PRMT5 inhibitor and application thereof

Abstract

The invention provides a compound with methyltransferase inhibitory activity, and particularly provides a compound with PRMT5 inhibitory activity. The compound can be used for preparing a pharmaceutical composition for treating diseases related to PRMT5 activity. # imgabs0 #

Description

A class of PRMT5 inhibitors and uses thereof Technical Field

[0001] The present invention relates to the field of pharmaceutical compounds. Specifically, the present invention provides a class of compounds for inhibiting PRMT5, and their use in pharmaceutical compositions. Background Art

[0002] Epigenetic regulation of gene expression is an important biological determinant of protein production and cell differentiation and plays a significant pathogenic role in many human diseases.

[0003] Epigenetic regulation involves the heritable modification of genetic material without changing its nucleotide sequence. Typically, epigenetic regulation is mediated by selective and reversible modifications of DNA and proteins (e.g., histones), such as methylation, which control conformational transitions between transcriptionally active and inactive states of chromatin. These covalent modifications can be controlled by enzymes such as methyltransferases (e.g., PRMT5), many of which are associated with specific genetic changes that may cause human disease. PRMT5 plays a role in diseases such as proliferative diseases, metabolic diseases, and blood diseases.

[0004] PRMT5 is a known cellular essential gene, and conditional PRMT5 knockout and siRNA knockdown studies have shown that PRMT5 inhibition in normal tissues is associated with a range of diseases (e.g., pancytopenia, infertility, skeletal muscle loss, cardiac hypertrophy). Therefore, new strategies are needed to exploit this metabolic vulnerability and preferentially target PRMT5 in MTAP-null tumors while sparing PRMT5 in normal tissues (MTAPWT). Targeting PRMT5 with MTA-cooperated small molecule inhibitors can preferentially target the MTA-bound state of PRMT5, which is enriched in MTAP-null tumor cells, while providing a therapeutic index superior to normal cells with intact MTAP and low MTA levels.

[0005] Therefore, there is a need in the art to provide new small molecule compounds that target PRMT5 in MTAP-null tumors.

[0006] Summary of the Invention

[0007] The purpose of the present invention is to provide a new class of small molecule compounds targeting PRMT5 in MTAP-null tumors.

[0008] The first aspect of the present invention provides a compound represented by the following formula I, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof:

[0009] in,

[0010] Ra is

[0011] W is O or S;

[0012] X1 and X2 are each independently selected from the following group: CR, N; X3 is N;

[0013] The L1 is selected from the following group: chemical bond, -CHR-, -C(R)R-;

[0014] Ring A is selected from the following group: a substituted or unsubstituted 7-12 membered bridged ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 7-12 membered spirocyclic ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 8-12 membered fused bicyclic heterocyclic group (including a carbocyclic ring or a heterocyclic ring, preferably a pentahedral and hexacyclic ring), a substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl group (preferably a pentahedral and hexacyclic ring), or the ring A is a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, a substituted or unsubstituted 5-6 membered aromatic ring or heteroaromatic ring;

[0015] The E ring is selected from the following group: a substituted or unsubstituted 3-7 membered monocyclic heterocycle, a substituted or unsubstituted 7-12 membered bridged heterocycle, a substituted or unsubstituted 7-12 membered spiro heterocycle, a substituted or unsubstituted 8-12 membered fused polycyclic heterocyclyl (e.g., a fused bicyclic ring);

[0016] R8 is selected from the group consisting of H, deuterium, halogen, cyano, alkynyl, SF5, amino, nitro, hydroxyl, mercapto, aldehyde, carboxyl, substituted or unsubstituted or halogenated C1-C6 alkyl, substituted or unsubstituted or halogenated C1-C6 alkoxy, or R8 is

[0017] R8' is selected from the following groups: H, deuterium, halogen, cyano, amino, nitro, hydroxyl, thiol, aldehyde, carboxyl, unsubstituted or halogenated C1-C6 alkyl, substituted or unsubstituted phenyl ring, substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated), substituted or unsubstituted 3-12 membered heterocycle (including saturated or partially unsaturated), or R8' is

[0018] The L3 is selected from the following group: chemical bond, -O-, -CHR-, -C(R)R-, carbonyl, S, -NH-;

[0019] Ring B is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including saturated or partially unsaturated rings), and a substituted or unsubstituted 3-7 membered heterocyclic ring (including saturated or partially unsaturated rings).

