Pyridopyridone mat2a inhibitor and pharmaceutical composition comprising same, and medical use

Abstract

The present invention provides a compound as represented by formula I and an isomer or pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising same, and a pharmaceutical use as a MAT2A inhibitor. The present invention provides a MAT2A inhibitor compound which has a novel structure, high activity, good selectivity, and good oral absorption in rats. The compound has a good prospect of application in the prevention and / or treatment of indications related to MAT2A inhibition, such as cancer and autoimmune diseases.

Description

A pyridopyridone MAT2A inhibitor and its pharmaceutical composition and medical use Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to a pyridopyridone compound and its stereoisomers, or pharmaceutically acceptable salts thereof, as a MAT2A inhibitor, a preparation method, a pharmaceutical composition, and use thereof in preparing a drug for use in subjects suffering from diseases such as cancer. Background Art

[0002] Methionine adenosyltransferase (MAT) (also known as S-adenosylmethionine synthetase) is a cellular enzyme that catalyzes the synthesis of S-adenosylmethionine (SAM or AdoMet) from methionine and ATP; this catalysis is considered the rate-limiting step of the methionine cycle. SAM is an propylamino group donor in polyamine biosynthesis and the major methyl group donor for DNA methylation. It is involved in gene transcription and cell proliferation, as well as the production of secondary metabolites.

[0003] Human MAT consists of three isoforms: MAT1 and MAT3, which are expressed in liver tissue, and MAT2A, which is ubiquitously expressed in human cell types and the predominant form in human tumors. Crystal structure and mechanistic studies have revealed that despite their 85% amino acid sequence identity, their mechanisms of action differ significantly. In its purified active form, MAT2A forms a functional homodimer and binds to the regulatory protein MAT2B. MAT2B modulates MAT2A activity by increasing its sensitivity to inhibition by the ADOMet product, but without providing a significant rate enhancement. Cellular localization studies have demonstrated that MAT2A is present in both the cytoplasm and the nucleus. Nuclear condensation of MAT2A reportedly occurs during replication and the subsequent G2 phase, fulfilling the requirement for hypermethylation of DNA and histone methylation processes in the nucleus during S phase. ADOMet's activity in transmethylation reactions has been identified as a rate-limiting factor in lung cancer stem cell development, making MAT2A and related enzymes of the methionine cycle attractive targets for anticancer drugs. The methionine required for ADOMet production by MAT2A comes from dietary sources or the polyamine cycle, where 5-methylthio-D-ribose 1-phosphate, generated by S-methyl-5'-thioadenosine phosphorylase, is recycled to methionine. Increased expression of the MAT2A protein has been reported in cancers including colon, liver, stomach, blood, and liver cancers. Approximately 15% of human cancers display deletions in the MTAP (S-methyl-5'-thioadenosine phosphorylase) gene, thereby lacking the methionine recycling pathway for polyamine synthesis. Loss of MTAP at chr9p21 often includes deletion of the CDKN2a tumor suppressor gene locus. Comprehensive genetic lethality studies of MATP- / - cancer cells have demonstrated increased sensitivity to inhibition of MAT2A, PRMT5, and PRMT1.

[0004] In hepatocellular carcinoma (HCC), downregulation of MAT1A and upregulation of MAT2A occur, which is called MAT1A:MAT2A switch. The switch, accompanied by upregulation of MAT2B, results in lower SAM content, which provides a growth advantage for liver cancer cells. Because MAT2A plays a crucial role in promoting the growth of liver cancer cells, it is a target for anti-tumor therapy. Recent studies have shown that silencing by using small interfering RNA substantially inhibits the growth of liver cancer cells and induces apoptosis. See, for example, T. Li et al., J. Cancer 7(10)(2016)1317-1327.

[0005] Some MTAP-deficient cancer cell lines are particularly sensitive to inhibition of MAT2A, Marjon et al. (Cell Reports 15(3)(2016)574–587). MTAP (methylthioadenosine phosphorylase) is an enzyme widely expressed in normal tissues that catalyzes the conversion of methylthioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate. Adenine is salvaged to produce adenosine monophosphate, and 5-methylthioribose-1-phosphate is converted to methionine and formate. Due to this salvage pathway, MTA can serve as an alternative purine source when de novo purine synthesis is blocked, for example, by antimetabolites such as L-alanosine.

[0006] MAT2A is dysregulated in other cancers lacking MTAP deletion, including hepatocellular carcinoma and leukemia. J. Cai et al., Cancer Res. 58 (1998) 1444-1450; TS Jani et al., Cell Res. 19 (2009) 358-369. Silencing MAT2A expression by RNA interference produces antiproliferative effects in various cancer models. H. Chen et al., Gastroenterology 133 (2007) 207-218; Q. Liu et al., Hepatol Res. 37 (2007) 376-388.

[0007] Many human and mouse malignant cells lack MTAP activity. MTAP deficiency is not only found in tissue culture cells, but also in primary leukemias, gliomas, melanomas, pancreatic cancer, non-small cell lung cancer (NSCLC), bladder cancer, astrocytomas, osteosarcomas, head and neck cancer, myxoid chondrosarcomas, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcomas, non-Hodgkin lymphoma, and mesothelioma. The gene encoding human MTAP is located in region 9p21 on human chromosome 9p. This region also contains the tumor suppressor genes p16INK4A (also known as CDKN2A) and p15INK4B. These genes encode p16 and p15, inhibitors of the cyclin D-dependent kinases cdk4 and cdk6, respectively.

[0008] Alternatively, the p16INK4A transcript can be an alternative reading frame (ARF) spliced ​​into a transcript encoding p14ARF. p14ARF binds to MDM2 and prevents the degradation of p53 (Pomerantz et al. (1998) Cell 92: 713-723). The 9p21 chromosome region is of interest because it is often homozygous deleted in a variety of cancers (including leukemia, NSLC, pancreatic cancer, glioma, melanoma and mesothelioma). Deletion usually inactivates more than one gene. For example, Cairns et al. ((1995) Nat. Gen. 11: 210-212) reported that after studying more than 500 primary tumors, almost all of the deletions identified in these tumors involved a 170 kb region containing MTAP, p14ARF and P16INK4A. Carson et al. (WO99 / 67634) reported a correlation between the stage of tumor progression and the loss of homozygosity for the gene encoding MTAP and the gene encoding p16. For example, it was reported that the loss of the MTAP gene, but not p16INK4A, predicted cancer in the early stages of development, while the loss of the genes encoding p16 and MTAP predicted cancer in the later stages of cancer development. In some osteosarcoma patients, the MTAP gene was present at diagnosis but was lost at a later time point (Garcia-Castellano et al., Clin. Cancer Res. 8 (3) 2002 782-787).

[0009] WO2018039972 discloses MAT2A enzyme inhibitors for treating cancer; WO2021158792 mentions MAT2A inhibitors for treating autoimmune diseases or inflammatory diseases; WO2018045071 describes MAT2A enzyme inhibitors for treating diseases or conditions mediated by MAT2A overexpression; CN202080014106.0 also discloses the compound 3-(cyclohex-1-en-1-yl)-6-(4-methoxyphenyl)-2-phenyl-5-(pyridin-3-ylamino)pyrazolo[1,5-a]pyrimidin-7(4H)-one for treating MTAP-deficient non-small cell lung cancer (NSCLC) in patients in need; CN114874207A discloses a MAT2A inhibitor with a 6-in-6 ring nucleus; CN113999232A discloses a polycyclic compound that inhibits MTAP-deficient cancer cells.

[0010] The discovery and search for MAT2A inhibitors with novel structures and good oral absorption in rats has become a hot topic in the development of drugs for treating MTAP-deficient tumors. Summary of the Invention

[0011] SUMMARY OF THE INVENTION

[0012] The present invention addresses the significant need for safe and effective compounds and methods for treating, preventing, and controlling cancer while reducing or avoiding the toxicity and / or side effects associated with conventional therapies. To address the above technical problems, the present invention adopts the following technical solutions:

[0013] In one aspect, the present invention provides a compound as shown in Formula I and its stereoisomers, or a pharmaceutically acceptable salt thereof,

[0014] wherein R1 is selected from H, C1-C6 alkyl, C3-C6 cycloalkyl;

[0015] R2 and R3 are independently selected from 5-12 membered monocyclic or bicyclic aromatic or heteroaryl rings, wherein the aromatic or heteroaryl groups are optionally substituted by one or more identical or different substituents R a replace;

[0016] The heteroaryl group contains 1-3 heteroatoms selected from N, O, and S, and the ring system contains a saturated or partially unsaturated ring system such as a spiro ring, a bridged ring, a fused ring, and a fused ring;

[0017] R a is selected from hydrogen, deuterium, halogen, cyano, -C(O)NH2, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl;

[0018] R4 and R5 are independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxyl, C1-C 12 Alkyl, 3-12 membered cycloalkyl or heterocycloalkyl, 3-12 membered halogenated cycloalkyl or halogenated heterocycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, -NH-(CH2) n -C1-C6 haloalkyl, -NH-(CH2) n -C3-C8 heterocycloalkyl, -NH-(CH2) n -NR b R c 、-NH-(CH2) n -O-C1-C6 alkyl, -O-C1-C6 alkyl, -O-C1-C6 haloalkyl, -O-(CH2) n -heterocyclic group, -O-(CH2) n -cycloalkyl, -O-(CH2) n -C3-C6 heterocycloalkyl, -O-(CH2) n -O-C1-C6 alkyl;

[0019] The C1-C6 monoalkylamino, C1-C6 dialkylamino, -NH-(CH2) n -C1-C6 haloalkyl, -NH-(CH2) n -C3-C8 heterocycloalkyl, -NH-(CH2) n -NR b R c 、-NH-(CH2) n -O-C1-C6 alkyl, -O-C1-C6 alkyl, -O-C1-C6 haloalkyl, -O-(CH2) n -heterocyclic group, -O-(CH2) n -cycloalkyl, -O-(CH2) n -C3-C6 heterocycloalkyl, -O-(CH2) n -O-C1-C6 alkyl is optionally substituted by one or more substituents from the group consisting of halogen, hydroxy, amino, oxo, C1-C6 alkylamino, C1-C6 alkyl, C1-C6 alkoxy, oxo, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl;

[0020] R b 、R c Each is independently selected from H, C1-C6 alkyl.

[0021] N is an integer from 0 to 3, including 0.

[0022] As a preferred technical solution, R1 is selected from hydrogen, methyl, ethyl, isopropyl, and cyclopropyl.

[0023] As a preferred technical solution, R2 is selected from one or more identical or different R a Substituted 5-12 membered bicyclic heteroaryl ring, more preferably

[0024] As a preferred technical solution, R3 is selected from one or more identical or different R a Substituted 5-12 membered monocyclic aromatic or heteroaryl ring, more preferably

[0025] As a preferred technical solution, R4 is selected from Cl, methyl, methoxy, trifluoroethylamino, difluoromethyl, amino, trifluoroethoxy, trifluoromethyl,

[0026] As a preferred technical solution, R5 is selected from F, hydrogen, and methoxy.

[0027] As a preferred technical solution, the compound provided by the present invention has the structure shown in the following formula (Ia), (Ib), (Ic), (Id), and (Ie):

[0028] in,

[0029] R6 is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl;

[0030] R7 and R8 are independently selected from hydrogen, deuterium, halogen, cyano, -C(O)NH2, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl.

[0031] As a preferred technical solution, R7 and R8 are independently selected from hydrogen, chlorine, fluorine, methoxy, trifluoromethoxy, cyano, trifluoromethyl, and methyl.

[0032] As a preferred technical solution, R6 is selected from methyl.

[0033] The present invention also provides the following compounds and stereoisomers thereof, or pharmaceutically acceptable salts thereof:

[0034] The present invention also provides a pharmaceutical composition comprising any one of the above compounds and stereoisomers thereof or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0035] The present invention also provides the use of any one of the compounds described above and stereoisomers thereof, or pharmaceutically acceptable salts and drug combinations thereof in the preparation of a medicament for treating a subject suffering from a disease or condition associated with MAT2a or MTAP protein activity or expression, wherein the disease or condition is preferably cancer or an autoimmune disease, and the cancer is preferably selected from lung cancer, pancreatic cancer, liver cancer, colorectal cancer, bile duct cancer, gallbladder cancer, brain cancer, gastric cancer, leukemia, lymphoma, melanoma, thyroid cancer, nasopharyngeal cancer, glioma, bladder cancer, astrocytoma, basal cell carcinoma, osteosarcoma, head and neck cancer, chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma and mesothelioma; the autoimmune disease is preferably selected from thyroiditis, inflammatory bowel disease, lupus erythematosus, fibrosis, myasthenia gravis, vasculitis, psoriasis, arthritis, scleroderma, dermatitis and the like.

[0036] The present invention also provides the following important intermediate compounds and stereoisomers or pharmaceutically acceptable salts thereof:

[0037] In another aspect, the present invention provides a method for preparing a compound represented by formula (1), characterized in that it comprises the following steps:

[0038] (1) The starting material (i-1) is subjected to a substitution reaction to obtain an intermediate compound (i-2);

[0039] (2) The intermediate compound (i-2) is subjected to substitution and decarboxylation reactions to obtain the intermediate compound (i-3);

[0040] (3) The intermediate compound (i-3) is subjected to an amine transesterification reaction to obtain the intermediate compound (i-4);

[0041] (4) The intermediate compound (i-4) is subjected to a halogenation reaction to obtain the intermediate compound (i-5);

[0042] (5) The intermediate compound (i-5) is subjected to a substitution reaction to obtain the intermediate compound (i-6);

[0043] (6) The intermediate compound (i-6) is subjected to an electrophilic substitution reaction to obtain the intermediate compound (i-7);

[0044] (7) The intermediate compound (i-7) is reacted with Buchwald coupling reaction to obtain compound (1).

[0045] Finally, the present invention also provides a method for preparing the compound represented by formula (II), which comprises the following steps:

[0046] The compound of formula (IIA) is subjected to a substitution reaction with other compounds to obtain a compound of formula (II);

[0047] in,

[0048] R6 is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl;

[0049] R7 and R8 are each independently selected from hydrogen, deuterium, halogen, cyano, -C(O)NH2, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl;

[0050] R1, R4 and R5 are as defined above.

[0051] As a preferred technical solution, R6 is selected from methyl.

[0052] As a preferred technical solution, R1 is selected from hydrogen, methyl, ethyl, isopropyl, and cyclopropyl.

[0053] As a preferred technical solution, wherein R4 is selected from Cl, methyl, methoxy, trifluoroethylamino, difluoromethyl, amino, trifluoroethoxy, trifluoromethyl,

[0054] As a preferred technical solution, R5 is selected from F, hydrogen, and methoxy.

[0055] As a preferred technical solution, R7 and R8 are independently selected from hydrogen, chlorine, fluorine, methoxy, trifluoromethoxy, cyano, trifluoromethyl, and methyl.

