A nitrogen-containing fused ring compound and pharmaceutical use thereof

Abstract

The present application provides a kind of compound as shown in formula I or its tautomer, meso, racemate, enantiomer, diastereoisomer, or its mixture form or its pharmaceutically acceptable salt, its pharmaceutical composition and as MAT2A inhibitor drug use.The present application provides a kind of MAT2A inhibitor compound with novel structure, strong activity and good drug property, and the compound has good application prospect in preventing and / or treating MAT2A inhibition related indications such as cancer, autoimmune diseases and the like.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a nitrogen-containing fused-ring compound or its tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, as a MAT2A inhibitor, its preparation method, pharmaceutical composition, and use in preparation for use in subjects suffering from diseases such as cancer. Background Technology

[0002] Methionine adenosine transferase (MAT) (also known as S-adenosylmethionine synthase) 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 in the methionine cycle. SAM is the alanine donor in polyamine biosynthesis and a major methyl donor for DNA methylation, and it is involved in gene transcription, cell proliferation, and the generation of secondary metabolites.

[0003] Three isoforms of human MAT include MAT1 and MAT3, expressed in liver tissue, while MAT2A is universally expressed in human cell types and is the predominant form in human tumors. Crystal structure and mechanistic studies have shown that despite 85% amino acid sequence similarity, their mechanisms of action differ significantly. MAT2A forms a functional homodimer in its purified active form and binds to the regulatory protein MAT2B. MAT2B modulates the activity of MAT2A by increasing its sensitivity to inhibition by ADOMet products, but does not provide a significant rate enhancement. Cell localization studies have shown that MAT2A is present in both the cytoplasm and nucleus. Nuclear condensation of MAT2A has been reported to occur during replication and the subsequent G2 phase, satisfying the high methylation requirements of DNA and histone methylation processes in the S-phase nucleus. The activity of ADOMet in transmethylation has been identified as a rate-limiting factor in lung cancer stem cell development, making MAT2A and related enzymes of the methionine cycle potential targets for anticancer drugs. Methionine for ADOMet production by MAT2A is derived from dietary sources or the polyamine cycle, in which 5-methylthio-D-ribose-1-phosphate produced by S-methyl-5'-thioadenosine phosphorylase is recycled to methionine. MAT2A protein expression has been reported to be increased in cancers such as colon cancer, liver cancer, gastric cancer, blood cancer, and liver cancer. Approximately 15% of human cancers show deletion of the MTAP (S-methyl-5'-thioadenosine phosphorylase) gene, thus lacking the methionine recycling pathway for polyamine synthesis. MTAP deletion in chr9p21 typically involves deletion of the CDKN2a tumor suppressor gene site. Comprehensive genetic lethality studies of MATP- / - cancer cells have shown that these cancer cells have increased sensitivity to inhibition of MAT2A, PRMT5, and PRMT1.

[0004] In hepatocellular carcinoma (HCC), a downregulation of MAT1A and upregulation of MAT2A occur, known as the MAT1A:MAT2A switch. This switch, accompanied by upregulation of MAT2B, results in lower SAM levels, providing a growth advantage for HCC cells. Since MAT2A plays a crucial role in promoting HCC cell growth, it is a target for anti-tumor therapy. Recent studies have shown that silencing MAT2A using small interfering RNA essentially inhibits HCC cell growth 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)

[0006] 15(3)(2016)574–587). MTAP (methionine phosphorylase) is an enzyme widely expressed in normal tissues that catalyzes the conversion of methionine (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-aranoxin.

[0007] MAT2A dysregulation is present in other cancers lacking MTAP (including hepatocellular carcinoma and leukemia). J. Cai et al., Cancer Res. 58 (1998) 1444-1450; T.S. Jani et al., Cell. Res. 19 (2009) 358-369. Silencing MAT2A expression via RNA interference has produced 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.

[0008] Many human and mouse malignant cells lack MTAP activity. MTAP deficiency is not only present in tissue culture cells, but also in primary leukemia, glioma, melanoma, pancreatic cancer, non-small cell lung cancer (NSCLC), bladder cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, 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, which are inhibitors of cyclin D-dependent kinases cdk4 and cdk6, respectively.

[0009] Alternatively, the p16INK4A transcript can be spliced ​​into a variable reading frame (ARF) encoding the p14ARF. The p14ARF binds to MDM2 and prevents the degradation of p53 (Pomerantz et al. (1998) Cell 92: 713-723). The 9p21 chromosomal region is of interest because it is frequently homozygous deleted in a variety of cancers, including leukemia, NSLC, pancreatic cancer, glioma, melanoma, and mesothelioma. Deletions often inactivate 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 deletions identified in these tumors involved a 170kb region containing MTAP, p14ARF, and p16INK4A. Carson et al. (WO99 / 67634) reported a correlation between tumor development stage and loss of homozygosity in genes encoding MTAP and p16. For example, the absence of the MTAP gene, but not p16INK4A, has been reported to predict cancer in the early stages of development, while the absence of genes encoding both p16 and MTAP has been reported to predict cancer in a more advanced stage of development. In some osteosarcoma patients, the MTAP gene is present at diagnosis but absent at a later time point (Garcia-Castellano et al., Clin. Cancer Res. 8(3) 2002 782-787).

[0010] MAT2A enzyme inhibitors for treating cancer are disclosed in WO2018039972; MAT2A inhibitors for treating autoimmune or inflammatory diseases are mentioned in WO2021158792; MAT2A enzyme inhibitors for treating diseases or conditions mediated by MAT2A overexpression are described in WO2018045071; 3-(cyclohexyl-1-en-1-yl)-6-(4-methoxyphenyl)-2-phenyl-5-(pyridin-3-ylamino)pyrazolo[1,5-a]pyrimidin-7(4H)-one, a compound for treating MTAP-deficient non-small cell lung cancer (NSCLC) in patients in need, is also disclosed in CN202080014106.0; a MAT2A inhibitor with a 6-6-cyclic parent nucleus is disclosed in CN114874207A; and a polycyclic compound for inhibiting MTAP-deficient cancer cells is disclosed in CN113999232A.

[0011] The discovery and search for novel MAT2A inhibitors with excellent drug-like properties has become a major focus in the development of drugs for treating MTAP-deficient tumors. Summary of the Invention

[0012] This 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 solve the aforementioned technical problems, this invention adopts the following technical solution:

[0013] On the one hand, the present invention provides compounds of formula (I) or tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof.

[0014]

[0015] in,

[0016] Ring A is selected from tricyclic heteroaryl and tricyclic heterocyclic groups, wherein the tricyclic heteroaryl or tricyclic heterocyclic group is optionally substituented by 1 to 3 identical or different substituents R. a replace;

[0017] R3 is selected from a 5-12 member aromatic ring or heteroaromatic ring, wherein the 5-12 member aromatic ring or heteroaromatic ring is optionally replaced by one or more identical or different substituents R. a replace;

[0018] R aSelected from hydrogen, deuterium, halogen, cyano, nitro, amide, sulfonamide, hydroxyl, amino, urea, phosphoryl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl, C1-C6 alkyl-S-, C1-C6 alkyl-SO-, C1-C6 alkyl-SO2-; or two of the above R a It can form 3-12 member saturated or partially unsaturated or aromatic ring systems through carbon chains or heteroatoms;

[0019] R1 and R2 are independently selected from hydrogen, C1-C6 alkyl, and C3-C4 alkyl groups, respectively. 12 cycloalkyl or heterocycloalkyl;

[0020] R4 and R5 are independently selected from hydrogen, deuterium, halogen, cyano, nitro, amide, sulfonamide, hydroxyl, amino, urea, phosphoryl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl; or R4 and R5 can form 3-12 membered saturated or partially unsaturated or aromatic ring systems through carbon atoms.

[0021] As a preferred technical solution, it is a compound represented by formula (II) or its tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or a pharmaceutically acceptable salt thereof:

[0022]

[0023] A, R3, R4, and R5 are defined as above.

[0024] As a preferred technical solution, the ring A is selected from 10-20 ternary heteroaryl groups and 10-20 ternary heterocyclic groups, wherein the 10-20 ternary heteroaryl group and 10-20 ternary heterocyclic group are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, or C1-C6 haloalkyl groups.

[0025] As a preferred technical solution, the ring A is selected from 12-14 member trifused heteroaryl groups and 12-14 member trifused heterocyclic groups, wherein the 12-14 member trifused heteroaryl groups and 12-14 member trifused heterocyclic groups are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, and C1-C6 haloalkyl groups.

[0026] As a more preferred technical solution, ring A is selected from 13-membered tricyclic heteroaryl and 13-membered tricyclic heterocyclic group, wherein the 13-membered tricyclic heteroaryl and 13-membered tricyclic heterocyclic group are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, and C1-C6 haloalkyl groups.

[0027] As a preferred technical solution, it is a compound represented by formula (III) or its tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or a pharmaceutically acceptable salt thereof:

[0028]

[0029]

[0030] in,

[0031] X is selected from N, O, and S;

[0032] Y is selected from N and CH;

[0033] R6 is selected from hydrogen, deuterium, halogen, cyano, nitro, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl;

[0034] Ring A is selected from The above-mentioned ring A may be optionally substituted with one or more hydrogens, deuteriums, halogens, cyano groups, nitro groups, hydroxyl groups, amino groups, C1-C6 alkyl groups, C1-C6 alkoxy groups, C1-C6 haloalkyl groups, C1-C6 haloalkoxy groups, C2-C4 alkenyl groups, C2-C4 alkynyl groups, C1-C6 monoalkylamino groups, C1-C6 dialkylamino groups, 3-8 membered cycloalkyl groups, or heterocyclic alkyl groups;

[0035] n is 0, 1, 2, or 3.

[0036] As a preferred technical solution, R1 and R2 are independently selected from hydrogen, C1-C6 alkyl, and C3-C4 alkyl groups, respectively. 12 Cycloalkyl.

[0037] As a preferred technical solution, it is a compound of formula (IV) or its tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or a pharmaceutically acceptable salt thereof:

[0038]

[0039] As a preferred technical solution, R6 is selected from hydrogen, halogen, C1-C6 alkyl, cyano, and C1-C6 alkoxy.

