, 2H), 2.63 - 2.36 (m,4H), 2.32 (m, IH), 2.15 (m, IH). MS: 563.1 (M+l)+.
Compound 23
Figure imgf000090_0001
H NMR (400 MHz, CDCI3): δ 7.78 - 7.49 (m, 2H), 7.39 (m, 4H), 7.24 - 6.82 (m, 4H), 6.38 (m, 3H), 5.94 (m, l H), 4.50 (m, l H), 4.22 (m, 1 H), 3. 10 - 2.59 (m, 3H), 2.59 - 1 .99 (m, 6H). MS: 556.2 (M+ l )+.
Example 3. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino) - 2-oxoethyl)-N-(3-fluorophenyl)-5-oxo-l-(thiazol-4-yl)pyrrolidine-2-carboxamide
Compounds 42 and 43 were prepared according to the following scheme, using the following rotocol.
Figure imgf000090_0002
Compound 43
A mixture (2S)-N-( \ -(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2 -oxoethyl)-N- (3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (200 mg, 0.417 mmol), 4-bromothiazole (0.045 mL, 0.626 mmol, 1 .5 eq), 3P04 ( 124 mg, 0.585 mmol, 1 .4 eq), Cul (8 mg, 0.1 eq) and trans- 1 ,2-diaminocyclohexane (0.24 eq) in dioxane (2 mL) was stirred at 1 10 °C under microwave for 30 min. The resulting mixture was filtered through a Celite pad. The filtrate was concentrated and the residue was purified by a standard method to give the desired product. (S)-N-((R)-l-(2-Chlorophenyl)-2-(3,3-difluorocycloburylamino)-2-oxoethyl)-N-(3- fluorophenyl)-5-oxo-l-(thiazoI-4-yl)pyrrolidine-2-carboxamide (Compound 42)
Figure imgf000091_0001
Ή NMR (400 MHz, CDC13): δ 8.68 (d, J = 2.1 Hz, 1 H), 7.65 (m, 5H), 7.30 - 6.90 (m, 4H), 6.47 (s, 1 H), 6.23 (s, 1 H), 4.88 (dd, J = 9.3, 3.0 Hz, 1 H), 4.20 (s, 1 H), 3.17 - 2.63 (m, 3H), 2.58 - 1.99 (m, 5H). MS: 563. 1 (M+ l )+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoe
fluorophenyl)-5-oxo-l-(thiazol-4-yl)pyrrolidine-2-carboxamide ( Compound 43)
Figure imgf000091_0002
1 H NMR (400 MHz, CDCI3): δ 8.60 (s, 1 H), 8.06 - 7.56 (m, 2H), 7.35 (s, 1 H), 7.22 - 6.79 (m, 5H), 6.42 (s, 1 H), 6.13 (s, 1 H), 4.96 (d, J= 7.8 Hz, 1 H), 4.25 (m, 1 H), 3.14 - 2.70 (m, 4H), 2.63 - 2.21 (m, 4H). MS: 563. 1 (M+ l )+.
Example 4. Preparation of (S)-N-((S)-l-(2-chlorophenyI)-2-(3,3-difluorocyclobutylamino)-2 -oxoethyI)-N-(3-fluorophenyl)-5-oxo-l-(pyridin-2-yImethyl)pyrroIidine-2-carboxamide
Compound 44 was prepared according to the following scheme, using the following protocol.
Figure imgf000091_0003
Compound 44. To a solution of (2S)-N-( l -(2-chlorophenyl)-2-(3,3-difluorocycIobutyl -amino)- 2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (200 mg, 0.42 mmol) in dry DMF (20 mL) was added NaH (20 mg, 0.84 mmol) at 0 °C. The mixture was stirred at this 0 °C for 0.5 h followed by addition of 2-(bromomethyl)pyridine ( 106 mg, 0.42 mmol). The mixture was then allowed warm to room temperature and stirred overnight. The resulting mixture was slowly added dropwise to 100 mL of water, and then extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with saturated aq. LiCl, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): 6 8.5 1 (s, 1 H), 7.88 - 7.37 (m, 3H), 7. 19 - 5.95 (m, 10H), 5.14 (m, 1 H), 4.34 (m, 1 H), 4. 10 (m, 2H), 3.00 (m, 2H), 2.81 - 1 .57 (m, 6H). MS: 571 .2 (M+l)+.
Example 5. Preparation of (S)-N-((S)-l-(2-chlorophenyI)-2-(3,3-difluorocyclobutylamino)- 2- oxoethyI)-N-(3-fluorophenyl)-3-hydroxy-2-(pyrimidin-2-ylamino)propanamide.
Compound 9 was prepared according to the following scheme, using the following protocol.
Figure imgf000092_0001
Step A: (S)-2-Oxooxazolidine-4-carboxylic acid. To a solution of NaOH (0.8 g, 20 mmol) in water (4 mL) was added (S)-2-(benzyloxycarbonylamino)-3-hydroxypropanoic acid ( 1 g, 4.2 mmol) portionwise at 0 °C over 3 min. The resulting solution was warmed to r.t and stirred for 2 h. After cooling to 0 °C, the solution was adjusted to pH= l -2 with 2 N HC1. The mixture was extracted with EtOAc (4 x 10 mL). The combined organic layers were dried over anhydrous Na2S04 and concentrated in vacuo to give the desired product as a white solid. Ή NMR (400 MHz, DMSO-d6): 6 13.93 - 12.30 (m, 1 H), 8. 15 (s, 1 H), 4.49 (t, J = 8.6 Hz, 1 H), 4.32 (m, 2H); MS: 130.0 (M- I V.
Step B: (4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluoro - phenyl)-2~oxooxazolidine-4-carboxamide. 2-Chlorobenzaldehyde ( 160 mg, 1 .14 mmol), 3- fluoroaniline ( 127 mg, 1 . 14 mmol), (5)-2-oxooxazolidine-4-carboxylic acid ( 150 mg, 1 .14 mmol) and l,l-difluoro-3-isocyanocyclobutane (181 mg, 90% ofpurity, 1.37 mmol) were used in the UGI reaction to give the desired product as a white solid. 1H MR (400MHz, CDC13): δ 8.15- 8.01 (m, 1H), 7.62-7.52 (m, 1H), 7.31-6.69 (m, 9H), 6.24 (s, 1H), 5.65-4.66 (m,4H), 2.60 (m, lH),2.20-2.05 (m,3H), 1.76-1.51 (m, 5H), 1.29-0.83 (m, 5H); MS: 482.1 (M+l)+.
Step C: (S)-N-((R)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3- - fluorophenyl )-2-oxo-3-(pyrimidin-2-yl )oxazolidine-4-carboxamide and(S )-N-( ( S )-l-(2-chloro ^h yl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fluorophenyl)-2-oxo-3-(pyrimidin- 2-yl)oxazolidine-4-carboxamide. A mixture of (4S -N-(l-(2-chlorophenyl)-2- (3,3- difiuorocyclo-butylamino)-2-oxoethyl)-N-(3-fiuorophenyl)-2-oxooxazolidine-4-carboxamide (350 mg, 0.73 mmol), 2-bromopyrimidine(150 mg, 0.94 mmol), Cs2C03 (500 mg, 1.52 mmol), Pd2(dba)3 (66 mg, 0.07 mmol) andXant-Phos (42 mg, 0.07 mmol) in 1,4-dioxane (15 mL) was stirred under N2 at 80 °C for 18 h and then filtered through a Celite pad. The filtrate was concentrated in vacuo and the residue was purified a standard method to give (5)-N-((i?)-l-(2- chlorophenyl)-2-(33-difluorocyclobutylamino)-2-oxoethyl)-N-(3-fiuorophenyI)-2-oxo-3- (pyrimidin-2-yl)oxazolidine-4-carboxamide (8). ¾ NMR (400 MHz, CDC13): δ 8.73 (d, J= 4.8 Hz, 2H), 7.95 (s, 0.8H), 7.74 (s, 02H), 7.41 (d, J= 7.5 Hz, 1.6H), 7.24 (t, J= 7.2 Hz, 1H), 7.17 - 6.94 (m, 4.3H), 6.73 (d, J= 6.7 Hz, 1H), 6.48 (d, J = 73.8 Hz, 2H), 4.93 (s, lH), 4.41 (dd, J= 8.6, 4.8 Hz, lH),4.29(t,J=8.6Hz, lH),4.14(m, lH),2.80(m, 2H), 2.21 (s, 1H), 2.18 - 2.07 (m, 1H);MS: 560.1 (M+l)+, and (5 -N-((5 -l-(2-chloro-phenyl)-2-(3,3-difiuorocyclo-butylamino)-2- oxoethyl)-N-(3-fluoro -phenyl)-2-oxo-3-(pyrimidin-2-yI)oxazolidine-4-carboxamide (9). !H NMR (400MHz, CDC13): δ 8.68 (d, J= 4.8 Hz, 2H), 7.65 (s, lH), 7.30 (s, 1H), 7.18 (s, 1H), 7.13- 6.86 (m, 5H), 6.50 (s, 1H),6.38 (m, 1H), 5.00 (m, 1H), 4.43 (dd,J= 8.7,4.8 Hz, 1H), 4.32(m, lH),4.20(m, 1H), 2.99 (m, 2H),2.50(m, 2H).MS: 560.1 (M+l)+.
Example 6. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl) - amino)-2-oxoethyl)-N-(3-lluorophenyl)-6-oxo-l-(pyrimidin-2-yl)piperidine-2-carboxamide Compounds 19 and20 were prepared accordingto the following scheme, usingthe following protocol.
Figure imgf000094_0001
Step A. (S)-6-Oxopiperidine-2-carboxylic acid. A solution of (S)-2-aminohexanedioic acid (470 mg, 2.9 mmol) in 20% AcOH (5 mL) was stirred at 1 10 °C overnight. The solvent was removed in vacuo and the residue was dissolved in EtOH ( 10 mL). The unreacted amino acid was precipitated and filtered off. The filtrate was concentrated to give the crude desired product which was used directly in the next step. MS: 142. 1 (M- l )'.
Step B. (S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-o^
-phenyl)-6-oxopiperidine-2-carboxamide. 3-Fluoroaniline (21 7 mg, 1.96 mmol), 2-chloroben- zaldehyde (274 mg, 1 .96 mmol), (5 -6-oxopiperidine-2-carboxylic acid (280 mg, 1.96 mmol) and l , l -difluoro-3-isocyanocyclobutane (280 mg, 1 .96 mmol) were used in the UGI reaction to give the desired product. MS: 494.1 (M+ l )+ .
Step C. (S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxo - fluowphenyl)-6-oxo-l-(pyrimidin-2-yl)piperidine-2-carboxamide and (S)-N-((R)-l-(2-chloro - phenyl)-2-((3,3-difluorocyclobutyl)aniino)-2-oxoethyl)-N-(3-fluorophenyl)-6-oxo-l- (pyrimidin-2-yl)piperidine-2-carboxamide A mixture consisting of ( l /?)-N-( l -(2-chlorophenyl) - 2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluorophenyl)-3-oxo-2-(pyrimidin-2- yl)cyclohexanecarboxamide (250 mg, 0.5 1 mmol), 2-bromopyrimidine (121 mg, 0.76 mmol), Cs2C03 (33 1 mg, 1 .01 mmol), Pd2(dba)3 (46 mg, 0.05 mmol) and Xant-Phos (29 mg, 0.04 mmol) in 1 ,4-dioxane ( 15 mL) was stirred under N2 at 80 °C overnight and then filtered. The filtrate was concentrated in vacuo and the residue was purified by a standard method to give the desired products.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difl orocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoro - phenyl)-6-oxo-l-(pyrimidin-2-yl)piperidine-2-carboxamide (Compound 19). Ή NMR (400 MHz, CDC13): δ 8.73 (m, 2H), 7.70 (s, 1 H), 7.26 - 6.95 (m, 6H), 6.87 (t, J = 7.2 Hz, 1 H), 6.53 (s, 1H), 6.33 (s, 1H), 4.77 (d, J= 5.3 Hz, 1H), 4.33 (s, 1H), 3.01 (d, J= 5.5 Hz, 2H), 2.85 - 2.28 (m, 4H), 2.05 (m, 2H), 1.81 (s, 2H). MS: 571.1 (M + l)+.
(S )-N-((R)-l-(2-Chloro-phenyl )-2-( ( 3,3-difluorocyclobutyl)amino)-2-oxoethyl )-N-( 3-fluoro - phenyl)-6-oxo-l-(pyrimidin-2-yl)piperidine-2-carboxamide (Compound 20). ΊΤ NMR (400 MHz, CDC13): δ 8.74 (d, J= 4.8 Hz, 2H), 7.99 (m, 1H), 7.56 - 7.32 (m, 1H), 727 - 6.85 (m, 6H),
6.72 (s, 1H), 6.51 (m, 1H), 4.67 -4.48 (m, 1H), 4.34 - 4.01 (m, 1H), 2.95 - 2.60 (m, 2H), 2.59 - 2.40 (m, lH), 2.40 - 2.19 (m, 2H), 2.15 -2.00 (m, 2H), 1.97 - 1.59 (m, 4H). MS: 571.1 (M + l)+.
Example 7. Preparation of (S )-N-((S )-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)- 2 -oxoethyl)-N-(3-fluorophenyl)-5-oxo-4-(pyrimidin-2-yl)morpholine-3-carboxamide
Compound 30 wasprφared according to the following scheme, using the following protocol.
Figure imgf000095_0001
Step A : (S)-3-Hydroxy-2-(4-methoxybenzylamino)propanoic acid. (5)-2-amino-3-hydroxy - propanoic acid (8.4 g, 80 mmol) was dissolved in a solution of NaOH (3.2 g, 80 mmol) in H20 (40 mL). After cooling to 10 °C, 4-methoxybenzaldehyde (21.7 g 160 mmol) was added dropwise over 10 min. The mixture was stirred at room temperature for 30 min and then cooled to 0°C. NaBH4 (1.67 g, 44 mmol) was added portionwise and the resulting mixture was warmed slowly to r.t and stirred for 2 h. The mixture was washed with Et20 (2 x50 mL). The aqueous phase was adjustedto pH 4.5 with 2 N HC1 at 0 °C. The precip itate was filtered, washed with petroleum ether (20 mL) and dried in vacuo to give the desired product as a white solid. M S: 226.1 (M+l)+.
StepB: (S)-Benzyl4-(4-methoxybenzyl)-5-oxornorpholine-3-carboxylate. (5 -3-Hydroxy-2- ((4-methoxybenzyl)amino)propanoic acid (5.0 g, 22 mmol) was dissolved in a solution of NaOH (1.15 g, 29 mmol) in H20 (60 mL). After coolingto 0 °C, 2-chloroacetyl chloride (3.6 mL, 44 mmol) was added drop wise followed by aq. NaOH (30% wt) to keep pH=13. After stkring for another 4 h, the reaction was cooled to 0 °C and acidified with 2 HClto adjust pH=2~3. The resulting mixture was extracted with EtO Ac (2 x30 mL). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was dissolved in acetone (150 mL) and then treated with BnBr (9.7 g, 51 mmol) and DIP EA (19 mL, 1 1 1 mmol). The reaction mixture was stirred for 24 h at room temperature and concentrated in vacuo. The residue was purified by column chromatography to afford the desired product as a white solid. M S: 356.1 (M+l)+.
Step C: (S Benzyl 5-oxomorpholine-3-carboxylate. To a solution of (5)-benzyl 4-(4- methoxybenzyl)-5-oxomorpholine-3-carboxy late (200 mg, 0.56 mmol) in CH3CN (5 mL) and H20 (5 mL) was added CAN (eerie ammonium nitrate) (1.5 g, 2.8 mmol) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h. DIP EA was added at 0 °C to adjust thepH to 6~7 and the mixture was concentrated in vacuo. The residue was purified by column chromatography to afford the desired product as a white solid M S: 236.1 (M+ l)+.
StepD: (S )-5-Oxomorpholine-3-carboxylic acid. To amixture of (5 -benzyl 5-oxomorpholine- 3-carboxylate (160 mg, 0.7 mmol) in M eOH (8 mL) was added 10% Pd/C (about 5 mg). The reaction was stirred under an atmosphere of hydrogen for 30 min at room temperature. The reaction mixture was filtered through a Celite pad and concentrated in vacuo to afford the desired product as awhite solid. MS: 146.1 (M + l)+.
Step E: ( S )-N-( ( S )-!-( 2-Chlorophenyl )-2-( ( 3,3-difluorocyclobutyl )amino)-2-oxoethyl )-N-( 3 - fluorophenyl)-5-oxomorpholine-3-carboxamide. 3-Chlorobenzaldehyde (104 mg, 0.74 mmol), 3-fiuoroaniline (83 mg, 0.74 mmol), (5 -5-oxomorpholine-3-carboxylic acid (108 mg, 0.74 mmol) and l,l-difiuoro-3-isocyanocyclobutane (248 mg, 1.48 mmol) were used in theUGI reaction to afford the desired product. M S: 496.1 (M+l)+.
Step F: Compound 30. A mixture of (5 -N-((5 -l-(2-chlorophenyl)-2-((3,3-difluoro - cyclobutyl)amino)-2-oxoethyl)-N-(3-fiuorophenyl)-5-oxomorpholine-3-carboxamide (100 mg, 0.2 mmol), 2-bromopyrimidine (36 mg, 022 mmol), Pd2(dba)3 (28 mg, 0.03 mmol), XantPhos (16 mg, 0.03 mmol) and Cs2C03 (160 mg, 0.5 mmol) in 1,4-dioxane (4 mL) was stirred at 100 °C for 3.5 h under N2. The reaction mixture was then cooled to room temperature and filtered. The solid was washed with DQVI (2 x20 mL). The filtrate was evaporated and the residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): δ 8.77 (m, 2H), 7.85 (m, 1 H), 7.41 (s, 1 H), 7.28 - 7.21 (m, 1 H), 7.21 - 7.10 (m, 2H), 7.09 - 6.90 (m, 3H), 6.87 (m, 1 H), 6.68 - 6.33 (m, 2H), 4.80 (m, 1 H), 4.43 - 4.22 (m, 2H), 4.13 (m, 2H), 3.94 (m, 1H), 2.99 (m, 1H), 2.86 (m, 1H), 2.63-2.26 (m, 2H). MS: 474.1 (M+l)+
Example 8.
The following analogs were synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods.
(2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclopentyl)amino)-2-oxoethyl)-N-(3-fluoro phenyl)-5-oxo-l-(pyrimidin-2-yI)pyrrolidine-2-carboxamide (racemic) - Compound 73
Figure imgf000097_0001
Ή NMR (400 MHz, CDCI3): δ 8.71 (d,J=4.8Hz, 2H), 7.72 (s, 1H), 7.37 (s, 1H), 7.18 (s, 1H), 7.11 -6.85 (m, 5H), 6.47 (s, 1H), 5.70 (d, J= 7.3 Hz, 1H), 4.86 (d, J= 7.0 Hz, 1H),4.53 (d,J = 6.3 Hz, 1 H), 3. 1 (s, 1 H), 2.95 - 2.88 (m, 1 H), 2.64 - 2.47 (m, 2H), 2.40 - 1.65 (m, 8H). MS: 572.2 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-fluorophenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 64
Figure imgf000097_0002
H NMR (400 MHz, CDCb): δ 8.74 (s, 1H), 8.52 (s, 1H), 7.72 (d, j=7.1 Hz, 1H), 7.43 m, 1H), 7.25 -7.17 (m, 1H), 7.13 - 6.81 (m, 4H), 6.43 (s, 1H), 6.12 (s, 1H),4.92 (d,J 1H), 4.37 - 4.28 (m, 1H), 3.10-2.82 (m, 3H), 2.59 - 2.49 (m, 2H), 2.42 - 2.36 (m, 1H), 2.31 - 2.22 (m, 1H), 2.06 - 1.88 (m, 2H). MS: 582.1 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyl)amino)-2-oxo-l-phenylethyl)- -(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 138
Figure imgf000098_0001
Ή NMR (400 MHz, CDC13): δ 8.78 (s, 1H), 8.44 (d, J= 4.9 Hz, 1H), 7.65 (s, 1H), 7.39 - 7.15 (m, 6H), 7.14-6.92 (m, 4H), 6.65 (m, 1H), 6.16 (s, 1H), 5.82 (s, 1H),4.86 (d,J=6.8 Hz, 1H), 4.31 (s, 1H), 3.15 -2.77 (m, 3H), 2.68 - 1.91 (m, 5H). MS: 548.2 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyl)amino)-l-(2-fluorophenyl)-2- oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 149
Figure imgf000098_0002
Ή NMR (400 MHz, CDC13): δ 8.74 (m, 1H), 8.50 (d, J= 4.2 Hz, 1 H), 7.65 (s, 1H), 7.45 - 7.14 (m.4H), 7.13 -6.69 (m, 5H), 6.25 (m, 2H), 4.88 (dd, J= 9.2, 3.1 Hz, 1H),4.33 (s, 1H), 3.21 - 2.72 (m, 3H), 2.65 - 1.88 (m, 5H).MS: 566.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri midin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 68
Figure imgf000098_0003
Ή NMR (400 MHz, CDC13): δ 8.95 (d, J= 4.7 Hz, 1H), 7.68 (s, 1H), 7.34 (d, J= 4.6 Hz, 2H), 7.16 (s, 1H), 7.04 (d, J= 3.6 Hz, 3H), 6.92 (s, 2H), 6.51 (s, 1H), 5.92 (s, 1H), 4.81 (d,J=9.5 Hz, 1H),4.33 (s, 1H), 2.91 (m, 3H), 2.64 - 2.26 (m, 4H), 2.01 (s, 1H). MS: 583.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopy rimidin-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 85
Figure imgf000099_0001
Ή NMR (400 MHz, CDC13): δ 8.98 (d, J= 4.7 Hz, 1H), 7.74 (s, 1H), 7.38 (dd, J= 11.2, 5.7 Hz, 2H), 7.06 (m, 5H), 6.52 (s, 1H), 5.47 (d, J= 7.7 Hz, 1H), 4.85 (d, J= 9.2 Hz, 1H), 3.99 (s, 1H), 2.93 (dd,J= 18.6, 8.9 Hz, 1H), 2.62 (d,J=9.5 Hz, 1H), 2.36 (s, 1H), 1.97 (m, 7H), 1.57-1.38 (m, 2H). MS: 611.2(M+1)+.
(S)-N-((S)-l-(2-ChlorophenyI)-2-((3,3-difluorocycIobutyl)amino)-2-oxoethyl)-N-(3,5- difluoro -phenyl)-5-oxo-l-(pyrimidin-2-yl)pyrrolidine-2-carboxamide (single enantiomer) - Compound 70
Figure imgf000099_0002
Ή NMR (400 MHz, CDC13): δ 8.70 (d, J= 4.8 Hz, 2H), 7.60 (s, 1H), 7.37 (d, J= 8.0 Hz, 1H), 7.26 - 7.19 (m, 1 H), 7.13 - 7.04 (m, 2H), 7.03 - 6.97 (m, 1 H), 6.86 (s, 1 H), 6.69 (dd, J= 9.8, 7.6 Hz, 1H), 6.46 (s, 1H), 6.07 (d,J=6.7Hz, 1H), 4.87 (dd, J= 9.1, 3.1 Hz, 1H), 4.36 (s, 1H), 3.11 -2.83 (m, 3H), 2.64-2.34 (m, 3H), 2.21 (m, 1H), 2.10 - 1.97 (m, 1H). MS: 576.1 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri din-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 71
Figure imgf000100_0001
Ή NMR (400 MHz, CDC13): δ 8.73 (d, J= 7.1 Hz, 1H), 8.60 - 8.46 (m, 1H), 7.56 (d, J= 7.7 Hz, 1H), 7.38 - 7.32 (m, 1H), 7.31 - 7.27 (m, 1H), 7.26 -7.18 (m, 1H), 7.14- 7.00 (m, 1H), 6.96 (m, 1H), 6.85 (s, 1H), 6.69 (m, 1H), 6.40 (s, 1H), 6.02 (d,J=6.6 Hz, 1H), 4.98 - 4.74 (m,lH), 4.39- 4.10 (m, 1H), 3.11 - 2.67 (m, 3H), 2.64 - 1.95 (m, 5H). MS : 600.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopy rimidin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 86
Figure imgf000100_0002
Ή NMR (400 MHz, CDC13): δ 8.98 (d, J= 4.8 Hz, 1H), 7.56 (s, 1H), 7.40 (m, 2H), 7.23 (t, J = 7.0 Hz, 1H), 7.08 (t, J- 7.6 Hz, 1H), 7.01 -6.84 (m, 2H), 6.71 (t,J=8.6 Hz, 1H), 6.51 (s, 1H), 6.00 (d, J= 6.7 Hz, 1 H), 4.85 (dd, J= 9.3, 2.7 Hz, 1 H), 4.36 (s, 1 H), 3.15 - 2.80 (m, 3H), 2.67 - 2.26 (m,4H), 2.08 (dt, J= 9.7, 8.1 Hz, 1H). MS: 601 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri - din-2-yl)-5-oxo-N-(3-sulfamoylphenyl)pyrrolidine-2-carboxamide (single enantiomer) - Compound 53
Figure imgf000100_0003
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1H), 8.50 (s, 1H), 7.73 (d, J= 7.5 Hz, 1H), 7.33 (d, J = 9.3 Hz, 1H), 7.25 -6.80 (m, 6H), 6.40 (s, 1H), 5.61 (d,J=6.9Hz, 1H), 4.91 (d,J=8.0Hz, 1H), 3.97 (s, 1H), 2.99-2.79 (m, 1H), 2.55 (dd, J= 13.7, 9.9 Hz, 1H),2.25 (t,J= 11.3 Hz, 1H),2.03 - 1.74 (m, 5H), 1.56- 1.36 (m, 2H). MS: 610.2 (M+l)+. (2S)-N-(l-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 81
Figure imgf000101_0001
'H NMR (400 MHZ, CDC13): δ 8.75 (s, 1H), 8.51 (d,J= 5.0 Hz, 1H), 7.62 (d,J= 9.0 Hz, 1H), 7.37 (d,J= 7.9 Hz, 1H), 7.27 (d,J= 5.1 Hz, 1H), 7.22 (t,J= 7.7Hz, 1H), 7.06 (t,J=7.5 Hz, 1H), 6.99 (d,J= 6.9 Hz, 1H), 6.88 (d, J= 7.4 Hz, 1H), 6.69 (t,J= 8.6 Hz, 1H), 6.41 (s, 1H), 5.69 (d,J= 7.8 Hz, 1H), 4.95 (dd, J= 9.3, 3.2 Hz, 1H), 3.98 (m, 1H), 2.95 - 2.84 (m, 1H), 2.65 - 2.55 (m, 1H), 2.30-2.20 (m, 1H), 2.05 - 2.12 (m, 1H), 2.03 (s, 2H), 1.94- 1.78 (m, 2H), 1.68 - 1.35 (m, 3H), 0.85 - 0.95 (m, 1H). MS: 628.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopy rimidin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 87
Figure imgf000101_0002
Ή NMR (400 MHz, CDC13): δ 8.97 (d, J= 4.8 Hz, 1 H), 7.60 (d, J= 8.7 Hz, 1 H), 7.46 - 7.34 (m, 2 H), 7.22 (t, J= 7.8 Hz, 1 H), 7.06 (t, J= 7.6 Hz, 1 H), 7.00 - 6.87 (m, 2 H), 6.70 (t, J= 8.6 Hz, 1 H), 6.48 (s, 1 H), 5.64 (d, J= 7.7 Hz, 1 H), 4.86 (dd, J= 9.3, 2.7 Hz, 1 H), 3.98 (d, J= 7.7 Hz, 1 H), 2.96 - 2.86 (m, 1 H), 2.63 - 2.55 (m, 1 H), 2.37 - 2.29 (m, 1 H), 2.15 - 1.99 (m, 5 H), 1.96 - 1.77 (m, 2 H), 1.61 - 1.34 (m, 2 H). MS: 629.2 (M+ 1 )+ .
