CN118164966A

CN118164966A - MAT2A inhibitor and application thereof

Info

Publication number: CN118164966A
Application number: CN202211569257.XA
Authority: CN
Inventors: 崔洪; 王文义; 魏钊
Original assignee: Hengyuan Biomedical Technology Suzhou Co ltd
Current assignee: Hengyuan Biomedical Technology Suzhou Co ltd
Priority date: 2022-12-08
Filing date: 2022-12-08
Publication date: 2024-06-11

Abstract

The present disclosure relates to novel compounds useful as MAT2A inhibitors, pharmaceutical compositions comprising these compounds and methods of treatment by administration of these compounds or the pharmaceutical compositions.

Description

MAT2A inhibitor and application thereof

Technical Field

The present disclosure relates generally to novel compounds useful as MAT2A inhibitors as well as pharmaceutical compositions comprising these compounds and methods of treatment by administration of these compounds or the pharmaceutical compositions.

Background

Methionine Adenosyltransferase (MAT), also known as S-adenosylmethionine synthetase, is an enzyme that catalyzes the synthesis of S-adenosylmethionine (SAM or AdoMet) by methionine and ATP. Non-liver endogenous SAM is synthesized by methionine adenosine transferase IIA (MAT 2A) enzyme. In organisms, the expression and activity of DNA, RNA and proteins can be regulated by methylation modification, thereby controlling the growth, differentiation and death of cells. Thus, intracellular SAM levels will be tightly regulated.

The protein arginine N-methyltransferase (PRMT 5) is a methylase that utilizes SAM as a methyl donor, SAM being necessary for PRMT5 activity, while PRMT5 is competitively inhibited by 5' Methylthioadenosine (MTA). During the initiation of the methi-adenosine phosphorylase (methylthioadenosine phosphorylase, MTAP), the MTA level of the cells remained low. MTAP is located near cyclin-dependent kinase inhibitor 2A (CDKN 2A) tumor suppressor genes, and is co-deleted with CDKN2A in about 15% of all cancers. Since MTAP is the only enzyme currently known to catalyze the degradation of MTA, it is believed that the loss of MTAP results in the accumulation of MTA in cancer cells. The absence of MTAP results in the accumulation of MTA, making MTAP-deficient cells (MTAP-DELETED CELLS) more dependent on SAM production and thus on MAT2A activity than MTAP-expressing cells.

Experiments show that after MAT2A gene is knocked out, the symmetrical arginine dimethyl marker (SYMMETRIC ARGININE dimethylation mark, SDMA) of the HCT116 isogenic pair is changed differently, and the SDMA level of MTAP deletion cells is obviously lower than that of MTAP wild type cells. Under the background of MTAP deficiency cells and metabolic change in a high MTA environment, inhibition of SAM level reduction caused by MAT2A activity can inhibit PRMT5 activity, so that tumor cell death is selectively induced, and a synthetic lethal effect is achieved.

With the elucidation of the mechanism of action of MAT2A protein in recent years, MAT2A inhibitors have begun to be developed continuously. However, no MAT2A inhibitor has been approved for clinical use so far, and thus, development of a highly potent inhibitor is necessary to bring new therapeutic means to related tumor patients in the future.

Disclosure of Invention

The present disclosure provides certain 2-oxoquinazoline derivatives that are inhibitors of methionine adenosyltransferase 2A (MAT 2A). Also disclosed herein are pharmaceutical compositions comprising such compounds and methods of treating diseases treatable by inhibition of MAT2A, such as cancer, including cancers characterized by reduced or inactive methicillin-p-phosphorylase (MTAP) activity.

In a first aspect, the present disclosure provides a compound of formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:

wherein:

w is CR ⁴ or N;

x is CR ⁵ or N;

y is CR ⁶ or N;

z is CR ⁷ or N;

v is CR ⁸ or N;

R ⁴、R⁵、R⁶、R⁷ are each independently selected from the group consisting of: hydrogen, alkyl, alkylcarbonyl, hydroxy, halogen, haloalkyl, cyano, alkynyl and-NR ^aR^b;

R ⁸ is selected from the group consisting of: hydrogen, alkyl, and hydroxy;

R ¹ is selected from the group consisting of: alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl and-NR ^cR^d;

R ² is selected from the group consisting of: hydrogen, alkyl, alkoxy, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein each of alkyl, alkoxy, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more R ^e;

R ³ is selected from the group consisting of: heterocyclyl, aryl, and heteroaryl, wherein each of heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more R ^e;

R ^a and R ^b are each independently selected from hydrogen or alkyl;

r ^c is selected from the group consisting of: hydrogen, alkyl, and cycloalkyl;

R ^d is selected from the group consisting of: alkyl, alkoxy, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein each of alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more R ^e;

Each R ^e is independently selected from the group consisting of: oxo, cyano, halogen, hydroxy, acyl, -NR ^fR^g、-OC(O)NR^fR^g, carbamoyl, carboxy, alkyl, alkenyl, alkynyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, cycloalkylalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

Each R ^f is independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, heteroaryl, wherein each of alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, and heteroaryl is optionally substituted with one or more groups independently selected from cyano, halo, hydroxy, or amino; and

Each R ^g is independently hydrogen or C _1-3 alkyl.

In one embodiment, the compound or stereoisomer, tautomer, or pharmaceutically acceptable salt thereof has a structure represented by the following formula (Ia) or (Ib):

in one embodiment, the R ¹ is-NR ^cR^d.

In one embodiment, the R ^c is hydrogen or alkyl.

In one embodiment, the R ^c is methyl.

In one embodiment, the R ^d is selected from the group consisting of: alkyl, alkoxy, and cycloalkyl, wherein each of alkyl, alkoxy, and cycloalkyl is optionally substituted with one or more R ^e.

In a preferred embodiment, each R ^e is independently selected from the group consisting of: cyano, hydroxy, -NR ^fR^g, carbamoyl, alkyl, alkenyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, cycloalkylalkyl, cycloalkyl and heterocyclyl.

In one embodiment, the R ^d is selected from the group consisting of: methyl group,

And methoxy.

In one embodiment, the R ² is selected from the group consisting of: hydrogen, alkyl, and aryl.

In one embodiment, the R ² is methyl.

In one embodiment, the R ² is phenyl.

In one embodiment, the R ³ is heterocyclyl.

In one embodiment, the R ³ is a five membered heterocyclyl.

In one embodiment, the R ³ is selected from

In one embodiment, the R ⁴ is hydrogen.

In one embodiment, the R ⁵ is hydrogen.

In one embodiment, the R ⁶ is selected from the group consisting of: alkylcarbonyl, halogen, haloalkyl, alkynyl and-NR ^aR^b.

In one embodiment, the R ⁶ is selected from the group consisting of: chlorine, trifluoromethyl,

In one embodiment, the R ⁷ is hydrogen.

In one embodiment, the R ⁸ is hydrogen.

In one embodiment, the compound has a formula selected from the group consisting of:

In a second aspect, the present disclosure provides a pharmaceutical composition comprising a compound as described in the first aspect above, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

In a third aspect, the present disclosure provides a method of treating a MAT 2A-mediated disease in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound as described in the first aspect above, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the second aspect above.

In one embodiment, the disease in the third aspect above is cancer.

In a fourth aspect, the present disclosure provides a method of treating MTAP-deficient cancers in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound as described in the first aspect above, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the second aspect above.

In a fifth aspect, the present disclosure provides a method for treating cancer in a patient, wherein the cancer is characterized by reduced or non-expression of MTAP gene, a deletion of MTAP gene, or reduced function of MTAP protein, the method comprising administering to the subject a therapeutically effective amount of a compound as described in the first aspect above, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the second aspect above.

In a sixth aspect, the present disclosure provides the use of a compound as described in the first aspect above, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the second aspect above, in the manufacture of a medicament for the treatment of cancer.

In one embodiment, the cancer in the third or fourth or fifth or sixth aspect above is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

Detailed Description

The definition of specific functional groups and chemical terms is described in more detail below. For purposes of this disclosure, chemical elements are identified according to the periodic Table of elements (Periodic Table of THE ELEMENTS), CAS version, handbook of physics and chemistry (CHEMISTRY AND PHYSICS), 75 th edition, inner cover, and specific functional groups are generally defined as described herein. In addition, the general principles of organic chemistry and specific functional moieties and reactivities are described in the following references: organic chemistry (Organic Chemistry), thomas Sorrell, 2 nd edition, university science book press (University Science Books), assailitot, 2006; smith and March, mach' S ADVANCED Organic Chemistry, 6 th edition, john wili father company (John Wiley & Sons, inc.), new york, 2007; larock, integrated organic transformation (Comprehensive OrganicTransformations), 3 rd edition, VCH Press (VCH Publishers, inc.), new York, 2018; carruthers, some modern methods of organic synthesis (Some Modern Methods of OrganicSynthesis), 4 th edition, cambridge university Press (Cambridge University Press), cambridge, 2004; the entire contents of each of the references are incorporated herein by reference.

Throughout this disclosure, linking substituents are described. It is particularly desirable that each linking substituent includes both the forward and reverse forms of the linking substituent. For example, -NR (CR 'R') -includes both-NR (CR 'R') -and- (CR 'R') NR-. In the case where a linking group is explicitly required for a structure, the Markush variable (Markush variable) listed for the group is understood to be the linking group. For example, if the structure requires a linking group and the markush group definition of the variables lists "alkyl", it is understood that "alkyl" means a linking alkylene.

Where a bond to a substituent is shown to intersect a bond connecting two atoms in a ring, the substituent may be bonded to any atom in the ring. Where a substituent is listed, but it is not specified through which atom the substituent is bonded to the remainder of a given formula compound, the substituent may be bonded through any atom in the formula. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

Where any variable (e.g., R ⁱ) occurs more than one time in any component or formula of a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0 to 2R ⁱ moieties, then the group may optionally be substituted with up to two R ⁱ moieties, and R ⁱ is independently selected at each occurrence from the definition of R ⁱ. Moreover, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

As used herein, the term "C _i-j" indicates a range of numbers of carbon atoms, where i and j are integers, and the range of numbers of carbon atoms includes the endpoints (i.e., i and j) and each integer point therebetween, and where j is greater than i. For example, C _1-6 indicates a range of one to six carbon atoms, including one carbon atom, two carbon atoms, three carbon atoms, four carbon atoms, five carbon atoms, and six carbon atoms. In some embodiments, the term "C _1-12" indicates 1 to 12 carbon atoms, especially 1 to 10 carbon atoms, especially 1 to 8 carbon atoms, especially 1 to 6 carbon atoms, especially 1 to 5 carbon atoms, especially 1 to 4 carbon atoms, especially 1 to 3 carbon atoms or especially 1 to 2 carbon atoms.

As used herein, the term "acyl" refers to-C (=o) -R, wherein R is a substituent such as hydrogen, alkyl, cycloalkyl, aryl, or heterocyclyl, wherein alkyl, cycloalkyl, aryl, and heterocyclyl are as defined herein.

As used herein, the term "alkyl", whether used as part of another term or independently, refers to a saturated straight or branched chain hydrocarbon group that may optionally be independently substituted with one or more substituents described below. The term "C _i-j alkyl" refers to an alkyl group having from i to i carbon atoms. In some embodiments, the alkyl groups contain 1 to 10 carbon atoms. In some embodiments, the alkyl groups contain 1 to 9 carbon atoms. In some embodiments, the alkyl group contains 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of "C _1-10 alkyl" include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Examples of "C _1-6 alkyl" are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2, 3-dimethyl-2-butyl, 3-dimethyl-2-butyl and the like.

As used herein, the term "alkylcarbonyl" refers to a group of formula R 'C (O) -, wherein R' is alkyl as defined above.

As used herein, the term "alkenyl", whether used as part of another term or independently, refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon double bond that may be optionally independently substituted with one or more substituents described herein and includes groups having a "cis" orientation and a "trans" orientation or alternatively an "E" orientation and a "Z" orientation. In some embodiments, alkenyl groups contain 2 to 12 carbon atoms. In some embodiments, alkenyl groups contain 2 to 11 carbon atoms. In some embodiments, alkenyl groups contain 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms, and in some embodiments, alkenyl groups contain 2 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl (ethylenyl or vinyl), propenyl (allyl), butenyl, pentenyl, 1-methyl-2-buten-1-yl, 5-hexenyl, and the like.

As used herein, the term "alkynyl", whether used as part of another term or independently, refers to a straight or branched chain hydrocarbon group having at least one carbon-carbon triple bond that may be optionally independently substituted with one or more substituents described herein. In some embodiments, alkenyl groups contain 2 to 12 carbon atoms. In some embodiments, alkynyl groups contain 2 to 11 carbon atoms. In some embodiments, alkynyl groups contain 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms, and in some embodiments, alkynyl groups contain 2 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and the like.

As used herein, the term "alkoxy", whether used as part of another term or independently, refers to an alkyl group, as previously defined, attached to the parent molecule through an oxygen atom. The term "C _i-j alkoxy" means that the alkyl portion of the alkoxy group has from i to i carbon atoms. In some embodiments, the alkoxy groups contain 1 to 10 carbon atoms. In some embodiments, the alkoxy groups contain 1 to 9 carbon atoms. In some embodiments, the alkoxy group contains 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of "C _1-6 alkoxy" include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), t-butoxy, neopentyloxy, n-hexyloxy, and the like.

As used herein, the term "alkoxyalkyl" refers to a group of formula-R "OR ', wherein R' and R" are independently alkyl as defined above.

As used herein, the term "amino" refers to the-NH ₂ group. The amino group may also be substituted with one or more groups such as alkyl, aryl, carbonyl, or other amino groups.

As used herein, the term "aryl", whether used as part of another term or independently, refers to mono-and polycyclic ring systems having a total of from 5 to 20 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains from 3 to 12 ring members. Examples of "aryl" include, but are not limited to, phenyl, biphenyl, naphthyl, anthracenyl, and the like, which may bear one or more substituents. As used herein, the term "aryl" also includes groups in which an aromatic ring is fused to one or more additional rings. In the case of a polycyclic system, only one ring need be aromatic (e.g., 2, 3-indoline), but all rings may be aromatic (e.g., quinoline). The second ring may also be fused or bridged. Examples of polycyclic aryl groups include, but are not limited to, benzofuranyl, indanyl, phthalimidyl, naphthalimidyl, phenanthridinyl, tetrahydronaphthyl, and the like. Aryl groups may be substituted at one or more ring positions with substituents as described above.