[0056] The inventors have discovered that these compounds are highly effective MAT2A inhibitors with extremely strong MAT2A inhibitory activity. They can be used to prepare drugs for the prevention and / or treatment of conditions associated with MAT2A inhibition, including cancers and autoimmune diseases caused by reduced or absent MTAP expression, MTAP gene deletion, or decreased MTAP protein function. The present invention is based on these findings.

[0057] Detailed Description of the Invention

[0058] Various aspects and features of the present invention are further described below.

[0059] The compounds according to the present invention may exist in stereoisomeric forms and the present invention therefore includes all stereoisomeric forms.

[0060] The compounds of the present invention possess asymmetric centers. Compounds of the present invention containing asymmetrically substituted atoms can be separated into optically active or racemic forms. Those skilled in the art will appreciate how to prepare optically active forms, such as by resolution of racemates or synthesis from optically active starting materials. Unless otherwise indicated with respect to specific stereochemistry or isomeric forms, the present invention encompasses all chiral, diastereoisomer, and racemic forms. Methods for preparing the compounds of the present invention and intermediates thereto are also intended to be included in the present invention. All stereoisomers of the compounds of the present invention are also intended to be included in the present invention.

[0061] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both occurring and not occurring.

[0062] "Alkyl" refers to a group of straight or branched saturated hydrocarbon groups having 1 to 10 carbon atoms ("C1-C10 alkyl"). In some embodiments, the alkyl group has 1 to 8 carbon atoms ("C1-C8 alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("C1-C6 alkyl"), the alkyl group has 1 to 5 carbon atoms ("C1-C5 alkyl"), the alkyl group has 1 to 4 carbon atoms ("C1-C4 alkyl"), the alkyl group has 1 to 3 carbon atoms ("C1-C3 alkyl"), the alkyl group has 1 to 2 carbon atoms ("C1-C2 alkyl"), and the alkyl group has 1 carbon atom ("C1 alkyl"). In some embodiments, the alkyl group has 2 to 6 carbon atoms ("C2-C6 alkyl"). Examples of C1-C6 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and the like. Each instance of an alkyl group may independently be optionally substituted, i.e., unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents; such as, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent ("substituted alkyl"). In certain embodiments, the alkyl group is an unsubstituted C1-C10 alkyl group (e.g., -CH3). In certain embodiments, the alkyl group is a substituted C1-C6 alkyl group. Common alkyl abbreviations include Me(–CH3), Et(–CH2CH3), iPr(–CH(CH3)2), nPr(–CH2CH2CH3), n–Bu(–CH2CH2CH2CH3), or i–Bu(–CH2CH(CH3)2).

[0063] The term "alkenyl" means a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and connected to the rest of the molecule by a single bond, such as, but not limited to, ethenyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl, and the like.

[0064] The term "alkynyl" means a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and connected to the rest of the molecule by a single bond, such as, but not limited to, ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-en-4-ynyl, and the like.

[0065] As used herein, the terms "halogen", "halo", "halo" and the like represent fluorine, chlorine, bromine or iodine, and particularly represent fluorine, chlorine, bromine, and particularly preferably fluorine and chlorine.

[0066] "Haloalkyl" refers to an alkyl group as described herein (e.g., C1-C6 alkyl) in which one or more hydrogen atoms are replaced by a halogen (e.g., monohaloalkyl, dihaloalkyl, and trihaloalkyl). Such groups include, but are not limited to, chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chloro-fluoroalkyl, chloro-difluoroalkyl, and 2-fluoroisobutyl.

[0067] "Alkoxy" refers to an alkyl group as described herein (e.g., C1-C6 alkyl) attached to the molecule through an oxygen atom. This includes groups in which the alkyl portion can be straight or branched, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0068] "Cycloalkyl" is a monocyclic saturated cycloalkyl group having 3 to 10 ring carbon atoms ("C3-10 cycloalkyl" or "C3-C10 cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms ("C3-8 cycloalkyl" or "C3-C8 cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("C3-6 cycloalkyl" or "C3-C6 cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("C5-6 cycloalkyl" or "C5-C6 cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms ("C5-10 cycloalkyl" or "C5-C10 cycloalkyl"). Examples of C5-C6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-C6 cycloalkyl groups include the aforementioned C5-C6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-C8 cycloalkyl groups include the aforementioned C3-C6 cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each example of a cycloalkyl group is independently unsubstituted ("unsubstituted cycloalkyl") or substituted ("substituted cycloalkyl") with one or more substituents. In certain embodiments, a cycloalkyl group is an unsubstituted C3-C10 cycloalkyl group. In certain embodiments, a cycloalkyl group is a substituted C3-C10 cycloalkyl group.

[0069] "Heterocycloalkyl" refers to a group in which one or more carbon atoms in the above-mentioned "cycloalkyl" are replaced by heteroatoms such as N, O, S, and P.

[0070] The term "alkylamino" refers to an -alkyl-NH2 structure or a substituted amino-NRdRe, wherein Rd and Re are each independently hydrogen or an alkyl group as described above. For example, the term "monoalkylamino" refers to a substituted amino-NRdRe, wherein one of Rd and Re is hydrogen and the other is an alkyl group as described above; "dialkylamino" refers to a substituted amino-NRdRe, wherein Rd and Re are each independently alkyl groups as described above.

[0071] The term "haloalkylamino" refers to an -alkyl-NH2 structure or a substituted amino-NRdRe, wherein Rd and Re are each independently hydrogen or a haloalkyl group as described above. For example, the term "monohaloalkylamino" refers to a substituted amino-NRdRe, wherein one of Rd and Re is hydrogen and the other is a haloalkyl group as described above; "dihaloalkylamino" refers to a substituted amino-NRdRe, wherein Rd and Re are each independently haloalkyl as described above.

[0072] The term "heterocyclyl" means a stable 3- to 20-membered non-aromatic cyclic group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur. Unless otherwise specified in this specification, a heterocyclyl group can be a monocyclic, bicyclic, tricyclic or more ring system, which can include a fused ring system, a bridged ring system or a spirocyclic system; the nitrogen, carbon or sulfur atoms in the heterocyclyl group can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl group can be partially or fully saturated. The heterocyclyl group can be connected to the rest of the molecule via a carbon atom or a heteroatom and by a single bond. In a heterocyclyl group containing a fused ring, one or more rings can be an aryl or heteroaryl group as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, the heterocyclic group is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclic groups include, but are not limited to, pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]nonan-7-yl, 2-oxa-6-aza-spiro[3.3]heptan-6-yl, 2,5-diaza-bicyclo[2.2.1]heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolane, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl, phthalimido, and the like.

[0073] The term "aryl" means a conjugated hydrocarbon ring system radical having 6 to 18 carbon atoms (preferably 6 to 10 carbon atoms). For the purposes of the present invention, an aryl group can be a monocyclic, bicyclic, tricyclic or more polycyclic ring system, and can also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the aryl group is connected to the rest of the molecule via a single bond via an atom on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1,4-benzoxazine-3(4H)-one-7-yl, and the like.

[0074] The term "heteroaryl" means a 5- to 16-membered conjugated ring system having 1 to 15 carbon atoms (preferably 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur within the ring. Unless otherwise specified in this specification, a heteroaryl group may be a monocyclic, bicyclic, tricyclic or higher ring system, and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is connected to the rest of the molecule via a single bond via an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. For the purposes of the present invention, a heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 5- to 10-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolyl, isoquinolyl, naphthazinyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothiophenyl, oxatriol, oxazolyl, cinnolinyl, quinazolinyl, phenylthio, indolizinyl, o-phenanthroline, isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thienyl, naphthopyridinyl, [1,2,4]triazolo[4,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1,2,4]triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine, etc.

[0075] Unless otherwise indicated, when referring to a specifically named aryl (e.g., phenyl), heterocyclyl (e.g., pyrrolidinyl, piperidinyl, and morpholinyl), or heteroaryl (e.g., tetrazolyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, and furanyl) group, the reference is intended to include rings optionally having 0 to 3, preferably 0 to 2, substituents selected from the substituents listed above for arylheterocyclyl and / or heteroaryl groups.

[0076] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is not only physiologically acceptable to a subject but also refers to a synthetic substance that has pharmaceutical uses, such as a salt formed as an intermediate in a chiral resolution. Although such an intermediate salt cannot be directly administered to a subject, the salt may be useful in obtaining the final product of the present invention. Specifically, this includes acid addition salts (organic and inorganic acids) or base addition salts (including organic and inorganic bases).

[0077] "Pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or veterinary use.

[0078] The "tumors" and "diseases related to abnormal cell proliferation" mentioned in the present invention include but are not limited to leukemia, gastrointestinal stromal tumors, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell lung cancer, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer and other diseases.

[0079] As described herein, the term "disease" refers to a physical condition of the subject, which is related to the disease described in the present invention, for example, peripheral arterial disease and neurodegenerative diseases described in the present invention.

[0080] Cancer treatments of the present invention include standard treatments such as surgery, radiation therapy, chemotherapy, and hormone therapy.

[0081] "Cancer" or "malignancy" refers to any of a variety of diseases characterized by uncontrolled abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other sites in the body (i.e., metastasis), and any of a number of characteristic structural and / or molecular features. "Cancer cell" refers to a cell that is undergoing an early, intermediate, or advanced stage of multistep neoplastic progression. Cancers include mesothelioma, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis and hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, urinary tract cancer, melanoma, brain tumors, head and neck cancer, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchogenic carcinoma, advanced solid tumors, small cell lung cancer, metastatic non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myeloma, liposarcoma, fibrosarcoma, Ewing sarcoma and plasmacytoma, lymphoma, pancreatic ductal adenocarcinoma.

[0082] The compound of the present invention or the pharmaceutical composition containing the same can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, intravenous drip, subcutaneous injection, nasal cavity, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0083] The dosage form can be a liquid dosage form, a solid dosage form, or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and multiple emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), and sprays; semisolid dosage forms can be ointments, gels, pastes, and the like.

[0084] To achieve the purpose of medication and enhance the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known method of administration.

[0085] The compound or composition of the present invention can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

[0086] Beneficial technical effects

[0087] The inventors have discovered that the compounds of the present invention exhibit excellent MAT2A inhibitory activity and selectivity, with IC50 values ​​lower than those of the positive control drug AG270 and compound A (compound 167 in WO2020123395). The present invention provides a class of MAT2A inhibitor compounds with novel structures, potent activity, and good oral absorption in rats. These compounds have promising applications in the prevention and / or treatment of indications related to MAT2A inhibition, such as cancer and autoimmune diseases. DETAILED DESCRIPTION

[0088] The following embodiments are intended to help those skilled in the art better understand the technical solutions of the present invention, but the scope of protection of the present invention includes but is not limited to these.

[0089] For all of the following examples, standard procedures and methods known to those skilled in the art can be used. Unless otherwise indicated, all temperatures are expressed in degrees Celsius. The structures of the compounds are determined by nuclear magnetic resonance spectroscopy (NMR) and / or mass spectroscopy (MS).

[0090] The structures of the compounds of the present invention were confirmed by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are expressed in parts per million (ppm). NMR measurements were performed using a Brukeravance-400 NMR spectrometer. The solvents were deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3), with tetramethylsilane (TMS) as the internal standard.

[0091] Liquid phase mass spectrometry LC-MS measurement The liquid phase part used ACQUITY UPLC ultra-high pressure liquid chromatography, and the mass spectrometry part used XevoG2-SQtof mass spectrometer.

[0092] The starting materials used in the examples of the present invention are known and can be purchased commercially, or can be synthesized using or according to methods known in the art.

[0093] Example 1: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 1)

[0094] Step 1: Synthesis of 6-chloro-2-[(4-chlorophenyl)amino]-5-fluoronicotinic acid

[0095] 4-Chloroaniline (20.00 g, 156.80 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (200 mL) and cooled to -78°C. Lithium bis(trimethylsilyl)amide (235.20 mL, 235.20 mmol, 3.0 eq) was slowly added dropwise. The reaction was continued for 1 hour. 2,6-Dichloro-5-fluoronicotinic acid (16.46 g, 78.40 mmol, 1.0 eq) was then dissolved in ultra-dry tetrahydrofuran (60 mL) and slowly added dropwise to the reaction system at -78°C. The mixture was then transferred to room temperature and allowed to react for 1 hour. After completion of the reaction, saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted with ethyl acetate (200 mL). The organic phases were combined, dried, concentrated, washed with an appropriate amount of methanol, and filtered to yield 6-chloro-2-[(4-chlorophenyl)amino]-5-fluoronicotinic acid (20.1 g, 85.0% yield) as a pale yellow solid. LCMS(TOF MS ES+)m / z[M+H]+:178. 1 H NMR (400MHz, DMSO) δ10.40(s,1H),8.27–8.17(m,1H),7.70–7.58(m,2H),7.44–7.33(m,2H).

[0096] Step 2: Synthesis of ethyl 3-{6-chloro-2-[(4-chlorophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate

[0097] Potassium 3-ethoxy-3-oxopropanoate (8.28 g, 48.66 mmol, 2.0 eq), magnesium chloride (6.73 g, 72.99 mmol, 3.0 eq) and triethylamine (9.85 g, 97.32 mmol, 4.0 eq) were dissolved in acetonitrile solution (150 mL) and stirred at room temperature for 4 hours. 6-Chloro-2-{(4-chlorophenyl)amine}-5-fluoronicotinic acid (7.3 g, 24.33 mmol, 1.0 eq) and ultra-dry dichloromethane (150 mL) were added to another reaction flask, and a small amount of N,N-dimethylformamide was added as a catalyst. Subsequently, oxalyl chloride (5.07 g, 39.99 mmol, 1.5 eq) was slowly added dropwise at 0°C. After the addition was complete, the mixture was transferred to room temperature and reacted for 2 hours. After TLC monitoring of the reaction completion, the reaction solution was directly concentrated to obtain a yellow solid, which was then dissolved in tetrahydrofuran solution (100 mL) and slowly added dropwise to the potassium 3-ethoxy-3-oxopropanoate reaction system at 0°C. The reaction was continued at room temperature for 2 hours. After the reaction, the reaction solution was concentrated and purified using a forward phase column to obtain a yellow solid, ethyl 3-{6-chloro-2-[(4-chlorophenyl)amine]-5-fluoropyridin-3-yl}-3-oxopropanoate (6.3 g, 70% yield). LCMS (TOF MS ES+) m / z [M+H]+: 371. 1 H NMR (400MHz, DMSO) δ10.76 (s, 1H), 8.54 (d, J = 9.2Hz, 1H), 7.77–7.60 (m, 2H), 7.50–7.25(m,2H),4.29(s,2H),4.14(m,J=7.1Hz,2H),1.21(t,J=7.1Hz,3H).