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-l-(2,4-dichlorophenyl)-2-((3,3-difluorocyclobutyl)amino) -2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 196
Figure imgf000102_0001
Ή NMR (400 MHz, CDCI3): 68.77 (s, 1H), 8.49 (d,J=5.1 Hz, 1H), 7.56 (s, 1H), 7.40 (d,J = 2.1 Hz, 1H), 7.30 (s, 1H), 7.08 (dd,J=8.4, 2.1 Hz, 1H), 6.97 (d,J=8.4 Hz, 1H), 6.90 (s, 1H), 6.79-6.72 (m, 1H), 6.35 (s, 1H), 5.99 (d,J=6.6 Hz, 1H),4.93 (dd,J=9.3, 3.1 Hz, 1H), 4.33 (s, 1H), 3.12-2.95 (m, 2H), 2.95-2.83 (m, 1H), 2.66-2.32 (m, 3H), 2.24 - 2.18 (m, 1H), 2.12 - 1.99 (m, 1H). MS: 634.1 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-l-(2,5-dichlorophenyl)-2-((3,3-difluorocyclobutyl)amino) -2-oxoethyI)-N-(3,5-difluorophenyI)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 201
Figure imgf000102_0002
Ή NMR (400 MHz, CDC13): δ 8.76 (s, 1H), 8.49 (dd,J= 5.0, 0.6 Hz, 1H), 7.58 (s, 1H), 7.30 (t, J= 5.2 Hz, 2H), 7.22 (dd, J= 8.6, 2.5 Hz, 1H), 7.02 (d, J= 2.4 Hz, 1H), 6.88 (s, 1H), 6.76 (tt, J = 8.6,2.3 Hz, 1H), 6.34 (s, 1H), 6.14(d,J=6.8 Hz, 1H),4.94 (dd, J= 9.3, 3.2 Hz, 1H),4.43- 4.28 (m, 1H), 3.09-3.02 (m, 2H), 2.93 - 2.84 (m, 1H), 2.65-2.32 (m, 3H), 2.27-2.16 (m, 1H), 2.14 -2.00 (m, 1H).MS: 634.1 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-l-(2,6-dichlorophenyl)-2-(3,3-difluorocyclobutylamino)- 2-oxoethyl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) -
Compound 63
Figure imgf000102_0003
'HN R(400 MHZ, CDCI3): δ 8.77 (s, 1H), 8.45 (t,J=5.6 Hz, 1H), 7.88 (t,J = 10.0 Hz, 1H), 7.40 - 7.32 (m, 1 H), 7.26 - 7.21 (m, 2H), 7.10- 7.05 (m, 2H), 6.92 (d, J= 2.4 Hz, 1 H), 6.62 (d, J=8.6Hz, 1H), 5.53 (d,J=5.3 Hz, 1H), 4.84-4.75 (m, 1H), 4.40 (s, 1H), 3.06-2.92 (m, 3H), 2.65-2.42 (m, 4H), 2.18 - 2.02 (m, 1H). MS: 616.1 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-l-(2,6-dichlorophenyl)-2-((3,3-difluorocyclobutyl) amino) -2-oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single
Figure imgf000103_0001
Ή NMR (400 MHz, CDC13): δ 8.78 (s, 1 H), 8.44 (d, J= 5.0 Hz, 1H), 7.80 - 7.22 (m, 5H), 6.91 (s, 1 H), 6.81 (tt, J= 8.7, 2.3 Hz, 1 H), 6.45 (d, J= 8.5 Hz, 1 H), 5.56 (d, J= 6.8 Hz, 1 H), 4.83 (dd, J= 9.4, 2.7 Hz, 1 H), 4.40 (d, J= 8.0 Hz, 1 H), 3.23 - 2.92 (m, 3H), 2.69 - 2.39 (m, 4H), 2.23 - 2.02 (m, 1H). MS: 634.2 (M+l)+.
(2S)-l-(4-Cyanopyridin-2-yl)-N-(l-(2,3-dichlorophenyI)-2-(3,3-difluoro-cyclobutyIamino)-2- oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound
Figure imgf000103_0002
Ή NMR (400 MHz, CDC13): δ 8.72 (s, 1H), 8.57 (s, 1H), 7.44 (d, J= 7.9, 1H), 7.32 - 7.29 (m, 1 H), 7.17 - 6.68 (m, 4H), 6.53 - 6.41 (m, 1 H), 6.32 - 6.12 (m, 1 H), 4.90 - 4.65 (m, 1 H), 4.41 - 4.05 (m, 1H), 3.13 -2.01 (m, 8H). MS: 634.1 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-(3,3-difluorocyclobutylamino)-l-(2-fluorophenyl)-2- oxoethyl)-N-(3,5-difluorophenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 208
Figure imgf000104_0001
Ή NMR (400 MHz, CDC13): δ 8.63 (s, 1H), 8.40 (d, J= 4.9 Hz, 1H), 7.43 (s, 1H), 7.20 (s, 1H), 7.16 (d, J= 5.0 Hz, 1H), 6.90 (t, J= 8.2 Hz, 3H), 6.62 (t, J= 8.7 Hz, 2H), 6.20 (s, 1H), 6.14 (d, J = 6.4 Hz, 1H), 4.81 (dd, J= 9.1, 2.9 Hz, 1H),4.25 (s, 1H),2.92 (s, 2H), 2.85 - 2.70 (m, 1H), 2.56-2.22 (m, 3H), 2.15 (m, 1H), 2.04-1.90 (m, 1H). MS: 584.2 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-(3,3-difluorocyclobutylamino)-2-oxo-l-phenylethyI)- N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound
Figure imgf000104_0002
Ή NMR (400 MHz, CDCI3): δ 8.77 (s, 1H), 8.41 (d,J=5.1 Hz, 1H), 7.49 (s, 1H), 7.27 (dd,J= 8.2, 5.0 Hz, 2H), 7.24 (d, J= 5.4 Hz, 2H), 7.04 (d, J= 6.7 Hz, 2H), 6.71 (t, J= 8.8 Hz, 1H), 6.44 (s, 1H), 6.15 (s, 1H), 5.70 (d,J=6.3 Hz, 1H), 4.86 (dd, J= 9.3, 2.8 Hz, 1H),4.29 (s, 1H), 2.99 (m,2H), 2.90 (m, 1H), 2.62- 2.52 (m, 1H),2.45 (m, 1H), 2.38 - 2.25 (m, 2H), 2.07 (m, 1H). MS: 566.2 (M+l)+.
(S)-N-((S)-l-(3-Chloropyridin-2-yl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 198
Figure imgf000104_0003
Ή NMR (400 MHz, CDCI3): δ 8.75 (s, 1H), 8.49 (d,J=5.0Hz, 1H), 8.31 (d,J=3.4 Hz, 1H), 7.65 - 7.56 (m, 2H), 7.27 (m, 1H), 7.19- 7.15 (m, 1H),6.98 (m, 1H), 6.76 - 6.56 (m, 2H), 6.11 (d, J= 6.8 Hz, 1H), 5.04 - 5.01 (m, 1H), 4.38 (m, 1H), 3.05 - 2.98 (m, 2H), 2.92 - 2.83 (m, 1H), 2.60 -2.52 (m, 1H), 2.51 - 2.37 (m, 2H), 2.37 - 2.27 (m, 1H), 2.07 - 2.02 (m, 1H). MS: 601.1 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri -din-2-yl)-5-oxo-N-(3-sulfamoylphenyl)pyrrolidine-2-carboxamide (single enantiomer) - Compound 84
Figure imgf000105_0001
Ή NMR (400 MHz, CDCI3): δ 8.73 (d, J = 10.0 Hz, 1H), 8.57 - 8.45 (d, J= 8.0 Hz, 1H), 8.12 (d, J=7.7Hz, 1H), 7.83 - 7.76 (m, 2H), 7.61 - 7.56 (m, 1H), 7.48 -7.32 (m, 1H), 7.19 (t,J= 7.1 Hz, 1H), 7.05 -6.87 (m, 2H), 6.82-6.81 (m, 1H), 6.55 - 6.43 (m, 1H), 6.27 (d,J=6.7 Hz, 1H), 5.24 (s, 1 H), 4.84 (d, J= 7.2 Hz, 1 H), 4.69 (s, 1 H), 4.33 (s, 1 H), 2.98 - 2.87 (m, 3H), 2.63 - 2.24 (m, 4H), 2.09- 2.00 (m, 1H). MS: 643.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyI)-N-(3-cyano- phenyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 128
Figure imgf000105_0002
Ή NMR (400 MHz, CDC13): δ 8.76 (s, 1H), 8.51 (s, 1H), 8.23 (m, 1H), 7.58 - 7.27 (m, 4H), 6.93 (m, 3H), 6.43 (s, 1H), 5.85 (s, lH),4.78(s, 1H),4.34 (s, 1H), 3.10 - 2.82 (m, 3H), 2.37 - 2.52(m, 3H), 2.21 -2.23 (m, 1H), 1.89- 1.99 (m, 1H). MS: 589.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-cyano phenyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 166
Figure imgf000106_0001
'HNMR(400 MHz, CDCI3): δ 8.77 (s, 1H), 8.49 (d,J= 13.9 Hz, 1H), 8.22- 8.32 (m, 1H), 7.61 -7.27 (m, 4H), 7.17 - 7.19 (m, 2H), 6.90 - 7.00 (m, 2H), 6.42 (s, 1H), 5.50 (s, 1H), 4.80 (d, J = 9.5 Hz, 1H), 3.97 (s, 1 H), 2.99 - 2.80 (m, 1H), 2.56 - 2.58 (m, 1H), 2.21 -2.24 (m, 1H), 1.70- 2.10 (m, 6H), 1.41 - 1.44 (m, 2H). MS: 617.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyano phenyl)-l-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 167
Figure imgf000106_0002
Ή NMR (400 MHz, CDC13): δ 8.91 - 9.00 (m, 1H), 8.33 - 8.17 (m, 1H), 7.62 - 7.32 (m, 5H), 7.20 (t,J= 7.0 Hz, 1H), 7.02 - 7.06 (m, 1H), 6.95 -6.83 (m, 1H), 6.55 (s, 1 H), 6.05 - 5.88 (m, 1H), 4.72 (d, J= 9.3 Hz, 1H), 4.37 (s, 1H), 2.91 - 3.05 (m, 3H), 2.70-2.25 (m, 4H), 2.13-1.92 (m, 1H). MS: 590.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyI)amino)-2-oxoethyl)-N-(3- cyanophenyl)-l-(4-cyanopyrimidin-2-yl)-5-oxopyrro!idine-2-carboxamide (single enantiomer) - Compound 178
Figure imgf000106_0003
Ή NMR (400 MHz, CDC13): δ 8.99 (s, 1H), 8.32 (s, 1H), 7.57 (m, 1H), 7.54 - 7.28 (m, 2H), 7.19 (t, J= 7.2 Hz, 3H), 7.04 (t, J= 6.8 Hz, 1H), 6.93 (d, J= 7.7 Hz, 1H), 6.53 (s, 1H), 5.64 - 5.44 (m, 1H), 4.74 (d,J= 9.3 Hz, 1H), 3.99 (s, 1 H), 2.94 (dd, J= 17.8,9.4 Hz, 1H), 2.62 (m, 1H), 2.41 -2.24 (m, 1H), 2.10 - 1.82 (m, 7H). MS: 618.2 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyI)-2-(3,3-difluorocycIobutyIamino)-2-oxoethyI)-N-(3-cyano-5- fluorophenyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer)
- Compound 177
Figure imgf000107_0001
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1H), 8.50 (s, 1H), 8.13 - 8.08 (m, 1H), 7.44 - 7.27 (m, 2H), 7.23 (dd, J= 12.6, 6.3 Hz, 2H), 7.07 (t, J= 7.3 Hz, 1H), 6.93 (t, J= 6.4 Hz, 1H), 6.43 (d, J = 6.1 Hz, 1H), 6.14(dd,J= 13.9, 6.7 Hz, 1 H), 4.81 (dd,J=9.0, 2.3 Hz, 1H), 4.42-4.28 (m, 1 H), 3.12- 2.94 (m, 2H), 2.94 - 2.80 (m, 1 H), 2.67 - 2.29 (m, 3H), 2.23 - 1.92 (m, 2H). MS: 607.1 (M+l)+.
(S)-N-((S)-l-(2-ChlorophenyI)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-N-(3-cyano-5- fluorophenyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 184
Figure imgf000107_0002
Ή NMR (400 MHz, CDC13): δ 8.77 (s, 1H), 8.50 (s, 1H), 8.25 - 8.03 (m, 1H), 7.52 - 7.28 (m, 2H), 7.22 (t,J=7.7 Hz, 2H), 7.01 (dt,J= 14.1, 10.1 Hz, 2H), 6.42 (d,J=6.9 Hz, 1H), 5.58 (t, J = 9.9 Hz, 1H),4.83 (dd, J= 9.1, 2.3 Hz, 1H), 4.05 - 3.86 (m, 1H), 3.04 - 2.81 (m, lH),2.59(m, 1 H), 2.36 - 1.70 (m, 7H), 1.58 - 1.31 (m, 3H). MS: 636.2 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyI)-2-((4,4-difluorocycIohexyI)amino)-2-oxoethyI)-N-(3-cyano-5- fluorophenyl)-l-(4-cyanopyrimidin-2-yl)-5-oxopyrrolidine-2-carboxamide (single
enantiomer) - Compound 185
Figure imgf000108_0001
Ή NMR (400 MHz, CDC13): δ 8.97 (d, J= 4.4 Hz, 1H), 8.12 (m, 1H), 7.50 - 7.32 (m, 3H), 7.23 (d, J= 6.7 Hz, 2H), 7.06 (m, 1H), 6.95 (s, 1H), 6.50 (d, J= 8.6 Hz, 1H), 5.60 (d, J= 7.5 Hz, 1H), 4.74 (d,J=8.8Hz, 1H), 3.98 (s, 1H),2.90 (m, 1 H), 2.72 - 2.49 (m, 1H),2.28 (s, 1H), 2.17- 1.67 (m, 7H), 1.43 (m, 2H). MS: 637.2 (M+l)+.
(S)-N-(3-Cyano-5-fluorophenyl)-l-(4-cyanopyridin-2-yI)-N-((S)-2-(3,3-difluorocyclobutyl - amino)-2-oxo-l-phenyIethyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 211
Figure imgf000108_0002
Ή NMR (400 MHz, CDC!3): δ 8.71 (d,J= 10.1 Hz, 1H), 8.38 (s, 1H), 8.02 (m, 1H), 7.23 (m, 5H),6.97 (d,J=7.3 Hz, 3H), 6.20 (s, 1H), 5.97 (s, 1H), 4.70 (dd, J= 9.2, 2.4 Hz, 1H),4.27 (s, 1 H), 2.93 (m, 2H), 2.85 (t, J= 8.9 Hz, 1 H), 2.59 - 2.48 (m, 1 H), 2.49 - 2.29 (m, 2H), 2.29 - 2.20 (m, 1H), 2.08- 1.99 (m, 1H). MS: 573.2 (M+l)+.
(S)-N-(3-Cyano-5-fluorophenyl)-l-(4-cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyl) amino)-l-(2-fluorophenyl)-2-oxoethyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 207
Figure imgf000108_0003
Ή NMR (400 MHz, DMSO-d6): δ 8.78 (s, 1H), 8.62 (d, J= 5.1 Hz, 1H), 8.48 (s, 1H), 8.04 - 7.83 (m, 1H), 7.78 (s, 1H), 7.57 (s, 1H), 7.23 (m, 2H), 7.14 (d,J= 9.9 Hz, 1H), 6.95 (t,J=7.5 Hz, 1H), 6.84 (s, 1H), 6.20 (s, I H), 4.72 (s, lH),4.04(s, 1H), 4.00 - 3.82 (m, 1H), 3.09 - 2.67 (m,2H), 2.33 (m, 1H), 1.91 (s, 2H), 1.83 (s, 1H), 1.27-1.05 (m, IH). MS: 591.2 (M+l)+ .
(2S)-N-(l-(2-ChIorophenyI)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyI)-l-(4-cyanopyri - din-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound
Figure imgf000109_0001
Ή NMR (400 MHz, CDC13): δ 9.10 - 8.03 (m, 4H), 7.47 - 7.39 (m, 2H), 7.27 - 6.84 (m, 3H), 6.51 -6.01 (m,2H), 4.84 -4.70 (m, 1 H), 4.36 - 4.20 (m, 1H), 3.25 - 1.86 (m, 8H). MS: 583.1 (M+l)+ .
(S)-N-((S)-l-(2-ChlorophenyI)-2-(3,3-difluorocyclobutyIamino)-2-oxoethyl)-l-(4-cyanopyri - din-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 176
Figure imgf000109_0002
Ή NMR (400 MHz, CDCI3): δ 8.95 - 8.70 (m, 1H), 8.49 (d,J= 4.7 Hz, 1H), 8.36 - 8.11 (m, 1H), 8.12 (d,J= 8.6 Hz, 1H), 7.33 (d, J= 8.0 Hz, 1H), 7.21 (t,J=7.8Hz, 1H), 7.04 (t,J=7.6 Hz, 1 H), 6.48 - 6.41 (m, 1 H), 6.30 - 6.21 (m, 1 H), 4.84 - 6.79 (m, 1 H), 4.38 - 4.30 (m, 1H), 3.11 - 2.74 (m, 3H), 2.65 - 1.91 (m, 5H). MS: 583.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-l-(4-cyanopyri - din-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) -
Compound 193
Figure imgf000110_0001
'H MR (400 MHZ, CDC13): δ 8.77 (s, 1H), 8.49 (d,J=5.2 Hz, 1H), 8.40-8.27 (m, 1H), 8.21 - 8.04 (m, 1H), 7.41 - 7.36 (m, 1H), 7.26 - 7.23 (m, 1H), 7.20 (t,J= 6.9 Hz, 1H), 7.04 (t, J= 7.2 Hz, 1H), 6.93 (m, 1H), 6.52 -6.34 (m, 1H), 5.49 (s, 1H), 4.84 (d, J= 7.4 Hz, 1H), 4.01 -3.94 (m, 1H), 2.99-2.91 (m, 1H), 2.62 - 2.54 (m, 1H), 2.22- 1.71 (m, 7H), 1.31 (s, 3H). MS: 611.2 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyl)amino)-2-oxo-l-phenylethyl)- N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound
Figure imgf000110_0002
'HNMR (400 MHz, CDC13): δ 8.86 (m, 1 H), 8.39 (m, 2H), 8.03 (m, 1 H), 7.28 (d, J= 5.9 Hz, 4H), 6.98 (m, 2H), 6.29 (s, 1H), 5.85 (s, 1H),4.85 (m, 1H),4.33 (s, 1H), 3.26 - 2.82 (m, 3H), 2.69 - 1.88 (m, 5H). MS: 549.2 (M+l)+.
(S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyl)amino)-l-(2-fluorophenyI)-2- oxoethyl)-N-(5-fluoropyridin-3-yI)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 148
Figure imgf000110_0003
Ή NMR (400 MHz, CDC13): δ 8.99 - 8.60 (m, 1H), 8.55 - 7.97 (m, 3H), 7.35 - 7.19 (m, 3H), 7.07 - 6.89 (m,3H), 6.36 (m, 1H), 6.12 (s, 1H), 4.80 (s, lH),4.35(s, 1H), 3.22 - 2.79 (m, 3H), 2.64 - 1.85 (m, 5H). MS: 567.2 (M+l)+. (S)-l-(4-Cyanopyridin-2-yl)-N-((S)-2-((3,3-difluorocyclobutyI)amino)-2-oxo-l-phenylethyl)- N-(5-isocyanopyridin-3-yl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 212
Figure imgf000111_0001
Ή NMR (400 MHz, CDCI3): δ 9.34 (s, 1 H), 8.87 - 8.56 (m, 4H), 8.41 (s, 2H), 8.27 (s, 1 H), 7.54 (s, 7H), 7.01 (d, J= 6.9 Hz, 3H), 6.35 (s, 2H), 5.73 (s, 2H), 4.66 (s, 2H), 4.35 (s, 2H), 2.99 (m, 5H), 2.73 - 2.20 (m, 7H), 2.07 (s, 2H). MS: 556.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(3-cyano - phenyl)-N-(lH-indazol-7-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 186
Figure imgf000111_0002
Ή NMR (400 MHz, CDC13): δ 8.72 - 8.71 (m, 1 H), 8.66 (s, 1 H), 8.08 (s, 1 H), 7.69 (s, 1 H), 7.67 (s, 1 H), 7.50 - 7.49 (m, 1 H), 7.36 - 7.34 (m, 1 H), 7.1 1-7.07 (m, 1 H), 7.00-6.96 (m, 1 H), 6.83- 6.76 (m, 2H), 6.48 (s, 1 H), 5.07 - 5.07 (m, 1 H), 4.38 - 4.33 (m, 1 H), 3.05 - 2.91 (m, 2H), 2.80 - 2.71 (m, 1 H), 2.65 - 2.60 (m, 1 H), 2.53 - 2.46 (m, 2H), 2.03- 1.99 (m, 1 H), 1.75 - 1.67 (m, 1 H). MS: 603.2 (M+l )+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-3-(3-cyano phenyl)-N-(lH-indazol-7-yl)-2-oxooxazolidine-4-carboxamide (single enantiomer) - Compound 142
Figure imgf000111_0003
H NMR (400 MHz, CDC13): δ 13.03 (s, 1H), 8.73 (s, 1H), 8.55 - 8.54 (m, 1H), 8.02 (s, 1H), 8.58- 8.56 (m, 1H), 8.50 - 8.48 (m, 1H), 7.27-7.24 (m, 2H), 7.03 - 6.99 (m, 1H), 6.91 -6.87 (m, 1 H), 6.80 - 6.78 (m, 1 H), 6.72 - 6.68 (m, 1 H), 6.33 (s, 2H), 5.70 - 5.69 (m, 1 H), 4.99 - 4.97 (m, 1 H), 4.05 - 4.03 (m, 1 H), 2.78 - 2.95 (m, 1 H), 2.47 - 2.40 (m, 1 H), 2.08 - 4.99 (m, 6H), 1.90-1.82 (m, 2H), 1.67-1.63 (m, 1H), 1.58- 1.62 (m, 1H). MS: 633.2 (M+l)+.
(2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopy
4-yl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 152
Figure imgf000112_0001
Ή NMR (400 MHz, DMSO-d6): δ 13.05 (m, 1H), 8.70 (m, 2H), 8.54 (d,J=6.7Hz, 1H), 8.21 (s, 1H), 7.80 (d, J= 6.9 Hz, 1H), 7.63 (d, J= 5.0 Hz, 1H), 7.36 (m, 2H), 7.24 (d, J- 8.0 Hz, 1H), 7.18-6.97 (m, 1H), 6.92 - 6.79 (m, 1H), 6.77-6.70 (m, 1H), 6.35 (d, 1H),4.66 (m, 1H),4.20- 4.01 (m, 1 H), 3.05 - 2.78 (m, 2H), 2.68 - 2.52 (m, 2H), 2.49 - 2.26 (m, 2H), 2.22 - 1.53 (m, 2H). MS : 604.2 (M+l)+.
(S)-N-(3-(lH-Pyrazol-4-yl)phenyl)-N-((S)-l-(2-chlorophenyl)-2-(3,3-dinuorocyclobutylami- no)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (single
enantiomer) - Compound 200
Figure imgf000112_0002
Ή NMR (400 MHz, MeOD): δ 8.73 - 8.54 (m, 2H), 8.14-7.91 (m, 1H), 7.71 (d, J= 7.6 Hz, 1H), 7.56 - 7.28 (m, 4H), 7.25 - 6.92 (m, 4H), 6.70 (d, J= 7.6 Hz, 1H), 6.54 - 6.39 (m, 1H), 5.03 (dd, J= 9.4, 2.9 Hz, 1 H), 4.31 - 4.05 (m, 1 H), 3.00 - 2.73 (m, 3H), 2.64 - 2.00 (m, 5H). MS: 630.2 (M+l)+. (2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyl)-l-(4-cyanopyri - midin-2-yl)-5-oxo-N-(3-(trifluoromethoxy)pheny])pyrrolidine-2-carboxamide (racemic) - Compound 180
Figure imgf000113_0001
Ή NMR (400 MHz, CDC13): δ 8.96 (t, J= 5.5 Hz, 1H), 7.88 (s, 1H), 7.44 - 7.32 (m, 2H), 7.21 (m,2H), 7.10 (t,J= 7.3 Hz, 1H), 7.04-6.95 (m, 1H), 6.91 (m, 1H), 6.52 (m, 1H), 6.18 (m, 1H), 4.89-4.67 (m, 1 H), 4.31 (m, 1H), 3.22 - 2.75 (m, 3H), 2.70- 1.92(m, 5H). MS: 649.1 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(3-(difluoromethoxy)phenyI)-5-oxopyrrolidine-2-carboxamide (single
181
Figure imgf000113_0002
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1H), 8.44 (m, lH), 7.76 (d,J= 9.0 Hz, 1H), 7.33 (m, 2H), 7.21 -6.83 (m, 6H), 6.44 (t, J= 8.8 Hz, 1H), 6.28 -6.13 (m, 1H), 4.91 (m, 1H), 4.34 (s, 1H), 3.10-2.66 (m, 3H), 2.65 - 1.84 (m, 5H). MS: 630.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyI)-l-(4-cyano pyrimidin-2-yl)-N-(3-(difluoromethoxy)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 194
Figure imgf000113_0003
Ή NMR (400 MHz, CDC13): δ 9.04 - 8.59 (m, 1H), 7.74 (s, 1 H), 7.43 - 7.26 (m, 4H), 6.96 (m, 3H), 6.36 (m,2H), 4.81 (t,J=9.3 Hz, 1H), 4.55 (m, 1H), 4.33 (s, 1H), 4.06-3.89 (m, 1H), 3.15 -2.69 (m, 2H), 2.69 - 1.86 (m, 5H). MS: 631.1 (M+l)+.
(S)-N-((S)-l-(2C)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N- (3-methoxyphenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 129
Figure imgf000114_0001
Ή NMR (400 MHz, CDC13): δ 8.75 (s, 1H), 8.51 (d,J=5.0Hz, 1H), 7.47 (m, 1H), 7.38-7.08 (m, 3H), 6.99 (d, J= 6.7 Hz, 3H), 6.89 - 6.66 (m, 2H), 6.41 (s, 1H), 6.09 (d, J= 6.6 Hz, 1H), 4.97 (dd, J= 9.3, 3.2 Hz, 1H), 4.34 (s, 1H), 3.72 (m, 3H), 3.01 (dd, J= 7.5, 4.0 Hz, 3H), 2.65 - 2.23 (m, 4H), 2.04 (d, J= 9.0 Hz, 1H). MS: 594.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-l-(4-cyanopyri midin-2-yl)-N-(3-methoxyphenyl)-5-oxopyrrolidine-2-carboxamid (single enantiomer) - Compound 164
Figure imgf000114_0002
Ή NMR (400 MHz, CDC13): δ 8.92 (s, 1 H), 7.48 - 7.39 (m, 1 H), 7.33 - 7.26 (m, 2H), 7.22 - 7.08 (m, 2H), 7.04 - 6.82 (m, 3H), 6.73 (s, 2H), 6.48 (d,J= 9.5 Hz, 1H), 6.18 (m, 1H), 4.88 - 4.85 (m, 1H), 4.32 (s, 1H), 3.78 (s, 1H), 3.62 (s, 2H), 3.01 - 2.81 (m, 3H), 2.58 - 2.49 (m, 2H), 2.42-2.30 (m, 2H), 2.09 - 1.98 (m, 1H). MS: 595 (M+l)+.