As used herein, the term "carbamoyl" refers to-C (O) NH ₂.

As used herein, the term "carboxy" refers to-COOH.

As used herein, the term "cycloalkyl", whether used as part of another term or independently, refers to monovalent non-aromatic saturated or partially unsaturated monocyclic and polycyclic systems wherein all ring atoms are carbon and the system contains at least three ring-forming carbon atoms. In some embodiments, cycloalkyl groups may contain 3 to 12 ring-forming carbon atoms, 3 to 10 ring-forming carbon atoms, 3 to 9 ring-forming carbon atoms, 3 to 8 ring-forming carbon atoms, 3 to 7 ring-forming carbon atoms, 3 to 6 ring-forming carbon atoms, 3 to 5 ring-forming carbon atoms, 4 to 12 ring-forming carbon atoms, 4 to 10 ring-forming carbon atoms, 4 to 9 ring-forming carbon atoms, 4 to 8 ring-forming carbon atoms, 4 to 7 ring-forming carbon atoms, 4 to 6 ring-forming carbon atoms, 4 to 5 ring-forming carbon atoms. Cycloalkyl groups may be saturated or partially unsaturated. Cycloalkyl groups may be substituted. In some embodiments, cycloalkyl groups may be saturated cyclic alkyl groups. In some embodiments, cycloalkyl groups may be partially unsaturated cyclic alkyl groups containing at least one double or triple bond in their ring system. In some embodiments, cycloalkyl groups may be monocyclic or polycyclic. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. Examples of polycyclic cycloalkyl groups include, but are not limited to, adamantyl, norbornyl, fluorenyl, spiro-pentadienyl, spiro [3.6] -decyl, bicyclo [1, 1] pentenyl, bicyclo [2, 1] heptenyl, and the like.

As used herein, the term "cycloalkylalkyl" refers to a radical of formula-R 'R ", wherein R' is alkyl as defined above, and R" is cycloalkyl as defined above.

As used herein, the term "alkoxycycloalkyl" refers to a group of formula-R 'OR "wherein R' is alkyl as defined above and R" is cycloalkyl as defined above.

As used herein, the term "cyano" refers to-CN.

As used herein, the term "halogen" refers to an atom selected from fluorine, chlorine, bromine and iodine.

As used herein, the term "haloalkyl" refers to an alkyl group as defined above substituted with one or more halogens as defined above. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, trichloromethyl, 2-trifluoroethyl, 1, 2-difluoroethyl, 3-bromo-2-fluoropropyl, 1, 2-dibromoethyl and the like.

As used herein, the term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur as well as any quaternized form of basic nitrogen (including N-oxides).

As used herein, the term "heteroaryl", whether used as part of another term or independently, refers to an aryl group having one or more heteroatoms in addition to carbon atoms. Heteroaryl groups may be monocyclic. Examples of monocyclic heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, benzofuranyl, and pteridinyl. Heteroaryl also includes polycyclic groups in which the heteroaryl ring is fused to one or more aryl, alicyclic, or heterocyclic rings, wherein the linking group or point of attachment is on the heteroaryl ring. Examples of polycyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, benzothienyl, benzofuranyl, benzo [1,3] dioxolyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated carbocyclic group in which one or more ring atoms are heteroatoms independently selected from oxygen, sulfur, nitrogen, phosphorus, and the like, the remaining ring atoms being carbon, wherein one or more ring atoms may be optionally independently substituted with one or more substituents. In some embodiments, the heterocyclyl is a saturated heterocyclyl. In some embodiments, a heterocyclyl is a partially unsaturated heterocyclyl having one or more double bonds in its ring system. In some embodiments, the heterocyclyl may contain any oxidized form of carbon, nitrogen or sulfur and any quaternized form of basic nitrogen. "heterocyclyl" also includes groups in which the heterocyclyl is fused to a saturated, partially unsaturated, or fully unsaturated (i.e., aromatic) carbocyclic or heterocyclic ring. The heterocyclic group may be carbon-linked or nitrogen-linked, where possible. In some embodiments, the heterocycle is carbon-linked. In some embodiments, the heterocycle is nitrogen-linked. For example, the groups derived from pyrrole may be pyrrol-1-yl (nitrogen-linked) or pyrrol-3-yl (carbon-linked). Furthermore, the group derived from imidazole may be imidazol-1-yl (nitrogen linked) or imidazol-3-yl (carbon linked).

In some embodiments, the term "3-to 12-membered heterocyclyl" refers to a 3-to 12-membered saturated or partially unsaturated monocyclic or polycyclic heterocyclic ring system having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Fused, spiro, and bridged ring systems are also included within the scope of this definition. Examples of monocyclic heterocyclyl groups include, but are not limited to, oxetanyl, 1-dioxothietanylpyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, thiazolyl, piperidinyl, piperazinyl, piperidinyl, morpholinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyridonyl, pyrimidinonyl, pyrazinonyl, pyrimidinonyl, pyridazinonyl, pyrrolidinyl, triazinonyl, and the like. Examples of fused heterocyclic groups include, but are not limited to, phenyl condensed rings or pyridyl condensed rings, such as quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, quinolizinyl, quinazolinyl, azaindolizinyl, pteridinyl, chroenyl, isochroenyl, indolyl, isoindolyl, indolizinyl, indazolyl, purinyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzothienyl, benzothiazolyl, carbazolyl, phenazinyl, phenothiazinyl, phenanthridinyl, imidazo [1,2-a ] pyridinyl, [1,2,4] triazolo [4,3-a ] pyridinyl, [1,2,3] triazolo [4,3-a ] pyridinyl, and the like. Examples of spiroheterocyclyl groups include, but are not limited to, spiropyranyl, spirooxazinyl, and the like. Examples of bridged heterocyclyl groups include, but are not limited to, morphinan, hexamethylenetetramine, 3-aza-bicyclo [3.1.0] hexane, 8-aza-bicyclo [3.2.1] octane, 1-aza-bicyclo [2.2.2] octane, 1, 4-diazabicyclo [2.2.2] octane (DABCO), and the like.

As used herein, the term "hydroxy" refers to-OH.

As used herein, the term "hydroxyalkyl" refers to an alkyl group as defined above substituted with one or more hydroxyl groups.

As used herein, the term "oxo" refers to an =o substituent.

As used herein, the term "partially unsaturated" refers to a group that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (i.e., fully unsaturated) moieties.

As used herein, the term "substituted", whether preceded by the term "optional", means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. It is to be understood that "substitution" or "substituted" includes implicit preconditions that such substitution is consistent with the permissible valences of the substituted atoms, and that the substitution results in stable or chemically feasible compounds, e.g., compounds that do not spontaneously undergo transformations such as rearrangement, cyclization, elimination, and the like. Unless otherwise indicated, an "optionally substituted" group may have the appropriate substituent at each substitutable position of the group, and where more than one position in any given structure may be substituted with more than one substituent selected from the specified group, the substituents may be the same or different at each position. It will be appreciated by those skilled in the art that the substituents themselves may be substituted, if appropriate. Unless specifically stated as "unsubstituted," references to chemical moieties herein are to be understood as including substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

As used herein, the term "about" is intended to define a modified numerical value, thereby representing the value as a variable within an error range. When a particular range of error is not recited (e.g., standard deviation of the average values given in a chart or data table), the term "about" is understood to mean that a range of 10%, preferably 5%, is encompassed by the term "about" and is included.

As used herein, the term "disease" is intended to be generally synonymous with the terms "disorder," "disorder," and "condition" (as in medical conditions), interchangeably used, as they each reflect abnormal conditions of the human or animal body or a portion thereof that impair normal function, often manifested as distinguishing signs and symptoms, and resulting in reduced life span or reduced quality of life of the human or animal.

"Patient" is generally synonymous with the term "subject" and as used herein includes all mammals (including humans). Examples of patients include humans, livestock (e.g., cattle, goats, sheep, pigs, and rabbits), and companion animals (e.g., dogs, cats, rabbits, and horses). Preferably, the patient is a human.

As used herein, "in need of treatment" refers to a determination made by a physician or other caregiver that a subject is in need of or will benefit from treatment. The determination is made based on a variety of factors within the expertise of the physician or caregiver.

"Administering," "administering," and the like, when used in, for example, a patient, cell, tissue, organ, or biological fluid, refers to contacting a compound of formula (I), a pharmaceutical composition or diagnostic agent comprising the same, with a subject, cell, tissue, organ, or biological fluid. In the case of cells, administration includes contact of the agent with the cells (e.g., in vitro or ex vivo), as well as contact of the agent with a fluid, wherein the fluid is in contact with the cells.

As used herein, a "therapeutically effective amount" refers to an amount of a compound of formula (I), formula (Ia), formula (Ib), or a sub-embodiment described herein and/or a pharmaceutically acceptable salt thereof, which when administered alone or as part of a pharmaceutical composition and in a single dose or as part of a series of doses to a patient for treatment of a disease, is sufficient to affect treatment of such disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, the age, weight, etc., of the mammal to be treated. The therapeutically effective amount can be determined by measuring the relevant physiological effects and can be adjusted in connection with dosing regimens, diagnostic analysis of the subject's condition, and the like. For example, measuring the serum level of a compound of formula (I) (or a metabolite thereof) at a particular time after administration may indicate whether a therapeutically effective amount has been used.

"Treating" or "treatment" of a disease includes:

(1) Preventing a disease, i.e., preventing the clinical symptoms of the disease from developing in a mammal that may be exposed to or susceptible to the disease but has not experienced or exhibited symptoms of the disease;

(2) Inhibiting the disease, i.e., preventing or alleviating the progression of the disease or its clinical symptoms; or alternatively

(3) Remit the disease, i.e., cause regression of the disease or its clinical symptoms.

"Inhibit", "decrease" or any variation of these terms with respect to MAT2A includes any measurable decrease or complete inhibition to achieve the desired result. For example, the decrease in MAT2A activity can be reduced by about, up to about, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any derivable range thereof, as compared to its normal activity.

Compounds of formula (I)

The present disclosure provides novel compounds of formula (I), formula (Ia) or formula (Ib), or stereoisomers, tautomers or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing the compounds or stereoisomers, tautomers or pharmaceutically acceptable salts thereof, and various uses of the disclosed compounds.

wherein:

w is CR ⁴ or N;

x is CR ⁵ or N;

y is CR ⁶ or N;

z is CR ⁷ or N;

v is CR ⁸ or N;

R ⁸ is selected from the group consisting of: hydrogen, alkyl, and hydroxy;

R ^a and R ^b are each independently selected from hydrogen or alkyl;

r ^c is selected from the group consisting of: hydrogen, alkyl, and cycloalkyl;

Each R ^g is independently hydrogen or C _1-3 alkyl.

in one embodiment, the R ¹ is-NR ^cR^d.

In one embodiment, the R ^c is hydrogen or alkyl.

In one embodiment, the R ^c is methyl.

And methoxy.

In one embodiment, the R ² is methyl.

In one embodiment, the R ² is phenyl.

In one embodiment, the R ³ is heterocyclyl.

In one embodiment, the R ³ is a five membered heterocyclyl.

In one embodiment, the R ³ is selected from

In one embodiment, the R ⁴ is hydrogen.

In one embodiment, the R ⁵ is hydrogen.

In one embodiment, the R ⁷ is hydrogen.

In one embodiment, the R ⁸ is hydrogen.

The compounds provided herein are described with reference to the general formula and specific compounds. Furthermore, the compounds of the present disclosure may exist in a variety of different forms or derivatives, including but not limited to prodrugs, soft drugs, active metabolic derivatives (active metabolites) and pharmaceutically acceptable salts thereof, all of which are within the scope of the present disclosure.

As used herein, the term "prodrug" refers to a compound or a pharmaceutically acceptable salt thereof that upon metabolism under physiological conditions or conversion by solvolysis yields the desired active compound. Prodrugs include, but are not limited to, esters, amides, carbamates, carbonates, ureides, solvates or hydrates of the active compounds. Typically, prodrugs are inactive or less active than the active compound, but may provide one or more advantageous handling, administration, and/or metabolic properties. For example, some prodrugs are esters of the active compound; during metabolic breakdown, the ester groups are cleaved to yield the active drug. In addition, some prodrugs are enzymatically activated to produce the active compound or compounds that produce the active compound upon further chemical reaction. The prodrug may be developed from a prodrug form to an active form in a single step, or may have one or more intermediate forms that may or may not be active themselves. The preparation and use of prodrugs is discussed in the following references: higuchi and v.stilla, "Pro-drug as novel delivery system (Pro-drugs as Novel DELIVERY SYSTEMS)", volume 14 of the a.c.s. seminar Series (a.c. symposium Series), bioreversible carrier in drug design (BioreversibleCarriers in Drug Design), editors Edward b.roche, american pharmaceutical society (AmericanPharmaceutical Association) and pegamon Press, 1987. Prodrug: challenge and return (Prodrugs: CHALLENGES AND REWARDS), edit V.Stella, R.Borchardt, M.Hageman, R.Oliyai, H.Maag, J.Tilley, new York schopleger press (SPRINGER VERLAG NEW York), 2007, all of which are hereby incorporated by reference in their entirety.

As used herein, the term "soft drug" refers to a compound that exerts a pharmacological effect but breaks down into inactive metabolite degradants such that the activity time is limited. See, for example, soft medicine: principles and methods of safe drug design (Soft drugs: PRINCIPLES AND methods for THE DESIGN of safe drugs), nicholas Bodor, drug research review (MEDICINAL RESEARCH REVIEWS), volume 4, stage 4, 449-469, 1984, which references are hereby incorporated by reference in their entirety.

As used herein, the term "metabolite", e.g., an active metabolite, overlaps with the prodrug as described above. Such metabolites are therefore pharmacologically active compounds, or compounds that are further metabolized to pharmacologically active compounds, which are derivatives produced by metabolic processes in the subject. For example, such metabolites may result from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc. of the administered compound or salt or prodrug. Wherein the active metabolite is such a pharmacologically active derivative compound. For prodrugs, the prodrug compounds are generally inactive or less active than the metabolite. For active metabolites, the parent compound may be an active compound or may be an inactive prodrug.

Prodrugs and active metabolites may be identified using conventional techniques known in the art. See, for example, bertolini et al, 1997, journal of pharmaceutical chemistry (J Med Chem) 40:2011-2016; shan et al, journal of pharmaceutical science (JPharm Sci) 86:756-757; bagshawe,1995, drug Dev Res 34:220-230; wermuth, supra.