[0040] As a preferred technical solution, R4 is selected from hydrogen, halogen, amino, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl, preferably hydrogen, amino, halogen, or C1-C6 alkyl, and more preferably hydrogen, amino, or chlorine.

[0041] As a preferred technical solution, R5 is selected from hydrogen, halogen, amino, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl, preferably hydrogen, amino, halogen, or C1-C6 alkyl, and more preferably hydrogen, fluorine, or methyl.

[0042] The present invention also provides compounds thereof, or tautomers, meso compounds, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, selected from:

[0043]

[0044] The present invention also provides a pharmaceutical composition comprising any of the compounds described above and their isomers or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0045] The present invention also provides the use of any of the compounds and isomers thereof, or pharmaceutically acceptable salts thereof, and pharmaceutical combinations thereof, as described above, in the preparation of a medicament for use in a subject suffering from a disease or condition associated with the activity or expression of MAT2a or MTAP proteins, wherein the disease or condition is preferably cancer or an autoimmune disease, wherein 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 carcinoma, glioma, bladder cancer, astrocytoma, basal cell carcinoma, osteosarcoma, head and neck cancer, chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, and mesothelioma; wherein the autoimmune disease is preferably selected from thyroiditis, inflammatory bowel disease, lupus erythematosus, fibrosis, myasthenia gravis, vasculitis, psoriasis, arthritis, scleroderma, dermatitis, etc.

[0046] The inventors discovered that these compounds are highly effective MAT2A inhibitors with extremely strong MAT2A inhibitory activity, and can be used to prepare for the prevention and / or treatment of indications related to MAT2A inhibition, including reduced or absent MTAP expression, deletion of the MTAP gene, cancer caused by reduced MTAP protein function, and autoimmune diseases. This invention is based on the above findings. Invention Details

[0048] The various aspects and features of the present invention will be further described below.

[0049] Since the compounds according to the present invention can exist in stereoisomeric forms, the present invention includes all stereoisomeric forms.

[0050] The compounds of this invention have an asymmetric center. Compounds containing asymmetric substitution atoms in this invention can be isolated into optically active or racemic forms. Those skilled in the art know how to prepare the optically active forms, for example, through racemic resolution or synthesis from optically active starting materials. Unless otherwise specified, this invention includes all chiral, diastereomers, and racemates. Methods for preparing the compounds of this invention and their intermediates are part of this invention. All stereoisomers of the compounds of this invention are also part of this invention.

[0051] "Stereoisomers" are compounds composed of identical atoms bonded by the same bonds, but with different three-dimensional structures. This invention will cover various stereoisomers and mixtures thereof.

[0052] A tautomer is an isomer formed when a proton is transferred from one atom of a molecule to another atom of the same molecule.

[0053] All tautomer forms of the compounds of this invention are also included within the scope of this invention.

[0054] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms, and thus may produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention aims to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention may select racemic, diastereomer, or enantiomer as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0055] The term “optional” or “optionally” means that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0056] "alkyl" refers to a group having 1 to 10 carbon atoms, either straight-chain or branched, saturated hydrocarbon groups ("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"), 1 to 5 carbon atoms ("C1-C5 alkyl"), 1 to 4 carbon atoms ("C1-C4 alkyl"), 1 to 3 carbon atoms ("C1-C3 alkyl"), 1 to 2 carbon atoms ("C1-C2 alkyl"), or 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), etc. Each alkyl group may be optionally substituted independently, i.e., unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents; for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or substituted with 1 substituent (“substituted alkyl”). In some embodiments, the alkyl group is an unsubstituted C1–C10 alkyl group (e.g., –CH3). In some embodiments, the alkyl group is a substituted C1–C6 alkyl group. Commonly used alkyl abbreviations include Me (–CH3), Et (–CH2CH3), iPr (–CH(CH3)2), nPr (–CH2CH2CH3), n–Bu (–CH2CH2CH2CH3) or i–Bu (–CH2CH(CH3)2).

[0057] The term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting only 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 connected to the rest of the molecule by single bonds, such as, but not limited to, vinyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-1,4-dienyl, etc.

[0058] The term "alkynyl" refers to a straight or branched hydrocarbon chain group consisting only 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 connected to the rest of the molecule by single bonds, such as, but not limited to, ethynyl, propynyl, butynyl, pent-1-en-4-ynyl, etc.

[0059] As described herein, the terms “halogen,” “halogenated,” “halogenated,” etc., refer to fluorine, chlorine, bromine, or iodine, particularly fluorine, chlorine, and bromine, with fluorine and chlorine being especially preferred.

[0060] "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.

[0061] "Alkoxy" refers to an alkyl group (e.g., C1-C6 alkyl) as described herein, which is attached to the molecule by an oxygen atom. This includes portions in which the alkyl moiety can be straight-chain or branched, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0062] "Cycloalkyl" is a monocyclic saturated cycloalkyl group having 3 to 10 ring carbon atoms ("C3-10 cycloalkyl" or "C3-C10 cycloalkyl"). In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms ("C3-8 cycloalkyl" or "C3-C8 cycloalkyl"). In some embodiments, the cycloalkyl group has 3 to 6 ring carbon atoms ("C3-6 cycloalkyl" or "C3-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms ("C5-6 cycloalkyl" or "C5-C6 cycloalkyl"). In some embodiments, the 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 with one or more substituents (“substituted cycloalkyl”). In some embodiments, the cycloalkyl group is an unsubstituted C3-C10 cycloalkyl group. In some embodiments, the cycloalkyl group is a substituted C3-C10 cycloalkyl group.

[0063] "Heterocyclic alkyl" refers to a group in which one or more C atoms in the above "cycloalkyl" are replaced by heteroatoms such as N, O, S, and P.

[0064] The term "alkylamino" refers to an amino-NRdRe with 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 an alkyl group as described above.

[0065] The term "haloalkylamino" refers to an amino-NRdRe with 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 a haloalkyl group as described above.

[0066] The term "heterocyclic group" refers to 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, the heterocyclic group can be a monocyclic, bicyclic, tricyclic, or more ring system, which may include fused ring systems, bridged ring systems, or spirocyclic systems; the nitrogen, carbon, or sulfur atoms in the heterocyclic group may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be connected to the remainder of the molecule via a carbon atom or a heteroatom through a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the connection point with the remainder of the molecule is a non-aromatic ring atom. For the purposes of this 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]nonane-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptane-2-yl, aziridine, pyranyl, tetrahydropyranyl, thiaranyl, tetrahydrofuranyl, oxazinyl, dioxocyclopentyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, quinazinyl, thiazoalkyl, isothiazyl, isoxazylalkyl, dihydroindolyl, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolyl, phthalimide, etc.

[0067] The term "aryl" refers to a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms (preferably 6 to 10 carbon atoms). For the purposes of this invention, the aryl group can be a monocyclic, bicyclic, tricyclic, or more cyclic system, and can be fused with cycloalkyl or heterocyclic groups as defined above, provided that the aryl group is connected to the rest of the molecule via single bonds through atoms on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthrene, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1,4-benzoxazine-3(4H)-one-7-yl, etc.

[0068] The term "heteroaryl" refers to a 5- to 16-membered conjugated cyclic group having 1 to 15 carbon atoms (preferably 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically indicated in this specification, a heteroaryl group can be a monocyclic, bicyclic, tricyclic, or more cyclic system, and can also be fused with cycloalkyl or heterocyclic groups as defined above, provided that the heteroaryl group is connected to the rest of the molecule via single bonds through atoms on the aromatic ring. The nitrogen, carbon, or sulfur atom in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. For the purposes of this invention, the 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, thiophene, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridinyl, pyrazinyl, pyrazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furanyl, pyrrolithyl, triazolyl, tetrazolyl, triazinyl, inazinyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolinyl, isoquinolinyl, diazonyl, naphthidyl, quinoxolinyl, pteridyl, carbazolyl, carbazolyl, phenanthridine, phenanthroxolinyl, acridineyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothiophene, oxadiazolyl, and oxadiazolyl. Azolyl, cyclolinyl, quinazolinyl, phenylthio, indene, o-diazaphenyl, isoxazolyl, phenoxazinyl, phenthiazinyl, 4,5,6,7-tetrahydrobenzo[b]thiophene, naphthopyridyl, [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.

[0069] Unless otherwise stated, when referring to a specifically named aryl (e.g., phenyl), heterocyclic (e.g., pyrrolyl, piperidinyl, and morpholinyl), or heteroaryl (e.g., tetrazolyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, and furanyl), the reference is intended to include a ring having, where appropriate, 0 to 3, preferably 0 to 2, substituents selected from the substituents listed above for aryl heterocyclic and / or heteroaryl groups.

[0070] The example of the three parallel rings is as follows: Includes, but is not limited to, the structures described above.

[0071] As described herein, the term "pharmaceutically acceptable salt" means that the salt is not only physiologically acceptable to the subject, but can also refer to a synthetic substance with pharmaceutical value, such as a salt formed as an intermediate during chiral resolution. Although such intermediate salts cannot be directly administered to the subject, they can play a role in obtaining the end product of this invention. Specifically, this includes acid (organic and inorganic acid) addition salts or base addition salts (including organic and inorganic bases).

[0072] "Pharmaceutically acceptable carriers" include, but are not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that is permitted by the relevant government regulatory authority to be acceptable for human or animal use.

[0073] The "tumor" and "diseases related to abnormal cell proliferation" mentioned in this 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 carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, colorectal cancer, nasopharyngeal carcinoma, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, and other diseases.

[0074] As described herein, the term "disease" refers to a physical condition of the subject that is related to the disease described in this invention. Examples include peripheral arterial diseases and neurodegenerative diseases described in this invention.

[0075] The cancers covered by this invention include standard treatments such as surgery, radiation therapy, chemotherapy, and hormone therapy.

[0076] The compounds or pharmaceutical compositions containing them in this invention can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, intravenous drip, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0077] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular 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, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0078] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0079] The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation.

[0080] Beneficial technical effects

[0081] The inventors have discovered that the compounds in this invention have good MAT2A inhibitory activity and drug potential.

[0082] This invention provides a class of novel, highly active, and more druggable MAT2A inhibitor compounds, which have promising applications in the prevention and / or treatment of MAT2A inhibition-related indications such as cancer and autoimmune diseases. Detailed Implementation

[0083] The following examples 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 examples.