(2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyrimi -din-2-yl)-N-(3-cyclopropoxyphenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 192
Figure imgf000115_0001
Ή NMR (400 MHz, CDC13): δ 9.06 - 8.88 (m, 1H), 7.61 - 7.30 (m, 4H), 7.27 - 7.22 (m, 1H), 7.18 (t, J = 7.4 Hz, 2H), 7.08-6.92 (m, 1H), 6.87 (dd, J= 8.7, 2.1 Hz, 1H), 6.78 (t, J= 9.5 Hz, 1H), 6.50 (s, 1H), 6.04 (m, 3H), 5.57 - 5.14 (m, 2H), 4.88 (m, 1H), 4.77-4.10 (m, 3H), 3.15 - 2.75 (m, 3H), 2.68 - 2.47 (m, 2H), 2.45 - 2.21 (m, 3H), 2.20 - 1.90 (m, 1H). MS: 621.1 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri - din-2-yl)-N-(3-(hydroxymethyl)phenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 131
Figure imgf000115_0002
1 H NMR (400 MHz, CDC13): δ 8.73 (s, 1 H), 8.53 (s, 1 H), 7.94 - 7.70 (m, 1 H), 7.31 (s, 1 H), 7.26 (dd,J=5.1, 1.3 Hz, 1H), 7.22 - 7.10 (m, 4H), 7.02 - 6.87 (m, 2H), 6.44 (d,J= 10.5 Hz, 1H), 6.12 (d,J= 6.4 Hz, 1H), 4.91 (dd, J= 9.3, 3.2 Hz, 1H), 4.69 (s, 1H), 4.48 (s, 1H), 4.42 - 4.26 (m, 1H), 3.07 - 2.85 (m, 3H), 2.65-2.17 (m, 4H), 2.01 (s, 2H). MS: 594.2 (M+l)+.
(S)-N-((S)-l-(2-Xhlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-(l-hydroxycyclopropyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 140
Figure imgf000115_0003
Ή NMR (400 MHz, CDC13): δ Ή NMR (400 MHz, CDC13) : 58.73 (s, 1H), 8.52 - 8.44 (m, 1H), 7.64 - 7.30 (m, 3H), 7.22 - 6.90 (m, 5H), 6.42 - 6.38 (m, 1H), 6.03 (m, 1H), 4.87 (m, 1H), 4.30 (m, 1H), 3.05 -2.82 (m, 3H), 2.60 - 1.88 (m, 5H), 1.21 (d,J=3.2 Hz, 4H). MS: 620.2 (M+l)+. (S)-N-((S)-l-(2C)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N- (3-(2-hydroxypropan-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 179
Figure imgf000116_0001
Ή NMR (400 MHz, CDC13): δ 8.69 (s, 1 H), 8.54 (d, J= 5.0 Hz, 1 H), 7.93 - 7.70 (m, 1 H), 7.40 - 7.19 (m, 4H), 7. 1 1 (m, 2H), 7.01 - 6.72 (m, 2H), 6.45 (m, 2H), 5.05 - 4.76 (m, 1 H), 4.33 (s, 1 H), 3.13 - 2.58 (m, 3H), 2.42 (m, 4H), 2.09 - 1.83 (m, 1 H), 1.33 (s, 6H). MS: 622.2 (M+ l )+.
(S)-N-((S)-l-(2-ChIorophenyl)-2-(3,3-difluorocycloburyIamino)-2-oxoeth-yI)-l-(4- cyanopyri-din-2-yI)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyI)-5-oxopyrrolidine-2- carboxamide (single enantiomer) - Compound 150
Figure imgf000116_0002
Ή NMR (400 MHz, CDC13): δ 8.66 (s, 1 H), 8.49 (d, J= 4.8 Hz, 1 H), 7.73 - 7.48 (m, 1 H), 7.26 - 6.83 (m, 7H), 6.53 - 6.42 (m, 2H), 4.91 (d, J= 6.4 Hz, 1 H), 4.32 (s, 1 H), 3.02 - 2.72 (m, 3H), 2.58 - 1 .85 (m, 6H), 1 .63 (s, 2H), 1 .5 1 (d, J = 7.0 Hz, 2H), 1 .29 (d, J = 8.6 Hz, 4H). MS: 640.2 (M+ l )+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyI)amino)-2-oxoethyl)-l-(4- cyanopyri din-2-yI)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-5-oxopyrrolidine-2- carboxamide (single enantiomer) - Compound 155
Figure imgf000116_0003
Ή NMR (400 MHz, CDC13): δ 8.80 (s, 1H), 8.43 (s, 1H), 7.51 (d, 1H), 7.24 (m, 4H), 7.06 (s, 3H), 6.64 (m, 1H), 6.15 (m, 1H), 5.73 (s, lH),4.86(s, 1H),4.32 (s, 1H), 3.01 (m, 3H), 2.68 - 2.27 (m,4H), 2.12 (s, 1H), 1.44 (s, 1H), 1.29 (d, J= 9.0 Hz, 6H). MS: 639.2 (M+l)+
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri -din-2-yl)-N-(3-(2-hydroxyethyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 160
Figure imgf000117_0001
Ή NMR (400 MHz, CDC13): δ 8.76 (s, 1H), 8.52 (d, J= 5.0 Hz, 1H), 7.74 (s, 1H), 7.32 - 7.36 (m, 1H), 7.27-7.11 (m, 2H), 7.09 - 6.87 (m, 4H), 6.39 - 6.45 (m, 1H), 6.05 (d,J=6.9 Hz, 1H), 4.33 (s, 1H), 3.82 (s, 1H), 3.59 (s, 1H), 3.12 -2.79 (m, 4H), 2.74 - 2.16 (m, 5H), 1.99 -2.07 (m, 1H). MS: 608.2 (M+l)+.
(S)-N-((S)-l-(2-ClorophenyI)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyI)-l-(4-cyano pyridin-2-yl)-N-(3-(2-hydroxyethoxy)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 130
Figure imgf000117_0002
Ή NMR (400 MHz, CDC13): δ 8.72 (s, 1H), 8.48 (d, J= 5.0 Hz, 1H), 7.54 - 7.28 (m, 2H), 7.18 - 7.21 (m, 2H), 7.01 - 6.94 (m, 2H), 6.75 - 6.77 (m, 2H), 6.39 (s, 1H), 5.99 (s, 1H), 4.94 (dd, J = 9.3,3.4 Hz, 1H), 4.31 (s, 1 H), 3.79 - 4.06 (m, 4H), 3.07 - 2.80 (m, 3H), 2.58 - 2.21 (m, 4H), 1.87 - 2.00 (m, 2H). MS: 624.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-fluoro-5-((S)-methylsulfinyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 190
Figure imgf000118_0001
'HNMR(400 MHZ, CDCI3): δ 8.75 (s, 1H), 8.54 (m, 1H), 8.02-7.78 (m, 1H), 7.33 (s, 3H), 7.21 (m 1H), 7.06 (t, J- 7.4 Hz, 1H), 6.96 (m, 1H), 6.45 (m, 1H), 6.27 (m, 1H), 4.86 (m, 1H), 4.35 (m, 1H), 3.16-2.82 (m, 3H), 2.71 (s, 1H), 2.65 - 2.47 (m, 2H), 2.41 (m, 3H), 2.22 (m, 1H), 2.09 (m, 1H). MS: 644.1 (M+l)+.
(2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-(methylsuIfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 96
Figure imgf000118_0002
Ή NMR (400 MHz, CDC13): δ 8.84 -8.11 (m, 3H), 7.93 - 7.35 (m, 4H), 7.25 - 6.75 (m, 2H), 6.64 - 5.94 (m, 2H), 4.89 - 4.69 (m, 1 H), 4.28 (d, J= 5.7 Hz, 1 H), 3.13- 2.74 (m, 6H), 2.68 - 2.48 (m, 2H), 2.46-2.15 (m, 3H), 2.04 (s, 1H). MS: 642.1 (M+l)+.
(2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyri midin-2-yI)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 102
Figure imgf000118_0003
Ή NMR (400 MHz, CDCI3): δ 8.93 (t, J= 5.3 Hz, 1H), 8.50 - 8.15 (m, 1H), 7.94 - 7.71 (m, 2H), 7.66-7.46 (m, 1 H), 7.38 (t, J= 6.4 Hz, 1H), 7.28 (t,J=3.6Hz, 1H), 7.20-7.07 (m, 1H), 7.05 - 6.87 (m, 2H), 6.74 (m, 1 H), 6.52 (m, 1 H), 4.72 (dd, J= 9.2, 2.5 Hz, 1 H), 4.34 (d, J= 6.4 Hz, 1 H), 3.00 (s, 3H), 2.90 - 2.75 (m, 3H), 2.56 -2.19 (m, 5H), 1.98 (m, 1 H). MS: 643.1 (M+ 1)+. (2S)-N-(l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 95
Figure imgf000119_0001
Ή NMR (400 MHz, CDC13): δ 8.87 - 8.13 (m, 3H), 8.02 - 7.37 (m, 4H), 7.24 - 6.87 (m, 2H), 6.51 -6.39 (m, 1H), 5.77 - 5.28 (m, 1H), 4.89-4.65 (m, 1H), 3.94 (d,J=5.2 Hz, 1H), 3.16- 2.73 (m, 4H), 2.68 - 2.53 (m, 1 H), 2.44 - 2.20 (m, 1 H), 2.03 (m, 8H), 1.44 (m, 2H). MS: 670.2 (M+l)+.
(S)-N-((S)-l-(2C)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopyri midin-2-yl)- N-(3-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) -
Compound 103
Figure imgf000119_0002
Ή NMR (400 MHz, CDC13): δ 8.94 (dd, J= 7.9, 4.8 Hz, 1H), 8.56 - 8.15 (m, 1H), 7.97 - 7.62 (m, 2H), 7.56 - 7.29 (m, 3H), 7.13 (t, J= 7.6 Hz, 1H), 7.06 - 6.84 (m, 2H), 6.51 (d, J= 4.2 Hz, 1H), 6.10 (dd,J= 3.2, 7.4 Hz, 1H),4.74 (d,J=6.6 Hz, 1H), 3.98 (s, 1H), 3.01 (s, 1H),2.93- 2.72 (m, 3H), 2.52 (d, J= 9.6 Hz, 1H), 2.37-2.20 (m, 1H), 2.13 - 1.78 (m, 7H), 1.63 - 1.40 (m, 2H). MS: 671 (M+l)+.
(2S)-N-(l-(2-ChlorophenyI)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyI)-l-(4-cyanopy ridin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide
(racemic) - Compound 110
Figure imgf000120_0001
Ή NMR (400 MHz, CDC13): δ 8.45-8.79 (m, 2H), 8.40 - 8.13 (s, 1H), 8.09 - 7.67 (m, 1H), 7.63 - 7.30 (m, 2H), 7.23 - 6.87 (m, 3H), 6.55 - 6.30 (m, 1H), 6.22 - 5.94 (m, 1H), 4.96-4.61 (m, 1 H), 4.26 (m, 4H), 3.16 - 1.87 (m, 7H), 1.27 (d, 1 H). MS: 660.1 (M+ 1 )+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri midin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carbox- amide (single enantiomer) - Compound 109
Figure imgf000120_0002
Ή NMR (400 MHz, CDC13): δ 8.96 (d, J= 4.6 Hz, 3H), 7.99 (d, J= 8.5 Hz, 2H), 7.75 (s, 2H), 7.52 (d, J= 7.0 Hz, 3H), 7.37 (d, J= 4.9 Hz, 5H), 7.19 (t, J= 7.7 Hz, 3H), 7.01 (dt, J= 7.1 Hz, 6H), 6.40 - 6.60 (m, 3H), 6.06 (d, J= 6.5 Hz, 3H), 4.76 (d, J= 9.2 Hz, 1H), 4.35 (m, 4H), 3.14 - 1.87 (m, 8H). MS: 661.1 (M+l)+.
(2S)-N-(l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyI)amino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 105
Figure imgf000120_0003
Ή NMR (400 MHz, CDC13): δ 8.96 (t, J= 4.6 Hz, 1H), 7.53 - 7.36 (m, 3H), 7.23 (m, J= 7.8, 1.5 Hz, 1H), 7.14-6.94 (m, 3H), 6.68 (m,J=8.6,2.3 Hz, 1H), 6.60 (d,J=3.1 Hz, 1H), 6.07 (d,J = 6.7 Hz, I H), 4.75 (q, J = 4.0, 2.1 Hz, I H), 4.38 (d, J = 6.7 Hz, I H), 3.78 - 3.67 (m, 2H), 3.39 (m, I H), 3.26 - 2.92 (m, 3H), 2.67 - 2.36 (m, 2H). MS: 688.1 (M+ l )+.
(2S)-N-(l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopyri midin-2-yl)-N-(3-fluoro-5-(methylsu]fonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single
08
Figure imgf000121_0001
Ή NMR (400 MHz, CDC13): δ 8.97 (s, I H), 8.20 - 8.60 (m, I H), 8.09 - 7.68 (m, I H), 7.63 - 7.32 (m,5H), 7.22 - 6.93 (m, 3H), 6.64 - 6.03 (m, 2H), 5.62 (s, I H), 4.60 - 4.85 (m, I H), 3.21 - 1.70 (m, 12H), 1.50 - 1 .14 (m, 2H). MS: 689.1 (M+ l )+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyano pyrimidin-2-yl)-N-(3-fluoro-5-(methylsulfonyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 168
Figure imgf000121_0002
Ή NMR (400 MHz, CDC!3): δ 9.0 (s, I H), 8.05 - 8.02 (m, 1 H), 7.80 (m, 1 H), 7.56 - 7.00 (m, 7H), 6.58 (m, I H), 5.65 (m, I H), 4.80 (m, I H), 4.14 (m, I H), 3.00-0.88 (m, 15H). MS: 689.1
(M+ l f.
(S)-N-((S)-l-(2-ChlorophenyI)-2-(3,3-difluorocyclobutyIamino)-2-oxoethyI)-l-(4-cyanopyri din-2-yl)-N-(3-(methylsulfonamido)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 159
Figure imgf000122_0001
Ή NMR (400 MHz, DMSO-d6): δ 9.78 (s, 1H), 8.84 - 8.61 (m, 2H), 8.56 (s, 1H), 7.66 (m, 2H), 7.49 - 7.15 (m, 3H), 7.15- 6.79 (m, 4H), 6.25 (m, 1 H), 4.89 - 4.74 (m, 1 H), 4.19 - 4.04 (m, 1 H), 3.03 - 2.83 (m, 3H), 2.72 - 2.59 (m, 3H), 2.54 (m, 2H), 2.44 - 2.28 (m, 1 H), 1.99 (m, 2H). MS : 657.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyri- din-2-yl)-N-(3-(dimethylamino)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 161
Figure imgf000122_0002
Ή NMR (400 MHz, CDC13): δ 8.71 (d, J= 9.9 Hz, 1H), 8.50 - 8.41 (m, 1H), 7.29 (d, J= 7.8 Hz, 1H), 7.22 (dd, J= 5.0, 1.3 Hz, 1H), 7.18 - 7.05 (m, 2H), 6.99 - 6.86 (m, 3H), 6.56 - 6.47 (m, 2H), 6.37 (d,J= 6.6 Hz, 1H), 6.11 (s, 1H), 5.01 (d,J=9.2 Hz, 1H), 4.34 -4.28 (m, 1H), 3.07 - 2.70 (m, 8H), 2.61 -2.42 (m, 2H), 2.35 - 2.25 (m, 2H), 2.01 - 1.97 (m, 1H). MS: 607.2 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyI)-l-(4- cyanopyri din-2-yl)-N-(2-fluorophenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 187
Figure imgf000122_0003
Ή NMR (400 MHz, CDC13): δ 8.74 (m, 1H), 8.48 (m, 1H), 7.96 - 7.92 (m, 1H), 7.40 (m, 1H), 7.28 - 6.72 (m, 7H), 6.59 - 5.79 (m, 2H), 4.86 - 4.78 (m, 1H), 4.28 (s, 1H), 3.04 - 2.90 (m, 3H), 2.66-2.01 (m, 5H). MS: 582.1 (M+l)+.
(S)-N-((S)-l-(2-Clorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyri din-2-yl)-N-(2,3-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 188
Figure imgf000123_0001
Ή NMR (400 MHz, CDC13): δ 8.73 (m, 1H), 8.47 (d, J= 5.0 Hz, 1H), 7.84 - 7.73 (m, 1H), 7.43 (d, J= 8.1 Hz, 1H), 7.28 - 7.20 (m, 2H), 7.13 (dd, J= 8.2, 4.4 Hz, 2H), 7.01 - 6.83 (m, 2H), 6.62 (s, 1H), 6.42 -5.85 (m, 1H), 4.85 - 4.77 (m, 1H), 4.20 (m, 1H), 3.13 - 2.78 (m, 3H), 2.68 - 2.28 (m, 4H), 2.25 - 2.04 (m, 1 H). MS: 600.1 (M+ 1 ) +.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri din-2-yl)-N-(2,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 197
Figure imgf000123_0002
Ή NMR (400 MHz, CDC13): δ 8.73 (m, 1 H), 8.54 - 8.41 (m, 1H), 7.83-7.78 (m, 1H), 7.44- 7.39 (m, 1H), 7.28-7.21 (m, 2H), 7.13 - 6.88 (m, 3H), 6.81-6.80 (m, 1H), 6.61 -6.31 (m, 1H), 5.91 (d,J=6.5 Hz, 1H), 4.86-4.79 (m,lH), 4.29 (dd, J= 8.2, 6.7 Hz, 1H), 3.51 (s, 1H), 3.12- 2.85 (m, 3H), 2.68 - 2.56 (m, 1 H), 2.54 - 2.45 (m, 1 H), 2.43 - 2.24 (m, 2H), 2.23 - 2.06 (m, 1H). MS: 600.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(lH-indazol-5-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 203
Figure imgf000124_0001
'H MR(400 MHZ, CDCI3): δ 8.78 (s, 1H), 8.56 (m, 1H), 8.39 (s, 1H), 8.13 - 7.88 (m, 1H), 7.44-7.32(m, 2H), 7.28 - 7.00 (m, 4H), 6.99 - 6.79 (m, 2H), 6.48 (m, 1H), 5.75 - 5.48 (m, 1H), 5.06-4.75 (m, I H), 4.00 (s, 1H), 3.10 -2.77 (m, 1H), 2.63 - 2.44 (m, 1H), 2.37 - 2.20 (m, 1H), 2.15 - 1.77 (m, 7H), 1.42 (m, 2H). MS: 632.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri din-2-yl)-N-(lH-indazol-6-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 205
Figure imgf000124_0002
Ή NMR (400 MHz, CDCI3): δ 8.78 (s, 1 H), 8.57 (t, J= 5.0 Hz, 1 H), 8.23 - 7.76 (m, 2H), 7.54 - 7.30 (m, 2H), 7.16 (s, 1H), 7.04 - 6.86 (m, 3H), 6.47 (d,J= 11.7 Hz, 1H), 6.02 (d,J=6.1 Hz, 1H),4.92 (m, lH),4.36(s, 1H), 2.97 (m, 3H), 2.65 - 2.20 (m, 4H), 1.99 (m, 1H). MS: 604.2
(M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopy ridin-2-yl)-N-(lH-indazol-6-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) -
Compound 136
Figure imgf000124_0003
'HNMR (400 MHz, CDC13):510.41 -9.94 (m, 1H), 8.79 (s,lH), 8.57 (t,J= 5.1 Hz, 1H), 8.28 -8.09 (m, 1H), 7.93 (m, lH),7.52(s, 1H), 7.40 (d,J=7.9 Hz, 1H), 7.33 (d,J=7.5Hz, 1H), 7.15 -6.98 (m, 1H), 6.46 (d,J= 12.7 Hz, 1H), 5.50 (d, J= 7.9 Hz, 1H), 5.06 - 4.76 (m, 1H), 4.02 (s, 1H), 2.92 (dd, 1H), 2.63 -2.49 (m, 1H), 2.31 (s, 1H), 2.03 (m, 6H), 1.45 (s, 2H). MS: 632.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri -din-2-yl)-N-(lH-indazol-5-yl)-5-oxopyrrolidine-2-carboxamide (single
Figure imgf000125_0001
Ή NMR (400 MHz, CDC13): δ 8.75 (s, 1H), 8.64 - 8.46 (m, 1H), 8.34 (s, 1H), 8.09 (s, 1H), 7.94 - 7.92 (m, 1H), 7.42 - 7.32 (m, 2H), 7.24 - 7.02 (m, 2H), 6.94 - 6.85 (m, 2H), 6.49 - 6.45 (m, 1H), 6.08 -6.06 (m, 1 H), 5.00 - 4.76 (m, 1H), 4.35 - 4.31 (s, 1H), 3.00 - 2.85 (m, 3H), 2.64 - 2.11 (m, 4H), 2.01 - 1.93 (m, 1H). MS: 604.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(lH-indol-5-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 206
Figure imgf000125_0002
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1H), 8.55 (m, 1H), 8.12 (d, J= 13.8 Hz, 2H), 7.52-7.29 (m, 2H), 7.18-6.80 (m, 5H), 6.46 (m, 2H), 5.83 (s, 1H), 5.83 (s, 1H), 5.08 - 4.81 (m, 1H), 4.33 (s, 1H), 2.92 (m, 3H), 2.64 -2.16 (m, 4H), 2.01 (m, 1H). MS: 603.2 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(l-methyl-lH-indol-5-yl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Com ound 209
Figure imgf000126_0001
Ή NMR (400 MHz, CDC13): δ 8.83 - 8.39 (m, 1H), 8.01 (m, 1H), 7.68 - 7.32 (m, 1H), 7.28 - 6.72 (m, 8H), 6.55 - 6.38 (m, 1H), 5.90 (m, 1H), 5.00 - 4.73 (m, 1H), 4.33 (s, 1H), 3.80 - 3.62 (m, 3H), 2.91 (m, 3H), 2.62 - 1.78 (m, 5H). MS: 617.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri -din-2-yl)-N-(3-cyclopropylphenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 173
Figure imgf000126_0002
Ή NMR (400 MHz, CDC13): 68.76 (s, 1H), 8.59 (d, J= 4.8 Hz, 1H), 7.50-7.60 (m, 1H), 7.41 (d, J= 7.8 Hz, 1H), 7.28 - 7.19 (m, 2H), 7.14 - 6.94 (m, 2H), 6.62 - 6.79 (m, 1H), 6.26 - 6.07 (m, 2H), 4.86 (dd, J= 9.3, 2.9 Hz, 1H), 4.16 - 4.19 (m, 1H), 3.02-2.76 (m, 3H), 2.57 - 2.59 (m, 1H), 2.40-2.16 (m, 3H), 2.02 - 2.12 (m, 1H), 1.28 - 1.29(m, 2H), 0.90 (t,J= 6.9 Hz, 2H). MS: 604.2
(M+l)+.
(S)-N-((S)-l-(2-cChlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri -midin-2-yl)-N-(3-cyclopropylphenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 182
Figure imgf000127_0001
Ή NMR (400 MHz, CDC13): δ 8.94 (d, J= 4.5 Hz, 1H), 7.57 - 7.49 (m, 1H), 7.43 - 7.28 (m, 2H), 7.19 - 7.14 (m, 2H), 7.05 -6.79 (m, 4H), 6.51-6.46 (m, 1H), 6.00 - 5.97 (m, 1H), 4.82 - 4.80 (m, 1 H), 4.32 - 4.33 (m, 1 H), 3.09 - 2.81 (m, 3H), 2.64 - 2.24 (m, 4H), 2.05 - 1.72 (m, 2H), 0.99 - 0.76 (m, 4H). MS: 605.2 (M+l)+.
(S)-N-(3-(tert-Butyl)phenyl)-N-((S)-l-(2-chIorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2- oxoethyI)-l-(4-cyanopyrimidin-2-yI)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 165
Figure imgf000127_0002
1 H NMR (400 MHz, CDCI3): δ 8.94 (d, J = 4.8 Hz, 1 H), 8.00 - 7.54 (m, 1 H), 7.41 - 7.32 (m, 2H), 7.24 - 7.15 (m, 2H), 7.14 - 7.02 (m, 2H), 6.97 - 6.81 (m, 2H), 6.53 (s, 1H), 6.20 (dd, J = 12.7,6.8 Hz, 1H), 4.86 (m, 1H), 4.34 (s, 1H), 3.15 -2.80 (m, 3H), 2.63 - 2.27 (m, 4H), 2.13- 1.92 (m, 1H), 1.29 (s, 9H). MS: 621.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyri -din-2-yl)-N-(3-cyclopropyl-5-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 204
Figure imgf000127_0003
Ή NMR (400 MHz, CDC13): δ 8.77 (s, 1 H), 8.50 (s, 1 H), 7.50 - 7.33 (m, 2H), 7.24 -7.17 (m, 1H), 7.01 (m, 2H), 6.68 (m, 2H), 6.39 (m, 1H), 6.00 (s, 1H), 4.93 (s, 1H), 4.34 (s, 1H), 3.15 - 2.83 (m, 3H), 2.59 - 2.53 (m, 2H), 2.40 - 2.37 (m, 2H), 2.07 (s, 1H), 1.27 (s, 1H), 1.05 (s, 1H), 0.91 (d,J=6.7Hz, 1H), 0.67 (s, 1H), 0.43 (m, 1H). MS: 622.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(3-cyclopropyl-5-fluorophenyl)-5-oxopyrroIidine-2-carboxamide (single enantiomer) - Compound 202
Figure imgf000128_0001
Ή NMR (400 MHz, CDC13): δ 8.79 (s, 1H), 8.50 (s, 1H), 7.40 (m, 2H), 7.15 (m, 1H), 7.01 (m, 3H), 6.84 - 6.56 (m, 2H), 6.38 (m, 1H), 5.50 (s, 1H), 4.94 (s, 1H), 3.99 (s, 1H), 2.90 (m, 1H), 2.57 (m, 1H), 2.28 (s, 1H), 2.05 (m, 5H), 1.92-1.77 (m, 2H), 1.30 (m,2H), 0.91 (t,J=6.7 Hz, 2H), 0.67 (s, 2H). MS: 650.2 (M+l)+.
((S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocycIobutylamino)-2-oxoethyI)-l-(4- cyanopyri-din-2-yl)-N-(3-(N-methylsulfamoyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 157
Figure imgf000128_0002
Ή NMR (400 MHz, CD3OD): δ 8.89 - 8.59 (m, 3H), 8.50 - 8.01 (m, 2H), 7.69 - 7.31 (m, 5H), 7.17 (t,J= 7.6 Hz, 2H), 7.03 (t, J= 7.6 Hz, 2H), 6.95 (t, J= 7.9 Hz, 2H), 6.51 (s, 1H), 4.98 (s, 1H),4.24 (s, 2H), 3.01 -2.45 (m, 7H), 2.35 (s, 3H), 2.10-2.05 (m, 1H). MS: 657.1 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutyIamino)-2-oxoethyl)-l-(4-cyanopyri- din-2-yl)-N-(3-(N,N-dimethylsulfamoyl)phenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 156
Figure imgf000129_0001
Ή NMR (400 MHz, CDC13): δ 8.70 (s, IH), 8.60 (d, J= 4.9 Hz, IH), 8.17 (d, J = 7.7 Hz, IH), 7.86 (s, IH), 7.63 -7.55 (m, IH), 7.49 (t,J=7.8 Hz, IH), 7.27 (s, IH), 7.20 -6.92 (m, 4H), 6.50 (d, J= 6.9 Hz, 2H), 4.79 (d, J= 7.0 Hz, IH), 4.32 (s, IH), 3.05-2.75 (m, 4H), 2.60 - 1.90 (m, 10H). MS: 671.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(3- cyanopyri -din-2-yl)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single
Figure imgf000129_0002
Ή NMR (400 MHz, CDC13): δ 8.14 (d, J= 8.0 Hz, IH), 7.93 (d, J= 4.0 Hz, IH), 7.92 (m, IH), 7.17-7.28 (m,4H), 6.91-7.04 (m, 4H), 6.42 (s, IH), 6.31 (s, IH), 4.87-4.91 (m, IH), 4.35 (m, IH), 2.97-3.02 (m, 2H), 2.79 - 2.86 (m, IH), 2.45-2.57 (m, 3H), 2.23 - 2.26 (m, IH) , 2.09 - 2.11 (m, IH). MS: 582.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difIuorocyclobutyI)amino)-2-oxoethyl)-l-(4-cyano-3- fluoropyridin-2-yI)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - C mpound 82
Figure imgf000129_0003
Ή NMR (400 MHz, DMSO-d6): δ 8.36 (d,J= 4.7 Hz, IH), 7.70 (s, IH), 7.39 (m, 2H), 7.25-6.63 (m, 5H), 6.39 (s, IH), 5.96 (s, IH), 4.85 (s, IH), 4.34 (s, IH), 3.12-2.69 (m, 3H), 2.64-2.01 (m, 5H). MS : 600.0 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyano-3- fluoropyridin-2-yI)-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 83
Figure imgf000130_0001
Ή NMR (400 MHz, DMSO-d6): δ 8.37 (d, J= 4.6 Hz, IH), 7.75 (s, IH), 7.39 (m, 2H), 7.24-6.89 (m, 4H), 6.87-6.65 (d, IH), 6.50-6.27 (m, IH), 5.59-5.40 (m, IH), 4.92-4.75 (m, IH), 4.05-3.87 (m, IH), 2.95-2.68 (m, IH), 2.62-2.43 (m, IH), 2.41-2.25 (m, IH), 2.25-2.09 (m, 2H), 2.05-1.74 (m,4H), 1.59-1.24 (m, 3H). MS : 628.0 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano-3- fluoropyridin-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 88
Figure imgf000130_0002
Ή NMR (400 MHz, DMSO-d6): δ 8.73 (s, IH), 8.49 (m, IH), 7.96 (s, IH), 7.59-7.30 (m, 3H), 7.26-6.68 (m, 6H), 6.52-6.12 (m, IH), 5.96 (d, J = 10.5 Hz, 1H),4.95 (s, IH), 4.63 (m, 1H),4.49 (m, lH),4.22(s, I H), 4.14 -4.02 (m, 1 H), 3.46-2.65 (m, 4H), 2.55-2.00 (m, 2H), 1.69-1.49 (m, 2H). MS : 618.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(4,4-dinuorocyclohexylamino)-2-oxoethyl)-N-(3- fluorophen yl)-5-oxo-l-(pyrazin-2-yl)pyrrolidine-2-carboxamide (single enantiomer) - Compound 58
Figure imgf000130_0003
Ή NMR (400 MHz, CDC13): δ 9.74 (d,J= 1.5 Hz, 1H), 8.32 (m, 2H), 7.71 (s, 1H), 7.36 (m, 1H), 7.16 (m, 1H), 6.97 (m, 4H), 6.41 (s, 1H), 5.44 (d,J=7.0 Hz, 1H), 4.85 (d,J=6.0Hz, 1H), 3.96 (m, 1H), 2.98-2.82 (m, 1H),2.61 -2.48 (m, 1H), 2.35 - 2.21 (m, 1H),2.02 (m, 5H), 1.88 (m, 2H), 1.47-1.19 (m, 2H). MS : 586.2 (M+l)+.