As used herein, the term "pharmaceutically acceptable" means that the substance or composition is chemically and/or toxicologically compatible with the other ingredients comprising the formulation and/or the subject being treated.

As used herein, the term "pharmaceutically acceptable salt" is meant to include salts of the active compounds prepared with relatively non-toxic acids or bases, depending on the particular substituents found on the compounds described herein. When the compounds disclosed herein contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base (neat or in a suitable inert solvent). Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, trivalent manganese, divalent manganese, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines and the like, such as arginine, betaine, caffeine, choline, N' -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydrabamine (hydrabamine), isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid (neat or in a suitable inert solvent). Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrocarbonic, phosphoric, monohydrophosphoric, dihydrogenphosphoric, sulfuric, monohydrosulfuric, hydroiodic or phosphorous acid and the like, and salts derived from relatively non-toxic organic acids such as acetic, propionic, isobutyric, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic and the like. Also included are salts of amino acids, such as arginine salts and the like, and salts of organic acids, such as glucuronic acid or galacturonic acid and the like (see, e.g., berge, s.m. and the like, "pharmaceutically acceptable salts", journal of pharmaceutical science, 1977, 66, pages 1-19). Certain specific compounds of the present invention contain both basic and acidic functionalities, which allow the compounds to be converted into base or acid addition salts.

The neutral form of the compound may be regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties (e.g., solubility in polar solvents), but for the purposes of the present invention these salts are equivalent to the parent form of the compound.

The present disclosure also includes protected derivatives of the compounds of the present disclosure. For example, when the compounds of the present disclosure contain groups such as hydroxyl, carboxyl, thiol, or any group containing one or more nitrogen atoms, these groups may be protected by suitable protecting groups. A complete list of suitable protecting groups can be found in T.W.Greene, protecting groups in organic synthesis, 5 th edition, john Wiley father-son Co., ltd (2014), the disclosure of which is incorporated herein by reference in its entirety. Protected derivatives of the compounds of the present disclosure may be prepared by methods well known in the art.

The compounds of the present disclosure may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.

As used herein, the term "solvate" or "solvated form" refers to a solvent addition form containing a stoichiometric or non-stoichiometric amount of solvent. Some compounds tend to trap a fixed molar ratio of solvent molecules in the crystalline solid state, forming solvates. If the solvent is water, the solvate formed is a hydrate; and if the solvent is an alcohol, the solvate formed is an alkoxide. Hydrates are formed by combining one or more water molecules with one molecule of a substance that holds it as a molecular state of H ₂ O. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

The compounds of the present disclosure may exist in a variety of crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present disclosure and are intended to fall within the scope of the present disclosure.

As used herein, the terms "crystalline form," "polymorphic form," and "polymorph" are used interchangeably and refer to a crystalline structure of a compound (or a salt or solvate thereof) that can crystallize in a different crystal packing arrangement, all of which have the same elemental composition. Different crystal forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability and solubility. Recrystallization solvent, crystallization rate, storage temperature, and other factors may dominate one crystal form. Polymorphs of a compound can be prepared by crystallization under different conditions.

The compounds of the present disclosure may have asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., individual enantiomers) are all intended to be included within the scope of the present invention. When a stereochemical description is shown, it refers to a compound in which one isomer is present and substantially free of the other isomer. By "substantially free" of the other isomer is meant that the ratio of the two isomers is at least 80/20, more preferably 90/10, or 95/5 or more. In some embodiments, one of the isomers will be present in an amount of at least 99%.

Those skilled in the art will appreciate that the compounds of the present disclosure may exist in different tautomeric forms, and that all such forms are contemplated as falling within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can be converted to each other by a low energy barrier. The existence and concentration of the isomeric forms will depend on the environment in which the compound is located and may vary depending, for example, on whether the compound is solid or in an organic or aqueous solution. For example, proton tautomers (also known as proton-metamorphosing tautomers) include interconversions by proton transfer, such as keto-enol, amide-imide, lactam-lactam, imine-enamine isomerisation, and cyclic forms where a proton may occupy two or more positions of a heterocyclic ring system. Valence tautomers include interconversions by recombination of some of the bond-forming electrons. Tautomers may be in equilibrium or sterically locked into one form by appropriate substitution. Unless otherwise indicated, compounds of the present disclosure identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms.

The compounds of the present disclosure may also contain unnatural amounts of isotopes on one or more of the atoms that make up such compounds. Unnatural amounts of isotopes can be defined as amounts found in nature of the atoms in question to amounts of 100%. Only when one or more isotopically enriched atoms are present. Exemplary isotopes that can be incorporated into compounds of the invention, e.g., compounds of formula (I), formula (Ia), formula (Ib) (and any of the embodiments thereof disclosed herein, including specific compounds), include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, e.g., ²H、³H、¹¹C、¹³C、¹⁴C、¹³N、¹⁵N、¹⁵O、¹⁷O、¹⁸O、³²P、³³P、³⁵S、¹⁸F、³⁶Cl、¹²³I and ¹²⁵ I, respectively. Isotopically-labeled compounds (e.g., those labeled with ³ H and ¹⁴ C) are useful in compound or substrate tissue distribution assays. Tritium (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) isotopes are useful for their ease of preparation and detectability. In addition, substitution with heavier isotopes such as deuterium (i.e., ² H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, in the compounds disclosed herein that are included in table 1 below, one or more hydrogen atoms are replaced with ² H or ³ H, or one or more carbon atoms are replaced with ¹³ C-or ¹⁴ C-enriched carbon. Positron emitting isotopes (e.g., ¹⁵O、¹³N、¹¹ C and ¹⁵ F) can be used in Positron Emission Tomography (PET) studies to examine occupancy of substrate receptors. Isotopically-labeled compounds can generally be prepared following procedures analogous to those disclosed in the schemes or examples herein by substituting an isotopically-labeled reagent for a non-isotopically-labeled reagent.

Representative compounds of the present disclosure are listed in table 1 below:

TABLE 1

Note that: compounds 8 and 9 in the above table are chiral isomers and Peak 1 and 2 are named according to the order of the peaks.

Synthesis of Compounds

The compounds provided herein may be prepared using any known organic synthesis technique and may be synthesized according to any of a number of possible synthetic routes.

The reactions for preparing the compounds of the present disclosure may be carried out in suitable solvents that may be readily selected by those skilled in the art of organic synthesis. Suitable solvents may be substantially unreactive with the starting materials (reactants), intermediates or products at the temperature at which the reaction is carried out, for example, a temperature which may range from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction may be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, the appropriate solvent for the particular reaction step may be selected by one skilled in the art.

The preparation of the compounds of the present disclosure may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the choice of appropriate protecting groups can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in the following references: T.W.Greene and P.GM.Wuts, protecting group in organic Synthesis (Protective Groups in Organic Synthesis), 3 rd edition, john Willi parent, new York (1999); kocienski, protecting group (Protecting Groups), george Tami Press (GeorgThieme Verlag), 2003; and Peter g.m. wuts, greene' sProtective Groups in Organic Synthesis, 5th edition, wiley,2014, all of which are incorporated herein by reference in their entirety.

The reaction may be monitored according to any suitable method known in the art. For example, product formation may be monitored by spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., 1H or 13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatographic means such as High Performance Liquid Chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or Thin Layer Chromatography (TLC). The compounds can be purified by a variety of methods including High Performance Liquid Chromatography (HPLC) ("preparative LC-MS purification: improved compound specific method optimization" Karl f. Blom, brian Glass, RICHARD SPARKS, andrew p. Combos journal of combinatorial chemistry (j. Combi. Chem.) "2004,6 (6), 874-883, which is incorporated herein by reference in its entirety) and normal phase silica gel chromatography.

Pharmaceutical composition

In another aspect, a pharmaceutical composition is provided comprising one or more molecules or compounds of the present disclosure or a pharmaceutically acceptable salt thereof.

In another aspect, a pharmaceutical composition is provided comprising one or more molecules or compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

The compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or pharmaceutically acceptable salts thereof may be in the form of compositions suitable for administration to a subject. Typically, such compositions are pharmaceutical compositions comprising a compound of formula (I), (Ia), (Ib) or a sub-embodiment thereof as described herein or a pharmaceutically acceptable salt thereof and one or more pharmaceutically or physiologically acceptable excipients. In certain embodiments, a compound of formula (I), (Ia), (Ib) or a sub-embodiment thereof, or a pharmaceutically acceptable salt thereof, as described herein, is present in a therapeutically effective amount. Pharmaceutical compositions can be used in the methods disclosed herein; thus, for example, the pharmaceutical compositions can be administered to a subject ex vivo or in vivo to practice the methods of treatment and uses described herein.

The pharmaceutical composition may be formulated to be compatible with the intended method or route of administration; exemplary routes of administration are set forth herein. Furthermore, the pharmaceutical compositions may be used in combination with other therapeutically active agents or compounds described herein to treat diseases, disorders, and conditions contemplated by the present invention.

Pharmaceutical compositions comprising an active ingredient (e.g., a compound of formula (I), (Ia), (Ib) or a sub-embodiment thereof, as described herein, a pharmaceutically acceptable salt thereof) may be in a form suitable for oral use, such as tablets, capsules, dragees (troches), lozenges (lozenges), aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, solutions, microbeads or elixirs. Pharmaceutical compositions for oral use may be prepared according to any method known in the art for manufacturing pharmaceutical compositions, and such compositions may contain one or more agents, such as sweeteners, flavoring agents, coloring agents and preservatives to provide pharmaceutically elegant and palatable preparations. Tablets, capsules, and the like contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets, capsules, and the like. These excipients may be, for example, diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc.

Tablets, capsules and the like suitable for oral administration may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. The tablets may also be coated by techniques known in the art to form osmotic therapeutic tablets for controlled release. Additional agents include biodegradable or biocompatible particles or polymeric materials such as polyesters, polyamino acids, hydrogels, polyvinylpyrrolidone, polyanhydrides, polyglycolic acid, ethylene vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or copolymers of lactide and glycolide, polylactides and glycolide, or ethylene vinyl acetate copolymers to control delivery of the applied composition. For example, the oral agents may be embedded in microcapsules prepared by coacervation techniques or by interfacial polymerization (by using hydroxymethyl cellulose or gelatin-microcapsules or poly (methyl methacrylate) microcapsules, respectively), or in colloidal drug delivery systems. Colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, microbeads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Methods of preparing the above formulations are known in the art.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate, kaolin or microcrystalline cellulose, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.

The aqueous suspension comprises the active substance in admixture with excipients which are suitable for the manufacture thereof. These excipients may be suspending agents, for example sodium carboxymethyl cellulose, methyl cellulose, (hydroxypropyl) methyl cellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersants or wetting agents, for example natural phospholipids (e.g. lecithin), or condensation products of alkylene oxides with fatty acids (e.g. polyoxyethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols (e.g. heptadecaethyleneoxycetyl alcohol), or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols (e.g. polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols (e.g. polyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweeteners and flavoring agents such as those described above may be added to provide a palatable oral preparation.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are described in this Wen Zhongju.

The pharmaceutical composition may also be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil (for example olive oil or arachis oil), or a mineral oil (for example liquid paraffin) or a mixture thereof. Suitable emulsifying agents may be natural gums, for example acacia or tragacanth; natural phospholipids, such as soy, lecithin, and fatty acid derived or partial esters; hexitoanhydrides such as sorbitan monooleate; and condensation products of partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate.

The pharmaceutical compositions generally comprise a therapeutically effective amount of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof as described herein, and one or more pharmaceutically acceptable excipients. Suitable pharmaceutically acceptable excipients include, but are not limited to, antioxidants (e.g., ascorbic acid and sodium bisulfate), preservatives (e.g., benzyl alcohol, methyl paraben, ethyl or n-propyl paraben), emulsifiers, suspending agents, dispersants, solvents, fillers, compatibilizers, detergents, buffers, vehicles, diluents and/or adjuvants. For example, a suitable vehicle may be a physiological saline solution or a citrate buffered saline, possibly supplemented with other substances common in pharmaceutical compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. Those skilled in the art will readily recognize a variety of buffers that may be used in the pharmaceutical compositions and dosage forms contemplated herein. Typical buffers include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. For example, the buffer component may be a water-soluble material such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and salts thereof. Acceptable buffers include, for example, tris buffer, N- (2-hydroxyethyl) piperazine-N' - (2-ethanesulfonic acid) (HEPES), 2- (N-morpholino) ethanesulfonic acid (MES), 2- (N-morpholino) ethanesulfonic acid sodium salt (MES), 3- (N-morpholino) propanesulfonic acid (MOPS), and N-Tris [ hydroxymethyl ] methyl-3-aminopropanesulfonic acid (TAPS).

After the pharmaceutical composition is formulated, it may be stored in sterile vials in the form of a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored in a ready-to-use form, a lyophilized form that requires reconstitution prior to use, a liquid form that requires dilution prior to use, or other acceptable form. In some embodiments, the pharmaceutical composition is provided in a disposable container (e.g., a disposable vial, ampoule, syringe, or auto-injector (similar to, for example)), while a multi-use container (e.g., a multi-use vial) is provided in other embodiments.

The formulation may also include a carrier to protect the composition from rapid degradation or clearance from the body, such as controlled release formulations, including liposomes, hydrogels, prodrugs, and microencapsulated delivery systems. For example, a time delay material (e.g., glyceryl monostearate or glyceryl stearate) may be employed alone or with a wax. Any drug delivery device may be used to deliver the compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof, including implants (e.g. implantable pumps) and catheter systems, slow injection pumps and devices, all of which are well known to the skilled person.

Depot injections, typically administered subcutaneously or intramuscularly, may also be used to release the compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof over a period of time. Depot injections are typically solid or oil based and typically include at least one of the formulation ingredients described herein. One of ordinary skill in the art is familiar with possible formulations and uses of depot injections.

The pharmaceutical composition may be in the form of a sterile injectable aqueous or oleaginous suspension. The suspensions may be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents as mentioned herein. The sterile injectable 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. Acceptable diluents, solvents and dispersion media that may be used include water, ringer's solution, isotonic sodium chloride solution, cremophor EL ^TM (basf, pasiponi, new jersey) or Phosphate Buffered Saline (PBS), ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycol) and suitable mixtures thereof. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Prolonged absorption of a particular injectable formulations can be brought about by the inclusion of agents which delay absorption, for example, aluminum monostearate or gelatin.