[0084] For all the following embodiments, standard operations and methods known to those skilled in the art were used. Unless otherwise stated, all temperatures are expressed in °C (degrees Celsius). The structures of the compounds were determined by nuclear magnetic resonance spectroscopy (NMR) and / or mass spectrometry (MS).

[0085] The structures of the compounds of this invention were determined 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 was determined using a Brukeravance-400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents and tetramethylsilane (TMS) as an internal standard.

[0086] Liquid chromatography-mass spectrometry (LC-MS) was performed using an ACQUITY UPLC ultra-high pressure liquid chromatograph for the liquid phase and a Xevo G2-SQtof mass spectrometer for the mass spectrometry phase.

[0087] The starting materials used in the examples of this invention are known and commercially available, and can also be synthesized using or according to methods known in the art.

[0088] Synthesis of intermediate: 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0089]

[0090] Step 1: Synthesis of 6-chloro-2-[(4-chlorophenyl)amine]-5-fluoronicotinic acid

[0091] 4-Chloroaniline (20.00 g, 156.80 mmol, 2.0 eq) was dissolved in ultra-dry tetrahydrofuran (200 mL), cooled to -78 °C, and bis(trimethylsilylaminolithium) (235.20 mL, 235.20 mmol, 3.0 eq) was slowly added dropwise. After the addition was complete, the reaction was continued for 1 hour. At -78 °C, a solution of 2,6-dichloro-5-fluoronicotinic acid (16.46 g, 78.40 mmol, 1.0 eq) in ultra-dry tetrahydrofuran (60 mL) was slowly added dropwise to the reaction system. After the addition was complete, the system was transferred to room temperature and reacted for 1 hour. After the reaction was completed, a saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL). The extract was then washed with saturated brine, concentrated to remove the solvent, and a light brown solid was obtained. The solid was then slurried with methanol to obtain a pale yellow solid, 6-chloro-2-[(4-chlorophenyl)amine]-5-fluoronicotinic acid (20.1 g, yield 85.0%).

[0092] Step 2: Synthesis of ethyl 3-{6-chloro-2-[(4-chlorophenyl)amine]-5-fluoropyridin-3-yl}-3-oxopropionic acid ester

[0093] Potassium monoethyl malonate (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 to obtain system one. 6-Chloro-2-[(4-chlorophenyl)amine]-5-fluoronicotinic acid (7.30 g, 24.33 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (150 mL). A small amount of N,N-dimethylformamide was added as a catalyst, and oxaloyl chloride (5.08 g, 39.99 mmol, 1.5 eq) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature for further reaction. TLC confirmed that the reaction of the starting materials was complete. The reaction solution was then concentrated to obtain a yellow solid. The yellow solid was dissolved in tetrahydrofuran solution (100 mL) and slowly added dropwise to system one at 0 °C. The reaction was continued at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain a yellow solid. Forward purification yielded the target product, ethyl 3-{6-chloro-2-[(4-chlorophenyl)amine]-5-fluoropyridin-3-yl}-3-oxopropionic acid ethyl ester (6.30 g, yield 70%). 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).

[0094] Step 3: Synthesis of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthidium-2(1H)-one

[0095] Ethyl ethyl 3-{6-chloro-2-[(4-chlorophenyl)amine]-5-fluoropyridin-3-yl}-3-oxopropionate (6.3 g, 17.02 mmol, 1.0 eq) was dissolved in anhydrous ethanol (120 mL), and potassium carbonate (4.71 g, 34.05 mmol, 2.0 eq) was added. The reaction was carried out at 70 °C for 3 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was concentrated and purified by forward filtration (dichloromethane:methanol = 85%:15%) to obtain the target product 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthidium-2(1H)-one (5.10 g, yield 92.5%). LCMS (TOF MS ES+) m / z [M+H]+: 325. 1H NMR (400MHz, DMSO) δ 12.28 (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).

[0096] Step 4: Synthesis of 4,7-dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthidium-2(1H)-one

[0097] 5.10 g (15.74 mmol, 1.0 eq) of 7-chloro-1-(4-chlorophenyl)-6-fluoro-4-hydroxy-1,8-naphthidine-2(1H)-one was added to a reaction flask, followed by 50 mL of phosphorus oxychloride. The mixture was heated to 80 °C and reacted for 16 hours. After the reaction was completed, the phosphorus oxychloride was removed by concentration under reduced pressure. The product was then slurried and filtered to obtain the target product, 4,7-dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthidine-2(1H)-one (4.50 g, yield 83.6%). LCMS (TOF MS ES+) m / z [M+H]+: 343. 1H 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).

[0098] Step 5: Synthesis of 7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0099] 4,7-Dichloro-1-(4-chlorophenyl)-6-fluoro-1,8-naphthidium-2(1H)-one (1000 mg, 2.91 mmol, 1.0 eq), methylamine hydrochloride (294 mg, 4.35 mmol, 1.5 eq), and N,N-diisopropylethylamine (1501 mg, 11.6 mmol, 4 eq) were dissolved in acetonitrile (20 mL) and reacted at 80 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain a pale yellow solid, which was then slurried and filtered with ethyl acetate to obtain a yellow solid 7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one (410 mg, yield 41.6%), which was directly used for the next reaction. LCMS (TOF MS ES+) m / z [M+H]+: 338.

[0100] Step 6: Synthesis of 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0101] 7-Chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one (400.0 mg, 1.18 mmol, 1.0 eq) and N-succinimide bromide (232.4 mg, 1.30 mmol, 1.1 eq) were added to acetonitrile solution (8 mL) and reacted at room temperature for 4 hours. The reaction solution was stirred with silica gel and then separated in the forward direction (dichloromethane:methanol = 20:1) to give a pale yellow solid 3-bromo-7-chloro-1-(4-chloromethyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one (350 mg, yield 70.85%). 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).

[0102] Example 1: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-1,8-naphthid-2(1H)-one (Compound 1) and 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-4-(methylamino)-1,8-naphthid-2(1H)-one (Compound 2)

[0103]

[0104] Step 1: Synthesis of tert-butyl 4-(4-bromo-2-nitrophenyl)-3-oxoperpiperazine-1-carboxylate

[0105] 10 g (50.0 mmol, 1.0 eq) of 3-oxoperazine-1-carboxylic acid tert-butyl ester, 4-bromo-1-iodo-2-nitrobenzene (18.0 g, 55.0 mmol, 1.1 eq), tris(dibenzylacetone)dipalladium (2.29 g, 2.5 mmol, 0.05 eq) and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (2.89 g, 5.0 mmol, 0.1 eq) were added to toluene (100 mL) and reacted at 100 °C for 18 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filtrate was concentrated, and the filtrate was slurried with methyl tert-butyl ether and petroleum ether to give a yellow solid 4-(4-bromo-2-nitrophenyl)-3-oxoperazine-1-carboxylic acid tert-butyl ester (15.66 g, yield 78.3%). LCMS(TOF MS ES+)m / z[M+H]+:400.

[0106] Step 2: Synthesis of tert-butyl 8-bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0107] 4-(4-bromo-2-nitrophenyl)-3-oxopiperazine-1-carboxylic acid tert-butyl ester (15.0 g, 37.59 mmol, 1.0 eq), iron powder (10.5 g, 187.96 mmol, 5.0 eq), and ammonium chloride (30.16 g, 563.85 mmol, 15.0 eq) were added to ethanol (150 mL) and water (50 mL), and reacted at 80 °C for 16 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth, the filtrate was concentrated to remove ethanol, and the residue was filtered to give a yellow solid 8-bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (7.5 g, yield 56.9%). LCMS (TOF MS ES+) m / z [M+H]+: 352. 1 H NMR(400MHz,Chloroform-d)δ7.87(d,J=1.7Hz,1H),7.39(m,J=8.6,1.7Hz,1H),7.22(d ,J=8.5Hz,1H),4.93(s,2H),4.13(t,J=5.5Hz,2H),4.01(d,J=5.5Hz,2H),1.50(s,9H).

[0108] Step 3: Synthesis of tert-butyl 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0109] The following were added: 8-bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (7.5 g, 21.30 mmol, 1.0 eq), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (6.5 g, 25.56 mmol, 1.2 eq), 1,1-bis(diphenylphosphine)diberberine palladium dichloride (1.55 g, 2.13 mmol, 0.1 eq). Potassium acetate (5.21 g, 53.25 mmol, 2.5 eq) was added to 1,4-dioxane (80 mL), nitrogen was purged, and the reaction was carried out at 100 °C for 18 hours. After the reaction was completed, the mixture was filtered, the filtrate was concentrated, and purified by forward separation to give a yellow solid 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (6.06 g, yield 71.3%). LCMS (TOF MS ES+) m / z [M+H]+: 400.

[0110] Step 4: Synthesis of tert-butyl 8-(7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0111] The compound 8-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (6.0 g, 15.03 mmol, 1.0 eq), 3-bromo-7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one ( 6.89 g (16.54 mmol, 1.1 eq), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (1.09 g, 1.50 mmol, 0.1 eq), and potassium carbonate (6.22 g, 45.09 mmol, 3.0 eq) were dissolved in 1,4-dioxane (60.0 mL) and water (12.0 mL). The mixture was immediately purged with nitrogen three times and stirred at 90 °C for 18 hours. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated and purified by reverse-phase column chromatography to give a yellow solid 8-(7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (6.94 g, yield 75.8%). LCMS(TOF MS ES+)m / z[M+H]+:609. 1H NMR (400MHz, DMSO-d6) δ7.87(d,J=11.1Hz,1H),7.74(d,J=5.3Hz,1H),7.64–7.56(m,4H),7.41(d,J=8.2Hz,2H ), 7.23 (d, J = 8.4Hz, 1H), 4.81 (s, 2H), 4.22 (t, J = 5.2Hz, 2H), 3.94 (t, J = 5.4Hz, 2H), 2.56 (s, 3H), 1.48 (s, 9H).