[0098] Step 3: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one

[0099] Ethyl 3-{6-chloro-2-[(4-chlorophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate (6.30 g, 17.02 mmol, 1.0 eq) and potassium carbonate (4.70 g, 34.05 mmol, 2.0 eq) were dissolved in anhydrous ethanol (120 mL) and reacted at 70°C for 3 hours. After completion of the reaction, the mixture was filtered through celite, and the filtrate was purified by forward column chromatography to afford 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (5.1 g, 92.5% yield) as a solid. LCMS (TOF MS ES+) m / z [M+H]+: 325. 1H NMR (400MHz, DMSO) δ12.28 (s, 1H), 8.27 (d, J = 8.1Hz, 1H), 7.65–7.50 (m, 2H), 7.41–7.22 (m, 2H), 5.98 (s, 1H).

[0100] Step 4: Synthesis of 4,7-dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0101] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (5.1 g, 15.74 mmol, 1.0 eq) and phosphorus oxychloride (50 mL) were added to a reaction flask and reacted at 80°C for 16 hours. After completion of the reaction, the phosphorus oxychloride was removed by concentration under reduced pressure. Ethyl acetate was added to the slurry, and the product was filtered to yield a yellow solid, 4,7-dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (4.5 g, 83.6% yield). LCMS (TOF MS ES+) m / z [M+H]+: 343. 1 H NMR (400MHz, DMSO) δ8.51 (d, J = 8.2Hz, 1H), 7.70–7.53 (m, 2H), 7.43–7.32 (m, 2H), 7.26 (s, 1H).

[0102] Step 5: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0103] 4,7-Dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (1.0 g, 2.91 mmol, 1.0 eq), methylamine hydrochloride (294 mg, 4.35 mmol, 1.5 eq), and N,N-diisopropylethylamine (1.5 g, 11.6 mmol, 4.0 eq) were dissolved in acetonitrile (20 mL) and reacted at 80°C for 2 hours. After completion of the reaction, the reaction solution was concentrated to obtain a pale yellow solid, which was then slurried and filtered with ethyl acetate to afford a yellow solid, 7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (410 mg, 41.6% yield), which was directly used in the next step. LCMS (TOF MS ES+) m / z [M+H]+: 338.

[0104] Step 6: Synthesis of 3-bromo-7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0105] 7-Chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (400 mg, 1.18 mmol, 1.0 eq) and N-bromosuccinimide (232.4 mg, 1.30 mmol, 1.1 eq) were added to an acetonitrile solution (8 mL) and reacted at room temperature for 4 hours. After completion of the reaction, the reaction solution was directly concentrated and purified by forward separation (dichloromethane / methanol = 20:1) to afford 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (350 mg, 70.85% yield) as a pale yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 416. 1 H NMR (400MHz, CDCl3) δ7.70 (d, J = 10.7Hz, 1H), 7.54–7.39 (m, 2H), 7.22–7.13 (m, 2H), 5.14 (s, 1H), 2.72–2.68 (m, 3H).

[0106] Step 7: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0107] 3-Bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (297.0 mg, 0.71 mmol, 1.0 eq), 2-methyl-2H-indazole-5-boronic acid (188.0 mg, 1.07 mmol, 1.5 eq) and potassium phosphate (379.8 mg, 1.79 mmol, 2.5 eq) were added to a mixed solvent of dioxane (6 mL) and water (0.6 mL), and finally 1,1-bis(diphenylphosphino)diphenylferric palladium dichloride (52.3 mg, 0.07 mmol, 0.1 eq) was added. Under nitrogen protection, the reaction was carried out at 80 ° C for 16 hours. After the reaction, the reaction solution was concentrated and purified to obtain a yellow solid, 7-chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (40.4 mg, 12.05% yield). LCMS (TOF MS ES+) m / z [M+H]+: 468.0816. 1H NMR (400MHz, DMSO) δ8.36(s,1H),7.86(d,J=11.2Hz,1H),7.72(d,J=5.3Hz,1H),7.69(s,1H),7.58 (t,J=8.3Hz,3H),7.38(d,J=8.3Hz,2H),7.18(d,J=8.9Hz,1H),4.18(s,3H),2.52(d,J=4.6Hz,3H).

[0108] Example 2: Synthesis of 7-chloro-1-(2-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 2)

[0109] Step 1: Synthesis of 6-chloro-2-[(2-chlorophenyl)amino]-5-fluoronicotinic acid

[0110] A mixture of 2-chloroaniline (24.3 g, 190.5 mmol, 2.0 eq) and tetrahydrofuran (120 mL) was added to a three-necked flask. After replacing the atmosphere with nitrogen, the flask was cooled to -78°C and slowly added dropwise with lithium bis(trimethylsilyl)amide (286 mL, 285.7 mmol, 3.0 eq). After stirring at -78°C for 1.5 hours, a solution of 2,6-dichloro-5-fluoronicotinic acid (20 g, 95.2 mmol, 1.0 eq) in tetrahydrofuran was added dropwise. The mixture was transferred to room temperature and allowed to react for 2 hours. After completion of the reaction, the reaction was quenched by addition of dilute hydrochloric acid in an ice bath. The mixture was extracted three times with water and ethyl acetate. The organic phases were combined, dried, concentrated, washed, and slurried with an appropriate amount of methanol. Filtered to obtain 6-chloro-2-[(2-chlorophenyl)amino]-5-fluoronicotinic acid (12 g, 42.02% yield) as a yellow powder. LCMS(TOF MS ES+)m / z[M+H]+:300.9.1H NMR (400MHz, DMSO) δ10.80(s,1H),8.42(dd,J=8.4,1.5Hz,1H),8.28(d,J=8.4Hz,1H) ,7.53(dd,J=8.0,1.5Hz,1H),7.38(m,J=7.9,1.5Hz,1H),7.09(m,J=7.7,1.5Hz,1H).

[0111] Step 2: Synthesis of ethyl 3-{6-chloro-2-[(2-chlorophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate

[0112] Potassium 3-ethoxy-3-oxopropanoate (8.48 g, 49.8 mmol, 1.5 eq), magnesium chloride (9.48 g, 99.6 mmol, 3.0 eq) and triethylamine (15.12 g, 149 mmol, 4.5 eq) were dissolved in acetonitrile solvent (180 mL) and stirred at room temperature for 4 hours. 6-Chloro-2-[(2-chlorophenyl)amino]-5-fluoronicotinic acid (10 g, 33.2 mmol, 1.0 eq), N,N-dimethylformamide (1.3 mL, 1.66 mmol, 0.05 eq) and dichloromethane (140 mL) were added to another reaction flask, and then oxalyl chloride (6.33 g, 49.8 mmol, 1.5 eq) was slowly added dropwise until no bubbles were generated. After stirring at room temperature for 2 hours, the reaction solution was directly concentrated and the obtained solid was dissolved in dichloromethane (60 mL). The solution was slowly added dropwise to the potassium 3-ethoxy-3-oxopropanoate system at 0°C. After the addition was complete, the mixture was moved to room temperature and stirred for 2 hours. After the reaction, the reaction solution was concentrated, extracted three times with ethyl acetate and water, and the organic phases were combined and purified by normal phase column to obtain a yellow solid 3-{6-chloro-2-[(2-chlorophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropionic acid ethyl ester (11.0 g, yield 89.5%). LCMS(TOF MS ES+)m / z[M+H]+:371.03.1H NMR (400MHz, DMSO) δ11.10(s,1H),8.61(d,J=9.2Hz,1H),8.37(dd,J=8.3,1.5Hz,1H),7.56(dd,J=8.0,1.5Hz,1H) ,7.41(m,J=7.9,1.5Hz,1H),7.15(m,J=7.7,1.5Hz,1H),4.32(s,2H),4.16(m,J=7.1Hz,2H),1.21(t,J=7.1Hz,3H).

[0113] Step 3: Synthesis of 7-chloro-1-(2-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one

[0114] Ethyl 3-{6-chloro-2-[(2-chlorophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate (11.5 g, 31.1 mmol, 1.0 eq) and potassium carbonate (8.59 g, 62.2 mmol, 2.0 eq) were added to anhydrous ethanol (250 mL) and reacted at 70°C for 3 hours. After completion of the reaction, the mixture was filtered, and the filtrate was separated by normal phase column chromatography to obtain 7-chloro-1-(2-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (7.56 g, 75.1% yield) as a yellow powdery solid. LCMS(TOF MS ES+)m / z[M+H]+:324.9.1H NMR (400MHz, DMSO-d6) δ8.09(d,J=8.5Hz,1H),7.61–7.52(m,1H),7.47–7.36(m,2H),7.34–7.25(m,1H),4.96(s,1H),4.47(s,1H).

[0115] Step 4: Synthesis of 4,7-dichloro-1-(2-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0116] 7-Chloro-1-(2-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (7.55 g, 23.3 mmol, 1.0 eq) was added to phosphorus oxychloride (80 mL) and refluxed at 70°C for 16 hours. After completion of the reaction, the reaction solution was concentrated, the pH was adjusted to alkaline with sodium bicarbonate, and extracted three times with ethyl acetate and water. The organic phases were combined, dried, and concentrated to afford 4,7-dichloro-1-(2-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (5.35 g, 67.2% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 342.9. 1H NMR (400MHz, DMSO) δ8.57 (d, J = 8.2Hz, 1H), 7.78–7.69 (m, 1H), 7.61–7.52 (m, 3H), 7.35 (s, 1H).

[0117] Step 5: Synthesis of 7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0118] 4,7-Dichloro-1-(2-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (1.5 g, 4.39 mmol, 1.0 eq), methylamine hydrochloride (441 mg, 6.59 mmol, 1.5 eq), and triethylamine (1.77 g, 17.6 mmol, 4.0 eq) were dissolved in acetonitrile (30 mL) and reacted at 80°C for 3 hours. After completion of the reaction, the mixture was concentrated, extracted with aqueous sodium bicarbonate and ethyl acetate, and separated by normal phase column chromatography to obtain 7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (1.4 g, 94.7% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 338.1.

[0119] Step 6: Synthesis of 3-bromo-7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0120] Dissolve 7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (1.54 g, 4.57 mmol, 1.0 eq) and N-bromosuccinimide (895 mg, 5.05 mmol, 1.1 eq) in acetonitrile (40 mL) and stir at room temperature for 4 hours. After the reaction, separate the mixture using a normal phase column to obtain 3-bromo-7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (1.5 g, 79.1% yield) as a yellow solid. LCMS(TOF MS ES+)m / z[M+H]+:415.9; 1H NMR (400MHz, DMSO) δ7.92–7.83(m,2H),7.68(m,J=7.2,3.7Hz,1H),7.50(dd,J=6.9,3.4Hz,2H),2.43(d,J=4.5Hz,3H).

[0121] Step 7: Synthesis of 7-chloro-1-(2-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0122] 3-Bromo-7-chloro-1-(2-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthyridin-2(1H)-one (1.00 g, 2.41 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (506 mg, 2.89 mmol, 1.2 eq), and potassium phosphate (1.28 g, 6.03 mmol, 2.5 eq) were dissolved in 1,4-dioxane (15 mL) and water (1.5 mL). Dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (176 mg, 0.24 mmol, 0.1 eq) was added and stirred at 80 °C under nitrogen for 12 hours. After the reaction, the mixture was filtered, concentrated, extracted with dichloromethane and water, and the organic phase was concentrated. The mixture was separated by a reverse phase column to give a white powdery solid 7-chloro-1-(2-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (670 mg, yield 59.6%). LCMS(TOF MS ES+)m / z[M+H]+:468.1.1H NMR (400MHz, DMSO) δ8.37(s,1H),7.88(d,J=11.1Hz,1H),7.75(d,J=5.0Hz,1H),7.72–7.63(m,2H),7.60 (d, J=9.0Hz, 1H), 7.50 (m, J=2.4Hz, 3H), 7.16 (dd, J=8.5, 1.6Hz, 1H), 4.18 (s, 3H), 2.47 (d, J= 4.5Hz, 3H).

[0123] Example 3: Synthesis of 1-(4-chlorophenyl)-6-fluoro-7-methoxy-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 3)

[0124] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-7-methoxy-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0125] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (60 mg, 0.13 mmol, 1.0 eq) prepared in Example 1 was added to methanol (1 mL), followed by sodium hydride (51.4 mg, 1.28 mmol, 10.0 eq), and stirred at 80°C for 5 hours. After the reaction, the insoluble material was removed by filtration, and the product was purified to obtain 1-(4-chlorophenyl)-6-fluoro-7-methoxy-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (8.2 mg, 13.8% yield) as a white solid. LCMS (TOF MS ES+) m / z [M+H] + :464.1202. 1 H NMR (400MHz, DMSO) δ8.32 (s, 1H), 7.76–7.68 (m, 2H), 7.55 (dd, J = 12.3, 8.5Hz, 4H), 7 .33(d,J=8.3Hz,2H),7.24(d,J=9.0Hz,1H),4.16(s,3H),3.43(s,3H),2.51(s,3H).

[0126] Example 4: Synthesis of 1-(4-chlorophenyl)-6-fluoro-7-methyl-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 4)

[0127] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-7-methyl-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 4)

[0128] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (50 mg, 0.11 mmol, 1.0 eq) obtained in Example 1, methylboric acid (13.2 mg, 0.22 mmol, 2.0 eq), tetrakis(triphenylphosphine)palladium (12.3 mg, 0.01 mmol, 0.1 eq) and potassium carbonate (44.2 mg, 0.33 mmol, 3.0 eq) were dissolved in 1,4-dioxane (1.5 mL) and water (0.3 mL), the system was replaced with nitrogen, and the mixture was stirred at 120°C for 2 hours. After the reaction, the insoluble material was removed by filtration and purified to obtain 1-(4-chlorophenyl)-7-cyclopropyl-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (13.2 mg) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 448.1288. 1 H NMR (400MHz, DMSO-d6) δ8.31 (s, 1H), 7.84 (d, J = 12.0Hz, 1H), 7.61–7.53 (m, 4H), 7.3 4–7.29(m,3H),7.10(dd,J=8.9,1.6Hz,1H),4.17(s,3H),2.49(s,3H),2.23(s,3H).

[0129] Example 5: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (Compound 5)

[0130] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one

[0131] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (60 mg, 0.128 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (127 mg, 1.28 mmol, 10.0 eq), methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (11.6 mg, 0.012 mmol, 0.1 eq) and sodium tert-butoxide (61.5 mg, 0.64 mmol, 5.0 eq) prepared in Example 1 were dissolved in 1,4-dioxane (2 mL), the system was purged with nitrogen three times, and the mixture was stirred at 90°C for 1 hour. After the reaction, the insoluble matter was removed by filtration, and the filtrate was concentrated and purified by preparative purification to give a yellow solid 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (11.2 mg, yield 16.4%). LCMS (TOF MS ES+) m / z [M+H] + :531.10. 1 H NMR (400MHz, DMSO) δ8.33(s,1H),7.78(d,J=12.5Hz,1H),7.66–7.53(m,3H),7.48–7.34(m ,4H),7.11(d,J=8.9Hz,1H),6.37(s,1H),4.21(s,3H),3.34(s,2H),2.42(d,J=4.6Hz,3H).