The compounds of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein may also be administered in the form of suppositories for rectal administration or sprays for nasal or inhalation. Suppositories may be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include, but are not limited to, cocoa butter and polyethylene glycols.

Methods of treating diseases

In another aspect, the present disclosure provides a method of treating a MAT 2A-mediated disease in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound provided herein or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition provided herein.

In some embodiments, the disease in the method of treating a MAT 2A-mediated disease in a patient is cancer, e.g., selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

In another aspect, the present disclosure provides a method of treating MTAP-deficient cancer in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound provided herein, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition provided herein.

In some embodiments, the cancer in the method of treating MTAP-deficient cancer in a patient is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

In yet another aspect, the present disclosure provides a method for treating cancer in a patient, wherein the cancer is characterized by reduced or no expression of MTAP gene, a deletion of MTAP gene, or reduced function of MTAP protein, comprising administering to the subject a therapeutically effective amount of a compound provided herein, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition provided herein.

In some embodiments, the cancer in the method for treating cancer in a patient is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

Pharmaceutical use

In another aspect, the present disclosure provides the use of a compound described herein, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, for the manufacture of a medicament for the treatment of cancer.

In some embodiments, the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

Route of administration

The compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof and compositions comprising them may be administered in any suitable manner. Suitable routes of administration include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implantation), intraperitoneal, intracisternal, intra-articular, intraperitoneal, intracerebral (intraparenchymal) and intraventricular), nasal, vaginal, sublingual, intraocular, rectal, topical (e.g., transdermal), buccal, and inhalation. Depot injections, usually administered subcutaneously or intramuscularly, may also be used to administer the compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof over a period of time. Particular embodiments of the present invention contemplate oral administration.

Combination therapy

The present invention contemplates the use of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof as described herein in combination with one or more active therapeutic agents (e.g. chemotherapeutic agents) or other prophylactic or therapeutic means (e.g. radiation). In such combination therapies, the various active agents often have different complementary mechanisms of action. Such combination therapies may be particularly beneficial by reducing the dosage of one or more agents thereby reducing or eliminating the adverse effects associated with the one or more agents. Furthermore, such combination therapies may have synergistic therapeutic or prophylactic effects on potential diseases, disorders or conditions.

As used herein, "combination" is meant to include therapies that can be administered singly, separately, e.g., therapies formulated separately for administration (e.g., as provided in a kit), as well as therapies that can be co-administered in a single formulation (i.e., a "co-formulation").

In certain embodiments, a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is administered or applied sequentially, e.g., one agent is administered prior to one or more other agents. In other embodiments, a compound of formula (I), (Ia), (Ib) or a sub-embodiment described herein, or a salt thereof, is administered simultaneously, e.g., two or more agents are administered simultaneously or about simultaneously; the two or more agents may be present in two or more separate formulations or combined into a single formulation (i.e., a co-formulation). Whether two or more agents are administered sequentially or simultaneously, they are considered to be administered in combination for purposes of this disclosure.

In this case, the compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof may be used in combination with at least one other (active) agent in any suitable manner. In one embodiment, treatment with at least one active agent and at least one compound of formula (I), (Ia), (Ib) or sub-embodiment or salt thereof described herein is maintained for a period of time. In another embodiment, the treatment of at least one active agent is reduced or discontinued (e.g., when the subject is stable), and the treatment of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is performed in a constant dosage regimen. In another embodiment, the treatment of at least one active agent is reduced or discontinued (e.g., when the subject is stable), while the treatment of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is reduced (e.g., lower dose, less frequent dosing, or a shorter treatment regimen). In yet another embodiment, the treatment of at least one active agent is reduced or discontinued (e.g., when the subject is stable), and the treatment of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is increased (e.g., higher doses, more frequent dosing, or longer treatment regimen). In yet another embodiment, treatment of at least one active agent is maintained, and treatment of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is reduced or discontinued (e.g., lower doses, less frequent dosing, or a shorter treatment regimen). In yet another embodiment, the treatment of at least one active agent and a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is reduced or discontinued (e.g., lower doses, less frequent dosing, or a shorter treatment regimen).

The present disclosure provides methods of treating cancer with a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof, as described herein, and at least one additional therapeutic or diagnostic agent.

In some embodiments, a compound of formula (I), (Ia), (Ib) or a fruiting embodiment or salt thereof described herein is administered in combination with at least one additional therapeutic agent selected from temozolomide, pemetrexed, pegylated liposomal doxorubicin (Doxil), eribulin (Halaven, sea Le Wei), ixabepilone (Ixempra), protein-bound paclitaxel (Abraxane), oxaliplatin, irinotecan, valnemulin (bcl 2 inhibitor), 5-azacytidine, anti-CD 20 therapeutic agents such as rituximab and obin You Tuozhu mab, hormonal agents (anatolzole, ai Kexi mad (exemestand), letrozole, norrad, leuprorelin acetate (lupon eligard CDK)), 4/6 inhibitors, palbociclib, abberamide, CPI (avermectin, cimepruno Li Shan-resistance (Cemiplimab-rwlc), and bevacizumab.

In certain embodiments, the present disclosure provides methods of treating cancer comprising administering a compound of formula (I), (Ia), (Ib) or a sub-embodiment described herein, or a salt thereof, in combination with a Signal Transduction Inhibitor (STI) to achieve additive or synergistic inhibition of tumor growth. As used herein, the term "signal transduction inhibitor" refers to an agent that selectively inhibits one or more steps in a signal pathway. Examples of Signal Transduction Inhibitors (STI) useful in the methods described herein include, but are not limited to: (i) bcr/abl kinase inhibitors (e.g., gleevec); (ii) Epidermal Growth Factor (EGF) receptor inhibitors, including kinase inhibitors and antibodies; (iii) Inhibitors of her-2/neu receptor (e.g., herceptin); (iv) Inhibitors of Akt family kinases or Akt pathways (e.g., rapamycin); (v) Cell cycle kinase inhibitors (e.g., fraapine); (vi) inhibitors of phosphatidylinositol kinase. Agents involved in immunomodulation may also be used in combination with one or more compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof to inhibit tumor growth in cancer patients.

In certain embodiments, the present disclosure provides methods of treating cancer comprising administering a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein in combination with a chemotherapeutic agent. Examples of chemotherapeutic agents include, but are not limited to, alkylating agents, such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, imperoshu, piposulfan; aziridines, such as benzodopa (benzodopa), carboquinone, methodol (meturedopa), and Wu Leiduo bar (uredopa); ethyleneimine and methyl melamines (METHYLAMELAMINE) include altretamine, triethylenemelamine, triethylenephosphoramide (trietylenephosphoramide), triethylenephosphoramide (triethylenethiophosphaoramide), and trimethylol melamine; nitrogen mustards, such as chlorambucil, napthalene mustards, chlorflufosamide (cholophosphamide), estramustine, ifosfamide, dichloromethyl diethylamine, mechlorethamine hydrochloride, melphalan, new enbixing, chlorambucil cholesterol, prednisone mustards, qu Luolin amine, uracil mustards; nitrosoureas such as carmustine, pimozide, fotemustine, lomustine, nimustine, and ramustine; antibiotics, such as doxorubicin, actinomycin, aflatoxin, diazoserine, bleomycin, C actinomycin, calicheamicin, karabin (carabicin), carminomycin, carcinomycin, chromomycin, D actinomycin, daunorubicin, ditubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, elubicin, nordaunorubicin (idambicin), marrubicin, mitomycin, mycophenolic acid, nolamycin, olivomycin, pelomycin, bofevernix Luo Mei (potfiromycin), puromycin, tri-iron doxorubicin, rodubicin, streptozocin, truffle, ubenimex, cilastatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as, for example, dimethyl folic acid, methotrexate, pterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thioadenine, thioguanine; pyrimidine analogs such as ambcitabine, azacytidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, deoxyfluorouridine, enocitabine, fluorouridine, 5-FU; androgens, such as carbosterone, drotasone propionate, cyclothioandrostane, emasculan, and testosterone; anti-adrenergic agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as She Wansuan; acetoglucurolactone; aldehyde phosphoramidate glycoside; aminolevulinic acid; amsacrine; bei Labu shake (bestrabucil); a specific group; idatroxas; ground phosphoramide (defofamine); colchicine; deaquinone; efluoroornithine (elfornithine); eli acetoamide; eggshell robust; gallium nitrate; hydroxyurea; mushroom polysaccharide; lonidamine; mitoguazone; mitoxantrone; mo Pai darol; diamine nitroacridine; prastatin; egg ammonia nitrogen mustard; pirarubicin; podophylloic acid; 2-ethyl hydrazide; procarbazine; carrying out a process of preparing the raw materials; sugar (sizofran) of the west adjuvant; germanium spiroamine; tenuazonic acid; triiminoquinone; 2,2',2 "-trichlorotriethylamine; uratam; vindesine; dacarbazine; mannitol; dibromomannitol; dibromodulcitol; pipobromine; adding cytosine; arabinocytidine (Ara-C); cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum and platinum coordination complexes such as cisplatin and carboplatin; vinblastine; trastuzumab; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; north vitamin; nux An Tuo; teniposide; daunomycin; aminopterin; proper roda; ibandronate; CPT11; topoisomerase inhibitors; difluoromethyl ornithine (DMFO); retinoic acid; lamycin; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. In particular embodiments, the compounds of the present disclosure are co-administered with a cytostatic compound selected from cisplatin, doxorubicin, paclitaxel (taxol), docetaxel, and mitomycin C. In a particular embodiment, the cytostatic compound is doxorubicin.

Chemotherapeutic agents also include anti-hormonal agents, such as antiestrogens, including, for example, tamoxifen, raloxifene, aromatase inhibiting 4 (5) -imidazole, 4-hydroxy tamoxifen, trawoxifene (trioxifene), raloxifene (keoxifene), onapristone (onapristone), and toremifene; antiandrogens, such as flutamide, nilutamide, bicalutamide, enzalutamide, apalutamide, abiraterone acetate, leuprorelin, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above. In certain embodiments, the combination therapy comprises administration of a hormone or related hormonal agent.

The present disclosure also contemplates the use of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof as described herein in combination with an immune checkpoint inhibitor. The large number of gene and epigenetic changes that characterize all cancers provide a diverse set of antigens that the immune system can use to distinguish tumor cells from their normal counterparts. In the case of T cells, the final magnitude (e.g., level of cytokine production or proliferation) and quality (e.g., type of immune response produced, e.g., pattern of cytokine production) of the response initiated by T Cell Receptor (TCR) antigen recognition is regulated by a balance between co-stimulatory and inhibitory signals (immune checkpoints). Under normal physiological conditions, immune checkpoints are critical for preventing autoimmunity (i.e., maintaining self-tolerance) and protecting tissue from damage when the immune system reacts to pathogen infection. Expression of immune checkpoint proteins may be deregulated by tumors as an important immune resistance mechanism. Examples of immune checkpoint inhibitors include, but are not limited to, CTLA-4, PD-1, PD-L1, BTLA, TIM3, LAG3, OX40, 41BB, VISTA, CD96, TGF beta, CD73, CD39, A2AR, A2BR, IDO1, TDO2, arginase, B7-H3, B7-H4. Cell-based anti-cancer immunomodulators are also contemplated herein. Examples of such modulators include, but are not limited to, chimeric antigen receptor T cells, tumor infiltrating T cells, and dendritic cells.

The present disclosure contemplates the use of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein in combination with an immune checkpoint receptor and a ligand inhibitor as described above (e.g., ipilimumab, abamectin, nivolumab, pembrolizumab, atuzumab, nivolumab, and divalizumab).

Other therapeutic modalities that may be combined with the compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof include radiation therapy, monoclonal antibodies to tumor antigens, complexes of monoclonal antibodies and toxins, T cell adjuvants, bone marrow transplants or antigen presenting cells (e.g., dendritic cell therapy).

The present disclosure contemplates the use of a compound of formula (I), (Ia), (Ib) or an embodiment or salt thereof described herein, alone or in combination with radiation therapy and/or Temozolomide (TMZ), avastin or lomustine, for the treatment of glioblastoma.

The present disclosure includes pharmaceutically acceptable salts, acids, or derivatives of any of the above.

Administration of drugs

The amount of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein that can be administered to a subject depends, for example, on the target of administration (e.g., resolution desired); age, weight, sex, and health and physical condition of the subject to whom the formulation is administered; route of administration; and the nature of the disease, disorder, condition or symptom. The dosage regimen may also take into account the presence, nature and extent of any side effects associated with the administered agent. Effective dosages and dosage regimens can be readily determined from, for example, safety and dose escalation assays, in vivo studies (e.g., animal models), and other methods known to those of skill in the art.

Generally the dosing parameters prescribe that the dose should be less than the amount likely to produce irreversible toxicity to the subject (maximum tolerated dose (MTD)) and not less than the amount required to produce a measurable effect on the subject. Such amounts are determined by, for example, pharmacokinetic and pharmacodynamic parameters associated with ADME, taking into account the route of administration and other factors.

An effective dose (EFFECTIVE DOSE, ED) is a dose or amount of an agent that produces a therapeutic response or desired effect in a portion of the subject taking the agent. A "median effective dose" or ED ₅₀ of an agent refers to the dose or amount of the agent that produces a therapeutic response or desired effect in 50% of the population to whom the agent is administered. Although ED ₅₀ is generally used as a reasonable predictor of the efficacy of an agent, it is not necessarily the appropriate dosage for a clinician to consider all relevant factors. Thus, in some cases, the effective amount is greater than the calculated ED ₅₀, while in still other cases, the effective amount is less than the calculated ED ₅₀, and in still other cases, the effective amount is the same as the calculated ED ₅₀.

Furthermore, an effective dose of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein may be an amount that produces a desired result relative to a healthy subject when administered to the subject in one or more doses. For example, for a subject experiencing a particular disorder, an effective dose may be a dose that improves a diagnostic parameter, measure, marker, etc., of the disorder by at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or greater than 90%, where 100% is defined as the diagnostic parameter, measure, marker, etc., exhibited by a normal subject.

In certain embodiments, a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein may be administered (e.g., orally) one or more times per day at a dosage level of about 0.01mg/kg to about 50mg/kg, or about 1mg/kg to about 25mg/kg of the subject's body weight per day to achieve the desired therapeutic effect.