[0112] Step 5: Synthesis of 8-{1-(4-chlorophenyl)-6-fluoro-7-[(4-methoxybenzyl)amino]-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl}-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester

[0113] 8-(7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (1.0 g, 1.64 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (449 mg, 3.28 mmol, 2.0 eq) and Sodium tert-butoxide (314 mg, 3.28 mmol, 2.0 eq) was added to dioxane solvent (20 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) (155 mg, 0.16 mmol, 0.1 eq) was added, replacing the nitrogen atmosphere, and the reaction was carried out at 80 °C. After the reaction was completed, the mixture was filtered through diatomaceous earth, the filtrate was concentrated, and reverse-phase separation was performed to obtain a yellow solid 8-{1-(4-chlorophenyl)-6-fluoro-7-[(4-methoxybenzyl)amino]-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl}-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (970 mg, yield 83.2%). LCMS (TOF MS ES+) m / z [M+H]+: 710.

[0114] Step 6: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-1,8-naphthidium-2(1H)-one

[0115] 8-{1-(4-chlorophenyl)-6-fluoro-7-[(4-methoxybenzyl)amino]-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl}-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (950 mg, 1.33 mmol, 1.0 eq) was added to trifluoroacetic acid (10 mL) and reacted at room temperature for 3 hours. After the reaction was complete, the reaction solution was concentrated, and the pH was adjusted to 12 by adding sodium hydroxide solution. Then, dichloromethane was added for extraction, followed by drying and concentration to obtain a yellow solid, 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-1,8-naphthidium-2(1H)-one(1H)-one (605 mg, yield 92.3%). LCMS (TOF MS ES+) m / z [M+H]+: 490.

[0116] Step 7: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0117] 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-1,8-naphthid-2(1H)-one(1H)-one (1H)-one (100.0 mg, 0.20 mmol, 1.0 eq), potassium carbonate (41 mg, 0.3 mmol, 1.5 eq), and iodomethane (31.2 mg, 0.22 mmol, 1.1 eq) were added to N,N-dimethylformamide (2 mL) and reacted at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered and purified by high-performance liquid chromatography (HPLC) to obtain a white solid 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-8-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (32.5 mg, yield 31.6%). LCMS (TOF MS ES+) m / z [M+H]+: 504. 1H NMR(400MHz, DMSO-d6)δ8.12(d,J=12.2Hz,1H),7.52–7.47(m,3H),7.43(s,1H),7.25(m,J=8.8,7.0Hz,3H),7.11( d,J=8.2Hz,1H),5.87(s,2H),4.14(t,J=5.5Hz,2H),3.76(s,2H),2.94(t,J=5.6Hz,2H),2.48(s,3H),2.46(s,3H).

[0118] Example 2: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-4-methylamino-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-1,8-naphthid-2(1H)-one (compound 3) and 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-4-(methylamino)-1,8-naphthid-2(1H)-one (compound 4)

[0119]

[0120] Step 1: Synthesis of tert-butyl 4-(5-bromo-2-nitrophenyl)-3-oxoperpiperazine-1-carboxylate

[0121] 10 g (50.0 mmol, 1.0 eq) of 3-oxoperazine-1-carboxylic acid tert-butyl ester, 4-bromo-2-iodo-1-nitrobenzene (18.0 g, 55.0 mmol, 1.1 eq), tris(dibenzylacetone)dipalladium (2.29 g, 2.5 mmol, 0.05 eq), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (2.89 g, 5.0 mmol, 0.1 eq) were added to toluene (100 mL) and reacted at 100 °C for 18 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filtrate was concentrated, and the filtrate was slurried with methyl tert-butyl ether / petroleum ether (5 / 5 mL) to give a yellow solid 4-(5-bromo-2-nitrophenyl)-3-oxoperazine-1-carboxylic acid tert-butyl ester (18.24 g, yield 91.2%). LCMS(TOF MS ES+)m / z[M+H]+:400. 1 H NMR (400MHz, DMSO-d6) δ8.06(d,J=2.1Hz,1H),8.00(d,J=8.7Hz,1H),7.85(m,J=8.7,2.1Hz,1H),4.10–4.06(m,2H),3.74(s,4H),1.47(s,9H).

[0122] Step 2: Synthesis of tert-butyl 7-bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0123] 4-(5-bromo-2-nitrophenyl)-3-oxopiperazine-1-carboxylic acid tert-butyl ester (15.0 g, 37.59 mmol, 1.0 eq), iron powder (10.5 g, 187.96 mmol, 5.0 eq), and ammonium chloride (30.16 g, 563.85 mmol, 15.0 eq) were added to ethanol (150 mL) and water (50 mL), and reacted at 80 °C for 16 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth, the filtrate was concentrated to remove ethanol, and the residue was filtered to give a yellow solid 7-bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (8.8 g, yield 67.8%). LCMS (TOF MS ES+) m / z [M+H]+: 352. 1 H NMR (400MHz, DMSO-d6) δ7.80(d,J=1.9Hz,1H),7.54(d,J=8.6Hz,1H),7.34(m,J=8.6 ,1.9Hz,1H),4.77(s,2H),4.17(t,J=5.5Hz,2H),3.89(t,J=5.5Hz,2H),1.44(s,9H).

[0124] Step 3: Synthesis of tert-butyl 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0125] 7-Bromo-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (7.5 g, 21.30 mmol, 1.0 eq), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (6.5 g, 25.56 mmol, 1.2 eq), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (1.55 g, 2.13 mmol, 0.1 eq) and potassium acetate (5.21 g, 53.25 mmol, 2.5 eq) were added to 1,4-dioxane (80 mL), nitrogen was purged, and the reaction was carried out at 100 °C for 18 hours. After the reaction was complete, the mixture was filtered, the filtrate was concentrated, and purified by forward separation to give a yellow solid 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (5.1 g, yield 60.0%). LCMS (TOF MS ES+) m / z [M+H]+: 400.1 H NMR(400MHz,DMSO-d6)δ7.80(d,J=4.1Hz,1H),7.60–7.49(m,2H),4.78(d,J=9.3Hz,2H), 4.19 (m, J = 22.5, 5.5 Hz, 2H), 3.90 (t, J = 5.5 Hz, 2H), 1.45 (s, 9H), 1.11 (d, J = 35.8 Hz, 12H).

[0126] Step 4: Synthesis of tert-butyl 7-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid

[0127] The compounds 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborin-2-yl)-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (6.0 g, 15.03 mmol, 1.0 eq) and 3-bromo-7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one (6.0 g, 15.03 mmol, 1.0 eq) were prepared. 89 g (16.54 mmol, 1.1 eq), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (1.09 g, 1.50 mmol, 0.1 equiv.) and potassium carbonate (6.22 g, 45.09 mmol, 3.0 eq) were dissolved in 1,4-dioxane (60.0 mL) and water (12.0 mL), and nitrogen gas was immediately purged three times. The mixture was stirred at 90 °C for 18 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filtrate was concentrated, and purified by normal-phase column chromatography to obtain a yellow solid, 7-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (5.61 g, yield 61.3%). LCMS (TOF MS ES+) m / z [M+H]+: 609. 1 HNMR(400MHz,Chloroform-d)δ7.85–7.74(m,2H),7.48(d,J=4.4Hz,2H),7.46(s,1H),7.38(d,J=8.4Hz,1H),7.24(d, J=8.4Hz,2H),5.13(s,1H),4.14(d,J=9.4Hz,2H),3.98(s,2H),2.74(d,J=4.8Hz,3H),1.50(s,9H),1.35–1.20(m,2H).

[0128] Step 5: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-4-methylamino-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-1,8-naphthidium-2(1H)-one

[0129] 7-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrazine-2(1H)-carboxylic acid tert-butyl ester (1.0 g, 1.64 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (449 mg, 3.28 mmol, 2.0 eq) and tert-butyl Sodium alkoxide (314 mg, 3.28 mmol, 2.0 eq) was added to a dioxane solvent (20 mL), followed by the addition of methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (155 mg, 0.16 mmol, 0.1 eq). Nitrogen gas was then purged, and the reaction was carried out at 80 °C for 3 hours. The mixture was cooled to room temperature, and then 10 mL of trifluoroacetic acid was added to the system. The reaction was continued at room temperature for another 3 hours. After the reaction was complete, the reaction solution was concentrated, and sodium hydroxide solution was added to adjust the pH to 12. Then, dichloromethane (40 mL) was added for extraction and separation. The solution was dried and concentrated to obtain a yellow solid 7-amino-1-(4-chlorophenyl)-6-fluoro-4-methylamino-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-1,8-naphthidium-2(1H)-one (202 mg, yield 30.8%). LCMS (TOF MS ES+) m / z [M+H]+: 490.

[0130] Step 6: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0131] 7-amino-1-(4-chlorophenyl)-6-fluoro-4-methylamino-3-(1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-1,8-naphthid-2(1H)-one (100.0 mg, 0.20 mmol, 1.0 eq), potassium carbonate (41 mg, 0.3 mmol, 1.5 eq), and iodomethane (31.2 mg, 0.22 mmol, 1.1 eq) were added to N,N-dimethylformamide (2 mL) and reacted at room temperature for 1 hour. After the reaction was completed, the mixture was filtered and purified by high-performance liquid chromatography (HPLC) to obtain a white solid, 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyrazin-7-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (26.2 mg, yield 25.5%). LCMS (TOF MS ES+) m / z [M+H]+: 504. 1 H NMR(400MHz, DMSO-d6)δ8.14(d,J=12.3Hz,1H),7.56(d,J=8.2Hz,1H),7.52–7.46(m,2H),7.38(s,1H),7.30–7.21(m,3H),7 .09(m,J=8.3,1.5Hz,1H),5.95(s,2H),4.12(t,J=5.7Hz,2H),3.76(s,2H),2.92(t,J=5.6Hz,2H),2.48(s,3H),2.45(s,3H).

[0132] Example 3: Synthesis of 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (compound 15)

[0133]

[0134] Step 1: Synthesis of 2-{[(5-bromo-2H-indazol-3-yl)methyl](methyl)amino}ethanol-1-ol

[0135] 5-Bromo-2H-indazole-3-carboxaldehyde (10.0 g, 44.64 mmol, 1.0 eq) and 2-(methylamino)ethane-1-ol (3.4 g, 44.64 mmol, 1.0 eq) were dissolved in methanol (100 mL) and reacted at room temperature for 0.5 h. Then, sodium cyanoborohydride (3.37 g, 53.57 mmol, 1.2 eq) was added at room temperature, and the reaction was continued for another 3 h. After the reaction was complete, the mixture was extracted with ethyl acetate (100 mL), washed with saturated brine, and concentrated to remove the solvent, yielding a yellow solid 2-{[(5-bromo-2H-indazole-3-yl)methyl](methyl)amino}ethanol-1-ol (11.1 g, yield 88.1%), which was used directly in the next reaction. LCMS (TOF MS ES+) m / z [M+H] + :284.