[0132] Example 6: Synthesis of 1-(4-chlorophenyl)-7-(difluoromethyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 6)

[0133] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-vinyl-1,8-naphthalen-2(1H)-one

[0134] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (200 mg, 0.427 mmol, 1.0 eq) obtained in Example 1, potassium vinyl trifluoroborate (85.9 mg, 0.641 mmol, 1.5 eq), tetrakis(triphenylphosphine)palladium (49.3 mg, 0.042 mmol, 0.1 eq) and potassium carbonate (176.9 mg, 1.28 mmol, 3.0 eq) were dissolved in 1,4-dioxane (4 mL) and water (0.8 mL) and reacted at 110°C. After the reaction, dichloromethane and water were added for extraction, and the product was purified by normal phase column chromatography to obtain a yellow solid 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-vinyl-1,8-naphthalen-2(1H)-one (181 mg, yield 92.1%). LCMS (TOF MS ES+) m / z [M+H] + :460.1. 1 H NMR (400MHz, DMSO) δ8.32(s,1H),7.76(d,J=12.0Hz,1H),7.68–7.59(m,4H),7.48(d,J=5.0Hz,1H),7.38–7.35(m,2H),7.12(dd,J=8 .7,1.7Hz,1H),6.57(dd,J=18.0,11.7Hz,1H),5.58(dd,J=11.7,1.4Hz,1H),5.45–5.37(m,1H),4.18(s,3H),2.53(d,J=2.0Hz,3H).

[0135] Step 2: Synthesis of 8-(4-chlorophenyl)-3-fluoro-6-(2-methyl-2H-indazol-5-yl)-5-(methylamino)-7-oxo-7,8-dihydro-1,8-naphthyridine-2-carbaldehyde

[0136] 1-(4-Chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-vinyl-1,8-naphthalen-2(1H)-one (180 mg, 0.39 mmol, 1.0 eq) and potassium osmate dihydrate (24.3 mg, 0.078 mmol, 0.2 eq) were added to tetrahydrofuran (8 mL) and water (2 mL), and sodium periodate (252.7 mg, 1.17 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 5 hours. After the reaction, the reaction mixture was concentrated, extracted with dichloromethane and water, and concentrated to afford a brown solid, 8-(4-chlorophenyl)-3-fluoro-6-(2-methyl-2H-indazol-5-yl)-5-(methylamino)-7-oxo-7,8-dihydro-1,8-naphthyridine-2-carbaldehyde (173 mg, 96.0% yield). LCMS (TOF MS ES+) m / z [M+H]+: 462.1. 1 H NMR (400MHz, DMSO) δ9.68 (s, 1H), 8.44 (s, 1H), 8.20 (d, J = 12.4Hz, 1H), 7.61–7.54 (m, 5H), 7.42–7.36 (m, 3H), 4.22 (s, 3H), 2.53 (s, 3H).

[0137] Step 3: Synthesis of 1-(4-chlorophenyl)-7-(difluoromethyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0138] 8-(4-chlorophenyl)-3-fluoro-6-(2-methyl-2H-indazol-5-yl)-5-(methylamino)-7-oxo-7,8-dihydro-1,8-naphthyridine-2-carbaldehyde (170 mg, 0.368 mmol, 1.0 eq) was dissolved in dichloromethane (8 mL), and diethylaminosulfur trifluoride (207.3 mg, 1.288 mmol, 3.5 eq) was added. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, the mixture was filtered, concentrated, and extracted with dichloromethane and water. The organic phase was concentrated and separated by reverse phase column chromatography to afford 1-(4-chlorophenyl)-7-(difluoromethyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridine-2(1H)-one (4.6 mg, 2.6% yield) as a white powdery solid. LCMS(TOF MS ES+)m / z[M+H]+:484.1141. 1H NMR (400MHz, DMSO) δ8.41 (s, 1H), 7.80 (d, J = 11.7Hz, 1H), 7.71–7.51 (m, 5H), 7.38 (s,2H),7.15(dd,J=8.9,1.6Hz,1H),6.91–6.53(m,1H),4.19(s,3H),2.49(s,3H).

[0139] Example 7: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 7)

[0140] Step 1: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0141] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (60.0 mg, 0.128 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (176 mg, 1.28 mmol, 10.0 eq) and sodium tert-butoxide (24.5 mg, 0.257 mmol, 2.0 eq) obtained in Example 1 were added to a dioxane solvent (3.0 mL), and finally methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (12.2 mg, 0.0128 mmol, 0.1 eq) was added, and the temperature was raised to 60° C. for 3 hours. After the reaction was complete, the reaction mixture was concentrated, trifluoroacetic acid (2.5 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was then concentrated and subjected to preparative chromatography to afford 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (10.3 mg, 17.98% yield) as a white solid. LCMS (TOF MS ES+) m / z [M+H]+: 449.1069. 1 H NMR (400MHz, DMSO) δ8.29 (s, 1H), 8.13 (d, J = 12.3Hz, 1H), 7.60–7.45 (m, 4H), 7. 29–7.21(m,3H),7.11(d,J=9.0Hz,1H),5.95(s,2H),4.17(s,3H),2.51(s,3H).

[0142] Example 8: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(3-oxocyclobutyl)methoxy]-1,8-naphthyridin-2(1H)-one (Compound 18)

[0143] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(3-oxocyclobutyl)methoxy]-1,8-naphthyridin-2(1H)-one

[0144] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (60.0 mg, 0.128 mmol, 1.0 eq), oxetane-3-ylmethylamine (111.5 mg, 1.28 mmol, 10.0 eq), and sodium tert-butoxide (24.7 mg, 0.257 mmol, 2.0 eq) obtained in Example 1 were added to dioxane solvent (3.0 mL), and finally methanesulfonic acid (2-dicyclohexylphosphine)-3,6- Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (12.2 mg, 0.0128 mmol, 0.1 eq) was heated to 60°C for 4 hours. The mixture was analyzed by LCMS and preparative chromatography to afford 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-[(3-oxocyclobutyl)methoxy]-1,8-naphthyridin-2(1H)-one as a white solid (19.5 mg, 29.55% yield). LCMS (TOF MS ES+) m / z [M+H]+: 519.1595. 1 H NMR (400MHz, CDCl3) δ7.87(d,J=5.7Hz,1H),7.72(t,J=9.3Hz,1H),7.67–7.51(m,2H),7.47–7.38(m,2H),7.29–7.19(m,2H),4.95(d,J=5 .7Hz,1H),4.62(dd,J=7.5,6.4Hz,2H),4.24–4.12(m,5H),3.41(t,J=6.0Hz,2H),2.93(m,J=13.0,7.5,5.7Hz,1H),2.71(d,J=4.9Hz,3H).

[0145] Example 9: Synthesis of 1-(4-chlorophenyl)-7-{[2-(dimethylamino)ethyl]amino}-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 21)

[0146] Step 1: Synthesis of 1-(4-chlorophenyl)-7-{[2-(dimethylamino)ethyl]amino}-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0147] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (50 mg, 0.106 mmol, 1.0 eq) obtained in Example 1, N,N-dimethylethane-1,2-diamine (128.2 mg, 0.534 mmol, 5.0 eq), methanesulfonic acid (4,5-bisdiphenylphosphino-9,9-dimethylxanthene) (2-amino-1,1'-biphenyl-2-yl) palladium (II) (10.1 mg, 0.01 mmol, 0.1 eq) and sodium tert-butoxide (20.5 mg, 0.212 mmol, 2.0 eq) were added to 1,4-dioxane (2 mL) and reacted at 100°C. After the reaction, the reaction solution was filtered and separated by preparative chromatography to obtain a yellow solid 1-(4-chlorophenyl)-7-{[2-(dimethylamino)ethyl]amino}-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (12.7 mg, yield 22.9%). LCMS (TOF MS ES+) m / z [M+H] + :520.1914. 1 H NMR (400MHz, DMSO) δ8.29(s,1H),8.06(d,J=12.4Hz,1H),7.58–7.46(m,4H),7.33–7.23(m,3H),7.17(dd,J=8.8,1.6Hz ,1H),5.67(t,J=5.5Hz,1H),4.18(s,3H),2.78(q,J=6.3Hz,2H),2.55–2.50(m,3H),2.19(t,J=6.6Hz,2H),1.89(s,6H).

[0148] Example 10: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-(2-morpholinethoxy)-1,8-naphthyridin-2(1H)-one (Compound 23)

[0149] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-(2-morpholinethoxy)-1,8-naphthyridin-2(1H)-one

[0150] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (80 mg, 0.17 mmol, 1.0 eq) prepared in Example 1, methanesulfonic acid (9,9-dimethyl-4,5-bisdiphenylphosphinoxanthene) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (16 mg, 0.017 mmol), cesium carbonate (168 mg, 0.51 mmol, 3.0 eq) and 2-morpholinoethanol-1-ol (44 mg, 0.34 mmol, 2.0 eq) were dissolved in 1,4-dioxane (2 mL) and added to a penicillin bottle. After nitrogen replacement, the mixture was heated to 100° C. and stirred for 4 hours. After the reaction was complete, preparative separation and purification were performed to obtain a white solid 1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-7-(2-morpholinethoxy)-1,8-naphthyridin-2(1H)-one (12 mg, yield 21.58%). LCMS (TOF MS ES+) m / z [M+H] + :563.1862. 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),8.01(d,J=11.4Hz,1H),7.72(s,1H),7.56(dd,J=9.4,7.8Hz,3H),7.48(d,J=5.0Hz,1H),7.38–7.29(m, 2H), 7.26 (dd, J=8.9, 1.6Hz, 1H), 4.17 (s, 3H), 3.60 (t, J=5.2Hz, 2H), 3.42 (t, J=4.5Hz, 4H), 2.49 (s, 3H), 2.41 (t, J=5.2Hz, 2H), 2.20 (s, 4H).

[0151] Example 11: Synthesis of 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-7-methyl-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (Compound 35)

[0152] Step 1: Synthesis of 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-7-methyl-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one

[0153] 7-Chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (60 mg, 0.121 mmol, 1.0 eq), methylboric acid (10.9 mg, 0.182 mmol, 1.5 eq), tetrakis(triphenylphosphine)palladium (14.0 mg, 0.012 mmol, 0.1 eq) and potassium carbonate (41.9 mg, 0.303 mmol, 2.5 eq) were added to 1,4-dioxane (2 mL) and water (0.2 mL) and reacted at 110°C. After the reaction, the reaction solution was filtered and separated by preparative chromatography to obtain a white solid 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-7-methyl-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (20.8 mg, yield 36.1%). LCMS (TOF MS ES+) m / z [M+H] + :474.1496. 1 H NMR (400MHz, DMSO) δ8.32(s,1H),7.84(d,J=12.0Hz,1H),7.62–7.54(m,2H),7.54(dd,J=8.8,1.8Hz ,3H),7.37–7.28(m,2H),7.10(dd,J=8.8,1.6Hz,1H),4.18(s,3H),2.24(s,4H),0.40–0.29(m,4H).

[0154] Example 12: Synthesis of 1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (Compound 9)

[0155] Step 1: Synthesis of 7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0156] 4,7-Dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (800 mg, 2.32 mmol, 1.0 eq), ethylamine hydrochloride (569 mg, 6.98 mmol, 3.0 eq), and N,N-diisopropylethylamine (2.4 g, 18.6 mmol, 8.0 eq) were dissolved in acetonitrile (10 mL) and reacted at 80°C. After completion of the reaction, the mixture was concentrated, slurried with ethanol and water, and the filter cake was dried to obtain 7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (680 mg, 82.9% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H] + :352.0. 1 H NMR (400MHz, DMSO) δ7.85(t,J=5.8Hz,1H),7.76(d,J=11.0Hz,1H),7.60–7.52(m ,2H),7.34–7.24(m,2H),6.64(s,1H),3.00–2.89(m,2H),0.82(t,J=7.1Hz,3H).

[0157] Step 2: Synthesis of 3-bromo-7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0158] 7-Chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (670 mg, 1.90 mmol, 1.0 eq) and N-bromosuccinimide (372 mg, 2.09 mmol, 1.1 eq) were dissolved in acetonitrile (12 mL) and stirred at 35°C for 1 hour. After completion of the reaction, the mixture was concentrated, slurried with ethanol and water, and the filter cake was dried to afford 3-bromo-7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (770 mg, 93.9% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 429.9. 1 H NMR (400MHz, DMSO) δ7.94(t,J=5.7Hz,1H),7.83(d,J=11.1Hz,1H),7.62–7.54(m,2H),7.39–7.31(m,2H),2.99–2.88(m,2H),0.82(t,J=7.1Hz,3H).

[0159] Step 3: Synthesis of 7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one

[0160] 3-Bromo-7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (760 mg, 1.76 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (621 mg, 3.52 mmol, 2.0 eq), and potassium phosphate (1.12 g, 5.28 mmol, 3.0 eq) were dissolved in 1,4-dioxane (20 mL) and water (2 mL). Dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (129 mg, 0.176 mmol, 0.1 eq) was added to the mixture. The mixture was protected by nitrogen and stirred at 80 °C for 12 hours. After completion of the reaction, the product was filtered, concentrated, extracted with dichloromethane and water, and the organic phase was concentrated and separated using a reverse phase column to afford 7-chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (220 mg, 25.8% yield) as a white powdery solid. LCMS (TOF MS ES+) m / z [M+H]+: 482.0. 1 H NMR (400MHz, DMSO) δ8.38(s,1H),7.91–7.79(m,2H),7.70(t,J=1.2Hz,1H),7.60(t,J=9.1Hz,3H),7. 41–7.37(m,2H),7.19(dd,J=8.9,1.6Hz,1H),4.20(s,3H),3.05–2.94(m,2H),0.88(t,J=7.1Hz,3H).

[0161] Step 4: Synthesis of 1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one

[0162] 7-Chloro-1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (70 mg, 0.145 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (143.7 mg, 1.45 mmol, 10.0 eq), methanesulfonic acid (4,5-bisdiphenylphosphino-9,9-dimethylxanthene) (2-amino-1,1'-biphenyl-2-yl) palladium (II) (13.7 mg, 0.014 mmol, 0.1 eq) and sodium tert-butoxide (27.8 mg, 0.29 mmol, 2.0 eq) were added to 1,4-dioxane (2.5 mL) and reacted at 60°C. After the reaction, the reaction solution was filtered and separated by preparative chromatography to obtain a yellow solid 1-(4-chlorophenyl)-4-(ethylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (14.3 mg, yield 18.1%). LCMS (TOF MS ES+) m / z [M+H] + :545.1398. 1 H NMR (400MHz, DMSO) δ8.32(s,1H),8.12(d,J=12.5Hz,1H),7.64–7.55(m,2H),7.49(t,J=7.5Hz,3H),7.32–7.23(m,2H),7.11(dd, J=8.8,1.6Hz,1H),6.34(t,J=7.2Hz,1H),4.17(s,3H),3.28(dd,J=9.6,7.1Hz,2H),2.95(m,J=6.9Hz,2H),0.84(t,J=7.0Hz,3H).