[0020] R2 and R2' are each independently selected from the following groups: R7, -L2R7; wherein, L2 is selected from the following groups: -O-, -CHR-, -C(R)R-; wherein R7 is selected from the following groups: hydrogen or none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 aromatic ring, substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings), substituted or unsubstituted 3-10 membered heterocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings); n is 0, 1, 2 or 3;

[0021] R3 is selected from the group consisting of H, deuterium, halogen, cyano, substituted or unsubstituted C1-C6 alkyl;

[0022] R4 and R5 together with the ring atoms to which they are connected form a 5-12 membered saturated or unsaturated ring, and the ring may be substituted or unsubstituted;

[0023] R is H, deuterium, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, or substituted or unsubstituted C3-C6 cycloalkyl;

[0024] Unless otherwise specified, in the above formulae, the substitution refers to the substitution of hydrogen atoms on the corresponding groups by one or more substituents selected from the following groups: deuterium, tritium, halogen, hydroxyl, carboxyl, mercapto, benzyl, C1-C 12 Alkoxycarbonyl, C1-C6 aldehyde, amino, C1-C6 amide, nitro, cyano, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-O-, unsubstituted or halogenated C1-C6 alkylene-OH, unsubstituted or halogenated C3-C8 cycloalkyl, C2-C 10 Alkenyl, unsubstituted or halogenated C1-C6 alkoxy, C1-C6 alkyl-amino, C6-C 10 Aryl, five-membered or six-membered heteroaryl, five-membered or six-membered non-aromatic heterocyclic group, -O-(C6-C 10 aryl), -O-(five-membered or six-membered heteroaryl), C1-C 12 Alkylaminocarbonyl, unsubstituted or halogenated C2-C 10 Acyl, sulfonyl (-SO2-OH), phosphoryl (-PO3-OH), unsubstituted or halogenated C1-C4 alkyl-S(O)2-, unsubstituted or halogenated C1-C4 alkyl-SO-, -SF6.

[0025] In another preferred embodiment, the Ra is selected from the following group:

[0026] Wherein, the R9 is selected from the following group: deuterium, tritium, halogen, hydroxyl, carboxyl, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or substituted C1-C6 alkyl-OH, -NH(unsubstituted or halogenated C1-C6 alkyl), -N(unsubstituted or halogenated C1-C6 alkyl)2; m is selected from 0, 1, 2 or 3; preferably, the R9 is selected from the following group: deuterium, tritium, halogen, unsubstituted or halogenated C1-C6 alkyl.

[0027] In another preferred embodiment, L1 is -CHR-, -C(R)R-; Ring A is selected from the following groups: substituted or unsubstituted 8-12-membered fused bicyclic heterocyclic group, substituted or unsubstituted 7-10-membered fused bicyclic heteroaryl; R8 is selected from the following groups: H, halogen, cyano, amino, alkynyl, SF5, hydroxyl, thiol, aldehyde, carboxyl, unsubstituted or halogenated C1-C6 alkyl, The B ring is selected from the following group: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocycle, a substituted or unsubstituted 3-6 membered heterocycle; L3 is selected from the following group: a chemical bond, -O-, -CHR-, carbonyl, S, or -NH-.

[0028] In another preferred embodiment, the R2 is selected from the following groups: R7, -L2R7; wherein, the L2 is selected from the following groups: -O-, -CHR-, carbonyl, S, -NH-; wherein R7 is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 an aromatic ring, or a substituted or unsubstituted 5-12 membered heteroaromatic ring.