[0136] Step 2: Synthesis of 9-bromo-2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole

[0137] 2-{[(5-bromo-2H-indazole-3-yl)methyl](methyl)amino}ethanol-1-ol (10.0 g, 35.33 mmol, 1.0 eq) and triphenylphosphine (13.9 g, 53.0 mmol, 1.5 eq) were dissolved in an ultra-dry tetrahydrofuran solution (150 mL). The mixture was stirred at 0 °C for 0.5 h, followed by the addition of diisopropyl azodicarbonate (10.7 g, 53.00 mmol, 1.5 eq) and the reaction was brought to room temperature for 2 h. After the starting material was detected by TLC, the reaction solution was concentrated to obtain a yellow oily substance. This substance was purified by forward chromatography (petroleum ether: ethyl acetate = 85%: 15%) to obtain the target product 9-bromo-2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole (3.2 g, yield 34.2%). LCMS(TOF MS ES+)m / z[M+H]+:266.

[0138] Step 3: Synthesis of (2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)boronic acid

[0139] 9-Bromo-2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]inazole (3.0 g, 11.32 mmol, 1.0 eq), diboronol ester (3.5 g, 13.58 mmol, 1.2 eq), 1,1-bis(diphenylphosphine)diberberine palladium dichloride (827 mg, 1.132 mmol, 0.1 eq) and potassium acetate (3.3 g, 33.96 mmol, 3.0 eq) were dissolved in 1,4-dioxane (120 mL) and reacted at 90 °C for 3 hours. After the reaction was completed, the solution was filtered through diatomaceous earth. The filtrate was concentrated and then purified by forward filtration (dichloromethane:methanol = 90%:10%) to obtain the target product (2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole-9-yl)boronic acid (1.8 g, yield 68.9%), LCMS (TOF MS ES+) m / z [M+H]+: 232.12.

[0140] Step 4: Synthesis of 6-chloro-5-fluoro-2-[(2-methyl-1H-pyrrole-3-yl)amino]nicotinic acid

[0141] 2-Methyl-1H-pyrrole-3-amine (20.00 g, 208.30 mmol, 2.0 eq) was dissolved in (200 mL) ultra-dry tetrahydrofuran and cooled to -78 °C. Bistrimethylsilylaminolithium (312.50 mL, 312.5 mmol, 3.0 eq) was slowly added dropwise. After the addition was complete, the reaction was continued for 1 hour. At -78 °C, 60 mL of ultra-dry tetrahydrofuran containing 2,6-dichloro-5-fluoronicotinic acid (21.87 g, 104.2 mmol, 1.0 eq) was slowly added dropwise to the reaction system. After the addition was complete, the system was transferred to room temperature and reacted for 1 hour. After the reaction was complete, a saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted with ethyl acetate (200 mL), washed with saturated brine, and concentrated to remove the solvent, yielding a light brown solid. Methanol was added and the mixture was slurried to obtain a pale yellow solid, 6-chloro-5-fluoro-2-[(2-methyl-1H-pyrrolo-3-yl)amino]nicotinic acid (22.1 g, yield 78.8%). LCMS (TOF MS ES+) m / z [M+H]+: 270.

[0142] Step 5: Synthesis of ethyl 3-{6-chloro-5-fluoro-2-[(2-methyl-1H-pyrrolo-3-yl)amino]pyridin-3-yl}-3-oxopropionate

[0143] Potassium monoethyl malonate (8.3 g, 48.66 mmol, 2.0 eq), magnesium chloride (6.7 g, 72.99 mmol, 3.0 eq), and triethylamine (9.8 g, 97.32 mmol, 4.0 eq) were dissolved in acetonitrile solution (150 mL) and stirred at room temperature to obtain system one. 6-Chloro-5-fluoro-2-[(2-methyl-1H-pyrrolo-3-yl)amino]nicotinic acid (6544 mg, 24.33 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (150 mL). A small amount of N,N-dimethylformamide was added as a catalyst, and oxaloyl chloride (5076 mg, 39.99 mmol, 1.5 eq) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature for further reaction. TLC confirmed that the reaction of the starting materials was complete. The reaction solution was then concentrated to obtain a yellow solid. The yellow solid was dissolved in tetrahydrofuran solution (100 mL) and slowly added dropwise to system one at 0 °C. The reaction was continued at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain a yellow solid. Forward purification yielded the target product, ethyl 3-{6-chloro-5-fluoro-2-[(2-methyl-1H-pyrrolo-3-yl)amino]pyridin-3-yl}-3-oxopropionate (6.4 g, yield 78.1%). LCMS(TOF MS ES+)m / z[M+H]+:340.

[0144] Step 6: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthidium-2(1H)-one

[0145] Ethyl 3-{6-chloro-5-fluoro-2-[(2-methyl-1H-pyrrolo-3-yl)amino]pyridin-3-yl}-3-oxopropionate (6.3 g, 18.58 mmol, 1.0 eq) was dissolved in anhydrous ethanol (120 mL), and potassium carbonate (5.1 g, 37.16 mmol, 2.0 eq) was added. The reaction was carried out at 70 °C for 3 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth. The filtrate was concentrated and purified by forward filtration (dichloromethane:methanol = 85%:15%) to obtain the target product 7-chloro-6-fluoro-4-hydroxy-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthidium-2(1H)-one (4.8 g, yield 88.2%), LCMS (TOF MS ES+) m / z [M+H]+: 294.

[0146] Step 7: Synthesis of (4,7-dichloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthidium-2(1H)-one

[0147] 7-Chloro-6-fluoro-4-hydroxy-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthid-2(1H)-one (4.8 g, 16.4 mmol, 1.0 eq) was added to the reaction flask, followed by phosphorus oxychloride (50 mL). The mixture was heated to 80 °C and reacted for 1 hour. After the reaction was complete, the phosphorus oxychloride was removed by concentration under reduced pressure. The product was then slurried with ethyl acetate and filtered to obtain the target product (4,7-dichloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthid-2(1H)-one (4.2 g, yield 82.7%). LCMS (TOF MS ES+) m / z [M+H]+: 312.

[0148] Step 8: Synthesis of 7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0149] (4,7-Dichloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-1,8-naphthidin-2(1H)-one (908 mg, 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 the reaction was completed, the reaction solution was concentrated to give a pale yellow solid. The solid was slurried and filtered with ethyl acetate to give a yellow solid 7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidin-2(1H)-one (525 mg, yield 58.8%), which was directly used for the next reaction. LCMS (TOF MSES+) m / z [M+H]+: 307.

[0150] Step 9: Synthesis of 3-bromo-7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0151] 7-Chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidin-2(1H)-one (360 mg, 1.18 mmol, 1.0 eq) and N-succinimide bromide (232.4 mg, 1.30 mmol, 1.1 eq) were added to acetonitrile solution (8 mL) and reacted at room temperature for 4 hours. After the reaction was complete, the reaction solution was directly mixed with silica gel and separated in the forward direction (dichloromethane:methanol = 20:1) to give a pale yellow solid 3-bromo-7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidin-2(1H)-one (320 mg, yield 70.95%). LCMS (TOF MS ES+) m / z [M+H]+: 385.

[0152] Step 10: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0153] 3-Bromo-7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidin-2(1H)-one (276 mg, 0.73 mmol, 1.0 eq), (2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole-9-yl)boronic acid (254 mg, 1.10 mmol, 1.5 eq) and potassium phosphate (384 mg, 1.82 mmol, 2.5 eq) were added to a mixed solvent of dioxane (60 mL) and water (6 mL). Finally, 1,1-bis(diphenylphosphine)diberberine palladium dichloride (52.9 mg, 0.073 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, a reverse-phase column chromatography was used to prepare a yellow solid 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (152 mg, yield 43.3%). LCMS (TOF MS ES+) m / z [M+H]+: 492.

[0154] Step 11: Synthesis of 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0155] 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthid-2(1H)-one (63.0 mg, 0.128 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (176 mg, 1.28 mmol, 10.0 eq), and tert-butyl... Sodium alkoxide (24.5 mg, 0.257 mmol, 2.0 eq) was added to dioxane (3.0 mL), followed by (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). The reaction was carried out at 60 °C for 3 hours. After the reaction was completed, the reaction solution was concentrated, and trifluoroacetic acid (2.5 mL) was added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated directly, and preparative chromatography yielded a white solid 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole-9-yl)-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (18.3 mg, yield 30.21%). LCMS (TOF MS ES+) m / z [M+H]+: 473.21.

[0156] Example 4: Synthesis of 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (compound 16)

[0157]

[0158] Step 1: Synthesis of 6-chloro-5-fluoro-2-[(2-methylthiophene-3-yl)amino]nicotinic acid

[0159] 2-Methylthiophen-3-amine (20.00 g, 177.0 mmol, 2.0 eq) was dissolved in 200 mL of ultra-dry tetrahydrofuran and cooled to -78 °C. Bistrimethylsilylaminolithium (265.50 mL, 265.5 mmol, 3.0 eq) was slowly added dropwise. After the addition was complete, the reaction was continued for 1 hour. 2,6-Dichloro-5-fluoronicotinic acid (18.57 g, 88.5 mmol, 1.0 eq) was dissolved in 60 mL of ultra-dry tetrahydrofuran 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. The reaction was quenched with saturated ammonium chloride solution, extracted with 200 mL of ethyl acetate, washed with saturated brine, and concentrated to remove the solvent, yielding a light brown solid. Methanol was added and the mixture was slurried to obtain a pale yellow solid, 6-chloro-5-fluoro-2-[(2-methylthiophen-3-yl)amino]nicotinic acid (23.2 g, yield 84.8%). LCMS(TOF MS ES+)m / z[M+H]+:287.