For administration of oral medicaments, the compositions may be provided in the form of tablets, capsules and the like, containing from 1.0 to 1000 mg of the active ingredient, in particular 1.0、3.0、5.0、10.0、15.0、20.0、25.0、50.0、75.0、100.0、150.0、200.0、250.0、300.0、400.0、500.0、600.0、750.0、800.0、900.0 and 1.00.0 mg of the active ingredient.

In certain embodiments, the dosages of the compounds of formula (I), (Ia), (Ib) or sub-embodiments or salts thereof described herein are contained in a "unit dosage form". The phrase "unit dosage form" refers to physically discrete units, each unit containing a predetermined amount of a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein, alone or in combination with one or more additional agents, sufficient to produce the desired effect. It will be appreciated that the parameters of the unit dosage form will depend on the particular agent and the effect to be achieved.

Kit for detecting a substance in a sample

The invention also contemplates kits comprising compounds of formula (I), (Ia), (Ib), or sub-embodiments described herein, or salts thereof, and pharmaceutical compositions thereof. As described below, the kit is generally in the form of a physical structure containing the various components and may be used, for example, to carry out the above-described methods.

The kit may comprise one or more compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof (provided, for example, in a sterile container), which may be in the form of a pharmaceutical composition suitable for administration to a subject. The compounds of formula (I), (Ia), (Ib) or sub-embodiments described herein or salts thereof may be provided in a form ready for use (e.g. tablet or capsule) or in a form that is required, for example, to be reconstituted or diluted (e.g. powder) prior to administration. When a compound of formula (I), (Ia), (Ib) or a sub-embodiment or salt thereof described herein is in a form that requires reconstitution or dilution by a user, the kit may further include diluents (e.g., sterile water), buffers, pharmaceutically acceptable excipients, etc., packaged together or separately with the compound of formula (I), (Ia), (Ib) or sub-embodiment or salt thereof described herein. When combination therapies are contemplated, the kit may comprise several agents alone, or they may already be combined in the kit. Each component of the kit may be enclosed in a separate container, and all of the various containers may be in a single package. The kit of the present invention may be designed for proper maintenance of the conditions (e.g., refrigeration or freezing) necessary for the components contained therein.

The kit may comprise a label or package insert that includes therein identifying information of the components and their instructions for use (e.g., dosage parameters, clinical pharmacology of the active ingredient(s), including mechanism of action, pharmacokinetics and pharmacodynamics, adverse reactions, contraindications, etc.). The label or insert may include manufacturer information such as lot number and expiration date. The label or package insert may be, for example, integrated into the physical structure containing the components, contained within the physical structure alone, or attached to the components of the kit (e.g., ampoule, tube, or vial).

The label or insert may additionally include or be incorporated into a computer-readable medium, such as a magnetic disk (e.g., hard disk, card, memory disk), optical disk, such as CD-or DVD-ROM/RAM, DVD, MP, magnetic tape, or electronic storage medium (e.g., RAM and ROM), or a combination thereof, such as magnetic/optical storage medium, flash memory medium, or memory card. In some embodiments, no actual instructions are present in the kit, but a means is provided to obtain instructions from a remote source, such as through the internet.

Synthetic examples

The following examples and references (intermediates) are intended to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the invention and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the following experiments, or all experiments that can be performed, are performed. It should be understood that the exemplary descriptions written at the present time are not necessarily made, but rather that these descriptions may be made to generate data or the like as described herein. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for.

Example 1

First step

Reaction 1-1 (10 g,90.8 mmol), N, N-diisopropylethylamine (23 g,181 mmol) was dissolved in N, N-dimethylformamide (100 mL), 2- (trimethylsilyl) ethoxymethyl chloride (18.2 g,109 mmol) was added at 0deg.C, and after the addition was completed, the temperature was raised to 50deg.C for 16h. After the completion of the reaction, water (400 mL), ethyl acetate extraction (100 mL. Times.3), organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 1-2 (15 g, yield 68.8%).

Second step

Intermediate 1-2 (12 g,50.4 mmol), t-butylsulfinamide (9.2 g,75.6 mmol) was dissolved in tetrahydrofuran (300 mL), tetraethyltitanate (46 g,201.6 mmol) was added and stirred for 16h at 70℃under nitrogen. After the reaction was completed, cooled to room temperature, ethyl acetate (150 mL) was added, poured into ice water (300 mL), stirred for 15min, the solid was filtered off, the filtrate was extracted by liquid-liquid separation, the aqueous phase was extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 1-3 (12.5 g, yield 73.1%).

Third step

Intermediate 1-3 (12.5 g,36.4 mmol) was dissolved in tetrahydrofuran (120 mL) and lithium tri-sec-butylborohydride (54.7 mL,54.6 mmol) was added dropwise at-50℃under nitrogen. After the completion of the dropwise addition, the reaction was naturally warmed to room temperature and quenched for 1h, saturated aqueous ammonium chloride (50 mL) was added dropwise under ice bath, the reaction was quenched with stirring for 10min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 1-4 (9.6 g, yield 76.4%).

Fourth step

Intermediate 1-4 (5 g,14.5 mmol) was dissolved in 1, 4-dioxane (50 mL), hydrochloric acid-dioxane solution (50 mL) was added, reacted at room temperature for 1h, the solvent was removed by concentration under reduced pressure, dichloroethane (50 mL) was added to the residue, triethylamine (4 mL,29.0 mmol) was added to give crude IM-01, which was carried forward without purification.

Fifth step

Compounds 1-5 (2.5 g,14.45 mmol) were dissolved in 1, 2-dichloroethane (25 mL) and oxalyl chloride (1.65 mL,19.51 mmol) was slowly added at room temperature. Stirring at 55℃for 1h, then heating to 85℃and stirring for 16h. The reaction system was cooled to 0℃and IM-01 (3.5 g,13.00 mmol) was added dropwise thereto, followed by stirring at room temperature for 2 hours. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by column chromatography to give compound 1-6 (2.11 g, yield 33.2%) as a yellow powder.

Sixth step

Compounds 1-6 (2.11 g,4.79 mmol) were dissolved in toluene (20 mL) and nitrogen blanketed, lithium bis trimethylsilylamide (50 mL,7.19 mmol) was added dropwise at 0deg.C and stirred at 80deg.C for 16h. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (50 g), and concentrated by filtration. The crude product was purified by column chromatography to give compound 1-7 (1.56 g, yield 93.0%) as a white powder.

Seventh step

Compounds 1-7 (150 mg,0.36 mmol) were dissolved in anhydrous acetonitrile (5 mL) and 4-dimethylaminopyridine (88 mg,0.72 mmol), triethylamine (0.25 mL,1.80 mmol) and 2,4, 6-triisopropylbenzenesulfonyl chloride (545 mg,1.80 mmol) were added, respectively, at room temperature. And (3) stirring for 4 hours at 80 ℃ under the protection of nitrogen to obtain an IM-02 crude product solution, wherein the solution is directly used for the next step without purification.

Eighth step

Compound IM-02 was added 3-methylaminooxetane (622.2 mg,7.14 mmol) at room temperature and stirred for 2h at room temperature. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give 1-8 (50 mg, yield 27.6%) as pale yellow solid.

Ninth step

Compounds 1-8 (50 mg,0.10 mmol) were dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise at room temperature and stirred at room temperature for 1h. The solvent was removed by concentration under reduced pressure, dichloromethane (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 1 (5.0 mg, yield 13.4%) as a white solid.

¹H NMR(400MHz,DMSO-d6)δ12.09(s,1H),8.05(d,J＝8.4Hz,1H),7.62(s,1H),7.53(s,1H),7.28(d,J＝8.6Hz,1H),7.18(s,2H),6.39(s,1H),4.54(s,1H),4.11(dd,J＝13.0,6.7Hz,1H),3.91(d,J＝6.8Hz,1H),3.47(s,1H),2.87～2.77(m,1H),2.34(s,3H),1.81(d,J＝7.2Hz,3H).

MS calcd for C₁₇H₂₀ClN₅O₃[M+H]⁺378.2.

Example 2

First step

The compound IM-02 was added to 3-methylaminopropionitrile (293 mg,3.4 mmol) at room temperature, and the reaction mixture was stirred at room temperature for 2h. Extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give compound 2-1 (40 mg, yield 47.1%) as a transparent oil.

Second step

Compound 2-1 (40 mg,0.082 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The reaction solution was stirred at room temperature for 1h, and the obtained reaction solution was directly concentrated, and the crude product was purified by preparative chromatography to give white solid 2 (15 mg, yield 51.7%).

¹H NMR(400MHz,DMSO)δ12.01(s,1H),7.84(d,J＝8.8Hz,1H),7.58(s,1H),7.44(s,2H),7.12–7.02(m,2H),6.89(s,1H),6.21(q,J＝7.0Hz,1H),3.88–3.73(m,2H),3.24(s,3H),2.58–2.52(m,2H),1.73(d,J＝7.2Hz,3H).

MS calculated for C₁₇H₁₉O₂ClN₆[M+H]⁺374.123,found 375.60.

Example 3

First step

The compound IM-02 (130 mg,0.19 mmol) was added N, N, N' -trimethylethylenediamine (387 mg,3.79 mmol) at room temperature. The resulting mixture was stirred at room temperature under an atmosphere of N ₂ for 2h. The mixture was extracted with ethyl acetate (50 mL. Times.3). The organic phases were combined and dried over sodium sulfate, filtered and concentrated. The mixture was concentrated to give the crude product. The crude product was purified by column chromatography to give compound 3-1 (34 mg, yield 35.6%) as a transparent oil.

Second step

Compound 3-1 (34 mg,0.07 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under an atmosphere of N ₂ for 1h. Evaporation of the solvent gave the crude product. The crude product was purified by column chromatography to give compound 3 (10.1 mg, yield 40.1%) as a pale yellow powder.

¹H NMR(400MHz,DMSO)δ12.00(s,1H),7.90(d,J＝8.8Hz,1H),7.57(s,1H),7.48(s,1H),7.12–7.02(m,2H),6.20(q,J＝7.0Hz,1H),3.79–3.65(m,2H),3.25(s,3H),2.60(t,J＝6.5Hz,2H),2.18(s,6H),1.72(d,J＝7.2Hz,3H).

MS calcd for C₁₈H₂₃ClN₆O[M+H]⁺375.16,found 375.70.

Example 4

First step

Compound IM-02 (130 mg,0.19 mmol) was added N- (2-methoxyethyl) methylamine (387 mg,3.79 mmol) at room temperature. The resulting mixture was stirred at room temperature under an atmosphere of N ₂ for 2h. The mixture was extracted with ethyl acetate (50 mL. Times.3). The organic phases were combined and dried over sodium sulfate, filtered and concentrated. The mixture was concentrated to give the crude product. The crude product was purified by column chromatography to give 4-1 (26 mg, yield 28.0%) as a transparent oil.

Second step

Compound 4-1 (26 mg,0.05 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under an atmosphere of N ₂ for 1h. Spin-drying the solvent to give the crude product. The crude product was purified by column chromatography to give compound 4 (10.9 mg, yield 56.9%) as a pale yellow powder.

¹H NMR(400MHz,DMSO)δ11.99(s,1H),7.94(d,J＝8.7Hz,1H),7.57(s,1H),7.50(s,1H),7.08(d,J＝9.8Hz,2H),6.20(q,J＝6.9Hz,1H),3.83–3.76(m,2H),3.67(t,J＝5.4Hz,2H),3.29(s,3H),3.26(s,3H),1.73(d,J＝7.1Hz,3H).

MS calcd for C₁₇H₂₀ClN₅O₂[M+H]⁺362.13,found 362.50.

Example 5

Compound 2 (200 mg,0.53 mmol) was dissolved in tetrahydrofuran (1 mL) and Burgess reagent (191 mg,1.07 mmol) was added under an ice-water bath. The reaction was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate, followed by addition of water and extraction with ethyl acetate (15 mL. Times.3). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The crude product was purified by preparative chromatography to give 5 as a white solid (15 mg, yield 7.9%).

¹H NMR(400MHz,DMSO)δ12.00(s,1H),7.89(d,J＝8.8Hz,1H),7.58(s,1H),7.53(d,J＝1.9Hz,1H),7.11(dd,J＝8.8,2.0Hz,1H),7.07(s,1H),6.23(q,J＝6.9Hz,1H),3.88(ddd,J＝27.3,13.7,6.8Hz,2H),3.33(s,3H),2.99(d,J＝6.8Hz,2H),1.73(d,J＝7.2Hz,3H).

MS calculated for C₁₇H₁₇OClN₆[M+H]⁺357.12,found 375.00.

Example 6

First step

The compound IM-02 was added to N-methyl ring Ding Jiaan (178.0 mg,1.8 mmol) at room temperature and stirred at room temperature for 2h. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by column chromatography to give 6-1 (100 mg, yield 56.2%) as a pale yellow solid.

Second step

Compound 6-1 (60 mg,0.13 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 1h. The solvent was removed by concentration under reduced pressure, dichloromethane (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 6 as a white solid (20.0 mg, yield 27.0%).

¹H NMR(400MHz,DMSO-d6)δ11.98(d,J＝36.9Hz,1H),7.84(d,J＝8.8Hz,1H),7.61(s,1H),7.52(d,J＝1.9Hz,1H),7.11(dd,J＝8.9,2.1Hz,2H),6.23(q,J＝7.0Hz,1H),3.80(dd,J＝13.6,7.5Hz,1H),3.68(dd,J＝13.5,7.0Hz,1H),3.24(d,J＝8.7Hz,3H),2.79(dt,J＝15.3,7.7Hz,1H),2.11～2.00(m,2H),1.95～1.76(m,4H),1.75(d,J＝7.1Hz,3H).

MS calcd for C₁₉H₂₂ClN₅O[M+H]⁺372.0.

Example 7

First step

Reactant 7-1 (8.7 g,90.8 mmol), N, N-diisopropylethylamine (23 g,181.0 mmol) was dissolved in N, N-dimethylformamide (100 mL), 2- (trimethylsilyl) ethoxymethyl chloride (18.2 g,109.0 mmol) was added at 0deg.C, and after the addition was completed, the temperature was raised to 50deg.C for 16h. After the completion of the reaction, water (400 mL), ethyl acetate extraction (100 mL. Times.3), organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 7-2 (4.1 g, yield 68.8%) as a crude product by column chromatography.