[0029] In another preferred embodiment, R2 is an ortho-substituted 5- or 6-membered heteroaromatic ring, as shown in the following formula:

[0030] Ortho substituent R 10 Selected from the group consisting of hydrogen, deuterium, halogen, halogenated or unhalogenated C1-C3 alkyl, halogenated or unhalogenated C1-C3 alkoxy;

[0031] The D ring is selected from the following groups: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, preferably, the D ring is selected from the following groups:

[0032] In another preferred embodiment, the L1 is -CH2-, -CH(CH3)-; and the A ring is selected from the following group:

[0033] wherein the C ring is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including saturated or partially unsaturated rings), and a substituted or unsubstituted 3-6 membered heterocyclic ring (including saturated or partially unsaturated rings);

[0034] Or ring A is selected from the following group:

[0035] R8 is The B ring is selected from the following group: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocycle, a substituted or unsubstituted 3-6 membered heterocycle; L3 is selected from the following group: a chemical bond, -O-, -CHR-, carbonyl, S, or -NH-.

[0036] In another preferred embodiment, the L3 is a chemical bond.

[0037] In another preferred embodiment, the B ring is a substituted or unsubstituted benzene ring, or a substituted or unsubstituted 5-6 membered heteroaromatic ring.

[0038] In another preferred embodiment, R2 is selected from the following groups: R7, -(CHR)R7; wherein R7 is selected from the following groups: hydrogen or none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C8 carbocyclic ring (including saturated or partially unsaturated cases, including monocyclic, condensed, spirocyclic or bridged rings), substituted or unsubstituted 3-8 membered heterocyclic ring (including saturated or partially unsaturated cases, including monocyclic, condensed, spirocyclic or bridged rings).

[0039] In another preferred embodiment, the compound has a structure shown in the following formula:

[0040] Wherein, Q is O, NH, CH2, or a chemical bond (i.e., is a five-membered ring);

[0041] R8 is defined as above;

[0042] The R 8a and R 8b Each independently selected from the following group: H; or said R 8a and R 8b and the carbon atoms to which they are connected together form a 4-7 membered carbocyclic or heterocyclic ring;

[0043] And when the R 8a and R 8b When each independently is H, the R 8a or R 8bmay be optionally substituted with R8; when said R 8a and R 8b When the carbon atoms connected to them together form a 4-7 membered carbocyclic ring or heterocyclic ring, the R8 can be located on the carbocyclic ring or heterocyclic ring.

[0044] In another preferred embodiment, the R3 is selected from the following group: H, deuterium, halogen, cyano, substituted or unsubstituted C1-C6 alkyl.

[0045] In another preferred embodiment, R2 is a substituted or unsubstituted 5-7 membered heteroaromatic ring; and the A ring is selected from the following group: a substituted or unsubstituted 5-6 membered aromatic ring or heteroaromatic ring, a substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl group; and R8 is CF3.

[0046] The second aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as described in any of the preceding aspects, one or more of its pharmaceutically acceptable salts, racemates, optical isomers, stereoisomers or tautomers, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary materials and / or diluents.

[0047] The third aspect of the present invention provides a compound as described in any of the preceding aspects, its racemate, optical isomer or pharmaceutically acceptable salt for the preparation of a drug for treating or preventing diseases associated with abnormal gene levels or abnormal expression of PRMT5 (such as corresponding nucleic acid mutations, deletions, or ectopic or fusion or overexpression of the methyltransferase).

[0048] In another preferred embodiment, the disease is selected from the following group: the disease or condition is ovarian cancer, lung cancer, lymphoma, glioblastoma, colon cancer, melanoma, gastric cancer, pancreatic cancer or bladder cancer.

[0049] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0050] Through extensive and in-depth research, the present inventors unexpectedly discovered for the first time a class of compounds that have PRMT5 regulatory effects, and completed the present invention on this basis.

[0051] the term

[0052] In the present invention, the halogen is F, Cl, Br or I.

[0053] In the present invention, unless otherwise specified, the terms used have the ordinary meanings known to those skilled in the art. In the present invention, unless otherwise specified, all chemical formulae are intended to encompass any possible optical or geometric isomers (e.g., R-type, S-type or racemate, or cis-trans isomers of olefins, etc.).

[0054] In the present invention, the term "C1-C6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

[0055] In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like.