2-(((S)-l-(2-ChIorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3-fluorophenyl) carbamoyl)-4-hydroxypyrrolidine-l-carboxylate (single enantiomer) - Compound 74
Figure imgf000131_0001
Ή NMR (400 MHz, CDC13): δ 8.60 (s, 1H), 7.89 (s, 1H), 7.71 (s, 1H), 7.45 - 7.29 (m, 2H), 7.25 -6.86 (m, 5H), 6.41 (s, 1H), 5.54 (s, 1H),4.98 (s, 1H), 3.98 (s, 1H), 3.16 - 2.66 (m, 2H),2.51 (s, 1H), 2.26 (s, 1H), 1.98 (m, 7H), 1.55 (m, 3H). MS: 591.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-N-(3-fluoro - phenyl)-2-oxo-3-(pyrimidin-2-yl)oxazolidine-4-carboxamide (single enantiomer) - Compound 76
Figure imgf000131_0002
Ή NMR (400 MHz, CDC13): δ 8.70 (d, J= 4.7 Hz, 2H), 7.67 (d, J= 8.0 Hz, 1H), 7.43 - 7.31 (m, 1H), 7.19 (d,J= 7.3 Hz, 1H), 7.13-6.86 (m, 5H), 6.46 (s, 1H), 5.58 (d, J= 6.8 Hz, 1H), 5.02 (d, J=4.4 Hz, 1H), 4.47 (dd, J= 8.7, 5.0 Hz, 1H), 4.24-4.13 (m, 1H),3.98 (s, 1H), 2.14 - 1.79 (m, 6H), 1.57-1.41 (m, 2H). MS: 588.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyri - din-2-yl)-N-(3-fluorophenyl)-2-oxooxazolidine-4-carboxamide (single enantiomer) - Compound 77
Figure imgf000132_0001
Ή NMR (400 MHz, CDCI3): δ 8.48 (s, 1H), 7.62 (d, J= 7.9 Hz, 1H), 7.33 (d, J= 8.9 Hz, 1H), 7.19 (d, J = 7.2 Hz, 2H), 7.10 - 6.85 (m, 5H), 6.44 (d, J= 5.1 Hz, 1H), 6.20 - 6.08 (m, 1H), 5.01 (m, 1 H), 4.46 (dd, J= 8.7, 4.7 Hz, 1H), 4.31-4.20 (m, 2H), 3.09 - 2.91 (m, 2H), 2.58 - 2.30 (m, 2H). MS : 584.1 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyano - pyridin-2-yl)-N-(3-fluorophenyl)-2-oxooxazolidine-4-carboxamide (single enantiomer) -
Compound 78
Figure imgf000132_0002
Ή NMR (400 MHz, CDC13): δ 8.55 (s, 1H), 8.50 (t, J= 5.8 Hz, 1 H), 7.67 (d, J= 8.5 Hz, 1H), 7.43 - 7.29 (m, 2H), 7.20 (d, J= 7.6 Hz, I H), 7.15 - 6.89 (m, 4H), 6.43 (d, J= 4.4 Hz, 1H), 5.54 (d,J=7.9Hz, 1H), 5.06 (d,J=4.7 Hz, 1H), 4.51 (dd, J= 8.8, 5.0 Hz, 1H),4.25 (m, 1H), 3.98 (s, 1H), 2.19 - 1.74 (m, 6H), 1.49 (m, 2H). MS: 612.2 (M+l)+.
(S)-N-((S)-l-(2-ChIorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-N-(3-cyano-5- fluorophenyl)-3-(3-cyanophenyl)-2-oxooxazolidine-4-carboxamide (single enantiomer) - Compound 134
Figure imgf000132_0003
'HNMR (400 MHz, CDCI3): δ 8.51 - 8.47 (m, 1H), 8.39-8.37 (d, 0.5H), 8.07-7.99 (m, 1H), 7.38 (s, 0.5H).7.33-7.31 (m, 1H), 7.26-7.22 (m, 1H), 7.08-7.07 (m, 1H), 6.90-6.87 (m, 1H), 6.53- 6.46 (m, 2H), 4.94 - 4.91 (m, 1 H), 4.44 - 4.40 (m, 1 H).4.34 - 4.32 (m, 1 H), 4.28 - 4.23 (m, 3.00 -2.99(m, 2H), 2.50 -2.43(m, 2H). MS: 608.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4- cyanopyri -din-2-yl)-N-(5-fluoropyridin-3-yI)-2-oxooxazolidine-4-carboxamide (single enantiomer) - Compound 135
Figure imgf000133_0001
Ή NMR (400 MHz, CDC13): δ 8.58 - 8.28 (m, 3H), 8.08 (d, J= 8.5 Hz, IH), 7.32 (dd, J= 5.1, 1.0 Hz, 2H), 7.28-7.20 (m, IH), 7.07 (m, IH), 6.91 (m, IH), 6.66-6.22 (m, 2H), 5.05 - 4.85 (m, IH), 4.57-4.09 (m, 3H), 3.02 (m, 2H), 2.69-2.30 (m, 2H). MS: 585.1 (M+l)+.
(S)-N-((S)-l-(2-ChlorophenyI)-2-((4,4-dinuorocyclohexyl)amino)-2-oxoethyl)-3-(4- cyanopyri -din-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxooxazolidine-4-carboxamide (single enantiomer) - Compound 132
Figure imgf000133_0002
Ή NMR (400 MHz, CDC13): δ 8.91 (s, IH), 8.41 (m, 4H), 8.11 (s, IH), 7.23 (s, 1H),7.05 (s, IH), 6.91 (s, IH), 6.52 (m, IH), 6.05 (m, IH), 4.95 (m, IH), 4.37 (m, 2H), 3.95 (s, IH), 1.71 (m, 10H). MS: 613.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-N-(3-fluoro phenyl)-2-oxo-3-(thiazol-4-yl)oxazolidine-4-carboxamide (single enantiomer) - Compound
72
Figure imgf000133_0003
Ή NMR (400 MHz, CDC13): δ 8.70-8.47 (m, 1H), 7.69-7.52 (m, 1H), 7.49 (d, J= 2.0 Hz, 1H), 7.42-7.26 (m, 1H), 7.25-6.84 (m, 5H), 6.42 (s, 1H), 6.21-6.02 (m, 1H), 5.03 (d,J=4.6Hz, 1H), 4.42 (m, 1H), 4.38- 4.05 (m, 2H), 2.98 (m, 2H), 2.64-2.29 (m, 2H). MS : 565.1 (M+l)+.
(4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-2-oxooxazolidine-4- carboxamide (racemic) - Compound 145
Figure imgf000134_0001
Ή NMR (400 MHz, CDC13): δ 8.63 - 8.50 (m, 1H), 8.42 (m, 1H), 7.48 - 7.40 (m, 1H), 7.29 (d, J = 7.0 Hz, 2H), 7.25 - 7.19 (m, 2H), 7.14 - 6.95 (m, 3H), 6.89 (m, 1H), 6.67 (d, J= 6.9 Hz, 1H), 6.54-6.42 (m, 1H), 5.11 -4.96 (m, 1H), 4.51 -4.40 (m, 1H), 4.32 (d,J=9.1 Hz, 1H),4.24- 4.09 (m, 1H), 3.12 - 2.73 (m, 2H), 1.52 (m, 2H), 1.32 (d, J= 9.0 Hz, 4H). MS: 642.2 (M+l)+. (4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4- cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxo-l,3-oxazinane-4-carboxamide (racemic) - Compound 90
Figure imgf000134_0002
Ή NMR (400 MHz, CDC13): δ 8.57 (s, 1H), 8.40(s, 1H), 7.68 (d,J=8.0 Hz, 1H), 7.25 - 6.91 (m, 8H), 6.48 (s, 1H), 6.25 (s, 1H), 5.08 (s, 1H), 4.51 -4.46 (m, 1H), 4.31 (m, 2H), 3.01 (m, 2H), 2.53 -2.50 (m, 2H), 2.29 - 2.13 (m, 2H). MS: 598.1 (M+l)+.
(S)-N-((S)-l-(2-ChlorophenyI)-2-((3,3-difluorocycIobutyI)amino)-2-oxoethyl)-3-(4-cyanopy - ridin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-l,3-oxazinane-4-carboxamide (single enantiomer) - Compound 133
Figure imgf000135_0001
Ή NMR (400 MHz, CDC13): δ 8.55 (d, J= 5.0 Hz, 1H), 8.34 (s, 1H), 7.54 (d, J= 8.4 Hz, 1H), 7.31 (dd,J=5.0, 1.1 Hz, 1H), 7.26- 7.16 (m, 2H), 7.13 - 7.04 (m, 1H), 6.98 (t,J=6.6Hz, 2H), 6.72-6.63 (m, 1H),6.49 (s, 1H),6.44 (d,J=6.9 Hz, 1 H), 5.11 (dd, J= 6.4, 3.5 Hz, 1H),4.51 - 4.22 (m, 3H), 2.98-3.04 (m, 2H), 2.67 - 2.41 (m, 2H), 2.33 - 2.09 (m, 2H).MS: 627.2 (M+l)+. (S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocycIohexyl)amino)-2-oxoethyl)-3-(4-cyano pyridin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-l,3-oxazinane-4-carboxamide (single
enantiomer) - Compound 139
Figure imgf000135_0002
Ή NMR (400 MHz, CDC13): 88.64 (d, J= 5.0 Hz, 1H), 8.47 (s, 1H), 7.45 (d, J= 7.4 Hz, 1H), 7.38 - 7.30 (m, 2H), 7.24 (d,J=7.1 Hz, 1H), 7.15-7.12 (m, 1H), 6.81-6.77 (m, 1H), 6.06 (s, 1H), 5.51 (d,J=7.5 Hz, 1 H), 5.05 - 4.88 (m, 1H), 4.62-4.56(m, 1H), 4.42 -4.30 (m, 1H), 3.87 (s, 1H), 2.35 -2.15 (m,2H), 1.97-1.79 (m, 5H), 1.40 (m,2H). MS: 643.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-3-(4-cyano pyrimidin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-l,3-oxazinane-4-carboxamide (single enantiomer) - Compound 144
Figure imgf000135_0003
Ή NMR (400 MHz, CDC13): δ 8.96 (d,J=4.7Hz, 1H), 7.56 (d,J= 10.0 Hz, 1H), 7.41 (dd,J = 9.7, 6.4 Hz, 2H), 7.24-7.22 (m, 1H), 7.14 - 6.95 (m, 3H), 6.70 (t, J= 8.6 Hz, 1H), 6.52 (s, 1H), 5.53 (d,J=7.6 Hz, 1H), 4.96 (dd, J= 7.8, 4.0 Hz, 1H), 4.46 (d,J=8.8Hz, 1H), 4.31 (dd,J = 10.7,5.1 Hz, 1H), 3.99 (s, 1H), 2.49-2.31 (m, 1H), 2.29 - 2.01 (m, 5H), 1.98- 1.78 (m, 2H), 1.49(dd,J = 17.9,8.5 Hz, 1H). MS: 645.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-3-(4-cyanopy - rimidin-2-yl)-N-(3,5-difluorophenyl)-2-oxo-l,3-oxazinane-4-carboxamide (single
enantiomer) - Compound 154
Figure imgf000136_0001
Ή NMR (400 MHz, CDC13): δ 8.89 (d, J= 4.8 Hz, 1H), 7.52 (d, J- 8.9 Hz, 1H), 7.40 (d, J= 4.8 Hz, 1H), 7.22 (dd,J=8.0, 1.2 Hz, 1H), 7.16-7.15 (m, 1H), 7.08 - 6.97 (m, 2H), 6.94 (dd,J=7.7, 1.5 Hz, 1H), 6.66 (dd, J= 9.7, 7.4 Hz, 1H), 6.56 (s, 1H), 6.43 (d,J=6.8 Hz, 1H), 4.91 (dd,J = 8.3, 4.5 Hz, 1H), 4.41-4.33 (m, 2H), 4.24-4.20 (m, 1H), 3.06-2.86 (m, 2H), 2.66-2.42 (m, 2H), 2.39- 2.25 (m, 1H), 2.24 - 2.12 (m, 1H). MS: 617.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyI)-3-(4-cyano pyridin-2-yl)-N-(5-fluoropyridin-3-yI)-2-oxo-l,3-oxazinane-4-carboxamide (single enantiomer) - Compound 143
Figure imgf000136_0002
Ή NMR (400 MHz, CDCI3): δ 9.08 - 7.79 (m, 3H), 7.62 - 6.70 (m, 5H), 6.50 (m, 2H), 4.95 (m, 1H), 4.62-4.03 (m, 3H), 2.99 (s, 2H), 2.51 (s, 2H), 2.18 (m, 2H). MS: 599.1 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocycIohexyl)amino)-2-oxoethyl)-3-(4-cyano - pyridin-2-yl)-N-(5-fluoropyridin-3-yl)-2-oxo-l ,3-oxazinane-4-carboxam ide (single enantiomer) - Compound 137
Figure imgf000137_0001
Ή NMR (400 MHz, CDC13): δ 8.43-8.90 (m, 3H), 8.30 (s, IH), 7.49-8.13 (m, I H), 7.29-7.31 (m, 2H), 7.17-7.21 (m, I H), 6.94-7.08 (m, 2H), 6.45-6.53 (m, I H), 5.80-593 (m, IH), 4.96-5.00 (m, 1 H), 4.47-4.51 (m, 1 H), 4.30 - 4.33 (m, 1 H), 3.96-3.98 (m, 1 H), 2.09 - 2.28 (m, 6H) , 1.83 - 1.95 (m, 2H), 1.49 - 1.63 (m, 2H). MS: 627.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-3-(4-cyanopyri din-2-yl)-N-(3-fluoro-5-(2-hydroxypropan-2-yl)phenyl)-2-oxo-l,3-oxazinane-4-carboxamide (single enantiomer) - Compound 146
Figure imgf000137_0002
Ή NMR (400 MHz, CDC13): δ 8.56 (t, J= 6.0 Hz, I H), 8.36 (s, I H), 7.72 - 7.45 (m, IH), 7.23 -
7.16 (m, I H), 7.12 (t, J= 7.1 Hz, I H), 7.06 - 6.86 (m, 3H), 6.38 (s, IH), 6.28 (d, J= 6.9 Hz, I H),
5.17 - 5.01 (m, I H), 4.50 - 4.44 (m, I H), 4.30 (m, 2H), 2.99 (d, J= 7.8 Hz, 2H), 2.62 - 2.37 (m, 2H), 2.36 - 2.06 (m, 2H), 1 .49 (d, J= 6.2 Hz, 2H), 1.32 (m, 4H). MS: 656.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocycIobutyIamino)-2-oxoethyl)-l-(4-cyanopyri - din-2-yl)-N-(3-fluorophenyl)-6-oxopiperidine-2-carboxamide (single enantiomer) - Compound 55
Figure imgf000137_0003
Ή NMR (400 MHz, CDC13): δ 8.59 (s, I H), 8.28 (s, I H), 7.72 (d, J= 7.2 Hz, I H), 7.43 - 7.33 (m, 1 H)„ 7.26 - 7.12 (m, 2H), 7.1 1 - 6.96 (m, 2H), 6.89 (dd, J= 8.3, 2.2 Hz, IH), 6.46 (s, I H), 6.27 (s, 1H), 5.00 (t,J= 4.6 Hz, 1H), 4.37-4.28 (m, 1H), 3.13-2.95 (m, 2H), 2.78-2.69 (m, 1H), 2.62-2.35 (m, 3H), 2.15-2.09 (m, 1H), 2.05 - 1.92 (m, 1H), 1.89 - 1.70 (m, 3H). MS: 596.2 (M+l)+.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyano - pyridin-2-yI)-N-(3-fluorophenyl)-6-oxopiperidine-2-carboxamide (single enantiomer) - Compound 75
Figure imgf000138_0001
Ή NMR (400 MHz, CDC13): δ 8.60 (s, 1H), 8.31 (s, 1H), 7.73-7.75 (m, 1H), 7.30 (m, 1H), 7.00- 7.17 (m, 5H), 6.87-6.91 (m, 1H), 6.45 (s, 1H), 5.50 (d, J= 7.0 Hz, 1H), 5.00-5.02 (m, 1H), 3.99 (m, 1 H), 2.60 - 2.74 (m, 1 H), 2.58-2.60 (m, 1 H), 2.01 - 2.14 (m, 6H) , 1.83 - 1.92 (m, 4H), 1.42 - 1.46 (m, 3H). MS: 624.2 (M+l)+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yI)-4-fluoro-N-(3-fluorophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 151
Figure imgf000138_0002
Ή NMR (400 MHz, CDC13): δ 8.75 (s, 1H), 8.57 (s, 1H), 7.76 (s, 1H), 7.36 (m, 2H), 7.06 (m, 6H), 6.39 (s, 1H), 5.51 (d,J=6.6 Hz, 1H), 5.12 (m, 1H),4.82 (s, 1H), 3.91 (m, 1H), 2.69 - 2.26 (m, 2H), 2.05 (m, 6H), 1.53 - 1.38 (m, 2H). MS: 628.2 (M+l)+.
Example 9. Preparation of (2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2- oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 191
Figure imgf000139_0001
Step A: 5-Nitroisophthaloyl dichloride. To a solution of 5-nitroisophthalic acid (2.3 g, 1 1 mmol) in S0C12 (6 mL) was added a drop of DMF and the mixture was stirred at reflux for 3 hr. The resulting reaction mixture was concentrated to give the crude product which was used directly in the next step.
Step B: 5-Nitroisophthalamide. 5-Nitroisophthaloyl dichloride (2.7 g, 9.7 mmol) was added portionwise to a cold solution of NH3 'H20 (40 mL) at 0 °C. The reaction mixture was stirred overnight and a white precipitate formed. The mixture was then filtered, washed with excess of water, and dried at 1 10 °C to give the crude product which was used directly in the next step. Step C: 5-Aminoisophthalamide. To a solution of 5-nitroisophthalamide (2 g, 9.6 mmol) in MeOH (200 mL) was added Pd/C (200 mg). The reaction was stirred overnight under a hydrogen atmosphere. The suspension was filtered and the filtrate was concentrated to afford the desired product which was used directly in the next step.
StepD: 5-((2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)l-(4cyano pyridin-2-yl)-5-oxopyrrolidine-2-carboxamido)isophthaIamide. A mixture of 2- chlorobenzaldehyde ( 1 .0 mL, 7.3 mmol) and 5-aminoisophthalamide ( 1.3 g, 7.3 mmol) was stirred at room temperature for 30 min under N2, followed by addition of (S)- l -(4-cyanopyridin- 2-yl)-5-oxopyrrolidine-2-carboxylic acid ( 1.7 g, 7.3 mmol). After stirring for 10 min, 1 , 1 - difluoro-3-isocyanocyclobutane (854 mg, 7.3 mmol) was added. The mixture was then stirred overnight and filtered and purified by a standard method to give the title product.
Step E: (2S)-N-(l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoet yl)l-(4-cyano pyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide. To a mixture of 5-((2S)- N-( l -(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl) l -(4-cyano pyridin-2-yl)-5- oxopyrrolidine-2-carboxamido)isophthalamide (850 mg, 1.3 mmol) in pyridine (0.62 mL, 7.8 mmol) and DCM (10 mL) was added TFAA (0.9 mL, 6.5 mmol). The reaction solution was stirred at room temperature overnight. The resulting mixture was concentrated and the residue was purified by a standard method to afford the title product. Ή NMR (400 MHz, CDC13): δ 8.77 (s, 1H), 8.62-8.42 (m, 2H), 7.87 (s, 1H), 7.75 (s, 1H), 7.40 (d, J= 7.8 Hz, 1H), 7.31 (d,J = 4.2 Hz, 1H), 7.25 (d,J=8.1 Hz, 1H), 7.10 (t, J= 7.3 Hz, 1H), 6.92 (d,J=7.5Hz, 1H), 6.47 (s, I H), 6.11 (d, J= 6.6 Hz, 1 H), 4.73 (dd, J= 9.4, 2.7 Hz, 1 H), 4.35 (s, 1 H), 3.14 - 2.82 (m, 3H), 2.68-2.31 (m, 3H),2.19(m, 1H), 2.09-1.91 (m, 1H). MS: 614.1 (M+l)+.
The following analogs were synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods.
(S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difIuorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(3,5-dicyanophenyl)-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 153
Figure imgf000140_0001
Ή NMR (400 MHz, CDC13): δ 8.74 (s, 1H), 8.53 (m, 2H), 7.81 (m, 2H), 7.48 - 7.16 (m, 4H), 7.09 (t,J= 7.5 Hz, 1H), 6.90 (d, J= 7.6 Hz, 1H), 6.46 (s, 1H), 6.17 (d, J= 6.7 Hz, lH),4.72(dd, J= 9.1, 2.3 Hz, 1H), 4.35 (s, 1H), 3.18 -2.71 (m, 3H), 2.68 - 1.83 (m, 5H). MS: 614.1 (M+l)+.
Example 10. Preparation of (S)-tert-butyl 3-(((S)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl) amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-5-oxopiperazine-l- carboxylate (single enantiomer) - Compound 97
Compound 97 was synthesized via the UGI reaction procedure set forth herein, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring and purified via standard methods.
Figure imgf000141_0001
Ή NMR (400 MHz, CDC13): δ 8.75 - 8.44 (m, 2H), 7.81-7.41 (m, 1H), 7.46-7.35 (m, 2H), 7.24 (t, J= 7.2 Hz, 1H), 7.16-6.97 (m, 2H), 6.84-6.75 (m, 2H), 6.43-5.82 (m, 1H), 5.09-4.98 (m, 1H), 4.77-4.73 (m, 1H), 4.48 (d,J= 13.5 Hz, 1H), 4.27-4.07 (m, 2H), 3.45-2.76 (m, 4H), 1.54 (s, 9H). MS : 613.2 (M+l)+.
Example 11. Preparation of (3S)-tert-butyI 3-((l-(2-chIorophenyl)-2-((3,3- difluorocyclobutyl) amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4- cyanopyrimid -2-yl)-5-oxopiperazine-l-carboxylate (racemic) - Compound 98
Figure imgf000141_0002
A mixture of (3S)-tert-butyl3-((l -(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl) (3,5-difluorophenyl)carbamoyl)-5-oxopiperazine-l-carboxylate (200 mg, 0.326 mmol), 2- bromopyrimidine-4-carbonitrile (0.489 mmol), Pd2(dba)3 (30.2 mg, 0.0323 mmol), XantPhos (19.1 mg, 0.03 mmol) and Cs2C03 (148.7 mg, 0.46 mmol) in 1,4-dioxane (10 mL) was stirred at 80 °C for 3 hr under N2. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated and the residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): δ 8.97 (d, J= 4.3 Hz, 1H), 7.85-7.55 (d, 1H), 7.51-7.39 (m, 2H), 7.25 (t, J= 7.6 Hz, 1H), 7.13-6.26 (m, 6H), 5.91 (d, J= 7.6 Hz, 1H), 4.92-4.08 (m, 5H), 3.38 (t,J= 14.9 Hz, 1H), 3.02 (s, 2H), 2.83-2.22 (d, 2H), 1.61 (s, 9H). MS : 716.1 (M+l)+.
The following analogs were synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocynide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods. (S)-tert-Butyl 3-(((S)-l-(2-chlorophenyl)-2-((4,4-difluorocyclo exyl)amino)-2-oxoethyl)(3,5- difluorophenyl)carbanioyl)-4-(4-cyanopyrimidin-2-yl)-5-oxopiperazine-l-carboxyIate (chiral) - Compound 93
Figure imgf000142_0001
H NMR (400 MHz, CDC13): δ 8.96 (d, J= 4.3 Hz, IH), 7.83 (s, IH), 7.43 (m, 2H), 7.21 (t, J = 7.4 Hz, IH), 7.08-6.62 (m, 4H), 6.63 - 6.37 (m, IH), 5.93 (m, IH), 4.85 (t,J=3.6Hz, IH), 4.63-4.23 (m, 2H), 4.16 (m, IH), 3.93 (s, IH), 3.43 (m, IH), 2.24 - 1.91 (m, 5H), 1.79 (m, 3H), 1.60 (m, IH). MS : 744.2 (M+l)+.
(3S)-tert-Butyl 3-((l-(2-chlorophenyl)-2-((3,3-difIuorocyclobutyI)amino)-2-oxoethyI)(3- fluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-l-carboxylate (single enantiomer) - Compound 89
Figure imgf000142_0002
Ή NMR (400 MHz, DMSO-d6): δ 8.80-8.37 (m, IH), 8.05-7.57 (m, IH), 7.58-7.31 (m, 3H), 7.21 (s, IH), 7.16-6.89 (m, 3H), 6.90-6.68 (m, IH), 6.67-6.30 (m, IH), 6.22 -5.84 (m, IH), 5.09- 4.87(m, IH), 5.83-4.57 (m, IH), 4.50 (m, IH), 4.25 (s, IH), 4.08 (m, IH), 3.50-2.70 (m, 4H), 2.60-2.10 (m, IH), 1.70 (s, 2H), 1.54 (m, IH). MS : 697.2 (M+l)+.
Example 12. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3- difluorocycIobutyl)amino)-2-oxoethyi)-l-(4-cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-6- oxopiperazine-2-carboxamide (single enantiomer) - Compound 99
Figure imgf000143_0001
TFA (0.3 mL) was added to a solution of (S)-tert-butyl 3-(((R)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)- 5-oxopiperazine-l-carboxylate (60 mg, 0.08 mmol) in DCM (1.0 mL) at 0 °C. The mixture was warmed to room temperature and stirred for I hr, and then concentrated. The residue was purified by a standard method to give the desired product. Ή NMR (400 MHz, CDC13): δ 8.94 (t, J=4.6 Hz, 1H), 7.48-7.36 (m, 3H), 7.21 (m,J=7.8, 1.5 Hz, 1H), 7.12-6.94 (m, 3H), 6.71-6.55 (m, 2H), 6.05 (d,J=6.7 Hz, 1H), 4.73 (q,J=4.0, 2.1 Hz, 1H), 4.36 (d, J= 6.7 Hz, 1H), 3.77- 3.65 (m, 2H), 3.50-3.35 (m, 1H), 3.18 (m, 1H), 3.12-2.96 (m, 2H), 2.64-2.35 (m, 2H). MS : 616.1 (M+l)+.