[0163] Example 13: Synthesis of 7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (Compound 34)

[0164] Step 1: Synthesis of 7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0165] 4,7-Dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (500 mg, 1.46 mmol, 1.0 eq), cyclopropylamine (125 mg, 2.19 mmol, 1.5 eq), and N,N-diisopropylethylamine (752 mg, 5.83 mmol, 4.0 eq) were dissolved in acetonitrile (20 mL) and reacted at 80°C. After completion of the reaction, the reaction solution was concentrated and slurried with ethanol and water to obtain 7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (410 mg, 77.3% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H] + :364.0. 1 H NMR (400MHz, DMSO) δ7.97(d,J=3.6Hz,1H),7.79(d,J=11.1Hz,1H),7.60–7.55(m,2 H),7.36–7.31(m,2H),6.70(s,1H),2.26(m,J=7.6,3.9Hz,1H),0.41–0.36(m,4H).

[0166] Step 2: Synthesis of 3-bromo-7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one

[0167] 7-Chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (400 mg, 1.10 mmol, 1.0 eq) and N-bromosuccinimide (205 mg, 1.15 mmol, 1.05 eq) were dissolved in acetonitrile (15 mL) and stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated and slurried with ethanol and water to obtain 3-bromo-7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (405 mg, 83.1% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 441.9. 1 H NMR(400MHz,DMSO)δ8.05(d,J=3.2Hz,1H),7.86(d,J=11.1Hz,1H),7.63–7.5 5(m,2H),7.43–7.34(m,2H),2.25(dd,J=7.6,4.1Hz,1H),0.42–0.31(m,4H).

[0168] Step 3: Synthesis of 7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one

[0169] 3-Bromo-7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-1,8-naphthyridin-2(1H)-one (100 mg, 0.22 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (79 mg, 0.44 mmol, 2.0 eq) and potassium phosphate (144 mg, 0.66 mmol, 3.0 eq) were dissolved in 1,4-dioxane (2.5 mL) and water (0.25 mL), and then dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (16 mg, 0.022 mmol, 0.1 eq) was added and stirred at 80 ° C for 12 hours under nitrogen protection. After the reaction, the mixture was filtered and separated by reverse phase column to afford a beige solid, 7-chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (66.2 mg, 60.9% yield). LCMS (TOF MS ES+) m / z [M+H]+: 494.0148. 1 H NMR (400MHz, DMSO) δ8.37(s,1H),7.94–7.88(m,1H),7.86(d,J=11.2Hz,1H),7.69(s,1H),7.63–7.52(m,3H) ,7.39(d,J=8.6Hz,2H),7.18(dd,J=8.9,1.7Hz,1H),4.18(s,3H),2.26(d,J=7.0Hz,1H),0.42–0.32(m,4H).

[0170] Example 14: Synthesis of 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (Compound 10)

[0171] Step 1: Synthesis of 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one

[0172] 7-Chloro-1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (70 mg, 0.14 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (140.3 mg, 1.40 mmol, 10.0 eq), methanesulfonic acid (4,5-bisdiphenylphosphino-9,9-dimethylxanthene) (2-amino-1,1'-biphenyl-2-yl) palladium (II) (13.4 mg, 0.014 mmol, 0.1 eq) and sodium tert-butoxide (27.2 mg, 0.28 mmol, 2.0 eq) were added to 1,4-dioxane (2 mL) and reacted at 60°C. After the reaction, the reaction solution was filtered, and the filtrate was concentrated and separated by preparative chromatography to obtain a yellow solid 1-(4-chlorophenyl)-4-(cyclopropylamino)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (22.1 mg, yield 28.3%). LCMS (TOF MS ES+) m / z [M+H] + :557.1467. 1 H NMR (400MHz, DMSO) δ8.33(s,1H),8.13(d,J=12.5Hz,1H),7.64–7.54(m,3H),7.53–7.45(m,2H),7.33–7.25(m,2H),7.11 (dd,J=8.9,1.6Hz,1H),6.37(t,J=7.2Hz,1H),4.18(s,3H),3.31–3.22(m,2H),2.22(d,J=8.1Hz,1H),0.38–0.31(m,4H).

[0173] Example 15: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (Compound 28)

[0174] Step 1: Synthesis of 6-chloro-5-fluoro-2-(o-tolylamino)nicotinic acid

[0175] o-Toluidine (5.1 g, 47.6 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (200 mL) and cooled to -78°C. Lithium bis(trimethylsilyl)amide (71.4 mL, 71.4 mmol, 3.0 eq) was slowly added dropwise. The reaction was continued for 1 hour. 2,6-Dichloro-5-fluoronicotinic acid (5.0 g, 23.8 mmol, 1.0 eq) was then dissolved in ultra-dry tetrahydrofuran (50 mL) and slowly added dropwise to the reaction system at -78°C. After completion, the mixture was transferred to room temperature and allowed to react for 1 hour. After the reaction, the pH was adjusted to 4 with dilute hydrochloric acid. The mixture was then extracted three times with water and ethyl acetate. The organic phases were combined, dried, concentrated, and slurried (petroleum ether:ethyl acetate = 5:1). Filtered to yield 6-chloro-5-fluoro-2-(o-tolylamino)nicotinic acid (4.66 g, 69.9% yield) as a yellow solid. LCMS(TOF MS ES+)m / z[M+H]+:281.0320. 1 H NMR (400MHz, DMSO) δ13.77 (s, 1H), 10.26 (s, 1H), 8.21 (m, J = 8.5, 2.7, 1.2Hz, 1H) ,8.07(dd,J=8.2,1.3Hz,1H),7.26–7.17(m,2H),7.07–6.98(m,1H),2.29(s,3H).

[0176] Step 2: Synthesis of ethyl 3-[6-chloro-5-fluoro-2-(o-tolylamino)pyridin-3-yl]-3-oxopropanoate

[0177] Step 1: Dissolve 6-chloro-5-fluoro-2-(o-tolylamino)nicotinic acid (4.5 g, 16.1 mmol, 1.0 eq) and N,N-dimethylformamide (118 mg, 1.6 mmol, 0.1 eq) in dichloromethane (45 mL). Add oxalyl chloride (3.0 g, 24.1 mmol, 1.5 eq) dropwise until no bubbles form. Step 2: Dissolve potassium 3-ethoxy-3-oxopropanoate (4.1 g, 24.1 mmol, 1.5 eq), magnesium chloride (4.6 g, 48.2 mmol, 3.0 eq), and triethylamine (7.3 g, 72.45 mmol, 4.5 eq) in tetrahydrofuran (60 mL) and stir under ice. Concentrate the mixture from Step 1, dissolve it in tetrahydrofuran (60 mL), and then slowly add it dropwise to the reaction mixture from Step 2 at 0°C. After the addition is complete, react at room temperature for 1.5 hours. After the reaction, the mixture was quenched with dilute hydrochloric acid and extracted three times with ethyl acetate and water. The organic phase was concentrated, mixed and purified by forward column separation using petroleum ether and ethyl acetate system. When the ethyl acetate content was 36%, a yellow solid 3-[6-chloro-5-fluoro-2-(o-tolylamino)pyridin-3-yl]-3-oxopropionic acid ethyl ester (4.8 g, yield 85.7%) was obtained. 1 H NMR (400MHz, DMSO) δ10.63(s,1H),8.55(d,J=9.3Hz,1H),7.96(dd,J=8.0,1.3Hz,1H),7.30(dd,J=7.5,1.6Hz,1H),7.26 (m,J=7.8,1.6Hz,1H),7.10(m,J=7.4,1.3Hz,1H),4.32(s,2H),4.17(m,J=7.1Hz,2H),3.30(s,3H)1.23(m,J=7.1Hz,3H).

[0178] Step 3: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one

[0179] Ethyl 3-[6-chloro-5-fluoro-2-(o-tolylamino)pyridin-3-yl]-3-oxopropanoate (4.8 g, 13.7 mmol, 1.0 eq) and potassium carbonate (3.78 g, 27.4 mmol, 2.0 eq) were dissolved in ethanol (100 mL) and reacted at 70°C for 1 hour. After the reaction was complete, the mixture was filtered, the filtrate was concentrated, and the mixture was separated and purified using a normal phase column to obtain 7-chloro-6-fluoro-4-hydroxy-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (2.1 g, 50% yield) as a yellow solid. 1H NMR (400MHz, DMSO) δ12.25(s,1H),8.29(d,J=8.0Hz,1H),7.43–7.31(m,3H),7.15(dd,J=7.5,1.5Hz,1H),6.03(s,1H),1.91(s,3H).

[0180] Step 4: Synthesis of 4,7-dichloro-6-fluoro-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one

[0181] 7-Chloro-6-fluoro-4-hydroxy-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (2.1 g, 6.9 mmol, 1.0 eq) was dissolved in phosphorus oxychloride (20 mL) and reacted at 70°C for 16 hours. After the reaction was complete, the mixture was concentrated, and sodium bicarbonate solution was added to quench the phosphorus oxychloride. The mixture was then extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate, concentrated, and slurried (ethyl acetate). The filter cake was retained to obtain 4,7-dichloro-6-fluoro-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (850 mg, 2.63 mmol, 38% yield) as a yellow solid. 1 H NMR (400MHz, DMSO) δ8.53 (d, J = 8.2 Hz, 1H), 7.44–7.34 (m, 3H), 7.30 (s, 1H), 7.23 (dd, J = 7.3, 1.5 Hz, 1H), 1.95 (s, 3H).

[0182] Step 5: Synthesis of 7-chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one

[0183] 4,7-Dichloro-6-fluoro-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (850 mg, 2.63 mmol, 1.0 eq), methylamine hydrochloride (534 mg, 7.92 mmol, 3.0 eq), and N,N-diisopropylethylamine (2.72 g, 21.11 mmol, 8.0 eq) were dissolved in anhydrous acetonitrile (10 mL) and reacted at 80°C for 2 hours. After the reaction was complete, the reaction solution was concentrated and slurried (water and ethanol), and the filter cake was retained and dried to obtain a crude yellow solid, 7-chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (922 mg). 1 H NMR (400MHz, DMSO) δ7.82–7.70(m,2H),7.43–7.28(m,3H),7.14(dd,J=7.3,1.7Hz,1H),6.67(s,1H),2.43(d,J=4.6Hz,3H),1.93(s,3H).

[0184] Step 6: Synthesis of 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one

[0185] 7-Chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (900 mg, 2.83 mmol, 1.0 eq) and N-bromosuccinimide (522 mg, 3.12 mmol, 1.1 eq) were dissolved in acetonitrile (10 mL) and reacted at 35°C for 2 hours. After the reaction was complete, the reaction solution was concentrated, slurried (ethanol and water), filtered, and the filter cake was retained and dried to obtain a yellow solid, 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (980 mg, 2.48 mmol, 87.7% yield). 1 H NMR (400MHz, DMSO) δ7.85 (d, J = 11.1 Hz, 2H), 7.45–7.31 (m, 3H), 7.18 (dd, J = 7.5, 1.6 Hz, 1H), 2.43 (d, J = 3.8 Hz, 3H), 1.93 (s, 3H).

[0186] Step 7: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one

[0187] 3-Bromo-7-chloro-6-fluoro-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (100 mg, 0.25 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (90 mg, 0.51 mmol, 2.0 eq), potassium phosphate (130 mg, 0.64 mmol, 2.5 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (18 mg, 0.03 mmol, 0.1 eq) were dissolved in water (0.5 mL) and 1,4-dioxane (2 mL) and reacted at 80°C for 2 hours. After the reaction was complete, preparative separation and purification were performed to obtain a light yellow solid 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(o-tolyl)-1,8-naphthyridin-2(1H)-one (33 mg, 30% yield). LCMS (TOF MS ES+) m / z [M+H] + :448.1327 / 450.1300. 1H NMR(400MHz,DMSO)δ8.36(s,1H),7.85(d,J=11.2Hz,1H),7.72–7.64(m,2H),7 .59(d,J=8.9Hz,1H),7.42–7.26(m,3H),7.22–7.14(m,2H),4.18(s,3H),2.44 0(d,J=4.6Hz,3H),1.98(s,3H).

[0188] Example 16: Synthesis of 7-chloro-6-fluoro-1-(4-methoxyphenyl)-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (Compound 29)

[0189] Step 1: Synthesis of 6-chloro-5-fluoro-2-(4-methoxyphenyl)amino)nicotinic acid

[0190] 4-Chloroaniline (5.86 g, 47.6 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (100 mL) and cooled to -78°C. Lithium bis(trimethylsilyl)amide (71.4 mL, 71.4 mmol, 3.0 eq) was slowly added dropwise. The reaction was continued for 1 hour. 4-Methoxyaniline (5 g, 23.8 mmol, 1.0 eq) was then dissolved in ultra-dry tetrahydrofuran (40 mL) and slowly added dropwise to the reaction system at -78°C. After the addition was complete, the mixture was transferred to room temperature and allowed to react for 1 hour. After the reaction was complete, dilute hydrochloric acid was added to quench the reaction, followed by extraction with ethyl acetate and water, followed by washing with saturated brine. The solvent was concentrated to remove the residue, yielding a light brown solid. Methanol was added to slurry to yield a light green solid of 6-chloro-5-fluoro-2-(4-methoxyphenyl)amino)nicotinic acid (6.0 g, 85.2% yield). LCMS(TOF MS ES+)m / z[M+H]+:297.02. 1 H NMR (400MHz, DMSO) δ10.16 (s, 1H), 8.18 (d, J = 8.3Hz, 1H), 7.53–7.45 (m, 2H), 6.98–6.89 (m, 2H), 3.75 (s, 3H).

[0191] Step 2: Synthesis of ethyl 3-{6-chloro-5-fluoro-2-[(4-methoxyphenyl)amino]pyridin-3-yl}-3-oxopropanoate

[0192] Potassium malonate (4.87 g, 28.6 mmol, 1.5 eq), magnesium chloride (5.45 g, 57.3 mmol, 3.0 eq), and triethylamine (8.69 g, 85.9 mmol, 4.5 eq) were dissolved in acetonitrile (80 mL) and stirred at room temperature to obtain Reaction System 1. 6-Chloro-5-fluoro-2-(4-methoxyphenyl)amino)nicotinic acid (5.65 g, 19.1 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (80 mL) and a small amount of N,N-dimethylformamide was added as a catalyst. Oxalyl chloride (3.64 g, 28.6 mmol, 1.5 eq) was added dropwise at room temperature. After TLC confirmed complete reaction of the starting materials, the reaction solution was concentrated to obtain a yellow solid. The yellow solid was dissolved in tetrahydrofuran (100 mL) and slowly added dropwise to Reaction System 1 at 0°C. The reaction was continued at room temperature for 2 hours. After the reaction, the mixture was acidified with dilute hydrochloric acid, extracted with ethyl acetate and water, dried, concentrated, and purified in the forward direction to obtain the target product, ethyl 3-{6-chloro-5-fluoro-2-[(4-methoxyphenyl)amino]pyridin-3-yl}-3-oxopropanoate (4.1 g, 58.57%). LCMS (TOF MS ES+) m / z [M+H]+: 367.06. 1 H NMR (400MHz, DMSO) δ10.62 (s, 1H), 8.48 (d, J = 9.2Hz, 1H), 7.55–7.46 (m, 2H), 6.98 –6.90(m,2H),4.27(s,2H),4.18–3.96(m,2H),3.75(s,3H),1.21(t,J=7.2Hz,3H).