[0160] Step 2: Synthesis of ethyl 3-{6-chloro-5-fluoro-2-[(2-methylthiophen-3-yl)amino]pyridin-3-yl}-3-oxopropionate

[0161] Potassium monoethyl malonate (11.9 g, 69.68 mmol, 2.0 eq), magnesium chloride (9.6 g, 104.37 mmol, 3.0 eq), and triethylamine (14.0 g, 138.19 mmol, 4.0 eq) were dissolved in acetonitrile solution (150 mL) and stirred at room temperature to obtain system one. 10.0 g (34.84 mmol, 1.0 eq) of 6-chloro-5-fluoro-2-[(2-methylthiophen-3-yl)amino]nicotinic acid was dissolved in 150 mL of ultra-dry dichloromethane. A small amount of N,N-dimethylformamide was added as a catalyst. Oxaloyl chloride (7.2 g, 56.78 mmol, 1.5 eq) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature for further reaction. TLC confirmed complete reaction of the reactants. The reaction solution was concentrated to obtain a yellow solid. The yellow solid was dissolved in 100 mL of tetrahydrofuran solution and slowly added dropwise to system one at 0 °C. The reaction was continued at room temperature for 2 hours. After the reaction was complete, the solution was concentrated to obtain a yellow solid. Forward purification yielded the target product, ethyl 3-{6-chloro-5-fluoro-2-[(2-methylthiophen-3-yl)amino]pyridin-3-yl}-3-oxopropionate (9.4 g, yield 75.6%). LCMS(TOF MS ES+)m / z[M+H]+:357.

[0162] Step 3: Synthesis of 7-chloro-6-fluoro-4-hydroxy-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0163] Ethyl 3-{6-chloro-5-fluoro-2-[(2-methylthiophen-3-yl)amino]pyridin-3-yl}-3-oxopropionate (8.0 g, 22.40 mmol, 1.0 eq) was dissolved in anhydrous ethanol (120 mL), and potassium carbonate (6.1 g, 44.20 mmol, 2.0 eq) was added. The reaction was carried out at 70 °C for 3 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was concentrated and purified by forward filtration (dichloromethane:methanol = 85%:15%) to give the target product 7-chloro-6-fluoro-4-hydroxy-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (4.8 g, yield 69.0%), LCMS (TOF MS ES+) m / z [M+H]+: 311.

[0164] Step 4: Synthesis of 4,7-dichloro-6-fluoro-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0165] 4.5 g (14.5 mmol, 1.0 eq) of 7-chloro-6-fluoro-4-hydroxy-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one was added to phosphorus oxychloride (50 mL) and reacted at 80 °C for 16 hours. After the reaction was completed, the phosphorus oxychloride was removed by concentration under reduced pressure, and the mixture was slurried with ethyl acetate and filtered to obtain the target product 4,7-dichloro-6-fluoro-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (3.9 g, yield 81.9%). LCMS (TOF MS ES+) m / z [M+H]+: 329.

[0166] Step 5: Synthesis of 7-chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0167] 4,7-Dichloro-6-fluoro-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (957 mg, 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 eq) were dissolved in acetonitrile (20 mL) and reacted at 80 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated to give a pale yellow solid. The solid was slurried and filtered with ethyl acetate to give a yellow solid 7-chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (475 mg, yield 50.4%), which was directly used for the next step of the reaction. LCMS (TOF MS ES+) m / z [M+H]+: 324.

[0168] Step 6: Synthesis of 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0169] 7-Chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (400 mg, 1.23 mmol, 1.0 eq) and N-succinimide bromide (264.6 mg, 1.48 mmol, 1.1 eq) were added to acetonitrile solution (8 mL) and reacted at room temperature for 4 hours. After the reaction was complete, the reaction solution was stirred with silica gel and separated in the forward direction (dichloromethane:methanol = 20:1) to give a pale yellow solid 3-bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (317 mg, yield 63.9%). LCMS (TOF MS ES+) m / z [M+H]+: 402.

[0170] Step 7: Synthesis of 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0171] 3-Bromo-7-chloro-6-fluoro-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (300 mg, 0.75 mmol, 1.0 eq), (2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole-9-yl)boronic acid (260 mg, 1.13 mmol, 1.5 eq) and potassium phosphate (395 mg, 1.88 mmol, 2.5 eq) were added to a mixed solvent of dioxane (60 mL) and water (6 mL). Finally, 1,1-bis(diphenylphosphine)diberberine palladium dichloride (54.5 mg, 0.075 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, a reverse-phase column chromatography was used to prepare a yellow solid 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (186 mg, yield 49.1%). LCMS (TOF MS ES+) m / z [M+H]+: 508.

[0172] Step 8: Synthesis of 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one

[0173] 7-chloro-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (100 mg, 0.197 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (270 mg, 0.0.197 mmol, 10.0 eq), and sodium tert-butoxide were administered. (37.8 mg, 0.395 mmol, 2.0 eq) was 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) (18.7 mg, 0.0197 mmol, 0.1 eq) was added. The mixture was reacted at 60 °C for 3 hours. After the reaction was completed, the reaction solution was concentrated, and trifluoroacetic acid (2.5 mL) was added. The mixture was stirred at room temperature for 1 hour. The reaction solution was then directly concentrated and dissolved in N,N-dimethylformamide (2.0 mL). Preparative chromatography yielded a white solid 7-amino-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1-(2-methylthiophen-3-yl)-1,8-naphthidin-2(1H)-one (31.5 mg, yield 32.64%). LCMS (TOF MS ES+) m / z [M+H]+: 490.17.

[0174] Example 5: Synthesis of 7-amino-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidium-2(1H)-one (compound 17)

[0175]

[0176] Step 1: Synthesis of tert-butyl 4-(4-bromo-2-nitrophenyl)-3-oxo-1,4-diazacycloheptan-1-carboxylate

[0177] 20 g (93.34 mmol, 1 eq.) of tert-butyl 3-oxo-1,4-diazacycloheptan-1-carboxylate, 30.5 g (93.34 mmol, 1 eq.) of 4-bromo-1-iodo-2-nitrobenzene, 38.7 g (280.02 mmol, 3 eq.) of potassium carbonate, 4.27 g (4.67 mmol, 0.05 eq.) of tris(dibenzylacetone)dipalladium, and 5.4 g (9.33 mol, 0.1 eq.) of 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene were added to toluene (200 mL). The reaction was refluxed at 100 °C for 18 h under nitrogen protection. After the reaction was complete, the reaction solution was concentrated, diluted with water (200 mL), and extracted three times with ethyl acetate (150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and separated by normal phase separation (ethyl acetate / petroleum ether = 1 / 5) to give a yellow solid product, tert-butyl 4-(4-bromo-2-nitrophenyl)-3-oxo-1,4-diazacycloheptan-1-carboxylate (18.66 g, yield 48.26%). LCMS: [M+H] = 414.06.

[0178] Step 2: Synthesis of 1-(4-bromo-2-nitrophenyl)-1,4-diazepam-2-one

[0179] 18 g (93.34 mmol, 1 eq.) of tert-butyl 4-(4-bromo-2-nitrophenyl)-3-oxo-1,4-diazacycloheptan-1-carboxylate was added to a mixed solution of trifluoroacetic acid (20 mL) and dichloromethane (60 mL), and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, the crude product 1-(4-bromo-2-nitrophenyl)-1,4-diazepam-2-one (17.42 g) was concentrated and used directly in the next step. LCMS: [M+H] = 414.06.

[0180] Step 3: Synthesis of 1-(4-bromo-2-nitrobenzene)-4-ethyl-1,4-diazepam-2-one

[0181] 1-(4-bromo-2-nitrophenyl)-1,4-diazepam-2-one (15 g, 47.75 mmol, 1.0 eq), acetaldehyde (15 mL, 50% wt), and sodium cyanoborohydride (4.50 g, 71.63 mmol, 1.5 eq) were dissolved in methanol (120 mL) and reacted at room temperature for 16 hours. After the reaction was completed, the reaction solution was concentrated and purified by forward purging (dichloromethane:methanol = 85%:15%) to give the target product 1-(4-bromo-2-nitrophenyl)-4-ethyl-1,4-diazepam-2-one (8.8 g, yield 53.86%). LCMS (TOF MS ES+) m / z [M+H]+: 341.14.

[0182] Step 4: Synthesis of 9-bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0183] 1-(4-bromo-2-nitrobenzene)-4-ethyl-1,4-diazepam-2-one (8.8 g, 25.71 mmol, 1.0 eq), reduced iron powder (14.40 g, 257.1 mmol, 10 eq), and ammonium chloride (13.75 g, 257.1 mmol, 10 eq) were added to a mixed solution of ethanol (200 mL) and water (200 mL), and reacted at 80 °C for 16 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and normal-phase column chromatography was used to separate the target product 9-bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphene (4.2 g, yield 55.51%). LCMS (TOF MS ES+) m / z [M+H]+: 295.20.

[0184] Step 5: Synthesis of (2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)boronic acid

[0185] 9-Bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (3.3 g, 11.32 mmol, 1.0 eq), diboronpinacol ester (3.5 g, 13.58 mmol, 1.2 eq), 1,1-bis(diphenylphosphine)diberberine palladium dichloride (827 mg, 1.132 mmol, 0.1 eq), and potassium acetate (3.3 g, 33.96 mmol, 3.0 eq) were dissolved in 1,4-dioxane (20 mL) and reacted at 90 °C for 3 hours. After the reaction was completed, the solution was filtered through diatomaceous earth. The filtrate was concentrated and purified by forward filtration (dichloromethane:methanol = 80%:20%) to obtain the target product (2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)boronic acid (1.1 g, yield 37.84%). LCMS (TOF MS ES+) m / z [M+H]+: 260.15.

[0186] Step 6: Synthesis of 7-chloro-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidine-2(1H)-one

[0187] 3-Bromo-7-chloro-6-fluoro-1-(2-methyl-1H-pyrrolo-3-yl)-4-(methylamino)-1,8-naphthid-2(1H)-one (276 mg, 0.73 mmol, 1.0 eq), (2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)boronic acid (285 mg, 1.10 mmol, 1.5 eq) and potassium phosphate (384 mg, 1.82 mmol, 2.5 eq) were added to a mixed solvent of dioxane (60 mL) and water (6 mL). Finally, 1,1-bis(diphenylphosphine)diferropalladium dichloride (52.9 mg, 0.073 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, a reverse-phase column chromatography was used to prepare a yellow solid 7-chloro-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidium-2(1H)-one (166 mg, yield 45.48%). LCMS (TOF MSES+) m / z [M+H]+: 551.15.