Second step

Intermediate 7-2 (11.4 g,50.4 mmol), t-butylsulfinamide (9.2 g,75.6 mmol) was dissolved in tetrahydrofuran (300 mL), tetraethyltitanate (46 g,201.6 mmol) was added and stirred for 16h at 70℃under nitrogen. After the reaction was completed, cooled to room temperature, ethyl acetate (150 mL) was added, poured into ice water (300 mL), stirred for 15min, the solid was filtered off, the filtrate was extracted by liquid-phase extraction, the aqueous phase was extracted with ethyl acetate (100 ml×3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate (50 g), filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 7-3 (12.1 g, yield 73.1%).

Third step

Intermediate 7-3 (12.1 g,36.4 mmol) was dissolved in tetrahydrofuran (120 mL) and lithium tri-sec-butylborohydride (54.7 mL,54.6 mmol) was added dropwise at-50℃under nitrogen. After the completion of the dropwise addition, the reaction was naturally warmed to room temperature and quenched for 1h, saturated aqueous ammonium chloride (50 mL) was added dropwise under ice bath, the reaction was quenched with stirring for 10min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 7-4 (9.2 g, yield 76.4%).

Fourth step

Intermediate 7-4 (4.8 g,14.5 mmol) was dissolved in 1, 4-dioxane (50 mL), hydrochloric acid-dioxane solution (50 mL) was added, reacted at room temperature for 1h, and the solvent was removed by concentration under reduced pressure to give a residue. Dichloroethane (50 mL) and triethylamine (4 mL,29.0 mmol) were then added, the pH was adjusted to 8, and the residue 7-5 was obtained after concentration and carried forward without purification.

Fifth step

Compounds 1-5 (2.5 g,14.45 mmol) were dissolved in 1, 2-dichloroethane (25 mL) and oxalyl chloride (1.65 mL,19.51 mmol) was slowly added at room temperature. Stirring at 55℃for 1h, then heating to 85℃and stirring for 16h. The reaction system was cooled to 0℃and 7-5 (3.4 g,13.00 mmol) was added dropwise thereto at 0℃and stirred at room temperature for 2 hours. To the reaction system was added 20mL of water, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give compound 7-7 (2.11 g, yield 33.0%) as a yellow powder.

Sixth step

Compound 7-7 (2.24 g,4.79 mmol) was dissolved in toluene (20 mL), and lithium bis (trimethylsilylamide) (50 mL,7.19 mmol) was added dropwise at 0deg.C under nitrogen and stirred at 80deg.C for 16h. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by column chromatography to give compound 7-8 (1.56 g, yield 80.0%) as a white powder.

Seventh step

Compound 7-8 (146 mg,0.36 mmol) was dissolved in anhydrous acetonitrile (5 mL) and 4-dimethylaminopyridine (88 mg,0.72 mmol), triethylamine (0.25 mL,1.80 mmol) and 2,4, 6-triisopropylbenzenesulfonyl chloride (545 mg,1.80 mmol) were added, respectively, at room temperature. The reaction solution was stirred at 80℃for 4h under nitrogen protection and was used directly in the next step without treatment.

Eighth step

Compounds 7-9 1-cyclopropyl-N-methyl methylamine (607.91 mg,7.14 mmol) was added at room temperature and stirred for 2h at room temperature. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by column chromatography to give 7-10 (100 mg, yield 58.7%) as a pale yellow solid.

Ninth step

Compounds 7-10 (100 mg,0.21 mmol) were dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise at room temperature and stirred at room temperature for 1h. The solvent was removed by concentration under reduced pressure, dichloromethane (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 7 as a white solid (43 mg, yield 58.0%).

¹H NMR(400MHz,DMSO-d6)δ11.91(d,J＝42.6Hz,1H),7.95(d,J＝8.8Hz,1H),7.79(d,J＝1.7Hz,1H),7.59(s,1H),7.20(dd,J＝8.8,1.8Hz,1H),6.99(s,1H),5.16(s,2H),3.55(d,J＝6.7Hz,2H),1.21(t,J＝5.9Hz,1H),0.67～0.49(m,2H),0.32(q,J＝4.9Hz,2H).

MS calcd for C₁₇H₁₈ClN₅O[M+H]⁺344.3.

Examples 8&9

First step

1-Cyclopropyl-N-methyl methylamine (607.91 mg,7.14 mmol) was added to the compound IM-02 at room temperature, and the mixture was stirred at room temperature for 2h. Water (20 mL) was added to the reaction system, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by column chromatography to give 8-1 (100 mg, yield 57.5%) as a pale yellow solid.

Second step

Compound 8-1 (100 mg,0.21 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 1h. The solvent was removed by concentration under reduced pressure, dichloromethane (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 8-2 as a white solid (43 mg, yield 58.0%).

Third step

Chiral resolution of compound 8-2 (43 mg) with 0.1% diethyl amine in the mobile phase gave compound 8 (22 mg) and compound 9 (14 mg) with ee >98%.

¹H NMR(400MHz,DMSO-d6)δ11.99(d,J＝32.5Hz,1H),7.89(d,J＝8.7Hz,1H),7.61(s,1H),7.52(s,1H),7.12(d,J＝10.6Hz,2H),6.23(q,J＝6.9Hz,1H),3.65～3.44(m,2H),3.33(s,3H),1.75(d,J＝7.2Hz,3H),1.20(d,J＝6.6Hz,1H),0.57(q,J＝5.2Hz,2H),0.32(d,J＝4.2Hz,2H).

MS calcd for C₁₈H₂₀ClN₅O[M+H]⁺358.2.

Example 10

First step

Compound 10-1 (2.0 g,7.8 mmol) was dissolved in tetrahydrofuran (20 mL) and sodium hydride (625 mg,15.6 mmol) was added at 0deg.C. The reaction solution was stirred at room temperature for 1h. Tert-butyl methylcarbamate (2.1 g,15.6 mmol) was then added and stirred at room temperature for 2h. Ethyl acetate was added thereto, water was added thereto, and extraction was performed with ethyl acetate (30 ml×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 10-2 (1.0 g, yield 57.1%) as a colorless oily compound.

Second step

Compound 10-2 (1.0 g,4.4 mmol) was dissolved in dichloromethane (20 mL) and 1, 4-dioxane (4M, 5 mL) was added at 0deg.C. The reaction solution was stirred at room temperature for 1h. Stirring for 1h at room temperature. Direct concentration gave 10-3 (550 mg) as a white solid.

Third step

Compound 10-3 (400 mg,3.6 mmol) was added IM-02 at room temperature and stirred at room temperature for 2h. The reaction mixture was diluted with ethyl acetate, and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 10-4 (120 mg, yield 65%) as a transparent oily compound.

Fourth step

Compound 10-4 (120 mg,0.23 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The reaction solution was stirred at room temperature for 1h, and the obtained reaction solution was directly concentrated, and the crude product was prepared by high performance liquid chromatography to give 10 (30 mg, yield 33.7%) as a white solid.

¹H NMR(400MHz,DMSO)δ12.01(s,1H),7.92(d,J＝8.7Hz,1H),7.54(d,J＝29.9Hz,2H),7.08(d,J＝10.1Hz,2H),6.21(q,J＝7.1Hz,1H),3.77(d,J＝4.2Hz,4H),3.24(s,3H),1.73(d,J＝7.1Hz,3H),0.44(dd,J＝9.0,6.7Hz,4H).

MS calculated for C₁₇H₁₈ClN₅O[M+H]⁺388.15,found 388.80.

Example 11

First step

Raw material 11-1 (10.0 g,90.8 mmol) was dissolved in N, N-dimethylformamide (100 mL), N-diisopropylethylamine (23.5 g,181.6 mmol) and 2- (trimethylsilyl) ethoxymethyl chloride (24.2 g,145 mmol) were added sequentially to the above reaction system, and the reaction was heated to 50℃for 16h. The reaction was cooled to room temperature, diluted with ethyl acetate, extracted with water (400 mL), three times with ethyl acetate (100 mL. Times.3), and the combined organic phases were washed with saturated brine (400 mL), dried, filtered, concentrated, and the crude product purified by column chromatography to give 11-2 (13.0 g, yield 55.3%) as a white solid.

Second step

Intermediate 11-2 (13.0 g,57.5 mmol) was dissolved in tetrahydrofuran (300 mL), and tert-butylsulfinamide (10.4 g,86.2 mmol) and Ti (OEt) ₄ (52.5 g,230.0 mmol) were added sequentially, and the reaction was stirred under nitrogen at 70deg.C for 16h. After the reaction was completed, the reaction mixture was cooled to room temperature, ethyl acetate (200 mL) was added, the mixture was poured into ice water (400 mL) and stirred for 15min, the filtrate was extracted with ethyl acetate (200 mL. Times.3), the combined organic layers were washed with saturated brine (400 mL), dried, filtered and concentrated, and the crude product was purified by column chromatography to give intermediate 11-3 (14.5 g, yield 80.3%).

Third step

Intermediate 11-3 (14.5 g,46.2 mmol) was dissolved in tetrahydrofuran (150 mL) and methyl Grignard reagent (55.4 mL,55.4 mmol) was slowly added dropwise to the reaction solution at 0deg.C under nitrogen. Then the reaction system was naturally warmed to room temperature, stirred for 3 hours, after the reaction was completed, the reaction was quenched with saturated aqueous ammonium chloride solution (100 mL) at 0 ℃, extracted three times (100 ml×3) with ethyl acetate after stirring for 10 minutes, the combined organic layers were washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography to give intermediate 11-4 (11.2 g, 77.1%).

Fourth step

Intermediate 11-4 (5.0 g,14.5 mmol) was dissolved in 1, 4-dioxane (50 mL), 1, 4-dioxane solution (50 mL) was added and reacted at room temperature for 1h, after the reaction was completed, the obtained hydrochloride was concentrated, dichloroethane (50 mL) was added, triethylamine (4 mL,29.0 mmol) was added under ice-water bath, and free 11-5 (3.6 g, crude product) was concentrated, and the next step was directly fed without purification.

Fifth step

4-Chloro-2-fluorobenzamide (2.0 g,11.6 mmol) was dissolved in anhydrous 1, 2-dichloroethane (20 mL) and oxalyl chloride (1.6 mL,12.7 mmol) was slowly added at room temperature. The reaction system was stirred at 80℃for 16h under an N ₂ atmosphere. The reaction mixture was cooled to room temperature, and after concentration, the crude product was dissolved in anhydrous 1, 2-dichloroethane (20 mL), to which 11-5 (3.0 g,12.7 mmol) was added dropwise at 0℃and stirred at room temperature for 2h. After completion of the reaction, the reaction mixture was poured into water and extracted with methylene chloride (100 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, which was purified by column chromatography to give 11-6 (546 mg, 11.8%) as a white solid.

Sixth step

Compound 11-6 (546 mg,1.8 mmol) was dissolved in toluene (20 mL) and potassium bistrimethylsilylamino (9.0 mL,9.0 mmol) was added dropwise at 0deg.C. The reaction solution was stirred at 80℃for 16h under an atmosphere of N ₂. The reaction was cooled to room temperature, and the reaction mixture was poured into a saturated ammonium chloride (50 mL) solution and extracted with ethyl acetate (50 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 11-7 (264 mg, 48.3%).

Seventh step

Compound 11-7 (264 mg,0.91 mmol) was dissolved in dichloromethane (3.0 mL), 4-dimethylaminopyridine (22 mg,0.18 mmol), triethylamine (0.18 mL,1.82 mmol) and di-tert-butyl dicarbonate (294 mg,1.37 mmol) were added separately at room temperature, and the reaction was stirred at room temperature for 2h. After the reaction was completed, the reaction mixture was poured into water (50 mL) and extracted with methylene chloride (20 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 11-8 (271mg, 76.3%).

Eighth step

Compound 11-8 (97 mg,0.24 mmol) was dissolved in anhydrous acetonitrile (3.0 mL) and 4-dimethylaminopyridine (58.6 mg,0.48 mmol), triethylamine (0.07 mL,0.72 mmol) and 2,4, 6-triisopropylbenzenesulfonyl chloride (362 mg,1.2 mmol) were added, respectively, at room temperature. The reaction mixture was stirred at 80℃for 4h under N ₂ atmosphere and the reaction was used directly in the next step.

Ninth step

1-Cyclopropyl-N-methyl methylamine (204 mg,2.4 mmol) was added to the reaction solution at room temperature, and stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was poured into water (20 mL) and extracted with methylene chloride (20 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 11-9 (54 mg, 39.4%).

Tenth step

Compound 11-9 (54 mg,0.12 mmol) was dissolved in dioxane (2 mL), and hydrochloric acid-dioxane solution (4M, 2 mL) was added dropwise at room temperature. The reaction solution was stirred at room temperature for 1h, after completion of the reaction, the crude product was concentrated to give 11 (13.6 mg, yield 31.1%) as a white solid by purification through high performance liquid chromatography.

¹H NMR(400MHz,MeOD)δ7.92(d,J＝8.6Hz,1H),7.63(s,1H),7.24(s,1H),7.13(d,J＝8.5Hz,1H),6.41(q,J＝7.1Hz,1H),6.23(s,1H),3.73–3.58(m,2H),3.42(s,3H),1.88(d,J＝7.2Hz,3H),1.37–1.17(m,1H),0.62(q,J＝5.6Hz,2H),0.35(q,J＝4.9Hz,2H).

MS calculated for C₁₈H₂₀ClN₅O[M+H]⁺358.14,found 358.00.

Example 12

First step

Compound 12-1 (150 mg,1.07 mmol) was added at room temperature to intermediate IM-02 and stirred at room temperature for 2h. The reaction mixture was diluted with ethyl acetate, and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 12-2 (55 mg, 42.3%) as a clear oil.

Second step

Compound 12-2 (55 mg,0.10 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The reaction was stirred at room temperature for 1h to give a direct concentration of the reaction solution, and the crude product was purified by preparative chromatography to give 12 (20 mg, 47.9%) as a white solid.

¹H NMR(400MHz,DMSO)δ8.21(s,1H),7.91(d,J＝8.8Hz,1H),7.52(s,1H),7.36(s,1H),7.18–7.12(m,1H),6.22–6.05(m,1H),4.67–4.56(m,2H),3.20(s,3H),3.10(d,J＝7.3Hz,1H),2.43(s,3H),2.17(s,3H),1.76(d,J＝7.0Hz,3H).