[0193] Step 3: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one

[0194] Dissolve ethyl 3-{6-chloro-5-fluoro-2-[(4-methoxyphenyl)amino]pyridin-3-yl}-3-oxopropanoate (4.0 g, 10.9 mmol, 1.0 eq) in anhydrous ethanol (10 mL), add potassium carbonate (3.02 g, 21.9 mmol, 2.0 eq), and heat to 70°C for 3 hours. After completion of the reaction, filter through celite and concentrate to obtain the desired product, 7-chloro-6-fluoro-4-hydroxy-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one (4.1 g, 115% yield). LCMS (TOF MS ES+) m / z [M+H]+: 321.04. 1 H NMR (400MHz, DMSO) δ8.04 (d, J = 8.3Hz, 1H), 7.15–6.83 (m, 4H), 4.93 (s, 1H), 3.80 (s, 3H).

[0195] Step 4: Synthesis of 4,7-dichloro-6-fluoro-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one

[0196] To a reaction flask, 7-chloro-6-fluoro-4-hydroxy-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one (4.0 g, 12.5 mmol, 1.0 eq) and phosphorus oxychloride (10 mL) were added. The temperature was raised to 70°C and the reaction was allowed to react overnight. The phosphorus oxychloride was removed by concentration under reduced pressure. Saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate. The mixture was dried and concentrated, slurried with ethyl acetate, and filtered to obtain the desired product, 4,7-dichloro-6-fluoro-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one (2.9 g, 69.1% yield). LCMS (TOF MS ES+) m / z [M+H]+: 338.98. 1 H NMR (400MHz, DMSO) δ8.48 (d, J = 8.1Hz, 1H), 7.26–7.17 (m, 3H), 7.12–7.04 (m, 2H), 3.84 (s, 3H).

[0197] Step 5: Synthesis of 7-chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0198] 4,7-Dichloro-6-fluoro-1-(4-methoxyphenyl)-1,8-naphthyridin-2(1H)-one (900 mg, 2.66 mmol, 1.0 eq), methylamine hydrochloride (450 mg, 6.66 mmol, 2.5 eq), and N,N-diisopropylethylamine (2.06 g, 15.9 mmol, 6.0 eq) were dissolved in acetonitrile (20 mL) and reacted at 80°C for 2 hours. After the reaction, the mixture was extracted with ethyl acetate and water, and the organic phase was concentrated to obtain a pale yellow solid. After filtration and slurrying with ethanol, 7-chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (740 mg, 83.5% yield) was obtained. LCMS (TOF MS ES+) m / z [M+H] + :334.06. 1 H NMR (400MHz, DMSO) δ7.78–7.66(m,2H),7.15(d,J=8.5Hz,2H),7.03(d,J=8.8Hz,2H),6.62(s,1H),3.81(s,3H),2.49(s,3H).

[0199] Step 6: Synthesis of 3-bromo-7-chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0200] 7-Chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (730 mg, 2.19 mmol, 1.0 eq) and N-bromosuccinimide (429 mg, 2.41 mmol, 1.1 eq) were added to an acetonitrile solution (8 mL) and reacted at room temperature for 2 hours. After completion of the reaction, the mixture was concentrated, slurried with an acetonitrile-water solution, and filtered to afford 3-bromo-7-chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (749 mg, 83.5% yield) as a pale yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 413.97. 1 H NMR (400MHz, DMSO) δ7.79 (d, J = 7.6 Hz, 2H), 7.25–7.13 (m, 2H), 7.08–6.99 (m, 2H), 3.81 (s, 3H), 2.51 (d, J = 2.1 Hz, 3H).

[0201] Step 7: Synthesis of 7-chloro-6-fluoro-1-(4-methoxyphenyl)-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one

[0202] 3-Bromo-7-chloro-6-fluoro-1-(4-methoxyphenyl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (60 mg, 0.15 mmol, 1.0 eq), 2-methyl-2H-indazole-5-boronic acid (52.5 mg, 0.30 mmol, 1.5 eq) and potassium phosphate (79.6 mg, 0.375 mmol, 2.5 eq) were added to a mixed solvent of dioxane (50 mL) and water (0.5 mL), and finally 1,1-bis(diphenylphosphino)diphenylferric palladium dichloride (11.0 mg, 0.015 mmol, 0.1 eq) was added. The reaction was carried out at 80 ° C for 16 hours under nitrogen protection. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated and purified by preparative chromatography to obtain 7-chloro-6-fluoro-1-(4-methoxyphenyl)-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one as a yellow solid (25 mg, 35.2% yield). LCMS (TOF MS ES+) m / z [M+H]+: 464.1015. 1H NMR(400MHz, DMSO)δ8.36(s,1H),7.83(d,J=11.2Hz,1H),7.70–7.62(m,2H),7.58(d,J=8.9Hz ,1H),7.25–7.18(m,2H),7.07–6.99(m,2H),4.18(s,3H),3.80(s,3H),2.52(d,J=4.6Hz,3H).

[0203] Example 17: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (Compound 30)

[0204] Step 1: Synthesis of 6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}nicotinic acid

[0205] 4-(Trifluoromethoxy)aniline (8.43 g, 47.6 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (200 mL) and cooled to -78°C. Lithium bis(trimethylsilyl)amide (71.4 mL, 71.4 mmol, 3.0 eq) was slowly added dropwise. The reaction was continued for 1 hour. 2,6-Dichloro-5-fluoronicotinic acid (5.0 g, 23.8 mmol, 1.0 eq) was then dissolved in ultra-dry tetrahydrofuran (50 mL) and slowly added dropwise to the reaction system at -78°C. The mixture was then transferred to room temperature and allowed to react for 1 hour. After completion of the reaction, saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted with ethyl acetate (200 mL). The organic phases were combined, dried, concentrated, washed, and slurried with an appropriate amount of ethyl acetate. Filtering afforded 6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}nicotinic acid (4.8 g, 60.0% yield) as a pale yellow solid. LCMS(TOF MS ES+)m / z[M+H]+:351.0. 1 H NMR (400MHz, DMSO) δ14.18(s,1H),10.46(s,1H),8.26(dd,J=8.6,1.1Hz,1H),7.79–7.68(m,2H),7.37(d,J=8.6Hz,2H).

[0206] Step 2: Synthesis of ethyl 3-(6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}pyridin-3-yl)-3-oxopropanoate

[0207] Step 1: Dissolve 6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}nicotinic acid (4.75 g, 13.6 mmol, 1.0 eq) and N,N-dimethylformamide (50 mg, 0.68 mmol, 0.05 eq) in dichloromethane (50 mL). Add oxalyl chloride (2.6 g, 20.4 mmol, 1.5 eq) dropwise and react for 1 hour until no bubbles form. Step 2: Dissolve potassium 3-ethoxy-3-oxopropanoate (3.46 g, 20.4 mmol, 1.5 eq), magnesium chloride (3.88 g, 40.7 mmol, 3.0 eq), and triethylamine (6.2 g, 61.07 mmol, 4.5 eq) in tetrahydrofuran (30 mL) and stir under ice. The mixed solution of step 1 was concentrated, and then tetrahydrofuran (5 mL) was added, and then slowly added dropwise to the reaction solution of step 2 at 0°C. After the reaction, the mixed solution was extracted, and the organic phase was concentrated, mixed, and separated and purified by forward column to obtain a yellow solid 3-(6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}pyridin-3-yl)-3-oxopropionic acid ethyl ester (4.07 g, yield 71%). 1 H NMR (400MHz, DMSO) δ10.81 (s, 1H), 8.57 (d, J = 9.2Hz, 1H), 7.81–7.71 (m, 2H), 7. 40(d,J=8.6Hz,2H), 4.32(s,2H), 4.17(q,J=7.1Hz,2H), 1.23(t,J=7.1Hz,3H).

[0208] Step 3: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one

[0209] Ethyl 3-(6-chloro-5-fluoro-2-{[4-(trifluoromethoxy)phenyl]amino}pyridin-3-yl)-3-oxopropanoate (4.0 g, 9.5 mmol, 1.0 eq) and potassium carbonate (2.63 g, 19 mmol, 2.0 eq) were dissolved in ethanol (40 mL) and reacted at 70°C for 1 hour. After the reaction was complete, the mixture was cooled, filtered, and the filtrate was concentrated to obtain a crude yellow solid, 7-chloro-6-fluoro-4-hydroxy-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (4.05 g). 1 H NMR (400MHz, DMSO) δ8.10 (d, J = 8.4Hz, 1H), 7.44 (d, J = 8.3Hz, 2H), 7.36–7.24 (m, 2H), 4.98 (s, 1H).

[0210] Step 4: Synthesis of 4,7-dichloro-6-fluoro-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one

[0211] 7-Chloro-6-fluoro-4-hydroxy-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (4.0 g, 10.6 mmol, 1.0 eq) was dissolved in phosphorus oxychloride (35 mL) and reacted at 70°C for 16 hours. After the reaction was complete, the mixture was concentrated, and sodium bicarbonate solution was added to quench the phosphorus oxychloride. The mixture was then extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate, concentrated, and slurried (ethyl acetate). The filter cake was retained to obtain 4,7-dichloro-6-fluoro-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (2.9 g, 7.3 mmol, 69.8% yield) as a yellow solid. 1 H NMR (400MHz, DMSO) δ8.53 (d, J = 8.2Hz, 1H), 7.62–7.56 (m, 2H), 7.53–7.48 (m, 2H), 7.30 (s, 1H).

[0212] Step 5: Synthesis of 7-chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one

[0213] 4,7-Dichloro-6-fluoro-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (900 mg, 2.3 mmol, 1.0 eq), methylamine hydrochloride (466 mg, 6.8 mmol, 3.0 eq), and N,N-diisopropylethylamine (2.36 mg, 18.4 mmol, 8.0 eq) were dissolved in anhydrous acetonitrile (10 mL) and reacted overnight at 80°C. After the reaction was complete, the reaction solution was concentrated and slurried (with water and ethanol), and the filter cake was retained and dried to obtain 7-chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (833 mg, 2.15 mmol, 93.5% yield) as a yellow solid. 1 HNMR (400MHz, DMSO) δ7.82–7.75(m,2H),7.52(d,J=8.4Hz,2H),7.44(d,J=8.5Hz,2H),6.68(s,1H),2.48(d,J=4.5Hz,3H).

[0214] Step 6: Synthesis of 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one

[0215] 7-Chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (800 mg, 2.06 mmol, 1.0 eq) and N-bromosuccinimide (403 mg, 2.27 mmol, 1.1 eq) were dissolved in acetonitrile (8 mL) and reacted at 35°C for 3 hours. After the reaction was complete, the reaction solution was concentrated, slurried (ethanol and water), filtered, and the filter cake was retained and dried to obtain a yellow solid, 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (830 mg, 1.78 mmol, 86.6% yield), namely ALKZJ001-S131-6. 1 H NMR (400MHz, DMSO) δ7.89–7.82(m,2H),7.54(d,J=8.5Hz,2H),7.48(d,J=9.0Hz,2H),2.47(d,J=4.6Hz,3H).

[0216] Step 7: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one

[0217] Dissolve 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (60 mg, 0.13 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (45 mg, 0.26 mmol, 2.0 eq), potassium phosphate (690 mg, 0.325 mmol, 2.5 eq), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (9.5 mg, 0.013 mmol, 0.1 eq) in water (0.2 mL) and 1,4-dioxane (2 mL) and react at 80°C for 16 hours. After the reaction is complete, filter and prepare. A yellow solid, 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (40.2 mg, 0.077 mmol, 30% yield), was obtained. LCMS (TOF MS ES+) m / z [M+H] + :518.0528 / 520.0504. 1H NMR (400MHz, DMSO) δ8.37(s,1H),7.86(d,J=11.2Hz,1H),7.72(d,J=5.1Hz,1H),7.69(s,1H),7.59( d,J=8.9Hz,1H),7.54–7.47(m,4H),7.18(dd,J=9.0,1.6Hz,1H),4.18(s,3H),2.49(d,J=1.7Hz,3H).

[0218] Example 18: Synthesis of 3-(7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl)benzonitrile (Compound 31)

[0219] Step 1: Synthesis of 6-chloro-2-[(3-cyanophenyl)amino]-5-fluoronicotinic acid

[0220] 3-Aminobenzonitrile (6.5 g, 55.26 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (200 mL) and cooled to -78°C. Lithium bis(trimethylsilyl)amide (83 mL, 82.9 mmol, 3.0 eq) was slowly added dropwise. The reaction was continued for 1 hour. 2,6-Dichloro-5-fluoronicotinic acid (5.8 g, 27.63 mmol, 1.0 eq) was then dissolved in ultra-dry tetrahydrofuran (50 mL) and slowly added dropwise to the reaction system at -78°C. The mixture was then transferred to room temperature and allowed to react for 1 hour. After completion of the reaction, dilute hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate (400 mL). The organic phases were combined, dried, concentrated, washed, and slurried with an appropriate amount of ethyl acetate. Filtering afforded 6-chloro-2-[(3-cyanophenyl)amino]-5-fluoronicotinic acid (5.4 g, 67.16% yield) as a pale yellow solid. LCMS(TOF MS ES+)m / z[M+H]+:292. 1 H NMR (400MHz, DMSO-d6) δ14.23(s,1H),10.53(s,1H),8.29(m,J=8.5,2.3Hz,1H),8.22–8.17(m,1H),7.91(m,J=8.2,1.9Hz,1H),7.61–7.48(m,2H).

[0221] Step 2: Synthesis of ethyl 3-{6-chloro-2-[(3-cyanophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate

[0222] Step 1: Dissolve 6-chloro-2-[(3-cyanophenyl)amino]-5-fluoronicotinic acid (5.35 g, 18.4 mmol, 1.0 eq) and N,N-dimethylformamide (1 mL) in dichloromethane (50 mL). Add oxalyl chloride (3.5 g, 27.58 mmol, 1.5 eq) dropwise and react for 1 hour until no bubbles form. Step 2: Dissolve potassium 3-ethoxy-3-oxopropanoate (4.7 g, 27.58 mmol, 1.5 eq), magnesium chloride (5.2 g, 55.2 mmol, 3.0 eq), and triethylamine (8.36 g, 82.8 mmol, 4.5 eq) in tetrahydrofuran (30 mL) and stir under ice. The mixed solution from step 1 was concentrated, and tetrahydrofuran (5 mL) was added. The mixture was then slowly added dropwise to the reaction solution from step 2 at 0°C. After the reaction, the mixed solution was extracted, and the organic phase was concentrated, mixed, and purified using a forward column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain ethyl 3-{6-chloro-2-[(3-cyanophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate (2.1 g, 31.82% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 362.