[0056] In the present invention, the term "C2-C6 alkenyl" refers to a straight chain or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

[0057] In the present invention, the term "C2-C6 alkynyl" refers to a straight chain or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

[0058] In the present invention, the term "C3-C10 cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms "C3-C8 cycloalkyl," "C3-C7 cycloalkyl," and "C3-C6 cycloalkyl" have similar meanings.

[0059] In the present invention, the term "C3-C10 cycloalkenyl" refers to a cyclic alkenyl group having 3 to 10 carbon atoms in the ring, including but not limited to cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl and cyclodecyl. The term "C3-C7 cycloalkenyl" has a similar meaning.

[0060] In the present invention, the term "C1-C12 alkoxycarbonyl" refers to an alkoxycarbonyl group having 1 to 12 carbon atoms in the alkyl chain, including but not limited to methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl and the like.

[0061] In the present invention, the term "C1-C12 alkylaminocarbonyl" refers to an alkylaminocarbonyl group having 1 to 12 carbon atoms in the alkyl chain, including but not limited to methylaminocarbonyl, ethylaminocarbonyl, propylaminocarbonyl, isopropylaminocarbonyl, tert-butylaminocarbonyl, benzylaminocarbonyl, dimethylaminocarbonyl and the like.

[0062] In the present invention, the terms "aromatic ring" and "aryl group" have the same meaning. Preferably, "aryl group" is "C6-C12 aryl group" or "C6-C10 aryl group." The term "C6-C12 aryl group" refers to an aromatic ring group having 6 to 12 carbon atoms and no heteroatoms in the ring, such as phenyl and naphthyl. The term "C6-C10 aryl group" has a similar meaning.

[0063] In the present invention, the terms "aromatic heterocycle" or "heteroaryl" have the same meaning and refer to a heteroaromatic group containing one to multiple heteroatoms. The heteroatoms referred to herein include oxygen, sulfur, and nitrogen. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0064] In the present invention, the term "3-12 membered heterocyclic group" refers to a saturated or unsaturated 3-12 membered ring group containing 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen, such as dioxolanyl. The term "3-7 membered heterocyclic group" has a similar meaning.

[0065] In the present invention, the term "substituted" refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in the embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. A cyclic substituent, such as a heterocycloalkyl, may be connected to another ring, such as a cycloalkyl, to form a spirobicyclic system, for example, the two rings having a common carbon atom. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. The substituents include, but are not limited to, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl, 3- to 12-membered heterocyclic groups, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C1-8 aldehyde, C2-10 acyl, C2-10 ester, C1-C12 alkoxycarbonyl, amino, alkoxy, C1-10 sulfonyl, etc.

[0066] When a term such as "C1-8" or similar is used, it means that the group can have 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms.

[0067] When, for example, "3-12 membered" or similar expressions are used, it means that the group may have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms or heteroatoms as ring atoms.

[0068] PRMT5 modulator compounds

[0069] The present invention provides a class of compounds having PRMT5 regulatory activity:

[0070] in,

[0071] Ra is

[0072] W is O or S;

[0073] X1 and X2 are each independently selected from the following group: CR, N; X3 is N;

[0074] The L1 is selected from the following group: chemical bond, -CHR-, -C(R)R-;

[0075] Ring A is selected from the following group: a substituted or unsubstituted 7-12 membered bridged ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 7-12 membered spirocyclic ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 8-12 membered fused bicyclic heterocyclic group (including a carbocyclic ring or a heterocyclic ring, preferably a pentahedral and hexacyclic ring), a substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl group (preferably a pentahedral and hexacyclic ring), or the ring A is a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, a substituted or unsubstituted 5-6 membered aromatic ring or heteroaromatic ring;

[0076] The E ring is selected from the following group: a substituted or unsubstituted 3-7 membered monocyclic heterocycle, a substituted or unsubstituted 7-12 membered bridged heterocycle, a substituted or unsubstituted 7-12 membered spiro heterocycle, a substituted or unsubstituted 8-12 membered fused polycyclic heterocyclyl (e.g., a fused bicyclic ring);

[0077] R8 is selected from the group consisting of H, deuterium, halogen, cyano, alkynyl, SF5, amino, nitro, hydroxyl, mercapto, aldehyde, carboxyl, halogenated or unhalogenated C1-C6 alkyl, halogenated or unhalogenated C1-C6 alkoxy, or R8 is When the A ring is a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, a 5-6 membered aromatic ring or heteroaromatic ring, the R8 is