The following compound was synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods.
(S)-N-((S)-l-(2-ChIorophenyI)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyl)-l-(4-cyano pyridin-2-yl)-N-(3,5-difluorophenyl)-6-oxopiperazine-2-carboxamide (single enantiomer) - Compound 100
Figure imgf000143_0002
Ή NMR (400 MHz, CDC13): δ 8.68 - 8.28 (m, 1H), 7.61 - 7.28 (m, 2H), 7.20 (dd, J= 7.9, 1.3 Hz, OH), 7.02 - 6.90 (m, 1H), 6.66 (tt, J= 8.6, 2.3 Hz, 1H), 6.49 (d, J= 2.7 Hz, OH), 6.09 (m, 1H),4.90 (dd,J=3.8,2.0Hz, 1H), 4.42 -4.16 (m, 1H), 3.71 (m, 1H), 3.50 - 3.23 (m, 1H), 3.18 -2.78 (m, 2H), 2.63-2.13 (m, 2H). MS : 615.2 (M+l)+. Example 13. (S)-4-Acetyl-N-((S)-l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxo ethyl)-l-(4-cyanopyridin-2-yl)-N-(3-fluorophenyI)-6-oxopiperazine-2-carboxamide (single enantiomer) - Compound 92
Figure imgf000144_0001
To a solution of (3S)-tert-butyl 3-(( l -(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2- oxoethyl)(3-fluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine-l -carboxylate ( 100 mg, 0.14 mmol) in DCM (3 mL) was added TFA dropwise ( 1 mL) at 0 °C. The mixture was stirred at room temperature for 2 hr and then concentrated. The residue was dissolved in DCM and cooled to 0 °C. DIPEA (0.055 mL, 0.34 mmol) was added to the mixture followed by Ac20 (0.03 1 mL, 0.34 mmol) at 0°C. Then the mixture was stirred at room temperature for 2 hr. The solution was concentrated and the residue was purified by a standard method to afford the desired product. Ή N R (400 MHz, CDC13): δ 8.54 (s, 2H), 7.70-744 (m , 2H), 7.36 (m, 2H), 7.20 (t, J = 7.2 Hz, 1 H), 7.14 - 6.99 (m, 2H), 6.94 (t, J= 7.4 Hz, 1 H), 6.80 (s, 1 H), 6.66 (d, J = 7.8 Hz, 1 H), 6.58 - 6.42 (m, 1 H), 5.09 (dt, J = 5.2, 3.1 Hz, 1 H), 4.93 (m, 1 H), 4.63 (m, 1 H), 4.54 - 4.41 (m, 1 H), 4.35-4.3 1 (m, 1 H), 3. 16 (s, 1 H), 3. 12 - 2.96 (m, 3H), 2.86 (s, 1 H), 2.25 (s, 3H). MS: 639.2 (M+ l )+.
Example 14. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-4-cyclopropyl-N-(3,5- difluorophenyl -6-oxopiperazine-2-carboxamide (single enantiomer) - Compound 106
Figure imgf000144_0002
TFA (0.3 mL) was added to a solution of (S)-tert-butyl 3-(((R)- l -(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)- 5-oxopiperazine-l-carboxylate (60 mg, 0.084 mmol) in DCM (1.0 mL) at 0 °C. The mixture was stirred at room temperature for 1 hr then concentrated. The residue was dissolved in MeOH (2 mL) followed by addition of (l-ethoxycyclopropoxy)trimethylsilane (88 mg, 0.50 mmol), AcOH (50 mg, 0.84 mmol) and NaBH3(CN)(27 mg, 0.42 mmol). The resulting suspension was stirred at 80 °C under N2 for 1.5 hr. The reaction mixture was partitioned between EtOAc and H20. The organic layer was separated, washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): δ 7.64 (d, J= 7.8 Hz, 1H), 7.30 (d, J= 5.3 Hz, 2H), 7.19 (s, 1H), 7.07 (s, 3H), 6.66 (s, 1H), 6.32 (s, 1H), 6.09 (m, 1H), 5.09 (s, 1H), 4.28 (s, 1H), 3.76 - 3.59 (m, 1H), 3.46- 3.33 (m, 1 H), 3.08-2.89 (m, 4H), 2.59 - 2.31 (m, 2H), 0.94 (s, 1 H), 0.61-0.37 (m, 4H). MS: 655.2 (M+l)+.
Example 15. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4- methyl-6-oxopiperazine-2-carboxamide (single enantiomer) - Compound 101
Figure imgf000145_0001
TFA (0.6 mL) was added to a solution of (3S)-tert-butyl 3-((l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)- 5-oxopiperazine-l-carboxylate (30 mg, 0.042 mmol) in DCM (2 mL) at 0 °C. The mixture was stirred at room temperature for 1 hr and then concentrated. The residue was dissolved in MeCN (4 mL) followed by addition of K2CO3(10 mg, 0.072 mmol) and iodomethane (2 mL). The resulting mixture was stirred at room temperature for 2 hr and then concentrated. The residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13) : δ 8.60 (m, 2H), 7.80 (s, 1H), 7.45 (d, J= 7.9 Hz, 1H), 7.33 (m, 1H), 7.07 (d, J= 4.3 Hz, 2H), 6.74 (t,J=8.6Hz, 1H), 6.48-5.91 (m, 3H), 4.92 (t, J= 4.7 Hz, 1H),4.20 (m, 1 H), 3.61 -3.40(m, 1 H), 3.14 (m, 1 H), 3.02 - 2.77 (m, 3H), 2.71 (m, 1 H), 2.42 - 2.26 (m, 5H), 2.04 (d, J= 9.0 Hz, 1H). MS: 629 (M+l)+. Example 16. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-(2- hydroxyethyl)-6-oxopiper azine-2-carboxamide (single enantiomer) - Compound 107
Figure imgf000146_0001
To a solution of (S)-tert-butyl 3-(((S)- l -(2-chlorophenyl)-2-((3,3-difluorocyclobutyl) amino)-2- oxoethyl)(3,5-difluorophenyl)carbamoyl)-4-(4-cyanopyridin-2-yl)-5-oxopiperazine- l -carboxyl ate (30 mg, 0.04 mmol) in DCM (3 mL) was added TFA ( 1 mL) at 0 °C. The mixture was stirred at room temperature for 1 hr and concentrated in vacuo. The residue was dissolved in EtOH (3 mL) followed by addition of TBA1 ( 16 mg, 0.04 mmol), Et3N ( 10 mg, 0.1 mol) and 2- bromoethanol (7 mg, 0.056 mmol). The resulting mixture was stirred at 85 °C for 3 hr and then filtered. The filtrate was concentrated and the residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): δ 8.96 (t, J = 4.6 Hz, 1 H), 7.53-7.36 (m, 3H), 7.23 (m, J = 7.8, 1 .5 Hz, 1 H), 7.14-6.94 (m, 3H), 6.68 (m, J = 8.6, 2.3 Hz, 1 H), 6.60 (d, J = 3. 1 Hz, 1 H), 6.07 (d, J = 6.7 Hz, 1 H), 4.75 (q, J= 4.0, 2.1 Hz, 1 H), 4.38 (d, J = 6.7 Hz, 1 H), 3.78-3.67 (m, 2H), 3.39 (m, 1 H), 3.26-2.92 (m, 3H), 2.67-2.36 (m, 2H). MS : 659.2 (M+ l )+.
The following compound was synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods.
Compound 104
Figure imgf000146_0002
Ή NMR (400 MHz, CDC13): δ 8.60-8.56 (m, 2H), 7.47-7.28 (m, 3 H), 7.22-7.01 (m, 4H), 6.72- 6.67 (m, 1 H), 6.54-6.44 (m, 2H), 5.24 (m, 1 H), 4.37-4.13 (m, 3H), 3.63-2.97 (m, 8H), 2.44-2.06 (m, 2H), 1 .34- 1 .28 (m, 3H). MS : 701.2 (M+ l )+.
Example 17. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)-l-(5-cyanooxa-zol-2-yl)-N-(3,5-difluoroph oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 162
Figure imgf000147_0001
Step A .- Oxazole-5-carboxamide. Ethyl oxazole-5-carboxylate (2 g, 14.2 mmol) was dissolved in NH3 solution (7 M in MeOH, 25 mL). The solution was stirred at room temperature for 2 hr and filtered. The solid was dried to give the desired product ( 1 .5 g, 92% yield) as a white powder which was used directly in the next step.
Step B : 2-Iodooxazole-5-carboxamide. Oxazole-5-carboxamide (560 mg, 5.0 mmol) was dissolved in anhydrous THF (7.5 mL) and flushed with N2. The solution was cooled to -42 °C and treated with fresh LiHMDS ( 15 mL, 1 M in THF). The solution became dark yellow was stirred for 20 min and fol lowed by the addition of a solution of ZnCl2 (30 mL, 0.5 M in THF). The reaction was warmed to 0 °C for 1 hr. After solid iodine ( 1 .65 g, 6.5 mmol) was added, the reaction mixture was stirred at room temperature for another 1 hr and then poured into saturated Na2S203 solution containing 25% aq. NH3 solution. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous a2S04 and concentrated. The resulting residue was purified by a standard method to give the desired product. MS : 239.0 (M+ l )+.
Step C: 2-((S)-2-(((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)a
difluorophenyl)carbamoyl)-5-oxopyrrolidin-l-yl)oxazole-5-carboxamide. The product was prepared by the general procedure for the Buchwald reaction. Ή NMR (400 MHz, CDC13): δ 7.59 (s, 1 H), 7.53 (s, 1 H), 7.37 (d, J - 7.9 Hz, 1 H), 7.20 (t, J = 7.0 Hz, 1 H), 7.04 (t, J = 7.6 Hz, 1 H), 6.96 (d, J = 7.9 Hz, 2H), 6.68 (t, J = 8.7 Hz, 1 H), 6.46 (s, 1 H), 6.36 (d, J = 6.4 Hz, 1 H), 5.68 (s, 1H), 4.82 (dd, J= 9.3, 2.6 Hz, lH), 4.33 (s, 1H), 4.16 -4.09 (m, 1H), 3.03-3.00 (m, 2H), 2.90 - 2.77 (m, 1H), 2.62 -2.35 (m, 3H), 2.29-228 (m, 1H), 2.19 - 2.08 (m, 1H). M S: 608.1 (M+l)+.
Step D : ( S)-N-( ( S)-l-( 2-Chloroph enyl )-2-( ( 3,3-difluomcyclobutyl )amino)-2-oxoethyl )-!-( 5- cyanooxazol-2-yl)-N-(3,5-difluorophenyl)-5-oxopyrrolidine-2- rboxamide.
2-((S)-2-(((S)-l-(2 hlorophenyl>2-((3,3-difiuorocyclobutyl)amino)-2 xoethyl)(3,5-difluoro- phenyl)carbamoyl)-5-oxopyrrolidin-l-yl)oxazole-5-carboxamide (100 mg, 0.16 mmo l) was dissolved in DCM (3 mL) and dry pyridine (0.8 mL). TFAA (0.1 mL) was added and the reaction solution was stirred for 25 min at room temperature and then concentrated in vacuo. The residue was dissolved in EtOAc and washed with H20, saturated aq. NaHC03 and brine. The organic phase was separated, dried over anhydrous Na2S04, and concentrated. The residue was purified by a standard method to give the desired product.1!! NMR (400MHz, CDC13): δ 7.63 (s, 1H), 7.55 (d, J= 7.0 Hz, 1H), 7.41 (d, J= 7.1 Hz, 1H), 7.25 (td, J= 7.8, 1.5 Hz, 1H), 7.08 (t, J = 7.6 Hz, 1H), 6.98 - 6.91 (m, 1H), 6.80 (d, J= 6.7 Hz, lH), 6.71 (dd, J= 9.7, 7.4 Hz, lH), 6.49 (s, 1H), 5.97 (d, J = 6.8 Hz, 1H), 4.80 (dd, J = 9.3, 2.8 Hz, 1H), 4.36 (s, 1H), 3.06-3.03 (m, 2H), 2.92 - 2.79 (m, 1H), 2.62 -2.29 (m, 4H), 2.18-2.12 (m, 1H). MS: 590.1 (M + l)+.
Example 18. Preparation of (2S ,4R)-N-(l-(2-chlorophenyl)-2-((3,3- difluorocyclobutyl)amino)-2-oxoethyl)-N-(3-cyano-phenyl)-l-(4-cyanopyridin-2-yl)-4- hydroxy-5-oxopyrrolidine-2-carboxamide (racemic) - Compound 170
Figure imgf000149_0001
Step A: (2S,4R)-l-tert-Butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)pyrrolidine-l,2- dicarboxylate. Imidazole (2.8 g, 40.8 mmol) was added to a solution of (2S,4R)- l -tert-butyl 2- methyl 4-hydroxypyrrolidine- l ,2-dicarboxylate (5.0 g, 20.4 mmol) and TBSC1 (4.6 g, 30.6 mmol) in anhydrous DMF ( 100 mL). The mixture was stirred at room temperature overnight and then partitioned between EtOAc and H20. The organic layer was separated, washed with aq. LiCl ( 10%) and brine, dried over anhydrous a2S04, and then concentrated. The residue was purified by column chromatography to afford the desired product as a colorless oil. MS: 360.2 (M+ l )+. Step B : (2S,4R)-l-tert-Butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-l,2- dicarboxylate. To a solution of NaI04 (7.5 g, 35.0 mmol) in water (80 mL) was added Ru02 (370 mg, 2.8 mmol) under the atmosphere of nitrogen. The resulting green-yellow solution was stirred for 5 min followed by addition of (2S,4R)- l -tert-butyl-2-methyl4-((tert-butyldimethyl silyl)oxy)pyrrolidine- l ,2-dicarboxylate (5.0 g, 14.0 mmol) in EtOAc (44 mL) in one portion. The mixture was stirred at room temperature overnight. The resulting mixture was then diluted with EtOAc and filtered through a pad of Celite. The organic layer was separated and washed with saturated aq. NaHSOs, which resulted in precipitation of Ru black. The organic layer was then washed with brine and dried over anhydrous Na2S04. Evaporation of the solvent gave the desired product as a colorless oil. MS: 374.2 (M+ l )+. Step C: ( 2S,4R )-4-(( tert-Butyl dimethyl 'silyl )oxy)-5-oxopyrrolidin e-2-carboxylic acid. TF A (6 mL) was added to a solution of (2S,4R)- l -tert-butyl 2-methyl 4-((tert-butyldimethylsilyl)oxy)-5- oxopyrrolidine- l ,2-dicarboxylate (2.5 g, 6.68 mmo l) in DCM (18 mL) at 0 °C. The mixture was stirred at room temperature for 1 hthen concentrated. The residue was dissolved in M eOH/THF (1 OmL/1 OmL) followed by addition of a solution of LiOH (842 mg, 20.1 mmol) in water (5 mL).
The resulting mixture was stirred at room temperature for 1 h and then partitioned between EtOAc and H20. The aqueous layer was separated and then adjustedtopH=6 with 1 NHC1 aq. and extracted with EtOAc (3 x 20 mL). Combined organic layers were washed with brine, dried over anhydrous a2S04, and then concentrated to afford the desired product. !H NMR (400 MHz, DMSO-d6): 5 12.87 (s, lH), 8.17 (s, 1H), 4.21 (t, J = 8.0Hz, 1H), 4.02 (d, J= 8.4 Hz, 1H), 2.39 - 2.23 (m, 1H), 2.09 (m, 1H), 0.84 (s, 9H), 0.07 (s, 6H). M S: 260.1 (M +l)+.
Step D : The same as general procedure for UGI reaction set forth herein.
Step E : The same as general procedure for Buchwald reaction set forth herein.
Step F : (2S,4R )-N-( l-(2-Chlorophenyl )-2-( ( 3,3-difluorocyclobutyl )amino)-2-oxoethyl )-N-(3- cyano-ph enyl )-!-( 4-cyan opyridin-2-yl )-4-hydroxy-5 -oxopyrrolidin e-2-carboxamide. TB AF in THF (IN, 0.3 mL) was added to a solution of (2S,4R)-4-((tert-buty ldimethylsily l)oxy)-N-(l-(2- chlorop henyl)-2-((3,3-difiuorocy clobuty l)amino)-2-o xoethy l)-N-(3-cy anophenyl)- 1 -(4-cy ano pyridin-2-yl)-5-oxopyrrolidine-2-carboxamide (0.15 mmol) in THF at 0 °C and the reaction solution was stirred at this temperature for 20 min. The resultingmixture was concentrated and the residue was purified by a standard method to afford the desired product. ΊΤ NM R (400 MHz, CDC13): δ 8.82 - 8.43 (m, 2H), 8.40 - 8.17 (m, 1H), 7.63 - 7.30 (m, 3H), 7.26 - 6.66 (m, 4H), 6.68 - 6.34 (m, 2H), 6.65 - 6.31 (m, 2H), 4.87 - 4.56 (m, 2H), 4.23 (m, 1H), 4.01 - 3.76 (m, 1H), 3.15 - 1.96 (m, 6H). M S: 605.1 (M+ l)+.
The following analogs were synthesized via the procedure set forth herein, using the appropriate aldehyde, amine, carboxylic acid, isocynide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ringusing the reagents and solvents set forth herein, and purified via various standard methods .
(2S ,4R)-N-((S )-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino>2-oxoethyl)-l-(4-cyano - yridin-2-yl)-N-(3-fluorφhen3l)-4-hydroxy-5-oxo yrrolidine-2-carboxamide (single enantiomer) - Compound 113
Figure imgf000151_0001
Ή NMR (400 MHz, CDC13): δ 8.70 (m, 1H), 8.53 (s, 1H), 7.72 (d, J= 7.5 Hz, 1H), 7.32 (d, J = 4.9 Hz, 2H), 7.18 (d, J= 6.0 Hz, 1H), 7.09-6.85 (m, 4H), 6.43 (s, 1H), 6.20 (d, J= 5.3 Hz, 1H), 4.89 (s, 1H), 4.74 (t,J=9.2 Hz, 1H), 4.37-4.32 (m, 1H), 3.40 (s, 1H), 3.11 - 2.87 (m, 2H), 2.77 -2.14 (m,3H), 2.03-1.91 (m, 1H). MS: 598.1 (M+l)+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide - Compound 120
Figure imgf000151_0002
Ή NMR (400 MHz, CDC13): δ 8.98 (d, J= 4.4 Hz, 1H), 7.70 (s, 1H), 7.39 (d, J= 4.9 Hz, 2H), 7.20 - 6.86 (m, 4H), 6.50 (s, 1H), 5.75 (s, 1H), 5.35 (s, 1H), 4.92 - 4.63 (m, 2H), 4.34 (s, 1H), 2.91 (m, 3H), 2.21 (m, 4H). MS: 599.1 (M+l)+.
(2S,4R)-N-((S)-l-(2-ChIorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4- cyano pyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 121
Figure imgf000151_0003
Ή NMR (400 MHz, CDC13): δ 8.78 (s, 1H), 8.54 (s, 1H), 7.77 (d,J=8.1 Hz, 1H), 7.45-7.30 (m, 2H), 7.25 - 6.83 (m, 5H), 6.42 (s, 1H), 5.49 (d, J= 7.4 Hz, 1H), 4.83 (m„ 2H), 4.00 (s, 1H), 3.02 (s, 1H), 2.74 (m, 1H), 2.25 - 1.74 (m, 7H), 1.56- 1.33 (m, 2H). MS : 626.2 (M+l)+. (2S,4R)-N-((R)-l-(2-ChIorophenyl)-2-((4,4-difluorocycIohexyl)amino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 122
Figure imgf000152_0001
Ή NMR (400 MHz, CDC13): δ 9.00 (d, J= 4.8 Hz, 1H), 7.83 (m, 1H), 7.42 (t, J= 6.6 Hz, 2H), 7.22 (m, 2H), 7.18 - 7.08 (m, 1H), 7.08 - 6.67 (m, 2H), 6.17 (m, 1H), 5.70 (d, J= 7.6 Hz, 1H), 4.93-4.66 (m, 2H), 3.88 (d, J= 7.7 Hz, 1H), 3.37 (s, 1 H), 2.71 (m, 1H),2.03 (m, 5H), 1.88- 1.64 (m, 4H). MS : 627.2 (M+l)+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-dinuorocyclohexyl)amino)-2-oxoethyl)-l-(4- cyano pyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrroIidine-2-carboxamide - Compound 123
Figure imgf000152_0002
Ή NMR (400 MHz, CDC13): δ 8.99 (d, J= 4.4 Hz, 1 H), 7.74 (d, J= 7.9 Hz, 1H), 7.47 - 7.29 (m, 3H), 7.08 (m, 6H), 6.51 (s, 1H), 5.61 (s, 1H), 4.81 (m, 2H), 4.02 (d,J=7.2 Hz, 1H), 3.38 (s, 1H), 2.89-2.65 (m, 1H), 2.23 - 1.81 (m, 8H), 1.58-1.48 (m, 1H). MS : 627.2 (M+l)+.
(2S,4R)-N-((R)-l-(2-ChlorophenyI)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4-cya nopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 114
Figure imgf000152_0003
' H NM (400 MHz, CDCI3): δ 8.71 (d, J= 5.8 Hz, 1 H), 8.64 - 8.50 (m, 1 H), 7.94 - 7.56 (m, 1 H), 7.47 - 7.31 (m, 2H), 7.29 (d, J = 2.2 Hz, 1 H), 7.26 - 7.18 (m, 1 H), 7.16- 6.95 (m, 2H), 6.88-6.65 (m, 1H), 6.44 - 6.35 (m, 1H), 6.29 (s, 1H), 6.11 (d,J=6.7Hz, 1H), 4.77 (m, 2H), 4.40-4.08 (m, 1H), 3.27 (s, 1H), 3.09-2.58 (m, 3H), 2.54-2.12 (m, 2H), 2.10 - 1.95 (m, 1H) MS: 616 (M+l)+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyl)-l-(4-cya nopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single
15
Figure imgf000153_0001
1 H NMR (400 MHz, MeOD): δ 8.65 - 8.50 (m, 2H), 7.54 (d, J= 9.5 Hz, 1H), 7.43 - 7.32 (m, 1H), 7.22 - 7.12 (m, 2H), 7.03 (m, 1H), 6.97 - 6.87 (m, 1H), 6.84 - 6.75 (m, 2H), 6.36 (d, J = 8.5 Hz, 1H), 4.89 (d, J= 8.6 Hz, 1H), 4.65 - 4.49 (m, 2H), 4.13 (m, 1H), 2.93 - 2.72 (m, 2H), 2.57 -2.26 (m, 3H), 1.85 (m, 1H). MS: 616.1 (M+l)+.
(2S,4R)-N-((R)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 116
Figure imgf000153_0002
Ή NMR (400 MHz, CDC13): δ 8.98 (t, J= 5.0 Hz, 1 H), 7.88 (s, 1 H), 7.88 (s, 1H), 7.50 - 7.37 (m, 2H), 7.33 - 7.20 (m, 2H), 7.19- 7.06 (m, 2H), 6.83 - 6.66 (m, 1 H), 6.48 (m, 2H), 6.27 (s, 1H), 4.23 (s, 1H).3.32 (s, 1H), 2.87 (m, 2H), 2.66 (m, 1H), 2.35 - 2.02 (m, 3H). MS: 617.1 (M+l)+. (2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3,5-difluorophenyi)-4-hydroxy-5-oxopyrrolidine-2-carboxamide - Compound 117
Figure imgf000154_0001
Ή NMR (400 MHz, MeOD): 68.88 (d, J= 4.9 Hz, 1H), 7.56 (m, 2H), 7.34 (dd, J= 8.0, 1.1 Hz, 1H), 7.16 (td,J= 7.8, 1.6 Hz, 1H), 7.09 - 7.00 (m, 1H), 6.98-6.85 (m, 2H), 6.81 (m, 1H), 6.42 (s, 1H), 4.87 (d,J= 8.8 Hz, 1H), 4.59 -4.42 (m, 2H), 4.27-4.09 (m, 1H), 2.98 - 2.74 (m, 2H), 2.46 (m, 3H), 2.02 - 1.76 (m, 1H). MS: 617.1 (M+l)+.
(2S,4R)-N-((R)-l-(2-Chlorophenyl)-2-(4,4-difluorocyclohexylamino)-2-oxoethyl)-l-(4- cyanopyridin-2-yI)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 124
Figure imgf000154_0002
'HNMR (400 MHz, CDCI3): δ 8.71 (s, 1H), 8.64 (d, J= 5.0 Hz, 1H), 7.79 (s, 1H), 7.45 (d,J = 7.8 Hz, 1H), 7.35 (dd,J= 5.0, 1.0 Hz, 1 H), 7.30 - 7.24 (m, 1H), 7.16 (d,J= 6.3 Hz, 1H), 7.14- 7.05 (m, 1H), 6.79-6.68 (m, 2H), 6.27 (s, 1H), 5.87 (d, J= 7.5 Hz, 1H), 4.82 (t,J=6.9 Hz, 1H), 4.74 (t,J=9.2 Hz, 1H), 3.90-3.71 (m, 1H),3.27 (s, 1H),2.65 (m, 1H), 2.15 - 1.72 (m, 8H), 1.57- 1.43 (m, 1 H). MS: 644.2 (M+ 1 )+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-(4,4-dinuorocyclohexylamino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 125
Figure imgf000154_0003
Ή NMR (400 MHz, CDC13): δ 8.83 - 8.47 (m, 2H), 7.62 (d, J= 8.0 Hz, 1 H), 7.38 (d, J= 8.0 Hz, 1H), 7.32(d,J=5.0Hz, 1H), 7.21 (t,J=7.1 Hz, 1H), 7.05 (t,J=7.5 Hz, 1H),6.98 (d,J=7.7 Hz, 1H), 6.85 (d,J=7.7Hz, 1H), 6.68 (t,J=8.6Hz, 1H), 6.40 (s, 1H), 5.62 (d,J=7.7 Hz, 1H), 4.96-4.70 (m, 2H),4.01 (d,J=7.6 Hz, 1H), 3.37 (s, 1H),2.70 (m, 1H), 2.14 - 1.74 (m, 8H), 155-1.41(m, 1H). MS: 644.2 (M+l)+.
(2S,4R)-N-((R)-l-(2-Chlorophenyl)-2-(4,4-difluorocycIohexylamino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 126
Figure imgf000155_0001
Ή NMR (400 MHz, CDC13): δ 8.98 (dd, J= 4.7, 2.1 Hz, 1H), 7.63 (d, J= 7.3 Hz, 1H), 7.50 - 7.33 (m, 2H), 7.28 - 6.87 (m, 3H), 6.84 - 6.38 (m, 2H), 6.19 (s, 1 H), 5.82 (d, J= 7.6 Hz, 1 H), 4.94-4.65 (m, 2H), 3.86 (d,J=7.5 Hz, 1H), 3.57-3.49 (m, 1H), 2.68 (m, 1H), 2.16 - 1.86 (m, 6H), 1.81-1.77 (m, 2H). MS: 645.2 (M+l)+.