[0223] Step 3: Synthesis of 3-[7-chloro-6-fluoro-4-hydroxy-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile

[0224] Ethyl 3-{6-chloro-2-[(3-cyanophenyl)amino]-5-fluoropyridin-3-yl}-3-oxopropanoate (2.1 g, 5.85 mmol, 1.0 eq) and potassium carbonate (1.6 g, 11.63 mmol, 2.0 eq) were dissolved in ethanol (40 mL) and reacted at 70°C for 1 hour. After the reaction was complete, the mixture was cooled and filtered, and the filtrate was concentrated and purified by normal phase separation (dichloromethane:methanol = 10:1) to afford 3-[7-chloro-6-fluoro-4-hydroxy-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (1.34 g) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 316. 1 H NMR(400MHz,DMSO-d6)δ8.30(d,J=8.0Hz,1H),7.94(dd,J=7.6,1.5Hz,1H),7.87( d, J=1.8Hz, 1H), 7.74 (t, J=7.8Hz, 1H), 7.68 (dd, J=8.2, 1.6Hz, 1H), 5.96 (s, 1H).

[0225] Step 4: Synthesis of 3-[4,7-dichloro-6-fluoro-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile

[0226] [7-Chloro-6-fluoro-4-hydroxy-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (1.34 g, 4.25 mmol, 1.0 eq) was dissolved in phosphorus oxychloride (30 mL) and reacted at 70°C for 5 hours. After the reaction was complete, the mixture was concentrated, and sodium bicarbonate solution was added to quench the phosphorus oxychloride. The mixture was then extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate. After normal phase separation and purification, the product was purified by column chromatography with pure dichloromethane to afford 3-[4,7-dichloro-6-fluoro-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (650 mg, 46.43% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 334. 1 H NMR (400MHz, DMSO-d6) δ8.54(d,J=8.2Hz,1H),8.00(m,J=7.6,1.5Hz,1H),7.93(t,J=1.8Hz,1H),7.83–7.71(m,2H),7.31(s,1H).

[0227] Step 5: Synthesis of 3-[7-chloro-6-fluoro-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile

[0228] 3-[4,7-Dichloro-6-fluoro-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (630 mg, 1.89 mmol, 1.0 eq), methylamine hydrochloride (319 mg, 4.73 mmol, 2.5 eq), and N,N-diisopropylethylamine (1.46 g, 11.34 mmol, 6.0 eq) were dissolved in anhydrous acetonitrile (10 mL) and reacted at 80°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated and slurried (water and ethanol). The filter cake was retained and dried to afford 3-[7-chloro-6-fluoro-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (523 mg, 84.22% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 329. 1 H NMR (400MHz, DMSO-d6) δ7.93(m,J=7.4,1.6Hz,1H),7.90(d,J=2.1Hz,1H),7.84–7.74(m,2H),7.74–7.67(m,2H),6.68(s,1H),2.46(d,J=4.5Hz,3H).

[0229] Step 6: Synthesis of 3-[3-bromo-7-chloro-6-fluoro-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile

[0230] 7-Chloro-6-fluoro-4-(methylamino)-1-[4-(trifluoromethoxy)phenyl]-1,8-naphthyridin-2(1H)-one (503 mg, 1.53 mmol, 1.0 eq) and N-bromosuccinimide (300 mg, 1.68 mmol, 1.1 eq) were dissolved in acetonitrile (10 mL) and reacted at 35°C for 3 hours. After completion of the reaction, the reaction solution was concentrated, slurried (ethanol and water), filtered, and the filter cake was retained and dried to afford 3-[3-bromo-7-chloro-6-fluoro-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile as a yellow solid (574 mg, 92.28% yield). LCMS (TOF MS ES+) m / z [M+H]+: 407. 1 HNMR (400MHz, DMSO-d6) δ7.97–7.92(m,2H),7.88(d,J=5.3Hz,1H),7.85(d,J=11.1Hz,1H),7.78–7.70(m,2H),2.44(d,J=4.6Hz,3H).

[0231] Step 7: Synthesis of 3-(7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl)benzonitrile

[0232] 3-[3-bromo-7-chloro-6-fluoro-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl]benzonitrile (88 mg, 0.216 mmol, 1.0 eq), (2-methyl-2H-indazol-5-yl)boronic acid (76 mg, 0.43 mmol, 2.0 eq), potassium phosphate (137 mg, 0.65 mmol, 3.0 eq), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (15.7 mg, 0.021 mmol, 0.1 eq) were dissolved in water (0.2 mL) and 1,4-dioxane (2 mL) and reacted at 80°C for 16 hours. After the reaction was complete, the product was separated and purified to give a yellow solid 3-(7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-2-oxo-1,8-naphthyridin-1(2H)-yl)benzonitrile (30 mg, yield 30.30%), LCMS (TOF MS ES+) m / z [M+H]+: 459.1124. 1H NMR(400MHz, DMSO-d6)δ8.37(s,1H),7.96(d,J=1.9Hz,1H),7.96–7.89(m,1H),7.88(d,J=11.2Hz,1H),7 .78–7.67(m,4H),7.60(d,J=8.9Hz,1H),7.18(dd,J=8.9,1.6Hz,1H),4.18(s,3H),2.51(d,J=1.9Hz,3H).

[0233] Example 19: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (Compound 33)

[0234] Step 1: Synthesis of 7-chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one

[0235] 4,7-Dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthyridin-2(1H)-one (500.00 mg, 1.45 mmol, 1.00 eq), isopropylamine (860.40 mg, 14.55 mmol, 10.0 eq), and N,N-diisopropylethylamine (751.0 mg, 5.82 mmol, 4.0 eq) were dissolved in acetonitrile (7.5 mL) and reacted at 80°C for 2 hours. The acetonitrile was concentrated to obtain a yellow solid. The mixture was slurried and filtered with a mixture of ethanol and water (1:1) to obtain a crude yellow solid, 7-chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one (440 mg, 81.36% yield). The crude product was used directly in the next step without further purification. LCMS (TOF MS ES+) m / z [M+H] + :366.

[0236] Step 2: Synthesis of 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one

[0237] 7-Chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one (440.00 mg, 1.15 mmol, 1.0 eq) and N-bromosuccinimide (225.30 mg, 1.26 mmol, 1.1 eq) were added to an acetonitrile solution (9 mL) and reacted at room temperature for 2 hours. The mixture was slurried (ethanol:water = 1:1) and filtered to afford 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one (320 mg, 62.86% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 444.

[0238] Step 3: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one

[0239] 3-Bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(isopropylamino)-1,8-naphthyridin-2(1H)-one (320.0 mg, 0.72 mmol, 1.0 eq), 2-methyl-2H-indazole-5-boronic acid (252.95 mg, 1.44 mmol, 2.0 eq), and potassium phosphate (383.3 mg, 1.80 mmol, 2.5 eq) were added to a mixed solvent of (6 mL) dioxane and water (0.6 mL), and finally 1,1-bis(diphenylphosphino)diphenylferric palladium dichloride (52.3 mg, 0.07 mmol, 0.1 eq) was added, and the temperature was raised to 80°C for 4 hours. Reverse column separation and purification yielded 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-1,8-naphthyridin-2(1H)-one (170 mg, 47.55% yield) as a white solid at an acetonitrile content of 50%. LCMS (TOF MS ES+) m / z [M+H]+: 496.1092 / 498.1067. 1 H NMR (400MHz, DMSO) δ8.36(s,1H),7.85(d,J=11.2Hz,1H),7.68(s,1H),7.57(q,J=9.7Hz,4H),7.37(d, J=8.5Hz,2H),7.17(dd,J=9.0,1.6Hz,1H),4.18(s,3H),3.50(h,J=6.5Hz,1H),0.97(d,J=6.5Hz,6H).

[0240] Example 20: Synthesis of 1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (Compound 11)

[0241] Step 1: Synthesis of 1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one

[0242] 7-Chloro-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1,8-naphthyridin-2(1H)-one (70 mg, 0.14 mmol, 1.0 eq) obtained in Example 19, methanesulfonic acid (9,9-dimethyl-4,5-bisdiphenylphosphinothrene) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (13 mg, 0.014 mmol, 0.1 eq), sodium tert-butoxide (27 mg, 0.28 mmol, 2.0 eq) and 2,2,2-trifluoroethane-1-amine (138 mg, 1.4 mmol, 10.0 eq) were dissolved in 1,4-dioxane (2 mL), replaced with nitrogen, heated to 60° C., and stirred for 3.5 hours. After the reaction, HPLC separation and purification were performed to obtain a white solid 1-(4-chlorophenyl)-6-fluoro-4-(isopropylamino)-3-(2-methyl-2H-indazol-5-yl)-7-[(2,2,2-trifluoroethyl)amino]-1,8-naphthyridin-2(1H)-one (36 mg, yield 45.57%). LCMS (TOF MS ES+) m / z [M+H] + :559.1569. 1 H NMR (400MHz, DMSO-d6) δ8.32 (s, 1H), 8.11 (d, J = 12.5Hz, 1H), 7.60 (m, J = 8.8 ,1.0Hz,1H),7.57(d,J=1.2Hz,1H),7.52–7.47(m,2H),7.30–7.25(m,2H),7 .16(d,J=7.1Hz,1H),7.11(dd,J=8.9,1.5Hz,1H),6.33(t,J=7.2Hz,1H),4. 17(s,3H),3.48(h,J=6.6Hz,1H),3.31–3.23(m,2H),0.96(d,J=6.5Hz,6H).

[0243] Example 21: Synthesis of 7-amino-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (Compound 51)

[0244] Step 1: Synthesis of 6-chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)nicotinic acid

[0245] 3-Amino-2-methylpyridine (10.28 g, 95.23 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (60 mL), cooled to -78 ° C, and lithium bistrimethylsilylamide (142.83 mL, 142.83 mmol, 3.0 eq) was slowly added dropwise. After the addition was complete, the reaction was continued for 1 hour. Subsequently, 2,6-dichloro-5-fluoronicotinic acid (10.00 g, 47.61 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (30 mL) and slowly added dropwise to the reaction system at -78 ° C. After the addition was complete, the mixture was transferred to room temperature and reacted for 1 hour. After the reaction, saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL). The organic phases were combined, dried, concentrated, and washed with an appropriate amount of ethyl acetate. After filtration, a pale yellow solid of 6-chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)nicotinic acid (11.9 g, 89.0% yield) was obtained. LCMS (TOF MS ES+) m / z [M+H]+: 282.

[0246] Step 2: Synthesis of ethyl 3-(6-chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)pyridin-3-yl-3-oxopropanoate

[0247] Potassium 3-ethoxy-3-oxopropanoate (8.28 g, 48.66 mmol, 2.0 eq), magnesium chloride (6.73 g, 72.99 mmol, 3.0 eq) and triethylamine (9.85 g, 97.32 mmol, 4.0 eq) were dissolved in tetrahydrofuran solution (100 mL) and stirred at room temperature for 4 hours. 6-Chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)nicotinic acid (6.8 g, 24.33 mmol, 1.0 eq) and ultra-dry dichloromethane (60 mL) were added to another reaction flask, and a small amount of N,N-dimethylformamide was added as a catalyst. Subsequently, oxalyl chloride (5.07 g, 39.99 mmol, 1.5 eq) was slowly added dropwise at 0°C. After the addition was complete, the mixture was transferred to room temperature and reacted for 2 hours. After TLC monitoring of the reaction completion, the reaction solution was directly concentrated to obtain a yellow solid, which was then dissolved in tetrahydrofuran solution (60 mL) and slowly added dropwise to the potassium 3-ethoxy-3-oxopropanoate reaction system at 0°C. The reaction was continued at room temperature for 2 hours. After the reaction, the reaction solution was concentrated and purified using a forward phase column to obtain ethyl 3-(6-chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)pyridin-3-yl-3-oxopropanoate (4.4 g, 52.0% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 352.

[0248] Step 3: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0249] Ethyl 3-(6-chloro-5-fluoro-2-((2-methylpyridin-3-yl)amino)pyridin-3-yl-3-oxopropanoate (4.40 g, 12.53 mmol, 1.0 eq) and potassium carbonate (3.46 g, 25.07 mmol, 2.0 eq) were dissolved in anhydrous ethanol (50 mL) and reacted at 70°C for 3 hours. After completion of the reaction, the mixture was filtered through celite, and the filtrate was purified by forward column separation to afford solid 7-chloro-6-fluoro-4-hydroxy-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.6 g, 93.9% yield). LCMS (TOF MS ES+) m / z [M+H]+: 306.

[0250] Step 4: Synthesis of 4,7-dichloro-6-fluoro-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0251] 7-Chloro-6-fluoro-4-hydroxy-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.6 g, 11.80 mmol, 1.0 eq) and phosphorus oxychloride (20 mL) were added to a reaction flask and reacted at 80°C for 16 hours. After completion of the reaction, the product was concentrated under reduced pressure to remove the phosphorus oxychloride, and ethyl acetate was added to slurry. The product was filtered to yield a yellow solid, 4,7-dichloro-6-fluoro-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.3 g, 86.8% yield). LCMS (TOF MS ES+) m / z [M+H]+: 324.

[0252] Step 5: Synthesis of 7-chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0253] 4,7-Dichloro-6-fluoro-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (1.0 g, 3.09 mmol, 1.0 eq), methylamine hydrochloride (313 mg, 4.64 mmol, 1.5 eq), and N,N-diisopropylethylamine (1.6 g, 12.36 mmol, 4.0 eq) were dissolved in acetonitrile (20 mL) and reacted at 80°C for 2 hours. After completion of the reaction, the reaction solution was concentrated to obtain a pale yellow solid. The solid was slurried with ethanol and water and filtered to obtain a yellow solid, 7-chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (792 mg, 80.7% yield), which was directly used in the next step. LCMS (TOF MS ES+) m / z [M+H]+: 319.

[0254] Step 6: Synthesis of 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0255] 7-Chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (500 mg, 1.57 mmol, 1.0 eq) and N-bromosuccinimide (307.8 mg, 1.73 mmol, 1.1 eq) were added to an acetonitrile solution (10 mL) and reacted at room temperature for 4 hours. After completion of the reaction, the reaction solution was directly concentrated and slurried with ethanol and water to obtain 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (573 mg, 92.4% yield) as a pale yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 396.