MS calculated for C₂₀H₂₁O₂ClN₆[M+H]⁺413.14,found 412.70

Example 13

First step

N-allylmethylamine (245 mg,3.5 mmol) was added to the intermediate IM-02 reaction solution at room temperature, and stirred at room temperature for 2h. After the reaction was completed, the reaction mixture was poured into water (20 mL) and extracted with methylene chloride (20 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to give 13-1 (14 mg, 23.5%).

Second step

Compound 13-1 (14 mg,0.03 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added dropwise at room temperature. The reaction solution was stirred at room temperature for 2 hours, and after completion of the reaction, the crude product was concentrated to give 13 (3.9 mg, 38.6%) as a white solid by purification through high performance liquid chromatography.

¹H NMR(400MHz,MeOD)δ7.84(d,J＝8.8Hz,1H),7.57(s,1H),7.32(s,1H),7.10(s,1H),7.06(dd,J＝8.8,1.4Hz,1H),6.26(q,J＝7.0Hz,1H),6.11–5.97(m,1H),5.39–5.34(m,2H),4.35–4.30(m,1H),4.22–4.17(m,1H),3.22(s,3H),1.83(d,J＝7.1Hz,3H).

MS calculated for C₁₇H₁₈OClN₅[M+H]⁺343.12,found 343.00.

Example 14

First step

Compound 14-1 (15 g,156 mmol), potassium carbonate (43 g,311 mmol) was dissolved in acetonitrile (150 mL), 2- (trimethylsilyl) ethoxymethyl chloride (28.6 g,171.6 mmol) was added, and the reaction was completed at 50℃for 16h. After the completion of the reaction, water (500 mL) was added, extraction was performed three times with ethyl acetate (3X 500 mL), and the organic layer was washed with saturated brine (400 mL), dried and concentrated, and the crude product was purified by column chromatography to give intermediate 14-2 (11 g, 31.4%).

Second step

Compound 14-2 (3 g,13.3 mmol), t-butylsulfinamide (2.4 g,19.8 mmol) was dissolved in tetrahydrofuran (60 mL), tetraethyltitanate (15 g,52.8 mmol) was added and the reaction stirred under nitrogen at 70deg.C for 16h. After the reaction was cooled to room temperature, ethyl acetate (40 mL) was added, poured into ice water (50 mL), stirred for 15min, the solid was filtered off, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (2X 20 mL), the organic phase was combined with saturated brine (30 mL) and dried and concentrated, and the crude product was purified by column chromatography to give intermediate 14-3 (3.1 g, 71%).

Third step

Compound 14-3 (2.5 g,7.59 mmol) was dissolved in tetrahydrofuran (40 mL), and phenylmagnesium bromide (16 mL,16 mmol) was added dropwise to the reaction solution at 0deg.C under nitrogen. After the reaction was completed, the reaction system was warmed to room temperature and quenched with water (15 mL) dropwise in ice bath, stirred for 10min, extracted with ethyl acetate (2X 50 mL), the combined organic layers were washed with saturated brine (30 mL), dried and concentrated, and the crude product was purified by column chromatography to give intermediate 14-4 (3.1 g, 83.6%).

Fourth step

Compound 14-4 (2 g,4.9 mmol) was dissolved in 1, 4-dioxane (20 mL), and after the addition of hydrochloric acid-dioxane solution (10 mL), the reaction was carried out at room temperature for 16h, after the completion of the reaction, the solvent was dried by spinning, the hydrochloride was added to dichloroethane (30 mL), triethylamine (4 mL,29.0 mmol) was added under ice bath, the solvent was dried by spinning to give the free amine, and the next step was directly fed without purification.

Fifth step

Raw material 14-6 (1 g,4.8 mmol) was dissolved in N, N-dimethylformamide (25 mL), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (2.74 g,7.2 mmol), amine chloride (0.77 g,14.4 mmol) and N, N-diisopropylethylamine (1.86 g,14.4 mmol) were added, and after completion of the reaction, water (100 mL) was added, and after stirring for 10min, suction filtration was performed, and the cake was dried by water washing to give compound 14-7 (0.6 g, 60.3%) as a white powder.

Sixth step

Compound 14-7 (1 g,4.83 mmol) was dissolved in anhydrous 1, 2-dichloroethane (15 mL) and oxalyl chloride (800 mg,6.28 mmol) was slowly added at room temperature. The resulting mixture was stirred under nitrogen at 55℃for 1 hour and then at 85℃for 16 hours. The mixture was stirred at 0℃for 2 hours at room temperature by adding intermediate 14-5 (1.46 g,4.83 mmol). LCMS indicated complete reaction. The mixture was quenched with ice water (15 mL) and extracted with ethyl acetate (3×20 mL). The organic phases were combined and dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography to give compound 14-8 (750 mg, 33.2%) as a pale yellow powder.

Seventh step

Compound 14-8 (630 mg,1.53 mmol) was dissolved in toluene (12 mL) and lithium bis trimethylsilylamide (4.58 mL,4.58 mmol) was added dropwise at 0deg.C. The resulting mixture was stirred under nitrogen at 80℃for 16 hours. LCMS indicated complete reaction. The mixture was quenched with water (10 mL) and extracted with ethyl acetate (3X 10 mL). The organic phases were combined and dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography to give compound 14-9 (580 mg, 73.5%) as a pale yellow powder.

Eighth step

Compound 14-9 (560 mg,1.08 mmol) was dissolved in anhydrous acetonitrile (10 mL) and benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (1.92 g,4.34 mmol), 1, 8-diazabicyclo [5.4.0] undec-7-ene (660 mg,4.34 mmol) and a solution of dimethylamine in tetrahydrofuran (21.6 mL,43.2 mmol) were added, respectively, at room temperature. The resulting mixture was stirred at room temperature under sealed conditions for 16 hours, and the reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (3X 10 mL). The organic phases were combined and dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography to give compound 14-10 (160 mg, 27.2%) as a pale yellow solid.

Ninth step

Compound 14-10 (25 mg,0.05 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. LCMS indicated complete reaction and evaporation of solvent gave the crude product. The crude product was purified by high performance liquid chromatography to give compound 14 (10 mg, 52.6%) as a white powder.

¹H NMR(400MHz,DMSO-d6)δ12.13(s,1H),8.07(d,J＝8.5Hz,1H),7.97(s,1H),7.68(s,1H),7.63(s,1H),7.29(d,J＝7.4Hz,3H),7.24(d,J＝6.7Hz,1H),7.20(d,J＝7.7Hz,2H),7.12(s,1H),3.29(s,6H).

MS calcd for C₂₁H₁₈OF₃N₅[M+H]+414.15,found 414.2.

Example 15

First step

N-methylcyclopropylamine (508.0 mg,7.14 mmol) was added to the compound IM-02 at room temperature, and the mixture was stirred at room temperature for 2h. The reaction mixture was quenched with 20mL of water, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give 15-1 (150 mg, yield 90.0%) as a pale yellow solid.

Second step

Compound 15-1 (150 mg,0.10 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 1 hour. The solvent was removed by concentration under reduced pressure, dichloromethane (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 15 (40 mg, 30.3% yield) as a white solid.

¹H NMR(400MHz,DMSO-d6)δ12.02(s,1H),8.29(d,J＝8.8Hz,1H),7.61(s,1H),7.51(s,1H),7.09(d,J＝8.8Hz,2H),6.29(q,J＝7.0Hz,1H),3.43(s,1H),3.19(s,3H),1.75(d,J＝7.2Hz,3H),0.86(d,J＝5.9Hz,2H),0.65(dd,J＝10.4,3.8Hz,1H),0.53(s,1H).

MS calcd for C₁₇H₁₈ClN₅O[M+H]⁺344.2.

Example 16

First step

A solution of compound IM-02 (244 mg,0.36 mmol) in acetonitrile was added N-methylcyclobutylamine hydrochloride (870 mg,7.2 mmol) and N, N-diisopropylethylamine (923 mg,7.2 mmol) at room temperature. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. LCMS indicated complete reaction. The mixture was extracted with ethyl acetate (3X 50 mL). The organic phases were combined and dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by prep. to give compound 16-1 (150 mg, 85.0%) as a pale yellow solid.

Second step

Compound 16-1 (200 mg,0.41 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. LCMS indicated complete reaction and evaporation of solvent gave the crude product. The crude product was purified by high performance liquid chromatography to give compound 16 (40 mg, 27.3%) as a white powder.

¹H NMR(400MHz,DMSO-d6)δ12.00(s,1H),7.70(d,J＝8.7Hz,1H),7.53(d,J＝32.3Hz,2H),7.20–7.01(m,2H),6.26–6.16(m,1H),4.55(p,J＝8.6Hz,1H),3.11(s,3H),2.36–2.14(m,4H),1.79–1.52(m,5H).

MS calcd for C₁₈H₂₀OClN₅[M+H]⁺358.14,found 358.3.

Example 17

First step

Compound IM-02 was added methoxymethyl amine (436.2 mg,7.14 mmol) at room temperature and stirred at room temperature for 2h. To the reaction system was added 20mL of water, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give 17-1 (50 mg, yield 30.3%) as a pale yellow solid.

Second step

Compound 17-1 (50 mg,0.11 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 1 hour. The solvent was removed by concentration under reduced pressure, DCM (5 mL) was added, the system was basified with N, N-diisopropylethylamine to ph=8 and concentrated under reduced pressure to give a residue. The crude product was purified by high performance liquid chromatography to give 17 (2.0 mg, yield 5.6%) as a white solid.

¹H NMR(400MHz,DMSO-d6)δ12.05(s,1H),8.38(d,J＝8.8Hz,

1H),7.60(d,J＝11.9Hz,2H),7.19(d,J＝8.8Hz,1H),7.12(s,1H),6.42(d,J＝7.1Hz,1H),3.80(s,3H),3.44(s,3H),1.78(d,J＝7.2Hz,3H).

MS calcd for C₁₅H₁₆ClN₅O₂[M+H]⁺334.2.

Example 18

First step

Compound 18-1 (2.5 g,14.45 mmol) was dissolved in dichloroethane (25 mL) and oxalyl chloride (1.65 mL,19.51 mmol) was slowly added at room temperature. After stirring at 55℃for 1 hour, the temperature was raised to 85℃and stirred for 16 hours. IM-01 (3.5 g,13.00 mmol) was added dropwise thereto at 0deg.C and stirred at room temperature for 2 hours. To the reaction system was added 20mL of water, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate (50 g), and concentrated by filtration. The crude product was purified by column chromatography to give compound 18-2 (2.11 g, yield 33.2%) as a yellow powder.

Second step

Compound 18-2 (2.11 g,4.79 mmol) was dissolved in toluene (20 mL), and lithium bis (trimethylsilylamide) (50 mL,7.19 mmol) was added dropwise at 0deg.C under nitrogen and stirred at 80deg.C for 16h. To the reaction system was added 20mL of water, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (50 g), and concentrated by filtration. The crude product was purified by column chromatography to give compound IM-03 (1.56 g, yield 93.0%) as a white powder.

Third step

Compound IM-03 (400 mg,0.88 mmol) was dissolved in anhydrous dioxane (5 mL) and tributyl (2-ethoxyvinyl) tin (480 mg,1.44 mmol), triethylamine (180 mg,1.80 mmol) and tribenzylideneacetone dipalladium (54 mg,0.08 mmol) were added at room temperature, respectively. Under the protection of nitrogen, stirring for 16 hours at 80 ℃, adding 2N hydrochloric acid aqueous solution after the reaction is finished, and stirring for 1 hour. To the reaction system was added 10mL of water, extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give 18-4 (110 mg, yield 29.1%) as a yellow oil.

Fourth step

Compound 18-4 (150 mg,0.36 mmol) was dissolved in anhydrous acetonitrile (5 mL) and 4-dimethylaminopyridine (88 mg,0.72 mmol), triethylamine (0.25 mL,1.80 mmol) and 2,4, 6-triisopropylbenzenesulfonyl chloride (545 mg,1.80 mmol) were added, respectively, at room temperature. The reaction solution was concentrated under nitrogen protection at 80℃for 4 hours to give 18-5 crude, which was used directly in the next step without purification.

Compound 18-5A solution of dimethylamine in tetrahydrofuran (7.1 mL,7.14 mmol) was added at room temperature and stirred at room temperature for 2h. To the reaction system was added 20mL of water, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give 18-6 (90 mg, yield 76.9%) as a pale yellow solid.

Sixth step

Compound 18-6 (90 mg,0.20 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 1 hour. The solvent was removed by concentration under reduced pressure. The crude product was purified by high performance liquid chromatography to give white solid 18 (7.1 mg, yield 11.0%).

¹H NMR(400MHz,DMSO-d6)δ9.00(s,1H),8.11(d,J＝8.4Hz,1H),7.81(s,1H),7.76～7.66(m,2H),6.27(d,J＝5.3Hz,1H),3.30(s,6H),2.60(s,3H),1.84(d,J＝6.8Hz,3H).

MS calcd for C₁₇H₁₉N₅O₂[M+H]⁺326.2.

Example 19

First step

Cyclopropylmethylamine (241 mg,3.4 mmol) was added to compound IM-02 at room temperature, and the mixture was stirred at room temperature for 2h. To the reaction system was added 20mL of water, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate (5 g), and concentrated by filtration. The crude product was purified by column chromatography to give compound 19-1 (20 mg, 25%) as a clear oil.

Second step

Compound 19-1 (20 mg,0.038 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1 mL) was added dropwise at room temperature. The reaction solution was stirred at room temperature for 1 hour, and the obtained reaction solution was directly concentrated, and the crude product was purified by high performance liquid chromatography to obtain white solid 19 (12 mg, 80.5%).

¹H NMR(400MHz,DMSO)δ11.99(s,1H),8.44(s,1H),8.09(d,J＝8.6Hz,1H),7.56(s,1H),7.40(s,1H),7.15(d,J＝8.0Hz,1H),7.05(s,1H),6.45(q,J＝6.8Hz,1H),1.71(d,J＝7.2Hz,3H),1.21–1.11(m,1H),0.51–0.44(m,2H),0.31–0.25(m,2H).

MS calculated for C₁₇H₁₈ClN₅O[M+H]⁺344.12,found 344.30.