(2S,4R)-N-((S)-l-(2-Chlorophenyl)-2-(4,4-dinuorocyclohexylamino)-2-oxoethyl)-l-(4- cyanopyrimidin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 127
Figure imgf000155_0002
Ή NMR (400 MHz, CDC13): δ 8.99 (d, J= 4.8 Hz, 1H), 7.62 (d, J= 8.7 Hz, 1H), 7.49 - 7.35 (m, 2H), 7.22 (td,J=7.8, 1.5 Hz, 1H), 7.07 (t,J=7.1 Hz, 1H), 6.98 (dd,J=7.8, 1.3 Hz, 1H), 6.91 (d,J=8.2 Hz, 1H), 6.72 (tt, J= 8.6, 2.2 Hz, 1H), 6.48 (s, 1H), 5.64 (d,J=7.7Hz, 1H), 4.94- 4.69 (m,2H), 4.11 -3.91 (m, 1H), 3.46 (s, lH),2.79(m, 1H), 2.19 - 1.85 (m, 7H), 1.61 -1.40 (m,2H). MS: 645.2 (M+l)+. (2S,4R)-N-(l-(2-Chlorophenyl)-2-(3,3-dinuorocycIobutylamino)-2-oxoethyl)-N-(3-cyano-5- fluorophenyI)-l-(4-cyanopyridin-2-yl)-4-hydroxy-5-oxopyrroIidine-2-carboxamide (racemic) - Compound 169
Figure imgf000156_0001
Ή NMR (400 MHz, CDC13): δ 8.87-8.72 (m, 1H), 8.67 - 8.48 (m, 1H), 8.26 - 8.01 (m, 1H), 7.56 - 7.30 (m, 4H), 7.27 - 7.17 (m, 1 H), 7.10 (m, 1 H), 6.95 (t, J = 7.3 Hz, 1 H), 6.52 - 6.28 (m, 1H), 6.21 -5.95 (m, 1H), 4.88-4.64 (m, 2H), 4.30 (m, 1H), 3.21 -2.81 (m, 3H), 2.74 - 2.19 (m, 3H), 2.13 - 1.91 (m, 1H). MS: 623.1 (M+l)+.
(2S,4S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyano- pyri-din-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 118
Figure imgf000156_0002
Ή NMR (400 MHz, CD3OD): δ 8.97 (d, J= 4.7 Hz, 1H), 7.81 - 7.62 (m, 2H), 7.41 - 7.35 (m, 2H), 7.26-6.96 (m, 5H), 6.46 (d,J = 12.0 Hz, 1H), 4.81 -4.75 (m, 1H), 4.37-4.28 (m, 1H),
4.25 -4.15 (m, 1H), 2.91 (s, 2H), 2.60 - 2.37 (m, 3H), 2.00 - 1.87 (m, 1H). MS: 598.1 (M+l)+. (2S,4S)-N-((S)-l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyI)-l-(4-cyano- pyrimidin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 119
Figure imgf000156_0003
Ή NMR (400 MHz, CD3OD): δ 8.97 (d, J= 4.7 Hz, 1H), 7.81 - 7.62 (m, 2H), 7.41 - 7.35 (m, 2H), 7.26-6.96 (m, 5H), 6.46 (d,J= 12.0 Hz, 1H), 4.81-4.75 (m, 1H), 4.37 - 4.28 (m, 1H), 4.25 -4.15 (m, 1H), 2.91 (s, 2H), 2.60 -2.37 (m, 3H), 2.00 - 1.87 (m, 1H). MS: 599.1 (M+l)+. (2S,4S)-N-((S)-l-(2-ChlorophenyI)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4- cyano -pyridin-2-yl)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrroIidine-2-carboxamide (single
Figure imgf000157_0001
Ή NMR (400 MHz, CDC13): δ 8.87 - 8.57 (m, 2H), 7.96 (s, 1 H), 7.50 - 7.30 (m, 3H), 7.26 - 7.12 (m, 2H), 7.09 - 6.96 (m, 2H), 6.28 (s, 1H), 5.67 (d, J= 7.6 Hz, 1H), 4.74 (dd, J= 8.1, 4.6 Hz, 1H), 4.42-4.36 (m, 1H), 4.04 (s, 1H), 3.87 (d, J= 7.8 Hz, 1 H), 2.54 - 2.41 (m, 1H),2.22- 1.76 (m, 8H), 1.50-1.32 (m,2H). MS: 626.2 (M+l)+.
(2S,4S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-dinuorocyclohexyl)aniino)-2-oxoethyl)-l-(4-cya- nopyrimidin-2-yI)-N-(3-fluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 189
Figure imgf000157_0002
Ή NMR (400 MHz, CDC13): δ 9.00 (d, J= 4.7 Hz, 1H), 7.76 (s, 1H), 7.47 - 7.30 (m, 2H), 7.24 - 6.88 (m, 6H), 6.47 (d,J=6.7Hz, 1H), 5.54 (s, 1H), 4.74 (s, 1H), 4.35 (s, 1H), 3.99 (s, 1H),3.72 (d,J=34.8Hz, 1H), 2.58 -2.18 (m,2H), 1.88 (m,4H), 1.56-1.42 (m, 2H). MS: 627.2 (M+l)+. (2S,4S)-N-((S)-l-(2-Chlorophenyl)-2-((3,3-difluorocycIobutyl)amino)-2-oxoethyl)-l-(4- cyano -pyridin-2-yl)-N-(3,5-difluorophenyI)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 171
Figure imgf000158_0001
Ή NMR (400 MHz, CDC13): δ 8.68 (s, 1H), 8.52 (d, J= 5.0 Hz, 1H), 7.60 (d, J= 8.2 Hz, 1H), 7.45 - 7.17 (m,4H), 7.15-6.91 (m, 2H), 6.84 (d,J=8.7 Hz, 1H), 6.69 (t,J=8.7Hz, 1H), 6.54 - 6.36 (m, 2H), 4.87 - 4.60 (m, 1 H), 4.31 (m, 2H), 3.99 - 3.77 (m, 1 H), 3.15 - 2.78 (m, 2H), 2.62-2.26 (m,3H), 2.26-2.08 (m, 1H). MS: 616.1 (M+l)+.
(2S,4S)-N-((S)-l-(2-Chlorophenyl)-2-((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3,5-difluorophenyl)-4-hydroxy-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 174
Figure imgf000158_0002
1 H NMR (400 MHz, CDCI3): δ 8.75 (s, 1 H), 8.53 (d, J= 4.5 Hz, 1 H), 7.62 (s, 1 H), 7.44 -7.18 (m, 3H), 7.09 -6.96 (m, 2H), 6.86 (s, 1H), 6.71 (t,J=8.7 Hz, 1H), 6.38 (s, 1H), 5.58 (d, J= 7.6 Hz, 1H), 4.80 (dd, J= 8.0, 5.2 Hz, 1H), 4.37 (d, J= 5.6 Hz, 1H), 3.96 (s, 1H), 3.61 (d, J= 7.7 Hz, 1H), 2.62-2.29 (m, 1H), 2.13 (m, 6H), 1.48 (m, 2H). MS: 644.2 (M+l)+.
Example 19. Preparation of (2S)-N-((R)-l-(2-chlorophenyl)-2-(3,3- difluorocycloburylamino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4- hydroxy-4-methyl-5-oxopyrrolidine-2-carboxamide (single enantiomer) - Compound 183
Figure imgf000158_0003
Step A. (2S)-1 -tert-Butyl 2-methyl 4-( tert-butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidine- 1 ,2-dicarboxylate. LiHM DS (1 M in THF, 22.6 mL, 22.6 mmol) was added into a mixture of (2S,4R)-l -tert -butyl 2-methyl 4-(tert-butyldimethylsi^loxy)-5-oxopyrrolidine- l,2-dicar-boxylate (6.5 g, 17.4 mmol) in THF (60 mL) at -78°C under N2. The mixture was stirred at -78°C for 1 hr. A solution of iodomethane (2.7 g, 19.1 mmol) in THF (10 mL) was added dropwise tothe above mixture over 30 min Then the solution was stirred at -78°C for another 25 min. The resulting mixture was allowed to warm to room temperature and stirred overnight. The mixture was quenched with NH4C1 and extracted by ethyl acetate (60 mL x 3). The co mbined organic layers were dried and concentrated. The residue was purified by column chromatography to give the desired product. MS: 388 (M+ l)+.
Step B. (2S,4S)-M ethyl 4-((tert-butyldimethylsilyl)oxy)-5-oxopyrrolidine-2-carboxylate. A solution of (2S)- l -tert-butyl 2-methyl 4-(tert-butyldimetbylsilyloxy)-4-methyl-5-oxopyrroli-dine- 1 ,2-dicarbo xylate (960 mg, 25 mmo l) in TFA/DQVI (V : V = 1 : 3) was stirred at room temperature for 1 h. The mixture was then concentrated to give the desired product which was used directly in the next step . M S: 288 (M+ l)+.
Step C. (2S)-4-(tert-Butyldimethylsilyloxy)-4-methyl-5-oxopyrrolidin e-2-carboxylic acid. To a solution of (2S)-methyl 4-(tert -bury ldimethylsiryloxy)-4-methyl-5-oxopyrrolidine-2-carbo-xy late (400 mg, 1.4 mmol) in M eOH/THF/H20 (V : V : V = 2 : 2 : 1) was added LiOH (50 mg 2.1 mmol). The mixture was stirred at room temperature for 1 hr and then concentrated. The residue was partitioned between ethyl acetate and water. The aqueous phase was sep arated and adjusted to pH=3-4 with IN HCl solution. The aqueous layer wasthen extracted with ethyl acetate (2x20 mL). The combined organic layers were dried over anhydrous Na2S04 and concentrated to give the desired product. M S: 274 (M +l)+.
Step D. (2S)-4-(tert-Butyldimeth^silyloxy)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobut- ylamino)-2-oxoethyl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidin e-2-carboxamide. A solution of 3-fiuoroaniline (83 mg 0.75 mmol) and 2- chlorobenzaldehyde (105 mg, 0.75 mmol) in MeOH (5 mL) was stirred for 30 min at room temperature, followed by addition of (2S)-4- (tert-butyldimethy]silyloxy)-4-methyl-5-oxopyrrolidine-2-carboxylic acid (205 mg, 0.75 mmol). The resulting mixture was stirred for 10 min and fo llowed by the addition of 1 , l-difiuoro-3- isocyanocyclobutane (105 mg, 0.9 mmol). The mixture was stirred at room temperature overnight and concentrated, and then the residue was purified by a standard method to give the desired product. M S: 624 (M+l)+ .
Step E. (2S)-4-(tert-Butyldimethylsilyloxy)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutyl- amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-4-methyl-5-oxopyrrolidine-2- carboxamide. A mixture consisting of (2S)-4-(tert-butyldimethylsilyloxy)-N-(l-(2- chlorophenyl)-2-(33-difiuorocy clobuty lamino)-2-oxoethyl)-N-(3-fiuorophenyI)-4-methy 1-5- oxopyrrolidine-2-carboxamide (200 mg, 0.32 mmo l), 2-bromoisonicotinonitrile (88 mg 0.48 mmol), Cs2C03 (146 mg, 0.45 mmo l), Pd2(dba)3 (29 mg, 0.032 mmol), Xant-Phos (19 mg, 0.032 mmol) and 1,4-dioxane (5 mL) was stirred under N2 at 80 °C overnight. After filtration, the filtrate was concentrated in vacuo and the residue was purified by a standard method to give desired product. MS: 726 (M+l)+ .
Step F. ( 2S)-N-( (R)-l-( 2-Ch loroph enyl )-2-( 3,3-dflu orocyclobutyl amino )-2-oxoethyl)-l-( 4- cyanopyridin-2-yl )-N-( 3 -fluorophenyl )-4-hydroxy-4-methyl-5-oxopyrrolidine-2-carboxamide. To a solution of (2S)-4-(tert-butyldimethylsilyloxy)-N-(l-(2-chlorophenyl)-2-(3,3- difiuorocy clobuty lamino)-2-oxo ethy 1)- 1 -(4-cy an py ridin-2-y l)-N-(3-fiuorophery l)-4-methy 1-5- oxopyrrolidine-2-arboxamide (50 mg, 0.07 mmol) in THF (2 mL) was added TBAF (36 mg, 0.14 mmol) at 0 °C. The solution was stirred at 0 °C for 30 min and then partitioned between water andEtOAc. Combined organic layers were separated, dried, and concentrated in vacuo. The resulting residue was purified by a standard method to give the desired product, !H NMR (400 MHz, CDC13): δ 8.57 (d, J= 5.0 Hz, 1H), 8.48 (d, J= 3.8 Hz, lH), 7.54 - 7.17 (m, 5H), 6.98 - 6.84 (m, 3H), 6.67 (dd, J= 8.6 Hz, 1H), 6.33 (d, J= 5.2 Hz, 1H), 6.08 - 6.01 (m, 1H), 4.55 - 4.48 (m, lH), 4.29 (s, 1H), 3.22 -2.35 (m, 6H), 1.93 - 1.80 (m, 1H), 1.27 (s, 3H). M S: 612.2 (M+l)+.
Example 20. Preparation of (2S )-N-(l-(2-chlorophenyi)-2-(3,3-difluorocyclobutylamino)-2- oxo-ethyl)-l-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-sulfamoylphenyl)-5-oxopyrrolidine-2- carboxamide (racemic) - Compound 158
Figure imgf000161_0001
Step A. Benzyl(3-fluoro-5-nitrophenyl)sulfane. To a solution of 1 , 3-difluoro-5-nitrobenzene ( 15.9 g, 100 mmol) in DMF ( 160 mL) was added 2C03 ( 15.8 g, 1 10 mmol) and
phenylmethanethiol ( 12.4 g. 100 mmol) at 0 °C. The reaction was stirred at room temperature for 2 hr and then quenched with H20. The resulting mixture extracted with EtOAc (3x 100 mL). The combined organic layers were dried over anhydrous Na2S04 and concentrated in vacuo to afford the crude product as a yellow oil which was used in the next step without further purification. Step B. 3-Fluoro-5-nitrobenzene-l-sulfonyl chloride. To a solution of benzyl(3-fluoro-5- nitrophenyl)sulfane (3.0 g) in DCM (30 mL) was added deionized water (30 mL). Then chlorine was bubbled slowly into the mixture until the complete consumption of the starting material was observed (monitored by TLC). The organic layer was separated, washed with sat. aq. Na2S203 solution, dried and concentrated to afford the crude product which was used in the next step without further purification.
Step C. N-tert-b tyl-3-fl oro-5-nitrobenzenes lfonamide. To a solution of 3-fluoro-5- nitrobenzene- l -sulfonyl chloride in dry dioxane (30 mL) was slowly added fer/-butylamine ( 10 mL) at 0 °C. The reaction was allowed to warm to room temperature and stirred for 2 hr. The mixture was then concentrated and the residue was purified by column chromatography to afford the desired product. 'H MR (400 MHz, DM SO -de): δ 8.43 (s, 1H), 8.40 - 8.32 (m, lH), 8.10 - 8.05 (m, lH), 7.99 (s, 1H), 1.12 (s, 9H).
Step D. 3-Amino-N-tert-butyl-5-fluorobenzen esulfonamide. N-tert-buryl-3-fiuoro-5-nitro- benzenesulfonamide (1.0 g 3.6 mmo l), iron powder (l .O g, 18 mmol) and H4Cl (1.0 g 18 mmol) were mixed in EtOH (95%, 10 mL). The mixture was refluxed for 16 hr then filtered. The filtrate was concentrated and the residue was purified by a standard method to afford the desired product. 'H NMR (400 MHZ, DM SO-d6): δ 7.45 (s, 1H), 6.88 - 6.85 (m, 1H), 6.66 - 6.62 (m, 1H), 6.48 - 6.42 (m, 1H), 5.89 (s, 2H), 1.11 (s, 9H).
Step E. The same as general procedures for UGI reaction set forth herein.
Step F. The same as general procedures for Buchwald reaction set forth herein.
Step G. (S )-N-( ( S )-!-( 2-CHorophenyl )-2-( 3,3-difluorocyclobutylamino)-2-oxoethyl )-!-( 4- cyanopyridin -2-yl )-N-( 3-fluoro-5-su Ifamoylph enyl )-5-oxopyrrolidin e-2-carb ox amide. T o a solution of (2S)-N 3-( -tert-butylsulfamoyl)-5-fluorophenyl)-N-(l -(2-chlorophenyl)-2-(3,3- difiuorocy clo-buty lamino)-2-oxo ethy 1)- 1 -(4-cy anopy ridin-2-y l)-5-oxopy rrolidine-2-carbo xamide (80 mg, 0.1 1 mmol) in DCM (1 mL) was added TFA (1 mL). The reaction was stirred at room temperature for 16 hr and neutralized with saturated aq. NaHC03. The mixture was then extracted with EtOAc (3 x 10 mL). The combined organic layers were dried and concentrated. The residue was purified by a standard method to afford the target compound. !H NMR (400 MHz, DM SO-d6): δ 8.90 - 8.84 (m, 1H), 8.67 - 8.62 (m, 1H), 8.55 (s, 1H), 8.19 (s, 1H), 7.87 - 7.76 (m, lH), 7.65 - 7.60 (m, 2H), 7.45 - 7.40 (m, 3H), 7.21 (d, J= 7.0 Hz, 2H), 7.1 1 - 7.04 (m, 1H), 6.93 - 6.86 (m, 1H), 6.33 - 6.26 (m, 1H), 4.83 (m, 1H), 4.13 (s, 1H), 2.94 (m, 2H), 2.63 - 2.53 (m, 3H), 2.42 - 2.32 (m, 1H), 1.97 (s, 2H). MS: 661 (M+ l)+.
Example 21. Preparation of (2S )-N-(lH-benzo[d]imidazol-7-yl)-N-(l-(2-chlorophenyl)-2- ((4,4-difluorocyclohexyl)amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2- carboxamide (racemic) - Compound 141
Figure imgf000163_0001
Step A: 7-Nitro-lH-benzo[d]imidazole. A solution of 3-nitrobenzene- l ,2-diamine (900 mg, 5.88 mmol) in AcOH ( 12 mL) was stirred at 100 °C overnight. The mixture was neutralized with aq. NaHCC^ to pH= 8 at 0 °C and the precipitate was collected by filtration. The precipitate was dried in vacuo to afford the desired product.
Step B: 7-Mtro^-((2-(trimethylsilyl)ethoxy)methyl)-lH-benzold]imidazole. NaH (331 mg, 8.28 mmol) was added to a solution of 7-nitro- l H-benzo[d]imidazole (900 mg, 5.52 mmol) in DMF (7 mL) at 0 °C under N2. After stirring at 0 °C for 1 hr, SEMC1 ( 1 .38 g, 8.28 mmol) was added and the resulting mixture was stirred at room temperature for 2 hr. The reaction mixture was quenched with H20 and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04, and concentrated. The residue was purified by column chromatography to afford the desired product as a yellow oil.
Step C: l-((2-(TrimethylsiIyl)ethoxy)methyl)-lH-benzo[d]imidazol-7-amine. To a solution of 7- nitro- l -((2-(trimethylsilyl)ethoxy)methyl)-l H-benzo[d]imidazole (600 mg, 2.05 mmol) in EtOH/EtOAc ( 10 mL/2 mL) was added Pd/C (60 mg). After stirring under a hydrogen
atmosphere at room temperature overnight, the reaction mixture was filtered and the filtrate was concentrated. The residue was purified by a standard method to afford the desired product.
Step D: The same as general procedure for UGI reaction set forth herein.
Step E: (2S)-N-(lH-Benzo[d]imidazol- 7-yl)-N-(l-(2-chlorophenyl)-2-((4,4-difluorocyclohexyl) amino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide. TBAF ( 1 M in THF, 3 mL) was added to a solution of (2S)-N-( l -(2-chlorophenyl)-2-((4,4-difluoro - cyclohexyl)amino)-2-oxoethyl)- l -(4-cyanopyridin-2-yl)-5-oxo-N-( l -((2-(trimethylsilyl)ethoxy) - methyl)- 1 H-benzo[d]imidazol-7-yl)pyrrolidine-2-carboxamide in THF (0.5 mL) at 0 °C under N . After stirring at room temperature for 7 hr, the reaction was quenched with water at 0 °C. The resulting mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated. The resulting residue was purified by a standard method to afford the desired product. Ή NMR (400 MHz, CDC13): δ 13.08 (s, 1 H), 8.92 - 8.39 (m, 2H), 8. 19 (m, 1 H), 7.82 (m, 1 H), 7.51 - 7.3 1 (m, 2H), 7.25 (d, J = 5.2 Hz, 1 H), 7.13 - 6.70 (m, 3H), 6.41 (m, 1 H), 6.20 - 5.29 (m, 1 H), 4.85 (m, 1 H), 3.86 (s, 1 H), 2.97 - 2.39 (m, 2H), 2.36 - 1 .70 (m, 9H), 1 .40 (m, 2H). MS: 632.2 (M+ l )+.
Example 22. Preparation of (4S)-N-(l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)- 2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-l-methyl-2-oxoimidazolidine-4- carboxamide (racemic) - Compound 79
Figure imgf000164_0001
Step A: (S)-3-(Benzyloxycarbonyl)-2-oxoimidazolidine-4-carbo -xylic acid. To a solution of 6.6 g of sodium hydroxide in 140 mL of water at 0 °C, 8.8 g of bromine was added dropwise, followed by addition of (S)-4-amino-2-(benzyloxycarbonylamino)-4-oxobutanoic acid (13.4 g, 50 mmol) portionwise over 3 min. The resulting yellow solution was heated to 50 °C for 1 hr and then cooled to room temperature. After addition of sodium thiosulfate (2.0 g), the reaction mixture was washed with ether (2 x 30 mL). The aqueous layer was acidified to pH 1 -2 with 6 N HCI. After the precipitate was formed, the suspension was filtered. The sticky material was collected and re-crystallized in MeOH to afford the desired product as a white solid.Ή NMR (400 MHz, DMSO-d6): δ 13.29 (s, 1 H), 7.57 (s, 1 H), 7.40 - 7.27 (m, 4H), 5.27 - 5.04 (m, 2H), 4.66 (dd, J = 10.2, 3.2 Hz, 1 H), 3.63 (t, J = 10.0 Hz, 1 H), 3.20 (dd, J = 9.7, 3.2 Hz, 1 H).
Step B: (S)-Dibenzyl 2-oxoimidazolidine-l,5-dicarboxylate. To a 500 mL-flask were added (S)- 3-(benzyloxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid (5.3 g, 20 mmol), BnBr (2.8 mL, 23 mmol), 2C03 (8.28 g, 60 mmol), and acetonitrile (250 mL). The reaction solution was heated to reflux for 6 hr, cooled and then filtered. The filtrate was concentrated in vacuo and the residue was purified by column chromatography to afford the desired product as white solid. 1H MR (400 MHz, CDC13): δ 7.41 - 7.25 (m, lOH), 6.36 (s, 1H), 5.30 - 5.05 (m, 4H), 4.80 (dd, J = 10.2, 3.6 Hz, 1H), 3.74 (t, J= 10.0 Hz, 1H), 3.41 (dd, J= 9.7, 3.7 Hz, 1H).
Step C: (S)-Dibenzyl 3-methyl-2-oxoimidazolidine-l,5-dicarboxylate. To a dry 100 mL- flask were added (S)-dibenzyl 2-oxoimidazolidine-l ,5-dicarboxy late (1.5 g, 4.24 mmol), K2CO3 (1.17 g, 8.47 mmol), M el (5.2 mL, 84.7 mmol), and acetone (50 mL). The reaction solution was heated to reflux and stirred overnight. The resulting reaction mixture was cooled and filtered. The filtrate was concentrated in vacuo and the residue was purified by column chromatography to afford the desired product as a white solid 1H NMR (400MHz, CDC13): δ 7.40 - 7.26 (m, 10H), 5.27 - 5.07 (m, 4H), 4.70 (dd, J= 10.2, 3.8 Hz, 1H), 3.63 (dd, J= 10.1 , 9.7 Hz, 1H), 3.31 (dd, J = 9.6, 3.8 Hz, 1H), 2.84 (s, 3H). MS : 369 (M + l)+.
StepD: (S)-l-Meiiyl-2-oxoimidazolidine-4-carboxylicacid. To a dry 50 mL-fiask were added (S)-dibenzyl 2-oxoimidazolidine-l,5-dicarboxylate (0.5 g 1.36 mmol), Pd/C (10%, 100 mg) and M eOH (15 mL). The suspension was stirred overnight at room temperature under a hydrogen atmosphere. The resultingreaction mixture was filtered. The filtrate was concentrated in vacuo to afford the desired product as an off-white solid. 1H NMR (400MHz, CD3OD): δ 4.21 (dd, J= 9.9, 4.8 Hz, 1H), 3.70 (t, J= 9.6 Hz, 1H), 3.55 (dd, J= 9.3, 4.8 Hz, 1H), 2.74 (s, 3H). MS : 145 (M+l)+.
Step E: (4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl) -N-(3-fluoro- phenyl)-l-methyl-2-oxoimidazolidne-4-carboxamide. A mixture of 2-chlorobenzaldehyde (165 mg, 1.18 mmol) and 3-fiuorobenzenamine (131 mg, 1.18 mmo 1) in M eOH (3 mL) was stirred at room temperature for 30 min. Then (S)-l-methyl-2-oxoimidazolidine-4-carboxylic acid (170 mg, 1.18 mmol) was added and the reaction mixture was stirred for another 15 min, followed by addition of l,l-difiuoro-3-isocyanocyclobutane (138 mg, 1.18 mmol). The reaction mixture was stirred overnight and concentrated in vacuo. The residue was purified by a standard method to give the desired product. MS : 495 (M + l)+.
Step F: The same as the Buchwald reaction procedure set forth herein. !H NM R (400 MHz, CDC13): δ 8.64 - 8.34 (m, 2H), 7.94 - 7.59 (m, 1H), 7.50 - 6.61 (m, 8H), 6.34 - 6.07 (m, 1H), 4.94 - 4.67 (m, 1 H), 4.3-4.2 (m, ] H), 3.49 (m, 1 H), 3.46 - 3.22 (m, 1 H), 3.02-2.83 (m, 2H), 2.87 (s, 3H), 2.5-2.2 (m, 2H). MS : 597 (M+ l )+.
Example 23. Preparation of (S)-N-((S)-l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)- 2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxoimi dazolidine-4-carboxamide (single enantiomer) - Com ound 80
Figure imgf000166_0001
Figure imgf000166_0002
Step A: (S)-3,4-Dibenzyl 1-tert-butyl 2-oxoimidazolidine-l,3,4-tricarboxylate. To a 25 mL-flask were added (S)-dibenzyl 2-oxoimidazolidine- l ,5-dicarboxylate (40 mg, 0.1 1 mmol), (BOC)20 (26 mg,0.12 mmol), TEtOAc (0.06 mL,0.3 mmol), DMAP (cat.) and CH2C12 (2 mL). The mixture was stirred overnight. The solvent was then removed in vacuo and the residue was purified by column chromatography to give the desired product. Ή NMR (400 MHz, CDCI3): δ 7.39 - 7.27 (m, 10H), 5.24(s, 2H), 5.16(s, 2H), 4.67 (dd, J = 10.2, 3.5 Hz, 1 H), 3.94 (dd, J = 1 1. 1 , 10.3 Hz, 1 H), 3.74 (dd, J = 1 1 .2, 3.5 Hz, 1 H), 1 .5 1 (s, 9H).
Step B: (S)-l-(tert-Butoxycarbonyl)-2-oxoimidazolidine-4-carboxylic acid. To a dry 50 mL- flask were added (S)-3,4-dibenzyl l -tert-butyl 2-oxoimidazolidine- l ,3,4-tricarboxylate ( 1 .24 g, 2.73 mmol), Pd/ C( 10%, 200 mg) and MeOH (30 mL). The suspension was stirred overnight at room temperature under a hydrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to afford the desired product. Ή NMR (400 MHz, DMSO-d6): δ 6.06 (s, 2H), 4.3 1 (s, 1 H), 4.25 - 3.94 (m, 2H), 1 .52 (s, 9H).
Step C: (4S)-tert-Butyl 4-((l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino) -2-oxoethyl)(3- fluorophenyl)carbamoyl)-2-oxoimidazolidine-l-carboxylate. A mixture of 2- chlorobenzaldehyde ( 122 mg, 0.87 mmol) and 3-fluorobenzenamine (97 mg, 0.87 mmol) in MeOH (2 mL) was stirred at room tem erature for 30 min. Then (S)-l-(tert-butoxycarbonyl)-2- oxoimidazolidine-4-carboxylic acid (200 mg, 0.87 mmol) was added and the reaction mixture was stirred for another 15 min followed by addition of l,l-difiuoro-3-isocyanocyclobutane (102 mg, 0.87 mmol). The reaction mixture was further stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by a standard method to give the desired product. 'HNMR (400 MHZ, CDC ): δ 7.46 - 6.59 (m, 8H), 6.45 (s, lH), 4.41 -4.04 (m, 2H), 4.01- 3.78 (m, 1H), 3.64 -3.30 (m, 1H),2.92 (m, 2H),2.71 -2.27 (m,2H), 1.46 (s, 9H).MS: 581 (M+l)+.