[0256] Step 7: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0257] 3-Bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (200.0 mg, 0.50 mmol, 1.0 eq), 2-methyl-2H-indazole-5-boronic acid (132.0 mg, 0.75 mmol, 1.5 eq) and potassium phosphate (265 mg, 1.25 mmol, 2.5 eq) were added to a mixed solvent of dioxane (6 mL) and water (0.6 mL), and finally 1,1-bis(diphenylphosphino)diphenylferric palladium dichloride (36.5 mg, 0.05 mmol, 0.1 eq) was added. Under nitrogen protection, the reaction was carried out at 80 ° C for 16 hours. After the reaction, the reaction solution was concentrated and purified by reverse phase chromatography to obtain a yellow solid, 7-chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (110.8 mg, 48.9% yield). LCMS (TOF MS ES+) m / z [M+H]+: 449.

[0258] Step 8: Synthesis of 7-amino-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one

[0259] 7-Chloro-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (110.8 mg, 0.24 mmol, 1.0 eq), p-methoxybenzylamine (65.7 mg, 0.48 mmol, 2.0 eq), methanesulfonic acid (4,5-bis(diphenylphosphino-9,9-dimethylxanthene)(2-amino-1,1'-biphenyl-2-yl)palladium(II)) (22.7 mg, 0.024 mmol, 0.1 eq), and sodium tert-butoxide (46.0 mg, 0.48 mmol, 2.0 eq) were added to 1,4-dioxane (3.0 mL) and reacted at 100°C. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated to obtain a crude intermediate. The crude intermediate was dissolved in dichloromethane (2.0 mL) and trifluoroacetic acid (2.0 mL) and allowed to react at room temperature for 1 hour. After the reaction, the reaction solution was concentrated, neutralized with sodium bicarbonate solution, extracted with dichloromethane, and the organic phase was concentrated. Preparative separation and purification were performed to obtain 7-amino-6-fluoro-3-(2-methyl-2H-indazol-5-yl)-4-(methylamino)-1-(2-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (31 mg, 30.1% yield) as a white solid. LCMS (TOF MS ES+) m / z [M+H] + :430. 1 H NMR (400MHz, DMSO) δ8.39(dd,J=8.8,1.6Hz,1H),8.29(s,1H),8.15(d,J=12.3Hz,1H),7.62–7.54(m,3H) ,7.47–7.31(m,2H),7.22(d,J=5.1Hz,1H),6.01(s,2H),4.17(s,3H),2.42(d,J=4.6Hz,3H),2.01(s,3H).

[0260] The compounds listed in Table 1 below were prepared by methods similar to those described in the Examples, with appropriate variations in reactants, amounts of reagents, protection and deprotection, solvents, and reaction conditions. Characterization data for the compounds are summarized in Table 1 below.

[0261] Table 1: Structure and characterization of some compounds

[0262] Experimental Example 1: Effect of the compounds of the present invention on HCT116 MTAP - / - Cell proliferation assay

[0263] Experimental purpose: The purpose of this test case is to test the effect of compounds on HCT116 MTAP- / - Cell proliferation effect.

[0264] Background: Methionine adenosyltransferase 2A (MAT2A) is considered a synthetic lethal target in cancers that lack the methylthioadenosine phosphorylase (MTAP) gene. The MTAP gene is adjacent to the CDKN2A tumor suppressor and is co-deleted with CDKN2A in approximately 15% of cancers. Therefore, we tested the effects of compounds on HCT116 MTAP. - / - The inhibition rate of cell proliferation is used to screen MAT2A protein inhibitors.

[0265] Specific experimental process:

[0266] Construct HCT116 MTAP knockout cells and screen single clones. - / - WT and WT cells were seeded into 96-well plates, with 90 μL per well, at a density of 1000 cells / well. The cells were incubated overnight at 37°C. The next day, 10 μL of compound at various concentrations (final DMSO concentration of 1%) was added and the cells were incubated at 37°C for 10 days. On day 10, the old medium was aspirated and 110 μL of medium (at a 100:10 ratio of medium to CCK8) was added. The cells were incubated at 37°C for 1-4 hours. The absorbance was measured at 450 nM, and the IC50 value was calculated using GraphPad software. Compounds were screened by comparing the results with those of active drugs.

[0267] The IC50 (half maximal inhibitory concentration) is the half-maximal inhibitory concentration of the antagonist being measured. It indicates the half-maximum inhibitory concentration of a drug or substance (inhibitor) in inhibiting a biological process (or substances involved in that process, such as enzymes, cell receptors, or microorganisms). AG270 from Agios Pharmaceuticals and Compound A (Compound 167 in WO2020123395) were used as positive reference compounds. Their structures are shown below, and the experimental results are shown in Table 2.

[0268] Table 2: Inhibitory effect test results on HCT116 MTAP- / - cells

[0269] The results showed that the representative compounds of the present invention had an effect on HCT116 MTAP - / - The cells have a good inhibitory effect, which is better than the positive control.

[0270] Experimental Example 2: Determination of the effect of the compounds of the present invention on MAT2A protease function

[0271] Experimental purpose: The purpose of this test example is to test the ability of the compound to inhibit the function of MAT2A protease.

[0272] Background and Rationale: The metabolic enzyme methionine adenosyltransferase 2A (MAT2A) plays a crucial role in metabolism and epigenetics as it is the primary producer of the universal methyl donor, S-adenosylmethionine (SAM). MAT2A reacts with ATP and L-Met to generate SAM and a phosphate group. Therefore, after compound incubation, the ability of a compound to inhibit MAT2A enzyme function is assessed by measuring SAM production, allowing for the screening of MAT2A protein inhibitors.

[0273] Specific experimental process:

[0274] MAT2A protein expression: Full-length MAT2A was cloned into the pET24N vector containing an N-terminal (His) 6x tag and a tobacco etch virus (TEV) protease cleavage site. The constructed vector was transformed into Escherichia coli BL21(DE3) and incubated at OD 0.6 with the addition of 1 mM IPTG. The cells were incubated at 18°C ​​for 16 hours. The cells were harvested, disrupted by sonication, and the supernatant was centrifuged. The protein was purified using Ni-NTA and dialyzed for determination of protein concentration and purity. SAM assay: To the reaction system, add 91 μL of 50 mM Tris HCl (pH 7.5), 1.5 μL (10 / 3 M) KCl, 1.5 μL (1 M) MgCl2, 1 μL (100 mM) ATP, 1 μL (80 mM) L-Met, 1 μL (30 mM) EDTA (pH 7.67), 1 μL 5% BSA, 2 μL of the above-mentioned DMSO-dissolved drug, and 1 μL of MAT2A protein. Experimental groups were prepared at different drug concentrations, and 2 μL (100 μL) of the solution was added to the reaction system. The reaction was incubated at 37°C for 18 h. A solvent control group and a blank control group were set up. After the reaction was terminated, 40 μL of the solution was removed and quenched with 4 μL of 10% SDS. IC50 values ​​were calculated using GraphPad software, and compounds were screened by comparison with positively charged drugs. The results are shown in Table 3.

[0275] Table 3: Results of assays of MAT2A protease function

[0276] The results showed that the compounds of the present invention have very strong inhibitory effects on MAT2A protease function, with IC50 values ​​reaching the nM level, equivalent to or lower than those of positive drugs. This strong inhibitory effect has important therapeutic significance for the treatment of conditions or diseases related to MAT2A inhibition.

[0277] Experimental Example 3: Pharmacokinetics of the compounds of the present invention in rats

[0278] Experimental purpose: To investigate the pharmacokinetic characteristics of the compound of the present invention by oral gavage (PO) to SD female rats.

[0279] Specific experimental process:

[0280] The compound of the present invention and the control compound A (preparation method refers to compound 167 in patent application WO2020123395) were administered by gavage (10 mg / kg), PO solvent: 30% PEG300 + 10% Tween 80 + 60% water, prepared on the day of administration. Blood was collected from the jugular vein 0h before administration and 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h after administration (gavage), and about 0.02mL of each sample was collected. K2-EDTA was used for anticoagulation and the samples were placed on ice after collection. Plasma sample processing: After blood sample collection, it was placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 6800g, 6 minutes, 2-8°C). Plasma samples were stored in a -80°C refrigerator before analysis. Pharmacokinetic parameters were calculated using Phoenix WinNonlin7.0 based on the blood drug concentration data at different time points. The experimental results are shown in Table 4 below:

[0281] Table 4: In vivo pharmacokinetic test results in female rats

[0282] The pharmacokinetic data of the compound of the present invention in rats show that, compared with the control compound A, the compound of the present invention is better absorbed after oral administration in rats.

[0283] The above examples are merely representative. It can be seen from the above examples that the compounds of the present invention are ideal and highly effective MAT2A inhibitors and can be expected to be used for treating or preventing conditions or diseases associated with MAT2A inhibition.

Claims

1. A compound as represented by formula (I) and its stereoisomers, or a pharmaceutically acceptable salt thereof, in, R1 is selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl; R2 and R3 are independently selected from 5-12 membered monocyclic or bicyclic aromatic or heteroaryl rings, wherein the aromatic or heteroaryl groups are optionally substituted by one or more identical or different substituents R a replace; The heteroaryl group contains 1-3 heteroatoms selected from N, O, and S, and the ring system contains a saturated or partially unsaturated ring system such as a spiro ring, a bridged ring, a fused ring, and a fused ring; R a R4 and R5 are independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl; R4 and R5 are independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl; 12 Alkyl, 3-12 membered cycloalkyl or heterocycloalkyl, 3-12 membered halogenated cycloalkyl or halogenated heterocycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, -NH-(CH2) n -C1-C6 haloalkyl, -NH-(CH2) n -C3-C8 heterocycloalkyl, -NH-(CH2) n -NR b R c 、-NH-(CH2) n -O-C1-C6 alkyl, -O-C1-C6 alkyl, -O-C1-C6 haloalkyl, -O-(CH2) n -heterocyclic group, -O-(CH2) n -cycloalkyl, -O-(CH2) n -C3-C6 heterocycloalkyl, -O-(CH2) n -O-C1-C6 alkyl; The C1-C6 monoalkylamino, C1-C6 dialkylamino, -N(H)-(CH2) n -C1-C6 haloalkyl, -NH-(CH2) n -C3-C8 heterocycloalkyl, -N(H)-(CH2) n -NR b R c 、-NH-(CH2) n -C3-C8 heterocycloalkyl, -NH-(CH2) n -O-C1-C6 alkyl, -O-C1-C6 alkyl, -O-C1-C6 haloalkyl, -O-(CH2) n -heterocyclic group, -O-(CH2) n -cycloalkyl, -O-(CH2) n -C3-C6 heterocycloalkyl, -O-(CH2) n -O-C1-C6 alkyl is optionally substituted by one or more substituents from the group consisting of halogen, hydroxy, amino, oxo, C1-C6 alkylamino, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl; R b 、R c Each independently selected from H, C1-C6 alkyl; n is 0, 1, 2, or 3.

2. The compound according to claim 1, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R1 is selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl.

3. The compound according to claim 1, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R2 is selected from the group consisting of one or more identical or different R a The substituted 5-12 membered bicyclic heteroaryl ring is preferably 4. The compound according to claim 1, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R3 is selected from the group consisting of one or more identical or different R a Substituted 5-12 membered monocyclic aromatic or heteroaryl ring, preferably 5. The compound according to claim 1, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R4 is selected from Cl, methyl, methoxy, trifluoroethylamino, difluoromethyl, amino, trifluoroethoxy, trifluoromethyl, 6. The compound according to claim 1, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R5 is selected from F, hydrogen, and methoxy.

7. The compound according to any one of claims 1 to 6, its stereoisomers, or its pharmaceutically acceptable salts, characterized in that: It has the structure shown in the following formula (Ia), (Ib), (Ic), (Id), (Ie), in, R6 is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, preferably methyl; R7 and R8 are independently selected from hydrogen, deuterium, halogen, cyano, -C(O)NH2, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl.

8. The compound according to claim 7, its stereoisomer, or its pharmaceutically acceptable salt, characterized in that: R7 and R8 are independently selected from hydrogen, chlorine, fluorine, methoxy, trifluoromethoxy, cyano, trifluoromethyl, and methyl.

9. A compound and a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is:

10. A pharmaceutical composition comprising the compound according to any one of claims 1 to 9, its stereoisomers or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

11. Use of a compound according to any one of claims 1 to 10 and a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition according to claim 10 in the preparation of a medicament for treating a subject suffering from a disease or condition associated with MAT2A or MTAP protein activity or expression, wherein the disease or condition is preferably cancer or an autoimmune disease, the cancer being preferably selected from lung cancer, pancreatic cancer, liver cancer, colorectal cancer, bile duct cancer, gallbladder cancer, brain cancer, gastric cancer, leukemia, lymphoma, melanoma, thyroid cancer, nasopharyngeal cancer, glioma, bladder cancer, astrocytoma, basal cell carcinoma, osteosarcoma, head and neck cancer, chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma and mesothelioma; the autoimmune disease being preferably selected from thyroiditis, inflammatory bowel disease, lupus erythematosus, fibrosis, myasthenia gravis, vasculitis, psoriasis, arthritis, scleroderma, and dermatitis.

12. The intermediate compound shown below and its stereoisomer, or its pharmaceutically acceptable salt, 13. A method for preparing a compound represented by formula (1), characterized in that: The following steps are involved: (1) The starting material (i-1) is subjected to a substitution reaction to obtain an intermediate compound (i-2); (2) The intermediate compound (i-2) is subjected to condensation and decarboxylation reactions to obtain the intermediate compound (i-3); (3) The intermediate compound (i-3) is subjected to an aminolysis cyclization reaction to obtain the intermediate compound (i-4); (4) The intermediate compound (i-4) is subjected to a halogenation reaction to obtain the intermediate compound (i-5); (5) The intermediate compound (i-5) is subjected to an amine substitution reaction to obtain the intermediate compound (i-6); (6) The intermediate compound (i-6) is subjected to aromatic ring halogenation reaction to obtain the intermediate compound (i-7); (7) The intermediate compound (i-7) is reacted to give compound (1) through coupling reaction.

14. A method for preparing a compound represented by formula (II), comprising the following steps: The compound of formula (IIA) is subjected to a substitution reaction to obtain a compound of formula (II); Wherein, R6 is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, preferably methyl; R7 and R8 are each independently selected from hydrogen, deuterium, halogen, cyano, -C(O)NH2, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C1-C6 monoalkylamino, C1-C6 dialkylamino, alkenyl, alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl; R1, R4 and R5 are as defined in claims 1 to 6.

15. The preparation method according to claim 14, wherein R1 is selected from hydrogen, methyl, ethyl, isopropyl, and cyclopropyl.

16. The preparation method according to claim 14, wherein R4 is selected from Cl, methyl, methoxy, trifluoroethylamino, difluoromethyl, amino, trifluoroethoxy, trifluoromethyl, The preparation method according to claim 14 , wherein R 5 is selected from F, hydrogen, and methoxy.

18. The preparation method according to claim 14, wherein R7 and R8 are independently selected from hydrogen, chlorine, fluorine, methoxy, trifluoromethoxy, cyano, trifluoromethyl, and methyl.

Images