[0188] Step 7: 7-Amino-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidium-2(1H)-one

[0189] 7-chloro-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidium-2(1H)-one (150 mg, 0.27 mmol, 1.0 eq), (4-methoxyphenyl)methylamine (377 mg, 2.72 mmol, 10.0 eq), and tert-butanol were added. Sodium (51 mg, 0.54 mmol, 2.0 eq) was added to a dioxane solvent (3.0 mL), followed by (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). The reaction was carried out under nitrogen protection at 60 °C for 3 hours. Subsequently, trifluoroacetic acid (2.5 mL) was added to the system, and the mixture was stirred at room temperature for another 1 hour. After the reaction was complete, the reaction solution was concentrated, and preparative chromatography yielded a white solid 7-amino-1-(4-chlorophenyl)-3-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-6-fluoro-4-methylamino-1,8-naphthidine-2(1H)-one (28.3 mg). LCMS (TOF MS ES+) m / z [M+H]+: 532.19.

[0190] Example 6: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1,8-naphthidin-2(1H)-one (compound 6) and 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1,8-naphthidin-2(1H)-one (compound 5).

[0191]

[0192] Step 1: Synthesis of 5-bromo-2H-indazole-3-carboxaldehyde

[0193] 5-Bromo-indole (25.0 g, 12.83 mol, 1.0 eq) was dissolved in a mixed solution of acetone (400 mL) and water (100 mL). Sodium nitrate (12.25 g, 37.58 mol, 2.0 eq) was added at -10 °C, followed by dropwise addition of hydrochloric acid (308 mL). The reaction was carried out at room temperature for 18 hours. After the reaction was complete, the pH was adjusted to neutral, the mixture was extracted with ethyl acetate, the organic phase was concentrated, and a suitable amount of dichloromethane was added and stirred for 30 minutes. The mixture was filtered and dried to obtain a yellow solid, 5-bromo-2H-indazole-3-carboxaldehyde (14.8 g, yield 51.28%). LCMS (ESI) [M+H]+: 224.97.

[0194] Step 2: Synthesis of 2-{[(5-bromo-2H-indazol-3-yl)methyl]amino}ethanol-1-ol

[0195] 14.8 g (66.07 mmol, 1.0 eq) of 5-bromo-2H-indazole-3-carboxaldehyde was dissolved in a mixture of methanol and tetrahydrofuran (100 mL), and acetic acid (7.93 g, 132.14 mmol, 2.0 eq) was added. The reaction was carried out at room temperature for 0.5 h, followed by the addition of sodium cyanoborohydride (4.91 g, 79.28 mmol, 1.2 eq). After the reaction was complete, the pH was adjusted to neutral, and the mixture was extracted with dichloromethane and purified by normal phase to obtain a yellow solid crude product, 2-{[(5-bromo-2H-indazole-3-yl)methyl]amino}ethanol-1-ol (21.2 g), which was used directly in the next step of the reaction. LCMS (ESI) [M+H]+: 270.02.

[0196] Step 3: Synthesis of (5-bromo-2H-indazol-3-yl)methyl)(2-hydroxyethyl)carbamate tert-butyl ester

[0197] 2-{[(5-bromo-2H-indazole-3-yl)methyl]amino}ethanol-1-ol (8.0 g, 29.6 mmol, 1.0 eq) was dissolved in ultradry dichloromethane (100.0 mL). Triethylamine (8.96 g, 88.8 mol, 3.0 eq) and di-tert-butyl dicarbonate (5.17 g, 23.7 mol, 0.8 eq) were added dropwise under ice bath conditions, and the reaction was carried out at room temperature for 18 hours. After the reaction was complete, the mixture was purified in normal phase (petroleum ether:ethyl acetate = 1:4) to give a yellow solid (5-bromo-2H-indazole-3-yl)methyl)(2-hydroxyethyl)carbamate tert-butyl ester (2.4 g, yield 22.02%). LCMS (ESI) [M+H]+: 370.07. 1H NMR(400MHz,DMSO-d6)δ13.11(d,J=10.5Hz,1H),7.96(d,J=1.9Hz,1H),7.52– 7.44(m,2H),4.72(s,2H),3.43–3.39(m,2H),3.22–3.04(m,2H),1.44(s,9H).

[0198] Step 4: Synthesis of tert-butyl 9-bromo-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid

[0199] (5-Bromo-2H-indazole-3-yl)methyl)(2-hydroxyethyl)carbamate tert-butyl ester (2.19 g, 5.94 mmol, 1.0 eq) was dissolved in ultradry tetrahydrofuran (40.0 mL). Under ice bath conditions, a tetrahydrofuran solution of triphenylphosphine (2.33 g, 8.91 mmol, 1.5 eq) and di-tert-butyl azodicarbonate (2.05 g, 8.91 mmol, 1.5 eq) was added dropwise. The reaction was carried out at room temperature for 2.5 hours. After the reaction was complete, the product was purified by normal phase (petroleum ether:ethyl acetate = 3:2) to give a white solid 9-bromo-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (1.41 g, yield 72.68%). LCMS (ESI) [M+H]+: 352.06. 1 H NMR (400MHz, DMSO) δ8.05(d,J=1.9Hz,1H),7.54(d,J=9.1Hz,1H),7.33(m,J=9.1,1 .9Hz, 1H), 4.93 (s, 2H), 4.42 (t, J = 5.4Hz, 2H), 3.92 (t, J = 5.5Hz, 2H), 1.46 (s, 9H).

[0200] Step 5: Synthesis of tert-butyl 9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid

[0201] 9-Bromo-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (450 mg, 1.27 mmol, 1.0 eq) was dissolved in ultra-dry 1,4-dioxane (100 mL), and then pinacol diboronate (649 mg, 2.55 mmol, 2.0 eq), potassium acetate (373 mg, 3.81 mmol, 3.0 eq), and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (92.9 mg, 0.127 mmol, 0.1 eq) were added sequentially. The mixture was reacted at 80 °C for 2 hours. After the reaction was complete, ethyl acetate was added for extraction and separation. The organic phase was concentrated and purified by reverse phase to obtain a white solid crude product, 9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (670 mg), which was directly used in the next reaction. LCMS (ESI) [M+H]+: 400.2.

[0202] Step 6: Synthesis of tert-butyl 9-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid

[0203] 9-(4,4,5,5-tetramethyl-1,3,2-dioxaborin-2-yl)-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (300 mg, 7.51 mmol, 1.0 eq) and 3-bromo-7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-1,8-naphthidium-2(1H)-one (313 mg, 7.51 mmol, 1.0 eq) were dissolved in 1,4-dioxane (10 mL) and water (2.5 mL), followed by the addition of potassium carbonate (3... 11 mg (22.5 mmol, 3.0 eq) and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (27.5 mg, 0.37 mmol, 0.05 eq) were reacted at 80 °C for 2 h. The mixture was purified by normal-phase chromatography (petroleum ether:ethyl acetate = 1:4) to give a yellow solid 9-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (480 mg). LCMS (ESI) [M+H]+: 609.15. 1H NMR (400MHz, DMSO) δ7.86(d,J=11.2Hz,1H),7.71(s,1H),7.57(d,J=8.4Hz,3H),7.21(m,J=6.0, 1.5Hz, 3H), 4.95 (s, 2H), 4.44 (t, J = 5.8Hz, 2H), 3.95 (d, J = 6.6Hz, 2H), 1.99 (s, 3H), 1.46 (s, 9H).

[0204] Step 7: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1,8-naphthidium-2(1H)-one

[0205] 9-[7-chloro-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-2-oxo-1,2-dihydro-1,8-naphthopyridin-3-yl]-3,4-dihydropyrazino[1,2-b]indazole-2(1H)-carboxylic acid tert-butyl ester (400 mg, 0.65 mmol, 1.0 eq) was dissolved in ultra-dry 1,4-dioxane (4 mL), and sodium cyanoborohydride (267 mg, 1.95 mmol, 3.0 eq), sodium tert-butoxide (187.2 mg, 1.95 mmol, 3.0 eq), and methanesulfonic acid (4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene)(2-amino-1,1'-biphenyl-2-yl)palladium (61.7 mg, 0.065 mmol, 0.1 eq) were added. The mixture was reacted at 100 °C for 2 hours. After the reaction was complete, the reaction solution was concentrated, and then trifluoroacetic acid (8 mL) was added, and the reaction was continued for 3 hours. After the reaction was completed, the mixture was extracted with ethyl acetate, and the organic phase was concentrated and dried to obtain a yellow solid crude product, 7-amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1,8-naphthidium-2(1H)-one (303 mg), which was directly used in the next reaction. LCMS (ESI) [M+H]+: 489.1.

[0206] Step 8: Synthesis of 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one

[0207] 7-Amino-1-(4-chlorophenyl)-6-fluoro-4-(methylamino)-3-(1,2,3,4-tetrahydropyrazino[1,2-b]indazol-9-yl)-1,8-naphthid-2(1H)-one (59.53 mg, 0.122 mmol, 1.0 eq) and potassium carbonate (50.7 mg, 0.367 mmol, 3.0 eq) were dissolved in ultradry N,N-dimethylformamide (2.5 mL). Iodimethane (13.9 mg, 0.98 mmol, 0.8 eq) was slowly added dropwise under ice bath conditions, followed by reaction at room temperature for 1.5 hours. After the reaction was complete, a white solid 7-amino-1-(4-chlorophenyl)-6-fluoro-3-(2-methyl-1,2,3,4-tetrahydropyrazino[1,2-b]indazole-9-yl)-4-(methylamino)-1,8-naphthidium-2(1H)-one (3.3 mg, yield 5.38%) was prepared. LCMS (ESI) [M+H]+: 504.1754. 1 H NMR (400MHz, DMSO) δ8.14(d,J=12.3Hz,1H),7.55–7.45(m,4H),7.27–7.23(m,3H),7.11(m,J=9.1,1. 4Hz,1H),5.96(s,2H),4.41(t,J=5.5Hz,2H),3.94(s,2H),2.98(t,J=5.6Hz,2H),2.49–2.47(m,6H).

[0208] The compounds listed in Table 1 below were prepared using methods similar to those described in the examples, with appropriate variations in the amounts of reactants and reagents, protection and deprotection, solvents, and reaction conditions. Characterization data for the compounds are summarized in Table 1 below.