StepD: (4S)-tert-Butyl 4-((l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino) -2-oxoethyl){3- fluorophenyl)carbamoyl)-3-(4-cyanopyridin-2-yl)-2-oxoimidazolidine-l-carboxylate. To a 25 mL flask charged with 1,4-dioxane (4.5 mL) were added(4S)-tert-butyl4-((l-(2-chlorophenyI)- 2-(33-difluoro cyclo butylamino)-2-oxoethyl)(3-fiuoro -phenyl)carbamoyl)-2-oxoimidazolidine-
1- carboxylate (250 mg, 0.43 mmol), 2-bromoisonicotinonitrile (122 mg 0.65 mmol), CS2CO3 (281 mg, 0.862 mmol), Xant-Phos (25 mg 0.043 mmol) and Pd2(dba)3 (40 mg, 0.043 mmol). The mixture was degassed and refilled with nitrogen, and then heated to 100 °C for 3 hr. The resulting mixture was cooled and filtered. The filtrate was concentrated in vacuo and the residue was purified by a standard method to give both epimers. The epimers were further separated by a standard method to give the desired product. 'HNMR (400 MHz, CDC13): δ 8.58 (s, 1H), 8.48 (t, J= 5.9 Hz, 1H), 7.71 (d, J= 8.4 Hz, 1H),7.37 - 7.16 (m, 4H), 7.15 - 6.76 (m, 4H), 6.56 - 6.31 (m, 2H), 4.95 - 4.75 (m, 1H), 4.31 (s, 1H), 3.86 (dd,J= 10.8, 5.1 Hz, 1H), 3.66 (m, 1H), 2.99 (m, 2H),2.61 -2.27 (m, 2H), 1.56 (s,9H).MS : 683(M + 1)+.
Step E: ( S )-N-( ( S )-l -(2-Chlorophenyl )-2-( 3,3-difluorocyclo butyl amino) -2 -oxo ethyl )-3-(4- cyanopyridin-2-yl)-N-(3-fluorophenyl)-2-oxoimi dazolidine-4-carboxamide. To a solution of 2N HClMeOH (2 mL) at 0°C was added50 mgof (S)-tert-butyl-4-(((S)-l-(2-chlorophenyl)-2- (3,3-difiuorocyclobutylamino)-2-oxoethyl)(3-fluoropheryl) carbamoyl)-3-(4-cyanopyridin-2-yl)-
2- oxoimidazolidine-l-carboxylate. The reaction mixture was warmed to room temperature and stirred for 5 hr. The solvent was removed in vacuo and the residue was purified by a standard method to give the desired pro duct.1H NMR (400 MHz, CD3OD): δ 8.50 (d, J= 4.5 Hz, 1H), 7.65(d,J=8.6 Hz, 1H), 7.50-6.81 (m, 8H), 6.47 (d,J= 11.6Hz, 1H), 5.04 -4.92 (m, 1H), 4.22 (m, 1H), 3.59-3.46 (m, 1 H), 3.39 (dd, J= 9.9, 4.5 Hz, 1H), 2.91 (m, 2H), 2.63 - 2.36 (m, 2H). MS : 583 (M+l)+.
Example 24. Preparation of (4S)-N-(l-(2-chlorophenyI)-2-(3,3-difluorocyclobutyl -amino)- 2-oxoethyl)-3-(4-cyanopyridin-2-yl)-N-(3-fluorophenyl)-l-(2-hydroxyet -yl)-2- oxoimidazoli-dine-4-carboxamide (racemic)
Figure imgf000168_0001
Step A: (S)-Dibenzyl 3-(2-ethoxy-2-oxoethyl)-2-oxoimidazolidine- 1,5- dicarboxylate. To a dry
50 mL-flask charged with DME (5 mL) were added (S)-dibenzyl 2-oxoimidazolidine-l,5- dicarboxylate (200 mg, 0.56 mmol), 2CO3 (156 mg,1.13 mmol), and ethyl 2-bromoacetate (0.13 mL, 1.13 mmol). The mixture was heated to reflux for 3 hr. The reaction mixture was cooled and filtered. The filtrate was concentrated in vacuo and the residue was purified by column chromatography to afford the desired product as a colorless oil.'H NMR (400 MHz, CDCI3): δ 7.45 - 7.25 (m, 10H), 5.41 - 5.05 (m, 4H), 4.80 (dd, J= 10.2, 3.5 Hz, 2H), 4.29-4.08 (m, 3H), 3.90(dd,J= 12.2, 7.2 Hz, 2H), 3.45 (dd, J = 9.2, 3.5 Hz, 1H), 1.28 (td, J= 7.1, 2.1 Hz, 3H). Step B: (S)-l-(2-Ethoxy-2-oxoethyl)-2-oxoimidazolidine-4-carboxylic acid. To a dry 50 mL- flask were added (S)-dibenzyl 3-(2-ethoxy-2-oxoethyl)-2-oxoimidazolidine-l,5-dicarboxylate (170 mg, 0.386 mmol), Pd/C(10%, 35 mg) and MeOH (4 mL). The suspension was stirred at room temperature overnight under a hydrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to afford the desired product as an off-white solid.Ή NMR (400 MHz, CD3OD): δ 4.30 (dd, J= 10.0, 4.8 Hz, 1H), 4.20 (q, J= 7.1 Hz, 2H), 4.05 - 3.91 (m,2H), 3.91 -3.85 (m.1H), 3.69 (dd, J=9.0, 4.8 Hz, 1H), 1.29 (t,J=7.1 Hz, 3H). Step C: Ethyl 2-((4S)-4-((l-(2-chlorophenyl)-2-(3,3-difhorocyclobu tylamino)-2 -oxoethyl)(3- fluorophenyl )carbamoyl )-2-oxoimidazolidin-l -yl )acetate. A mixture of 2-chlorobenz aldehyde (518 mg, 3.70 mmol) and 3-fiuorobenzenamine (41 1 mg, 3.7 mmo l) in MeOH (12 mL) was stirred at room temperature for 30 min. Then (S)-l -(2-ethoxy-2-oxoethyl) -2-oxoimidazolidine- 4-carboxylic acid (800 mg, 3.7 mmol) was added and the reaction mixture was stirred for another 30 min, followed by addition of l ,l -difiuoro-3- isocyanocyclobutane (600 mg, 3.7 mmol). The reaction mixture was stirred overnight and concentrated in vacuo. The residue was purified by a standard method to give the desired product. M S : 567 : (M + l)+.
Step D: Ethyl 2-((4S)-4-((l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)(3- fluoroph enyl )carbamoyl )-3-( 4-cyan opyridin -2-yl )-2-oxoimidazolidin -1-yl )acetate - Compoun d
94. To a 25 mL-fiask were added ethyl 2-((4S)-4 (l -(2-chlorophenyl)-2-(3,3- difiuorocyclobutylamino)-2-oxo ethyl) (3-fiuoro -phenyl)carbamoyl)-2-oxoimidazolidin- l- yl)acetate (50 mg, 0.0882 mmol), 2-bromoisonicotinonitrile (21 mg, 0.115 mmol), CS2CO3 (58 mg,0.176 mmol), Xant-Phos (5.2 mg, 0.009 mmol), Pd2(dba)3 (8.2 mg, 0.009 mmo l) and 1 ,4- dioxane (1 mL). The mixture was degassed and refilled with nitrogen, and then heatedto 100 °C for 3 hr. The resulting mixture was cooled and filtered and then the filtrate was concentrated in vacuo. The residue was purified by a standard method to give both ep imers. 'H MR (400 MHz, CDC13): δ 8.63-8.57 (S, 1H), 8.55 - 8.38 (m, 1H), 7.63 (s, 1H), 7.46 - 6.84 (m, 8H), 6.45-6.37 (m, 1H), 6.22 - 5.94 (m, lH), 5.06 - 4.77 (m, 1H), 4.43-4.37 (m, 1H), 4.32-4.20 (m, 1H), 4.21 (q, J= 7.1 Hz, 2H), 3.82 - 3.46 (m, 3H), 3.12 - 2.82 (m, 2H), 2.66 - 2.25 (m, 2H), 1.29 (t, J= 7.1 Hz, 3H). MS : 669 (M + l)+.
Step E: (4S)-N-(l-(2-Chlorophenyl)-2-(3,3-difluorocyclobutyl-amino)-2-oxoethyl)-3-(4- cyanopyridin-2-yl)-N-(3-flu orophenyl)-l-(2-hydroxyeth-yl)-2-oxoimidazolidine-4-carboxa - mide - Compound 112. To a solution of ethyl 2-((4S)-4-((l -(2-chlorophenyl)-2-(3,3- difiuorocyclobutylamino)-2-oxo ethyl)(3-fluorophenyl)carbamoyl)-2-oxo-3-(pyrimidin-2- yl)imidazolidin- 1 -yl) acetate (100 mg, 0.155 mmol) in DME (2 mL) at 0 °C was added LiBH4 (22 mg) in two portions. The mixture was stkred for 0.5 hr, then wanned to room temperature. The resulting mixture was stirred for another 2 hr and quenched with H20 (2 mL) at 0 °C. The resulting mixture was extracted with EtO Ac (2 x 10 mL). The combined organic layers were combined, washed with brine, dried over anhydrous Na2S04, and concentrated in vacuo. The residue was purified by a standard method to give the desired product. Ή NMR (400 MHz, CDC13): δ 8.62-8.55 (m, 2H), 7.63 (d, J = 8.1 Hz, 1 H), 7.40 - 6.85 (m, 8H), 6.47 - 6.2 (m, 2H), 4.90 - 4.69 (m, 1 H), 4.30 - 4.15 (m, 1 H), 3.87 - 3.72 (m, 2H), 3.71 - 3.19 (m, 5H), 3.08 - 2.85 (m, 2H), 2.63 - 2.35(m, 2H). MS : 603 (M+ l )+.
The following compound was synthesized via the procedure set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted aromatic ring or heterocyclic (heteroaromatic) ring using the reagents and solvents set forth above, and purified via standard methods.
Ethyl 2-((4S)-4-((l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyI)(3-fluoro phenyl)carbamoyI)-3-(4-cyanopyrimidin-2-yl)-2-oxoimidazolidin-l-yl)acetate (racemic) - Compound 111
Figure imgf000170_0001
Ή NMR (400 MHz, CDC13): δ 8.90-8.82 (m, 1 H), 7.62-7.57 (m, 1 H), 7.46 - 6.82 (m, 8H), 6.52- 6.48(m, 1 H), 6.15 - 5.85 (m, 2H), 4.88-4.82 (m, 1 H), 4.45-4.35 (m, 1 H), 4.32-4.13 (m, 2H), 3.86 - 3.46 (m, 3H), 3.05-2.85(m, 2H), 2.56-2.32 (m, 2H), 1.29 (t, J= 7.1 Hz, 3H). MS : 670 (M+ l )+.
Example 25. Preparation of Additional Compounds of Formula I
General procedu es for the UGI reaction:
A mixture of aldehyde (3.5 mmol) and aniline (3.5 mmol) in MeOH (8 mL) was stirred at room temperature for 30 min. Then the acid (3.5 mmol) was added and the reaction mixture was stirred for another 30 min, followed by addition of the isocyanide (3.5 mmol). The resulting mixture was then stirred at room temperature overnight and quenched with H20. The resulting mixture was partitioned between EtOAc and H20. The organic layer was washed with brine, dried over anhydrous Na2S04, and then concentrated. The resulting residue was purified by a standard method to afford the desired product.
General procedures for the Buchwald reaction: A mixture of amine (0.30 mmol), aryl halide (0.30 mmol), CS2CO3 (129 mg, 0.39 mmol), Pd2(dba)3 ( 18 mg, 0.02 mmol) and Xant-Phos (9.4 mg, 0.02 mmol) in 1 ,4-dioxane ( 10 mL) was stirred under N2 at 80 °C overnight. After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by a standard method to give the desired products.
The following analogs were synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted-aromatic ring or
heteroaromatic ring using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
2-(2-Chlorophenyl)-4-((S)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidin-2-yl)-N-(3,3- difluorocyclobutyl)-3-(lH-indazol-7-yl)-4-oxobutanamide - Compound 300
Figure imgf000171_0001
Ή NMR (400 MHz, CDC13) δ 13.18 (s, 1 H), 8.79 (s, 0.5H), 8.70 - 8.63 (m, 1 H), 8.53 (d, J= 5.0 Hz, 0.5H), 8.22 (s, 0.5H), 8.06 (s, 0.5H), 7.83 (d, J= 8.0 Hz, 0.5H), 7.61 (d, J= 8.0 Hz, 0.5H), 7.50 (d, J= 7.3 Hz, 0.5H), 7.38 (m, 3H), 7. 13 (t, J= 7.7 Hz, 1 H), 7.05 (t, J= 7.5 Hz, 1 H), 6.98 (d, J= 7.3 Hz, H), 6.83 (t, J= 7.7 Hz, 0.5H), 6.67 (d, J= 7.2 Hz, 0.5H), 6.24 (d, J= 6.5 Hz, 0.5H), 6.02 (s, 0.5H), 5.79 (d, J= 6.3 Hz, 0.5H), 5.12 (dd, J= 9.3, 3.4 Hz, 0.5H), 4.95 (s, 0.5H), 4.61 (dd, J= 9.2, 3.0 Hz, 0.5H), 4.22 (d, J= 7.4 Hz, 1 H), 3.00 - 2.71 (m, 3H), 2.59 - 2.05 (m, 4H), 2.04 - 1 .95 (m, 1 H). MS: 603.2 (M+ l )+.
(S)-3-(4-cyanopyridin-2-yI)-N-((R)-2-((3,3-difluorocyclobutyI)amino)-2-oxo-l-phenylethyl)- N-(5-fluoropyridin-3-yl)-2-oxo-l,3-oxazinane-4-carboxamide - Compound 301
Figure imgf000171_0002
Ή NMR (400 MHz, CDC13): 68.54 - 8.21 (m, 3H), 8.08-7.99 (m, 1H), 7.31 - 7.27 (m, 2H), 7.26 - 7.20 (m, 2H), 7.09 - 6.87 (m, 2H), 6.38 - 6.17 (m, 1 H), 5.89 - 5.60 (m, 1H), 4.93 - 4.70 (m, 1H), 4.62-4.46 (m, 1 H), 4.42 - 4.21 (m, 2H), 3.12 - 2.91 (m, 2H), 2.60 - 2.10 (m, 4H). MS: 565.2 (M+l)+.
(S)-3-(4-cyanopyridin-2-yl)-N-((R)-2-(3,3-difluorocyclobutylamino)-l-(2-fluorophenyl)-2- oxoethyl)-N-(5-fluoropyridin-3-yl)-2-oxo-l,3-oxazinane-4-carboxamide - Compound 302
Figure imgf000172_0001
Ή NMR (400 MHz, CDC13): 58.95 -7.77 (m, 5H),7.21 (s, 1H), 7.15 (s, 1H), 7.00 - 6.71 (m, 3H), 6.45 (m, 2H), 4.81 (m, 1H), 4.38 (d, J= 7.8 Hz, 1H), 4.23 (m, 2H), 2.92 (s, 2H), 2.45 (s, 2H), 2.08 (s, 2H). MS: 583 (M+l)+.
2-(2-chlorophenyl)-4-((S)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidin-2-yl)-N-(3,3- difluorocyclobutyl)-3-(2-fluoropyridin-4-yl)-4-oxobutanamide - Compound 303
Figure imgf000172_0002
1 HNMR (400 MHz, CDC13) δ 8.76 - 8.70 (m, 1 H), 8.59 - 8.49 (m, 1 H), 8.17 - 8.04 (m, 1 H), 7.43 - 7.27 (m, 3H), 7.24 - 6.95 (m, 3H), 6.44 - 6.25 (m, 1H), 6.10 - 6.5.96 (m, 1H), 4.94 - 4.75 (m, 3.1 Hz, 1H), 4.34 -4.18 (m, 1H), 3.03 (m, 1 H), 2.93 - 2.81 (m, 2H), 2.65 -2.09 (m, 5H). MS: 582.1 (M+l)+.
Compound 304
Figure imgf000173_0001
Ή NMR (400 MHz, CDC13): δ 8.77 - 8.45 (m, 2H), 8.46 - 7.84 (m, 2H), 7.61 (m, 1H), 7.31 - 7.22 (m, 2H), 7.09 (m, 1H), 6.89 (d, J= 4.3 Hz, 2H), 6.46 (d, J= 5.7 Hz, 1H), 6.07 (m, 1H), 4.86 (m, 1H), 4.34 (d,J= 6.1 Hz, 1 H), 3.19 - 2.76 (m, 3H), 2.63 - 2.08 (m, 4H), 1.93 (m, 1H). MS: 605.1 (M+l)+.
(S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(7-fluoroquinolin-5-yl)-5-oxopyrrolidine-2-carboxamide - Compound
Figure imgf000173_0002
Ή NMR (400 MHz, CDC13): δ 8.98 (s, 1H), 8.89 (s, 1H), 8.31 (m, 3H), 7.46 (s, 3H), 7.21 -7.06 (m, 3H).6.31 (s, 1H), 6.13 (s, 1H), 5.04 (d,J=6.9 Hz, 1H), 4.86 (d,J=6.5 Hz, 1H), 4.37 (s, 2H), 2.51 (m, 5H), 2.24 (s, 1H). MS: 633.2 (M+l)+.
Example 26. Preparation of (2S)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutyI amino)-2- oxoethyl)-l-(4-cyanopyridin-2-yI)-N-(3-fluoro-5-(pyridin-4-yl)phenyl)-5-oxopyrrolidine-2- carboxamide - Compound 306
Compound 306 was prepared according to the following scheme, using the following protocol.
Figure imgf000173_0003
Step A: (2S)-N-(3-bromo-5-fluorophenyl)-N-(l-(2-chlorophenyl)-2-(3,3-difluoro - cyclobutylamino)-2-oxoethyl)-l-(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxamide. 3-
Bromo-5-fluoroaniline ( 1 89 mg, 1 mmol), 2-chlorobenzaldehyde (140 mg, 1 mmol), (S)-l -(4- cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxylic acid (231 mg, 1 mmol) and l , l -difluoro-3- isocyanocyclobutane ( 1 1 8 mg, 1 mmol) were used in the general UGI reaction to give the desired product. MS: 660.1 (M+ l )+.
Step B: Compound 306. A mixture of (2S)-N-(3-bromo-5-fluorophenyl)-N-( l -(2-chlorophenyl)- 2-(3,3-difluorocyclobutylamino)-2-oxoethyl)- l -(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2- carboxamide ( 150 mg, 0.23 mmol), pyridin-4-ylboronic acid (42.4 mg, 0.35 mmol), Cs2C03 (224 mg, 0.69 mmol), Pd2(dba)3 ( 16.2 mg, 0.023 mmol) and Xant-Phos (27 mg, 0.046 mmol) in 1 ,4- dioxane (5 mL) was stirred under N2 at 100 °C overnight and then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by a standard method to give the desired products. Ή NMR (400 MHz, CDC13): δ 8.81 - 8.57 (m, 3H), 8.56 - 8.37 (m, 1 H), 8.06 - 7.92 (m, 1 H), 7.46 - 7.36 (m, 3H), 7.24 - 7.12 (m, 3H), 7.12 - 6.93 (m, 2H), 6.98 - 6.01 (m, 2H), 4.98 - 4.86 (m, 1 H), 4.37 - 4.1 8 (m, 1 H), 3.15 - 2.63 (m, 4H), 2.53 - 2.4 (m, 2H), 2.32 - 2.26 (m, 2H). MS: 659. 1 (M+ l )+.
The following analogs were synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and boronic acid using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
(S)-N-((R)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(pyridin-4-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 307
Figure imgf000174_0001
Ή NMR (400 MHz, CDC13): δ 8.76 - 8.58 (m, 3H), 8.47 (m, 1H), 8.08 (s, 1H), 7.79 (d, J= 8.7 Hz, 1 H), 7.49 - 7.38 (m, 1H), 7.35 - 7.24 (m, 2H), 7.16 (m, 3H), 7.06 - 6.91 (m, 2H), 6.71 (m, 1H), 6.44 (m, 1H),4.97 (dd, J= 9.2, 3.0 Hz, 1H), 4.32 (m, 1H), 3.14 - 2.75 (m, 3H), 2.36 (m, 4H), 2.15 - 1.66 (m, 3H). MS: 659.1 (M+l)+.
(2S)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutyIamino)-2-oxoethyl)-l-(4-cyanopyridin- -yl)-N-(3-fluoro-5-(pyridin-3-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 308
Figure imgf000175_0001
Ή NMR (400 MHz, CDC13): δ 8.56 - 7.98 (m, 4H), 7.83 - 7.36 (m, 2H), 7.35 - 7.26 (m, 2H), 7.21 - 6.89 (m, 5H), 6.47 - 6.17 (m, 2H), 4.99-4.87 (m, 1H), 4.36-4.16 (m, 1H), 3.12 - 2.67 (m, 4H), 2.51 -2.36 (m, 2H),2.31 -2.19 (m, 2H). MS: 659.1 (M+l)+.
(S)-N-((5)-l-(2-chlorophenyl)-2-(3,3-dinuorocyclobutylamino)-2-oxo-ethyl)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(isoxazol-5-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 309
Figure imgf000175_0002
Ή NMR (400 MHz, CDC13): δ 8.77 - 8.32 (m, 4H), 7.79 (m, 1H), 7.59 (m, 2H), 7.42 (m, 3H), 7.25-6.92 (m, 3H), 6.71 (d,J=6.7 Hz, 1H),6.45 (m, 1H),4.98 (dd, J= 9.1, 3.4 Hz, 1H), 4.35 (s, 1H), 3.10-2.74 (m, 3H), 2.64 - 2.19 (m, 4H), 2.17 -2.00 (m, 1H), 1.91 -1.73 (m, 1H).MS: 659.1 (M+l)+. (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-dinuorocyclobutyl)amino)-2-oxoethyI)-l-(4- cyanopyridin-2-yI)-N-(3-fluoro-4-(pyridin-4-yl)phenyl)-5-oxopyrroIidine-2-carboxamide - Compound 310
Figure imgf000176_0001
Ή NMR (400 MHz, CDC13): δ 8.74 - 8.51 (m, 3H), 7.85 (s, 1 H), 7.68 - 7.63 (m, 1 H), 7.54 - 7.45 (m, 2H), 7.36 - 7.35 (m, 2H), 7.20 (m, 2H),7.03 - 6.98 (m, 2H), 6.45 (m, 1 H), 6.17-6.16 (m, 1 H), 4.94 - 4.95 (m, 1 H), 4.34 (m, 1 H), 3.0 - 2.84 (m, 3H), 2.6 - 2.26 (m, 4H), 2.07 (m, 1 H). MS: 659.1 (M+ l )+.
(S)-N-((R)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-4-(pyridin-3-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 311
Figure imgf000176_0002
Ή NMR (400 MHz, CDC13): δ 8.65 (m, 4H), 7.79 (d, J= 7.9 Hz, 1 H), 7.36 (m, 5H), 7.25 - 7.03 (m, 3H), 6.46 (s, 1 H), 6.29 (s, 1 H), 4.93 (m, 1 H), 4.15 (s, 1 H), 3.10 - 2.70 (m, 3H), 2.57 (m, 1 H), 2.38 - 1 .99 (m, 4H). MS: 659. 1 (M+ l )+.
(S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyI)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-4-(pyridin-3-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 312
Figure imgf000177_0001
Ή NMR (400 MHz, CDC13): δ 8.80 - 8.46 (m, 4H), 7.96 - 7.73 (m, 2H), 7.35 (dd, J= 7.8, 4.9 Hz, 3H), 7.24 - 6.92 (m, 5H), 6.46 (s, 1 H), 6.27 (d, J = 5.4 Hz, 1 H), 4.98 (dd, J - 9.3, 3.2 Hz, l H), 4.35 (m, 1 H), 3.01 (m, 2H), 2.95 - 2.82 (m, 1 H), 2.64 - 2. 17 (m, 5H). MS: 659.1 (M+ l)+.
Example 27. (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxo -ethyl)- l-(4-cyanopyridin-2-yl)-N-(3-fluoro-4-(pyridin-2-yl)phenyl)-5-oxopyrrolidine-2- carboxamide - Compound 313
Compound 313 was prepared according to the following scheme, using the following protocol.
Figure imgf000177_0002
Step A: l-(4-Cyano-pyridin-2-yl)-5-oxo-pyrrolidine-2-carboxylic acid [(2-chloro- phenyl)-(3,3- difluoro-cyclobutylcarbamoyl)-methylJ-[3-fluoro-4-(4,4,5,5-tet methyl-[l,3,2]dwxaborolan-2- yl)-phenyl]-amide. To a solution of (S)-N-(4-bromo-3-fluoro -phenyl)-N-((S)- l -(2- chlorophenyl)-2-((3,3-dif^uorocyclobutyl)amino)-2-oxoethyl)^
oxopyrrolidine-2-carboxamide (500 mg, 0.758 mmol) in 1 .4- dioxane (10 mL) was added Pd(dppf)Cl2 ( l 10.8 mg, 0. 152 mmol), CH3COOK ( 185.6 mg, 1 .90 mmol), and
bis(pinacolato)diboron (384.7 mg, 1 .5 16 mmol). The mixture was stirred at 80 °C overnight and filtered. The filtrate was evaporated under reduced pressure and the residue was purified by a standard method to give the desired product. MS: 708.2 (M+ l )+. Step B: Compound 313. To a solution of l -(4-cyano -pyridin-2-yl)-5-oxo-pyrrolidine-2- carboxylic acid [(2-chloro-phenyl)- (3,3-difluoro -cyclobutylcarbamoyl)-methyl]-[3-fluoro-4- (4,4,5,5-tetramethyl-[ l ,3,2]dioxaborolan-2-yI)-phenyl]-amide (650 mg, 0.919 mmol) and 2- bromo-pyridine (159.7 mg, 1.01 mmol) in 1.4-dioxane/H20 (7 mL/3 drops) was added
Pd(dppf)Cl2 (67.3 mg, 0.092 mmol), Cs2C03 (449.5 mg, 1 .38 mmol). The mixture was stirred at 95 °C for 3 hr and filtered. The filtrate was evaporated under reduced pressure and the residue was purified by a standard method to give the desired product. Ή NMR (400 MHz, CDC13): δ 8.71 (d, J= 1 1.6 Hz, 2H), 8.51 (s, 1 H), 7.99 - 7.64 (m, 4H), 7.32 (s, 2H), 7.10 (m, 4H), 6.45 (m, 1 H), 6.1 5 (m, 1 H), 4.95 (d, J= 6.8 Hz, 1 H), 4.34 (m, 1 H), 3.09 - 2.81 (m, 3H), 2.61 - 2.20 (m, 4H), 2.12 - 2.00 (m, 1 H). MS: 659.1 (M+ l )+.
The following analog was synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted-aromatic ring or heteroaromatic ring using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
(2S)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxoethyl)-l-(4-cyanopyridin- -yI)-N-(3-fluoro-5-(pyridin-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 314
Figure imgf000178_0001
Ή NMR (400 MHz, DMSO-d6): δ 8.84 - 8.64 (m, 3H), 8.65 - 8.24 (m, 1 H), 8.03 - 7.60 (m, 3H), 7.53 - 7.3 1 (m, 3H), 7.26 - 6.95 (m, 3H), 6.80 (m, 1 H), 6.43 (m, 1 H), 5. 12 - 4.85 (m, 1 H), 4.28 (m, 1 H), 2.85 (m, 3H), 2.67 - 1.98 (m, 5H). MS: 659 (M+ l )+.
Example 28. (S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxo -ethyl)- l-(4-cyanopyridin-2-yl)-N-(3-fluoro-5-(isoxazol-5-yl)phenyl)-5-oxopyrrolidine-2- carboxamide - Compound 315 ompound 315 was prepared according to the following scheme, using the following protocol.