Example 20

First step

Compound IM-03 (210 mg,0.45 mmol), trimethylethynyl silicon (70 mg,0.71 mmol) was dissolved in dioxane (5 mL), and dichloro-bis (triphenylphosphine) palladium (35 mg,0.05 mmol), cuprous iodide (21 mg,0.11 mmol), triphenylphosphine (28 mg,0.11 mmol) and triethylamine (5 mL) were added, respectively. The resulting mixture was nitrogen-substituted three times and reacted at 90℃for 16 hours. LCMS indicated complete reaction. The reaction mixture was added with water (10 mL) and extracted with ethyl acetate (3X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product is directly fed to the next step without purification.

Second step

Compound 20-1 (217 mg,0.45 mmol) was dissolved in tetrabutylammonium fluoride in tetrahydrofuran (6 mL,6 mmol), stirred at room temperature for 2 hours, and the reaction mixture was concentrated to give a crude product. The crude product was purified by column chromatography to give compound 20-2 (115 mg, 62.3%) as a pale yellow solid.

Third step

Compound 20-2 (85 mg,0.21 mmol) was dissolved in anhydrous acetonitrile (5 mL) and 4-dimethylaminopyridine (45 mg,0.37 mmol), triethylamine (95 mg,0.94 mmol) and 2,4, 6-triisopropylbenzenesulfonyl chloride (300 mg,1.0 mmol) were added, respectively, at room temperature. The resulting mixture was stirred at 80℃for 4 hours under nitrogen protection and the reaction solution was used directly in the next step without treatment.

Fourth step

Compound 20-3 (142 mg,0.21 mmol) was dissolved in anhydrous acetonitrile (5 mL) and dimethylamine (2.1 mL,4.2 mmol) was added at room temperature. The resulting mixture was stirred at room temperature under nitrogen for 2 hours. LCMS indicated complete reaction. The mixture was added with water (10 mL) and extracted with ethyl acetate (3X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by column chromatography to give compound 20-4 (52 mg, 56.7%) as a pale yellow solid.

Fifth step

Compound 20-4 (63 mg,0.41 mmol) was dissolved in dichloromethane (1.5 mL) and trifluoroacetic acid (1.5 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. LCMS indicated complete reaction and evaporation of solvent gave the crude product. The crude product was purified by high performance liquid chromatography to give compound 20 (20 mg, 45.2%) as a white powder.

¹H NMR(400MHz,DMSO-d6)δ11.98(s,1H),7.85(d,J＝8.4Hz,1H),7.52(d,J＝33.7Hz,2H),7.09(d,J＝8.5Hz,1H),7.04(s,1H),6.21(d,J＝7.2Hz,1H),4.35(s,1H),3.22(s,6H),1.72(d,J＝7.2Hz,3H).

MS calcd for C₁₇H₁₇ON₅[M+H]⁺308.14,found 308.0.

Example 21

Compound IM-03 (110 mg,0.24 mmol) was dissolved in dioxane (10 mL), and tribenzylidene acetone dipalladium (46 mg,0.05 mmol), 2- (dicyclohexylphosphine) 3, 6-dimethoxy-2, 4, 6-triisopropyl-1, 1' -biphenyl (54 mg,0.1 mmol), potassium t-butoxide (38 mg,0.34 mmol) and ethylamine (0.5 mL,0.5 mmol) were added, respectively. The resulting mixture was nitrogen-substituted three times and reacted at 100℃for 16 hours. LCMS indicated complete reaction. The reaction mixture was added with water (10 mL) and extracted with ethyl acetate (3X 10 mL). The organic phases were combined and dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography to give compound 21-1 (67 mg, 65.9%) as a yellow powder.

Second step

Compound 21-1 (65 mg,0.15 mmol) was dissolved in anhydrous acetonitrile (10 mL) and benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (265 mg,0.6 mmol), 1, 8-diazabicyclo [5.4.0] undec-7-ene (91 mg,0.6 mmol) and dimethylamine (3 mL,6 mmol) were added separately at room temperature. The resulting mixture was stirred at room temperature under sealed conditions for 16 hours, and the reaction mixture was extracted with ethyl acetate (3X 10 mL) by adding water (10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by prep. to give compound 21-2 (45 mg, 65.3%) as a pale yellow solid.

Third step

Compound 21-2 (40 mg,0.41 mmol) was dissolved in dichloromethane (1.5 mL) and trifluoroacetic acid (1.5 mL) was added dropwise at room temperature. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. LCMS indicated complete reaction and evaporation of solvent gave the crude product. The crude product was purified by prep. to give compound 21 (13 mg, 45.5%) as a white powder.

¹H NMR(400MHz,DMSO-d6)δ11.92(s,1H),7.56–7.40(m,2H),6.94(d,J＝27.9Hz,1H),6.55–6.38(m,1H),6.37–6.23(m,3H),3.13(d,J＝8.7Hz,6H),2.87(q,2H),1.72(dd,J＝7.2,4.5Hz,3H),1.03(t,J＝7.1Hz,3H).

MS calcd for C₁₇H₂₂ON₆[M+H]⁺327.19,found 327.3.

The control compounds of the test examples of the present invention were prepared as described in reference patent WO2021252678, the structure of which is shown below:

Biological examples

Test example 1: enzymatic Activity assay

The malachite green fluorescence test described below was used to determine the ability of compounds of the present disclosure to inhibit MAT2a enzyme.

Experimental materials:

enzyme: MAT2A

HMAT2A:50nM, sipulex (Cepter), 10mg/mL (234. Mu.M), amino acids 1-395 reaction time: 1 hour

L-methionine substrate: alfa elsha (ALFA AESAR)

ATP substrate: alfa elsha

Malachite green detection reagent: milibo Sigma (Millipore Sigma)

Detection buffer: 50mM Hepes,pH 7.5;50mM KCl;10mM MgCl ₂; 10mM DTT.

Temperature: 25 DEG C

The steps are as follows:

mu.L of DMSO or 3 Xfinal concentration of test compound in DMSO is transferred to an appropriate well of a microtiter plate, which is centrifuged at 1000rpm for 1 minute. mu.L of 3 Xfinal concentration of MAT2A enzyme in assay buffer or assay buffer alone was transferred to the appropriate wells, which were centrifuged at 1000rpm for 1 min. The well plate was incubated at room temperature for 15 minutes, and then 5. Mu.L of a 3 Xmixture of L-methionine and ATP substrate in detection buffer was transferred to all test wells. The well plate was centrifuged at 1000rpm for 1 minute and then incubated at room temperature for 1 hour. To all test wells, 5. Mu.L of malachite green detection reagent was added and the well plate was centrifuged at 1000rpm for 1min and incubated at room temperature for 30 min. The absorbance of the well plate at 620nm was read on a microplate reader (INFINITE M1000,1000). A high control with high absorbance (DMSO) represents no inhibition of the enzymatic reaction, while a low control with low absorbance (no enzyme) represents complete inhibition of the enzymatic reaction. After the reaction was completed, 10. Mu.L of the kinase detection reagent was added to each well, and after 30 minutes of reaction at 25℃the chemiluminescence was read using a PERKINELMER NIVO-multiple-tag analyzer, and the integration time was 0.5 seconds.

Data analysis:

The raw data is converted to inhibition rate using the equation (Sample-Min)/(Max-Min) ×100%, and the value of IC ₅₀ can be obtained by curve fitting four parameters (log (inhibitor) vs. response-Variable slope mode in GRAPHPAD PRISM).

IC ₅₀ for the compounds in table 1 above is disclosed in table 2 below:

TABLE 2 results of enzymatic Activity assay of Compounds

Numbering of compounds	IC₅₀(nM)	Numbering of compounds	IC₅₀(nM)
				1	9323	12	>1000
2	290.6	13	50.26
				3	865.5	14	>1000
4	80	15	604
				5	77.67	16	71.7
6	98	17	77.67
				7	61.35	18	>1000
8	89.56	19	105.9
				9	63.19	20	68.04
10	58.91	21	>1000
				11	49.79	Control compounds	90.57

Experimental results show that the compound has good inhibition activity on MAT2A enzyme and is superior to a control compound.

Test example 2: in vitro cell proliferation assay

Experimental materials:

McCoy's 5A medium, purchased from VivaCell, penicillin/streptomycin antibiotics from source cultures and fetal bovine serum from Gibco. HCT116 MTAP KO cell line was purchased from Horizon. CellTiter-Glo (cell viability chemiluminescent detection reagent) reagent was purchased from Promega.

The experimental method comprises the following steps:

HCT116 MTAP KO cell antiproliferation assay:

Cells were seeded in white 384-well plates, 40 μl of cell suspension containing 500 HCT116 MTAP KO cells per well. Cell plates were placed in a carbon dioxide incubator overnight for culture. The test compounds were diluted 3-fold with a row gun to the 9 th concentration, i.e. from 200. Mu.M to 0.03. Mu.M, and a double multiplex well experiment was set up. 78. Mu.L of medium was added to the intermediate plate, and 2. Mu.L of the gradient diluted compound per well was transferred to the intermediate plate at the corresponding position, and after mixing, 10. Mu.L of the gradient diluted compound per well was transferred to the cell plate. The concentration of compound transferred into the cell plate ranged from 1. Mu.M to 0.15nM. The cell plates were placed in a carbon dioxide incubator for 6 days. A cell plate was also prepared and the signal value read on the day of dosing as the maximum value (Max value in the following equation) was used in the data analysis. To this plate, 10. Mu.L of cell viability chemiluminescent detection reagent was added per well and incubated at room temperature for 10 minutes to stabilize the luminescent signal. Multiple marker analyzer readings were used. After incubation of the cell plates with the compounds, 10 μl of cell viability chemiluminescent detection reagent per well was added to the cell plates and incubated at room temperature for 10 minutes to stabilize the luminescent signal. Multiple marker analyzer readings were used.

Data analysis:

The raw data is converted to inhibition rate using equation (Sample-Min)/(Max-Min) ×100%, and the value of IC ₅₀ can be obtained by curve fitting four parameters (obtained in "log (inhibitor) vs. response-Variable slope" mode in GRAPHPAD PRISM).

IC ₅₀ for some of the compounds in table 1 above is disclosed in table 3 below:

table 3 results of in vitro cell proliferation experiments with Compounds

Test example 3: pharmacokinetic studies in mice

18 Male BALB/c mice were used, the dose was 10mg/kg, the administration route was gastric lavage, the vehicle was 5% DMSO+45% PEG400+50% aqueous solution, and the blood collection time points were 5, 15, 30 minutes and 1,2, 4, 6, 8, 24 hours after administration. After blood sample collection, placing the blood sample in a marked ice bath centrifuge tube, and rapidly centrifuging to separate out plasma, wherein the centrifugation conditions are as follows: 4000 rpm, 10min, 4 deg.C, and plasma at-60 deg.C. After thawing the slurry sample at room temperature, 50. Mu.L of the internal standard (200 ng/mL, acetonitrile, terfenadine) was added, and after vortexing and mixing for 1min, centrifugation was performed for 10min at 4℃under 15400 g. Taking supernatant for LC-MS/MS quantitative analysis, and outputting data such as original map, concentration, accuracy and the like by using analysis 1.6.3 software for main pharmacokinetic parameters.

The pharmacokinetic parameters of compound 8 and the control compound in table 1 above are disclosed in table 4 below:

TABLE 4 essential pharmacokinetic parameters in mice

Experimental results show that the compound of the invention shows excellent pharmacokinetic properties in mice and is superior to control compounds.

The foregoing description is considered as illustrative only of the principles of the disclosure. In addition, since numerous modifications and variations will be readily apparent to those skilled in the art, it is not desired to limit the invention to the exact construction and process shown and described above. Accordingly, all suitable modifications and equivalents may be resorted to as falling within the scope of the invention as defined by the appended claims.

Claims

1. A compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:

wherein:

w is CR ⁴ or N;

x is CR ⁵ or N;

y is CR ⁶ or N;

z is CR ⁷ or N;

v is CR ⁸ or N;

R ⁸ is selected from the group consisting of: hydrogen, alkyl, and hydroxy;

R ^a and R ^b are each independently selected from hydrogen or alkyl;

r ^c is selected from the group consisting of: hydrogen, alkyl, and cycloalkyl;

Each R ^g is independently hydrogen or C _1-3 alkyl.

2. The compound of claim 1, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, having a structure according to formula (Ia) or (Ib) below:

3. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ¹ is-NR ^cR^d.

4. The compound of claim 3, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ^c is hydrogen or alkyl.

5. The compound of claim 4, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ^c is methyl.

6. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ^d is selected from the group consisting of: alkyl, alkoxy, and cycloalkyl, wherein each of alkyl, alkoxy, and cycloalkyl is optionally substituted with one or more R ^e.

7. The compound of claim 6, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein each R ^e is independently selected from the group consisting of: cyano, hydroxy, -NR ^fR^g, carbamoyl, alkyl, alkenyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, cycloalkylalkyl, cycloalkyl and heterocyclyl.

8. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ^d is selected from the group consisting of: methyl group, And methoxy.

9. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ² is selected from the group consisting of: hydrogen, alkyl, and aryl.

10. The compound of claim 9, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ² is methyl.

11. The compound of claim 9, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ² is phenyl.

12. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ³ is heterocyclyl.

13. The compound of claim 12, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ³ is five membered heterocyclyl.

14. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ³ is selected from the group consisting of

15. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁴ is hydrogen.

16. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁵ is hydrogen.

17. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁶ is selected from the group consisting of: alkylcarbonyl, halogen, haloalkyl, alkynyl and-NR ^aR^b.

18. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁶ is selected from the group consisting of: chlorine, trifluoromethyl,

19. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁷ is hydrogen.

20. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R ⁸ is hydrogen.

21. The compound of claim 1 or 2, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the compound has a formula selected from the group consisting of:

22. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

23. A method of treating a MAT 2A-mediated disease in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1 to 21, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 22.

24. The method of claim 23, wherein the disease is cancer.

25. The method of claim 24, wherein the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

26. A method of treating MTAP-deficient cancer in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1 to 21, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 22.

27. The method of claim 26, wherein the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

28. A method for treating cancer in a patient, wherein the cancer is characterized by reduced or no expression of the MTAP gene, a deletion of the MTAP gene, or reduced function of the MTAP protein, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 21, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 22.

29. The method of claim 28, wherein the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.

30. Use of a compound according to any one of claims 1 to 21, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 22, in the manufacture of a medicament for the treatment of cancer.

31. The method of claim 30, wherein the cancer is selected from the group consisting of: leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer, bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-hodgkin's lymphoma, and mesothelioma.