[0209] Table 1: Structure and characterization of some compounds

[0210]

[0211]

[0212] Experimental Example 1: The effect of the compound of the present invention on HCT116 MTAP - / - Assay for cell proliferation

[0213] Experimental Objective: The purpose of this test case is to test the effect of the compound on HCT116 MTAP. - / - Cell proliferation.

[0214] Background and Principle: Methionine adenosine transferase 2A (MAT2A) is considered a synthetic lethal target in cancers with methylthioadenosine phosphorylase (MTAP) gene deletion. The MTAP gene is adjacent to the CDKN2A tumor suppressor and is co-deleted with CDKN2A in approximately 15% of cancers. Therefore, the effect of compounds on HCT116 MTAP was investigated. - / - The inhibition rate of cell proliferation was used for screening MAT2A protein inhibitors.

[0215] Specific experimental procedure:

[0216] HCT116 MTAP knockout cells were constructed and single clones were screened. HCT116 MTAP cells in logarithmic growth phase were then... - / - WT cells were seeded in 96-well plates at 90 μL per well, 1000 cells / well, and incubated overnight at 37°C. The next day, 10 μL of different concentrations of the compound (DMSO final concentration 1%) were added, and the plates were incubated at 37°C for 10 days. On day 10, the old medium was aspirated, and 110 μL of medium (medium to CCK8 ratio 100:10) was added, and the plates were incubated at 37°C for 1–4 hours. Absorbance was measured at 450 nM, and IC50 was calculated using GraphPad software. Compounds were screened by comparison with positive control drugs. IC50 (half maximal inhibitory concentration) refers to the half-inhibitory concentration of the measured antagonist. It indicates the amount by which a drug or substance (inhibitor) inhibits certain biological processes (or substances contained in these processes, such as enzymes, cell receptors, or microorganisms). AG270 (compound 153 reported in CN109890822A) from Angios Pharmaceuticals Ltd. was used as a positive control compound.

[0217] The experimental results are shown in Table 2 below.

[0218] Table 2: IC50 Experimental Data

[0219] Compound numbering IC50 Compound numbering IC50 AG270 578.7 1 371.2 2 41.34 3 177.62 4 51.49 5 91.54 6 49.45 7 38.72

[0220] The results show that the compounds of the present invention are effective against HCT116 MTAP. - / - It has a good inhibitory effect on cells.

[0221] Experimental Example 2: Determination of the functional effect of the compounds of the present invention on MAT2A protease

[0222] Experimental objective: The purpose of this test case is to test the inhibitory effect of the compound on the function of the MAT2A protease.

[0223] Background and Principle: The metabolic enzyme methionine adenosyltransferase 2A (MAT2A) plays a crucial role in metabolism and epigenetics because it is the major producer of the universal methyl donor s-adenosylmethionine (SAM). ATP and L-Met generate SAM and phosphate groups under the action of MAT2A. Therefore, after compound incubation, the inhibitory effect of the compound on MAT2A enzyme function is evaluated by detecting the amount of SAM produced, which is used for screening MAT2A protein inhibitors.

[0224] Specific experimental procedure:

[0225] MAT2A protein expression: The full-length MAT2A protein was cloned into the pET24N vector containing an n-terminal (His) 6x marker and a tobacco corrosion virus (TEV) protease cleavage site. The constructed vector was transformed into *E. coli* BL21(DE3), and after shaking to an OD of 0.6, 1 mM IPTG was added and the culture was incubated at 18°C ​​for 16 h. The bacterial cells were collected, sonicated, centrifuged, and the supernatant was collected. The protein was purified by Ni-NTA, and the protein concentration and purity were determined after dialysis. SAM assay: Reaction system: 91-x μL of 50 mM TrisHCl (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 drug dissolved in DMSO, and x μL of MAT2A protein were added sequentially. Different drug concentrations were prepared for the experimental groups. 2 μL (100x) of each concentration was added to the reaction system, and the reaction was carried out at 37°C for 18 h. A solvent control group and a blank control group were also included. After the reaction was terminated, 40 μL of the system was removed, and 4 μL of 10% SDS was added to quench the reaction. IC50 was calculated using GraphPad software, and compounds were screened by comparison with positive control drugs.

[0226] The experimental results are shown in Table 2 below.

[0227] Table 2: IC50 Experimental Data

[0228] Compound numbering IC50 Compound numbering IC50 AG270 70.77 1 49.03 2 16.82 3 30.46 4 9.46 5 27.25 6 8.96 7 7.56

[0229] The results showed that the compound of the present invention has a good inhibitory effect on the function of MAT2A protease.

[0230] The above examples are merely representative. As can be seen from the above examples, the compounds of the present invention are ideal and highly effective MAT2A inhibitors, and are expected to be used for the treatment or prevention of conditions or diseases related to MAT2A inhibition.

Claims

1. A compound of formula (I) or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in, Ring A is selected from tricyclic heteroaryl and tricyclic heterocyclic groups, wherein the tricyclic heteroaryl or tricyclic heterocyclic group is optionally substituented by 1 to 3 identical or different substituents R. a replace; R3 is selected from a 5-12 member aromatic ring or heteroaromatic ring, wherein the 5-12 member aromatic ring or heteroaromatic ring is optionally replaced by one or more identical or different substituents R. a replace; R a Selected from hydrogen, deuterium, halogen, cyano, nitro, amide, sulfonamide, hydroxyl, amino, urea, phosphoryl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl, C1-C6 alkyl-S-, C1-C6 alkyl-SO-, C1-C6 alkyl-SO2-; or two of the above R a It can form 3-12 member saturated or partially unsaturated or aromatic ring systems through carbon chains or heteroatoms; R1 and R2 are independently selected from hydrogen, C1-C6 alkyl, and C3-C4 alkyl groups, respectively. 12 cycloalkyl or heterocycloalkyl; R4 and R5 are independently selected from hydrogen, deuterium, halogen, cyano, nitro, amide, sulfonamide, hydroxyl, amino, urea, phosphoryl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl; or R4 and R5 can form 3-12 membered saturated or partially unsaturated or aromatic ring systems through carbon atoms.

2. The compound according to claim 1, characterized in that, It is a compound of formula (II) or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof: Wherein, A, R3, R4, and R5 are as defined in claim 1.

3. The compound according to any one of claims 1-2, characterized in that, The ring A is selected from 10-20 denoted tricyclic heteroaryl groups and 10-20 denoted tricyclic heterocyclic groups, wherein the 10-20 denoted tricyclic heteroaryl group and 10-20 denoted tricyclic heterocyclic group are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, or C1-C6 haloalkyl groups.

4. The compound according to any one of claims 1-2, characterized in that, The ring A is selected from 12-14 membered trianneal heteroaryl and 12-14 membered trianneal heterocyclic group, wherein the 12-14 membered trianneal heteroaryl and 12-14 membered trianneal heterocyclic group are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, or C1-C6 haloalkyl groups.

5. The compound according to claim 4, characterized in that, The ring A is selected from 13-membered trianneal heteroaryl and 13-membered trianneal heterocyclic group, wherein the 13-membered trianneal heteroaryl and 13-membered trianneal heterocyclic group are optionally substituted by one or more halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, or C1-C6 haloalkyl groups.

6. The compound according to claim 1, characterized in that, It is a compound of formula (III) or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof: in, X is selected from N, O, and S; Y is selected from N and CH; R6 is selected from hydrogen, deuterium, halogen, cyano, nitro, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl. C1-C6 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C6 monoalkylamino, C1-C6 dialkylamino, 3-8 membered cycloalkyl or heterocycloalkyl; Ring A is selected from The above-mentioned ring A may be optionally substituted with one or more hydrogens, deuteriums, halogens, cyano groups, nitro groups, hydroxyl groups, amino groups, C1-C6 alkyl groups, C1-C6 alkoxy groups, C1-C6 haloalkyl groups, C1-C6 haloalkoxy groups, C2-C4 alkenyl groups, C2-C4 alkynyl groups, C1-C6 monoalkylamino groups, C1-C6 dialkylamino groups, 3-8 membered cycloalkyl groups, or heterocyclic alkyl groups; n is 0, 1, 2, or 3; R1, R2, R4, and R5 are as defined in claim 1.

7. The compound according to claim 6, characterized in that, R1 and R2 are independently selected from hydrogen, C1-C6 alkyl, and C3-C4 alkyl groups, respectively. 12 Cycloalkyl.

8. The compound according to claims 6-7, characterized in that, It is a compound of formula (IV) or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof:

9. The compound according to any one of claims 6-8, characterized in that, R6 is selected from hydrogen, halogen, C1-C6 alkyl, cyano, and C1-C6 alkoxy.

10. The compound according to any one of claims 1-9, characterized in that, R4 is selected from hydrogen, halogen, amino, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl, preferably hydrogen, amino, halogen, or C1-C6 alkyl, and more preferably hydrogen, amino, or chlorine.

11. The compound according to claims 1-10, characterized in that, R5 is selected from hydrogen, halogen, amino, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkyl, preferably hydrogen, amino, halogen, or C1-C6 alkyl, and more preferably hydrogen, fluorine, or methyl.

12. A compound or its tautomers, meso compounds, racemates, enantiomers, diastereomers, mixtures thereof, or pharmaceutically acceptable salts thereof, selected from:

13. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12, or a tautomer, meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

14. The use of the compound of any one of claims 1 to 12, or a tautomer, meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition of claim 13, in the preparation of a medicament for use in a subject suffering from a disease or condition associated with the activity or expression of MAT2A or MTAP protein, wherein the disease or condition is preferably cancer or an autoimmune disease, wherein the cancer is preferably lung cancer, pancreatic cancer, liver cancer, colorectal cancer, bile duct cancer, gallbladder cancer, brain cancer, gastric cancer, leukemia, lymphoma, melanoma, thyroid cancer, nasopharyngeal carcinoma, glioma, bladder cancer, astrocytoma, basal cell carcinoma, osteosarcoma, head and neck cancer, chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, and mesothelioma; wherein the autoimmune disease is preferably thyroiditis, inflammatory bowel disease, lupus erythematosus, fibrosis, myasthenia gravis, vasculitis, psoriasis, arthritis, scleroderma, or dermatitis.

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