Figure imgf000179_0001
Step A: 3-(dibenzylamino)-5-fluorobenzonitrile. A mixture of 3-amino-5-fluoro -benzonitrile ( 13.6 g, 0.1 mol), 2C03 (62.1 g, 0.3 mol), BnBr (5 1 .4 g, 0.3 mol) in CH3CN ( 150 mL) was stirred at 78 °C overnight. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by a standard method to give the desired product. MS:
3 1 7.1 (M+ l )+.
Step B: l-(3-(dibenzylamino)-5-fluorophenyl)ethanone. To a mixture of 3-(dibenzyl -amino)-5- fluorobenzonitrile ( 16 g, 0.05 mol) in anhydrous THF ( 150 mL) at room temperature was dropwise added CH3MgBr ( 1 N solution in THF, 60 mL, 0.06 mol). The resulting mixture was stirred at 80 °C for 4 hr and then cooled down. The mixture was poured into 2 N HC1 (68 mL), followed by addition of methanol (68 mL). The mixture was concentrated and the residue was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine ( 100 mL), dried over anhydrous Na2SC>4 and then concentrated in high vacuum. The residue was purified by a standard method to give the desired product. MS: 334.1 (M+ l )+.
Step C: (E)-l-(3-(dibenzylamino)-5-fluorophenyl)-3-(dimethylamino)prop-2-en -1-one. A mixture of l -(3-(dibenzylamino)-5-fluorophenyl)ethanone (2 g, 6 mmol), DMF-DMA (860 mg, 7.2 mmol) in toluene (30 mL) was stirred at 120 °C for 8 hr. The mixture was concentrated in high vacuum and the residue was purified by a standard method to give the desired product. MS:
389.2 (M+ l )+. StepD: N,N-dibenzyl-3-fluoro-5-(isoxazol-5-yl)aniline. A mixture of (E)-l-(3-(di - benzylamino)-5-fluorophenyl)-3-(dimethylamino)prop-2-en-l-one (1.5 g, 3.86 mmol), hydroxy lamine hydrochloride (534 mg, 7.73 mmol) and pyridine (611 mg, 7.73 mmol) in ethanol (20 mL) was stirred at 78 °C overnight. The resulting mixture was evaporated under reduced pressure and the residue was purified by astandard method to give the desired product. M S:
359.1 (M+l)+.
Step E: 3-Fluoro-5-isoxazol-5-yl-phenylamine. 10% Pd/C (360 mg) was addedto a solution of dibenzyl-(3-fluoro-5-isoxazol-5-yl-phenyl)-amine (200 mg, 0.559 mmol) in a mixed solvent composed by EtOAc (36 mL), M eOH (15 mL) and H20 (7.5 mL). 6 drops of aq. HC1 (6 N) was then added into the above suspension and the resulting reaction mixture was stirred at 25°C for 1 hr. The mixture was filtered through Celite. The filtrate was evaporated under reduced pressure and the residue was purified by a standard method to give the desired product. MS: 179.0 (M+l)+. Step F: (2S)-N-(l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)- N-(3- fluoro-5-(isoxazol-5-yl)phenyl)-5-oxopywolidine-2-carboxamide. 2-Chlorobenzaldehyde (104 mg, 0.74 mmol), 3-fluoro-5-isoxazol-5-yl-pheny lamine (132 mg 0.74 mmol), (S)-5- oxopyrrolidine-2-carboxylic acid (95 mg, 0.74 mmol) and l ,l-difiuoro-3-isocyanocyclobutane (87 mg, 0.74 mmol) were used in the general UGI reaction to afford the desired product. M S: 547.1 (M+l)+.
Step G: Compound315. (2S -N-(l-(2-chloro -phenyl)-2-(3,3-difluorocyclobutylamino)-2- oxoethyl)-N-(3-fiuoro-5-(isoxazol-5-yl)phenyl)-5-oxopyrrolidine-2-carboxamide (200 mg, 0.37 mmol), 2-bromopyrimidine (102 mg, 0.56 mmol), Cs2C03 (240 mg, 0.74 mmol), Pd2(dba)3 (37 mg, 0.04 mmol) andXant-Phos (22 mg, 0.03 mmol) in 1,4-dioxane (15 mL) were stirred under N2 at 80 °C overnight and then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by a standard method to give the desired pro duct. !H NMR (400 MHz, CDC13): δ 8.69-8.17 (m, 3H), 7.80-7.28 (m, 3H), 7.25-6.93 (m, 5H), 6.63-6.30 (m, 3H), 4.96- 4.92 (m, lH), 4.374.34 (m, 1H), 3.06-2.83 (m, 3H), 2.58-2.21(m, 4H), 2.08-2.02 (m, 1H). M S: 649.1 (M+l)+. Example 29. (2S)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-ox oethyl) -l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(oxazoI-5-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 316
Compound 316 was prepared according to the following scheme, using the following protocol.
Figure imgf000181_0001
Step A: 5-(3-fluoro-5~nitrophenyl)oxazole. To a solution of 3-fluoro-5-nitrobenz -aldehyde (340 mg, 2.0 mmol) and 2-tosylacetonitrile (430 mg, 2.2 mmol) in MeOH (25 mL) and DME (25 mL) was added Amberlyst A26 OH-resin (3.7 g). The mixture was heated under reflux for 1 hr and cooled to r.t.. The resin was filtered and rinsed with MeOH. The filtrate was concentrated under reduced pressure to give the crude product which was used directly in the next step. MS: 209. 1 (M+ l )+.
Step B: 3-fluoro-5-(oxazo/-5-yl) niIine. To a solution of 5-(3-fluoro-5-nitrophenyl) -oxazole (400 mg, 2.0 mmol) in TFA ( 10 mL) was added Zn powder (380 mg, 6.0 mmol). The mixture was stirred at room temperature for 2 hr and poured into ice. The resulting mixture was neutralized with NH .H20 to pH= 10 and then extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2S04 and then concentrated in high vacuum. The residue was purified by a standard method to afford the desired product. MS: 179.0 (M+ l )+.
Step C: Compoun 316. 2-Chlorobenzaldehyde ( 104 mg, 0.74 mmol), 3-fluoro-5-(oxazol-5- yl)aniline ( 1 32 mg, 0.74 mmol), (S)- l -(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxylic acid ( 171 mg, 0.74 mmol) and 1 , 1 -difluoro- 3-isocyanocyclobutane (87 mg, 0.74 mmol) were used in the general UGI reaction to afford the desired product. Ή NMR (400 MHz, CDC13): δ 8.63 (m, 2H), 8. 12 - 7.82 (m, 1 H), 7.78 - 7.30 (m, 2H), 7.26 (t, J= 7.4 Hz, 2H), 7.15 (m, 2H), 7.12 - 6.83 (m, 2H), 6.70 - 6.16 (m, 2H), 5.03 - 4.74 (m, 1 H), 4.24 (m, 1 H), 2.93 (m, 2H), 2.63 (m, 2H), 2.20 (m, 3H). MS: 649.1 (M+ l )+.
The following analog was synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted-aromatic ring or heteroaromatic ring using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
(S)-N-((S)-l-(2-chlorophenyl)-2-((3,3-difluorocyclobutyI)amino)-2-oxoethyl)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(oxazoI-5-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 317
Figure imgf000182_0001
Ή NMR (400 MHz, CDC13): δ 8.68 (s, 1 H), 8.60 - 8.45 (m, 1 H), 8.06 -7.68 (m, 2H), 7.43 (s, 1 H), 7.34 - 7.10 (m, 4H), 7.04 - 6.93 (m, 2H), 6.54 (d, J = 6.9 Hz, 1 H), 6.43 (d, J = 1 1.5 Hz, 1 H), 4.95 (t, J = 6.5 Hz, 1 H), 4.35 (m, 1 H), 3.1 1 - 2.74 (m, 3H), 2.66 - 2.14 (m, 4H), 2.09 - 1.97 (m, 1 H). MS: 649.1 (M+ l )+.
Example 30. (2S)-N-(l-(2-chlorophenyl)-2-(3,3-difluorocyclobutylamino)-2-oxo ethyl) -l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(oxazol-2-yl)phenyl)-5-oxopyrrolidine-2-carboxamide - Compound 318
Compound 318 was prepared according to the following scheme, using the following protocol.
Figure imgf000183_0001
Step A: 2-(3-fluoro-5-nitrophenyl)oxazole. A mixture of 3-fluoro-5-nitrobenz -aldehyde (5 1 mg, 0.3 mmol) and 2,2-dimethoxyethanamine (32 mg, 0.3 mmol) was stirred at 1 10 °C for 2 hr and cooled to give a crude intermediate of (£ -N-(3-fluoro-5-nitrobenzylidene)-2,2- dimethoxyethanamine. A solution of 20 mg of the above intermediate in 0.5 mL of H2S04 was added into a mixture of 1 8 mg of P205 in 0.1 mL of H2S04. The resulting mixture was heated to 180 °C for 20 min, cooled down and neutralized with NH4OH to give the crude product which was used directly without further purification. MS: 209.1 (M+ l )+.
Step B: 3-fluoro-5-(oxazol-2-yl)aniline. To a solution of 2-(3-fluoro-5-nitrophenyl) -oxazole (380 mg, 1 .8 mmol) in TFA (10 mL) was added Zn powder (358 mg, 5.5 mmol). The mixture was stirred at r.t. for 2 hr and poured into ice. The resulting mixture was neutralized with
N H3.H2O to pH= 10 and then extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2S04 and then concentrated in high vacuum. The residue was purified by a standard method to afford the desired product. MS: 179.0 (M+ l )+.
Step C: Compound 318. 2-Chlorobenzaldehyde ( 104 mg, 0.74 mmol), 3-fluoro-5-(oxazol-2- yl)aniline ( 132 mg, 0.74 mmol), (S)- l -(4-cyanopyridin-2-yl)-5-oxopyrrolidine-2-carboxylic acid ( 171 mg, 0.74 mmol) and l , l -difluoro-3-isocyano cyclobutane (87 mg, 0.74 mmol) were used in the general UG1 reaction to afford the desired product. Ή NMR (400 MHz, CDC13): δ 8.70 (m, 2H), 8.56 - 7.83 (m, 1 H), 7.65 (m, 2H), 7.47 - 7.27 (m, 2H), 7.25 - 7.09 (m, 2H), 7.10 - 6.91 (m, 2H), 6.76 - 6.1 8 (m, 2H), 5.06 - 4.71 (m, 1 H), 4.25 (m, 1 H), 3.12 - 2.64 (m, 3H), 2.43 (m, 3H), 2.09 (m, 2H). MS: 649. 1 (M+ l )+. The following analog was synthesized via the procedures set forth above, using the appropriate aldehyde, amine, carboxylic acid, isocyanide and halo-substituted-aromatic ring or heteroaromatic ring using the reagents and solvents set forth above or similar reagents and solvents thereof, and purified via standard methods.
(S)-N-((S)-l-(2-chlorophenyI)-2-((3,3-difluorocycIobutyI)amino)-2-oxoethyI)-l-(4- cyanopyridin-2-yl)-N-(3-fluoro-5-(oxazoI-2-yl)phenyI)-5-oxopyrrolidine-2-carboxamide - Compound 319
Figure imgf000184_0001
Ή NMR (400 MHz, CDC13): 5 8.82 - 8.26 (m, 2H), 7.94 - 7.46 (m, 3H), 7.38 - 6.81 (m, 6H), 6.44 (m, 1 H), 6.20 (dd, J = 14.9, 6.9 Hz, 1 H), 4.94 (m, l H), 4.36 (m, 1 H), 3.15 - 2.70 (m, 3H), 2.62 - 1 .86 (m, 5H).
Example 31. (2S)-N-((lS)-l-(2-chlorophenyl)-2-(6,6-difluorospiro[3.3]heptan-2-yl amino)-2- oxoethyl)-l-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2- carboxamide - Compound 320
Compound 320 was prepared according to the following scheme, using the following protocol.
CF3CHzOH, RT, 2 days
Figure imgf000184_0002
NHCHO NC
Figure imgf000184_0003
Step A N-(6,6-difluorospiro[3.3]heptan-2-yl)formamide. To a suspension of 6,6- difluorospiro[3.3]heptan-2-amine hydrochloride (500 mg, 2.73 mmol) in HCOOEt (5 mL) was added Et3 (552 mg, 5.47 mmol). The resulting mixture was stirred first at room temperature for 30 min in a sealed tank and then heated to 85 °C overnight. The mixture was concentrated and followed by addition of EtOAc (10 mL). The resulting suspension was stirred at room
temperature for 30 min and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by a standard method to afford the desired product. !H NMR (400MHz, CDC13): δ 8.06 (s, 1H), 5.71 (s, lH), 4.42 (m, lH), 2.74 -2.45 (m, 8H).
StepB: 2,2-difluoro-6-isocyanospiro[3.3]heptanes. A solution of N-(6,6- difiuorospiro[3.3]heptan-2-yl)formamide (390 mg, 2.23 mmol), PPh3 (642 mg, 2.45 mmol), CCI4 (339 mg, 2.23 mmol), andEt3N (225 mg, 2.23 mmol) in DCM (10 mL) was heated at 45 °C overnight. The mixture was concentrated under reduced pressure. The residue was suspended in Et20 (10 mL) and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by a standard method to afford the desired product. 1H NMR (400MHz, CDC13): δ 4.01 - 3.85 (m, 1H), 2.80 -2.37 (m, 8H).
Step C: (2S)-N-(l-(2-chlorophenyl)-2-(6,6-difluorospiro[3.3]heptan-2-ylamino)-2- oxoethyiy N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide. 2-Chlorobenz -aldehyde (104 mg, 0.74 mmol), 5-fiuoropyridin-3-amine (83 mg, 0.74 mmol), (S)-5-oxopyrrolidine-2-carboxylic acid (95 mg 0.74 mmol) and 2,2-dfluoro-6-isocyanospiro[3.3]heptane (l 16 mg, 0.74 mmo l) were used in the general UGI reaction to afford the desired product. M S: 521 (M+ l)+.
StepD: Compound 320 was synthesized via the general procedure for Buchwald reaction set forth above. 1H NMR (400MHz, CDC13): δ 8.97 (s, 0.5H), 8.70 (s, 1H), 8.48 (d, J= 4.6 Hz, 1H), 8.41 - 8.22 (m, 1.5H), 8.15 (d, J= 8.7 Hz, 1H), 7.31 (d, J= 7.1 Hz, lH), 7.25 (d, J= 4.9 Hz, 1H), 7.19 (t, J= 7.4 Hz, 1H), 7.03 (t, J= 7.3 Hz, 1H), 6.91 (m, 1H), 6.42 (m, 1H), 6.05 (m, 1H), 4.83 (m, 1H), 4.38 (m, 11H), 2.97 - 2.75 (m, 1H), 2.68 - 2.1 1 (m, 8H), 2.10 - 1.82 (m, 3H). M S:
623.1 (M+l)+.
Example A: In Vitro Assays for IDHlm (R132H or R132C) Inhibitors
A test compound isprepared as 10 mM stock in DM SO and diluted to 50X final concentration in DMSO, for a 50 μΐ reaction mixture. IDH enzyme activity converting alp ha- ketoglutarate to 2-hydroxyglutaric acid is measured using a NADPH depletion assay. In the assay the remaining cofactor is measured at the end of the reaction with the addition of a catalytic excess of diaphorase andresazurin, to generate a fluorescent signal inproportion to the amount of NADPH remaining. IDH1-R132 homodimer enzyme is diluted to 0.125
Figure imgf000186_0001
in 40 μΐ of Assay Buffer(150 mM NaCl, 20 mM Tris-ClpH 7.5, 10 mM M gCl2, 0.05% BSA, 2 mM b- mercaptoethanol); 1 μΐ of test compound dilution in DM SO is added and the mixture is incubated for 60 minutes at room temperature. The reaction is started withthe addition of 10 μΐ of
Substrate Mix (20 μΐ NADPH, 5 mM alp ha-keto glut ar ate, in Assay Buffer) and the mixture is incubated for 90 minutes at room temperature. The reaction is terminated with the addition of 25 μΐ of Detection Buffer (36 μg ml diaphorase, 30 mM resazurin, in IX Assay Buffer), and is incubated for 1 minute before reading on a SpectraM ax platereader at Ex544/Em590.
Compounds are assayed fortheir activity against IDH1 R132C following the same assay as above with the following modifications: Assay Buffer is (50 mM potassiumphosphate, pH 6.5; 40 mM sodium carbonate, 5 mM MgCl2, 10% glycerol, 2 mM b-mercaptoethanol, and0.03% BSA). The concentration of NADPH and alpha-ketoglutarate in the Substrate Buffer is 20 μΜ and 1 mM , resp ectively .
Representative compounds of formula I set forth in Table 1 were tested in this assay or a similar assay and the results are set forth below in Table 3. As used in Table 3, "A" refers to an inhibitory activity against IDH 1 R132H or IDH 1 R132Cwith an IC50 < 0.1 μΜ; "B" refers to an inhibitory activity against IDH 1 R132H or IDH 1 R132Cwith an IC50 between 0.1 μΜ and 0.5 μΜ ;'^" refers to an inhibitory activity against IDH1 R132H or lDHl R132C with an IC50 between 0.5 μΜ and 1 μΜ; "D" refers to an inhibitory activity against IDH1 R132H or IDH1
R132C with an ICa between 1 μΜ and 2 μΜ .
Table 3. Inhibitory Activities of Rep resent at ive Compounds of formulal
Figure imgf000186_0002
Figure imgf000187_0001
Figure imgf000188_0001
Figure imgf000189_0001
Example B: Cellular Assays for IDHlm (R132H or R132C) Inhibitors.
Cells (HT 1080 or U87M G) are grown in T125 flasks in DMEM containing 10% FBS, l x penicillin/streptomycin and 500ug/mL G418 (present inU87MG cells only). They are harvested by trypsin and seeded into 96 wellwhite bottomplates at a density of 5000 cell/well in 100 ul/well in DMEM with 10% FBS. No cells are placed in columns 1 and 12. Cells are incubated overnight at 37 ^ in 5% C02. The next day test compounds are made up at 2xthe final concentration and lOOul are addedto each cell well. The final concentration of DM SO is 0.2% and the DM SO control wells are plated in row G. The plates are then placed in the incubator for 48 hours. At 48 hours, lOOul of media is removed from each well and analyzed by LC-M S for 2- HG concentrations. The cell plate is placed back in the incubator for another 24 hours. At 72 hours post compound addition, 10 mL/plate of Promega Cell Titer Glo reagent is thawed and mixed. The cell plate is removed from the incubator and allowedto equilibrate to room temperature. Then lOOul of Promega Cell Titer Glo reagent is added to each well of media. The cell p late is then p laced on an orbital shaker for 10 minutes and then allowed to sit at room temperature for 20 minutes. The plate is then read for luminescence with an integration time of 500ms.
The IC50 for inhibition of 2-HG production (concentration of test compound to reduce 2HGproduction by 50% comparedto control) in these two cell lines forvarious compounds of formula I is set forth in Table 2 above. As used in Table 2, "A" refers to an IC50 for inhibition of 2-HG roduction < 0.1 μΜ; "B" refers to an IC5o for inhibition of 2-HGproduction between 0.1 μΜ and 0.5 μΜ ; "C" refers to an IC50 for inhibition of 2-HG production between 0.5μΜ and 1 μΜ ;'Ό" refers to an IC5o for inhibition of 2-HG production between Ι μΜ and 2μM .
Example C: Metabolic Stabilities of C ompounds of Formula I
M etabolic stabilities of compounds of formula I can be tested with the following assay and species specific liver microsomes (LM) extraction ratio (Eh) can be calculated:
1.Buffer A: 1.0 L of 0.1 M monobasic Potassium Phosphate buffer containing 1.0 mM
EDTA; Buffer B: 1.0 L of 0.1 M Dibasic Potassium Phosphate buffer containing 1.0 mM EDTA; Buffer C: 0.1 M Potassium Phosphate buffer, 1.0 mM EDTA, pH 7.4 by titrating 700 mL of buffer B with buffer A while monitoring with the pH meter.
2. Reference compounds (Ketanserin) andtest compounds spiking solution:
500 μΜ spiking solution: add 10 μί of 10 mM DM SO stock solution into 190 μΐ, CAN;
1.5 μΜ spiking solution in microsomes (0.75 mgAnL) : add 1.5 μL· of 500 μΜ spiking solution and 18.75 μL· of 20 mg mL liver microsomes ίηίο 479.75μΕ of Buffer C.
3. NADPH stock solution (6 mM ) is prepared by dissolving NADPH into buffer C.
4. Dispense 30 μL· 1.5X compound/liver microsome solution in 96-wellplate and immediately add 135 μί ACN containing I S before adding 15uL Buffer C to prepare real 0 minute samples.
5. Add 15 μL· of NADPH stock solution (6 mM ) to the wells designated as Time 30, and start timing.
6. At the end of incubation (0 min), add 135 μL· of ACN containing the internal standard
Osalmid) to all the wells (30 min, and 0 min). Then add 15 μL· of NADPH stock solution (6 mM ) to the wells designated as Time 0.
7. After quenching, centriiiige the reaction mixtures at 3220g for 10 min.
8. Transfer 50 μL· of the sup ernatant from each well into a 96-well sample plate containing 50 μL· of ultra pure water (M illip ore) for LC/M S analysis .

Claims

Claims
1. A compound of formula I or a pharmaceutically acceptable salt, tautomer, isotopologue or hydrate thereo
Figure imgf000191_0001
I
R1 is optionally substituted C4-C6 carbocyclyl;
each R 2 and R 3 i·s i·ndependently selected from optionally substituted aryl or optionally substituted heteroaryl;
R4 is alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aralkyl, or optionally substituted heteroaralkyl;
ring A is 4-6 membered non-aromatic ring having 0-1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups;
each R5 is independently halo; -CF3; -CN; -OR6;-N(R6)2; -C(0)d-C4 alkyl; C1-C4 haloalkyl; C1-C4 alkyl optionally substituted with -OR6 or -N(R6)2; -O-C1-C4 alkyl optionally substituted with halo, -OR6 or -N(R6)2; -S02N(R6)2; -S02(d-C4 alkyl); -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -0-(C3-C6 carbocyclyl optionally substituted with one or two R6 groups); 5-6 membered heteroaryl; -C1-C4 alkyl-C(0)0-Ci-C4 alkyl; or -C(0)0-Ci-C4 alkyl; or
each R6 is independently H or Ci-C3 alkyl.
2. The compound of claim 1, wherein:
1 7
R is C4-C6 carbocyclyl optionally substituted with one to three R groups;
each R 2 and R 3 i·s i·ndependently selected from aryl or heteroaryl, wherein said aryl or heteroaryl is independently optionally substituted with one to three R groups;
R4 is alkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl, wherein said aryl, heteroaryl, aralkyl, and heteroaralkyl are each independently optionally substituted with one to three R7 groups;
ring A is 4-6 membered non-aromatic ring having 0-1 additional heteroatoms selected from N, O or S, wherein ring A is optionally substituted with one or two R5 groups; each R5 and R7 is independently halo; -CF3; -CN; -OR6;-N(R6)2; -C(0)Ci-C4 alkyl; C1-C4 haloalkyl; C1-C4 alkyl optionally substituted with -OR6 or -N(R6)2; -O-C1-C4 alkyl optionally substituted with halo, -OR6 or -N(R6)2; -S02N(R6)2; -S(0)-Ci_4 alkyl; -S02(Ci-C4
alkyl); -NR6S02R6; C3-C5 carbocyclyl optionally substituted with one or two R6 groups; -0-(C3- C6 carbocyclyl optionally substituted with one or two R6 groups); 5-6 membered heteroaryl; -Q- C4 alkyl-C(0)0-Ci-C4 alkyl; or -C(0)0-Ci-C4 alkyl; or
each R6 is independently H or Ci-C4 alkyl.
3. The compound of claim 1 or 2, wherein each R2 and R3 is independently aryl optionally substituted with one to three R7 groups.
4. The compound of claim 1 having formula Il-a,
Figure imgf000192_0001
rmula Il-a
wherein R10 is CR11 or N; and R11 is -F, -S02NH2, -S02CH3, -S(0)CH3, -CN, methoxy, -OCH2OH, -CH2OH, -S02N(CH3)2, -S02NHCH3, -NHS02CH3, -CH2CH2OH, -N(CH3) 2, t-butyl, cyclopropyl, -C(OH)(CH3)2, -OCF3, -OCHF2, -O-cyclopropyl, -1-methyl-cyclopropyl, or pyrazolyl.
5. The compound of any one of claims 1-4, wherein R1 is C4 or C6 cycloalkyl optionally substituted with one to two R7 groups and R7 associated with R1 is halo.
6. The compound of claim 5, wherein R is
Figure imgf000193_0001
7. The compound of claim 5, wherein ring A is:
Figure imgf000193_0002
ent to the amide moiety of formula and ¾ denotes ring A's attachment to R ; and each member of ring A is optionally substituted with one or two R5 groups.
8. The compound of claim 7, wherein ring A is:
Figure imgf000193_0003
9. The compound of claim 7, wherein R4 is aryl or heteroaryl, each aryl or heteroaryl is optionally substituted with one to three R7 groups.
10. The compound of claim 9, wherein R4 is:
Figure imgf000194_0001
4 7 7
each member of R is optionally substituted with one or two R groups and each R is independently F, CI, methyl, CF3, CN, OMe, or N(R6)2.
11. The compound of claim 10, wherein R4 is:
Figure imgf000194_0002
methyl.
12. The compound is selected from any one of compounds from Table 1.
13. A pharmaceutical composition comprising a compound of any one of claims 1 to 12; and a pharmaceutically acceptable carrier.
14. The composition of claim 13, further comprising a second therapeutic agent useful in the treatment of cancer.
15. A method of treating a cancer characterized by the presence of an IDH1 mutation, wherein the IDH1 mutation results in a new ability of the enzyme to catalyze the
NAPH-dependent reduction of a-ketoglutarate to ii(-)-2-hydroxyglutarate in a patient, comprising the step of administering to the patient in need thereof a composition of claim 13.
16. The method of claim 15, wherein the IDH1 mutation is an IDH1 R132H or R132C mutation.
17. The method of claim 15, wherein the cancer is selected from glioma
(glioblastoma), acute myelogenous leukemia, melanoma, non-small cell lung cancer (NSCLC), cholangiocarcinomas, chondrosarcoma, myelodysplastic syndromes (MDS), myeloproliferative neoplasm (MPN), colon cancer in a patient.
18. The method of claim 17, further comprising administering to the patient in need thereof a second therapeutic agent useful in the treatment of cancer.
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WO2017162157A1 (en) * 2016-03-22 2017-09-28 正大天晴药业集团股份有限公司 Sultam compound and application method thereof
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US11247987B2 (en) 2017-10-06 2022-02-15 Forma Therapeutics, Inc. Inhibiting ubiquitin specific peptidase 30
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Publication number Priority date Publication date Assignee Title
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090163545A1 (en) * 2007-12-21 2009-06-25 University Of Rochester Method For Altering The Lifespan Of Eukaryotic Organisms
CN102659765A (en) * 2011-12-31 2012-09-12 沈阳药科大学 Pyrimidine and triazine compound preparation method and application

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2805669C (en) * 2010-07-16 2018-08-21 Agios Pharmaceuticals, Inc. Therapeutically active compositions and their methods of use
UA111503C2 (en) * 2011-09-27 2016-05-10 Новартіс Аг 3-pyrimidin-4-yl-oxazolidin-2-ones as inhibitors of mutant idh
CA2860858A1 (en) * 2012-01-19 2013-07-25 Agios Pharmaceuticals, Inc. Therapeutically active compounds and their methods of use
US9474779B2 (en) * 2012-01-19 2016-10-25 Agios Pharmaceuticals, Inc. Therapeutically active compositions and their methods of use

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090163545A1 (en) * 2007-12-21 2009-06-25 University Of Rochester Method For Altering The Lifespan Of Eukaryotic Organisms
CN102659765A (en) * 2011-12-31 2012-09-12 沈阳药科大学 Pyrimidine and triazine compound preparation method and application

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