[0078] R8' is selected from the following groups: H, deuterium, halogen, cyano, amino, nitro, hydroxyl, thiol, aldehyde, carboxyl, halogenated or unhalogenated C1-C6 alkyl, substituted or unsubstituted phenyl ring, substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated), substituted or unsubstituted 3-12 membered heterocycle (including saturated or partially unsaturated), or R8' is

[0079] The L3 is selected from the following group: chemical bond, -O-, -CHR-, -C(R)R-, carbonyl, S, -NH-;

[0080] Ring B is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including saturated or partially unsaturated rings), and a substituted or unsubstituted 3-7 membered heterocyclic ring (including saturated or partially unsaturated rings).

[0081] R2 and R2' are each independently selected from the following groups: R7, -L2R7; wherein, said L2 is selected from the following groups: -O-, -CHR-, -C(R)R-; wherein R7 is selected from the following groups: hydrogen or none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6- 10 aromatic ring, substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings), substituted or unsubstituted 3-10 membered heterocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings); n is 0, 1, 2 or 3;

[0082] R3 is selected from the group consisting of H, deuterium, halogen, cyano, substituted or unsubstituted C1-C6 alkyl;

[0083] R4 and R5 together with the ring atoms to which they are connected form a 5-12 membered saturated or unsaturated ring, and the ring may be substituted or unsubstituted;

[0084] R is H, deuterium, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, or substituted or unsubstituted C3-C6 cycloalkyl;

[0085] Unless otherwise specified, in the above formulae, the substitution refers to the substitution of hydrogen atoms on the corresponding groups by one or more substituents selected from the following groups: deuterium, tritium, halogen, hydroxyl, carboxyl, mercapto, benzyl, C1-C 12Alkoxycarbonyl, C1-C6 aldehyde, amino, C1-C6 amide, nitro, cyano, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-O-, unsubstituted or halogenated C1-C6 alkylene-OH, unsubstituted or halogenated C3-C8 cycloalkyl, C2-C 10 Alkenyl, unsubstituted or halogenated C1-C6 alkoxy, C1-C6 alkyl-amino, C6-C 10 Aryl, five-membered or six-membered heteroaryl, five-membered or six-membered non-aromatic heterocyclic group, -O-(C6-C 10 aryl), -O-(five-membered or six-membered heteroaryl), C1-C 12 Alkylaminocarbonyl, unsubstituted or halogenated C2-C 10 Acyl, sulfonyl (-SO2-OH), phosphoryl (-PO3-OH), unsubstituted or halogenated C1-C4 alkyl-S(O)2-, unsubstituted or halogenated C1-C4 alkyl-SO-, -SF6.

[0086] Pharmaceutical compositions and methods of administration

[0087] Since the compounds of the present invention have excellent methyltransferase inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate related diseases caused by abnormal activity or expression of methyltransferases (such as PRMT5).

[0088] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 5-200 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0089] "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0090] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0091] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0092] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0093] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0094] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0095] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0096] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0097] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0098] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds. In some preferred embodiments, the compounds of the present invention can be administered together with other small molecule compounds to form PROTACs, or together with other macromolecular compounds such as monoclonal antibodies to form ADCs.

[0099] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0100] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0101] The definitions of each abbreviation are as follows:

[0102] The starting materials can be obtained from commercial sources or prepared by methods known or disclosed in the art.

[0103] Purification of intermediates and compounds is performed using standard chemical laboratory procedures, such as normal-phase or reverse-phase chromatography or recrystallization. Normal-phase chromatography utilizes prepacked silica gel columns or preparative thin-layer chromatography. Silica gel columns are typically glass columns or flash chromatography instruments. For normal-phase chromatography, the mobile phase is selected from petroleum ether / ethyl acetate, dichloromethane / methanol, or other suitable solvents, and the resulting mixture is prepared and eluted. Reverse-phase preparative liquid chromatography utilizes a C18 column and is performed using a preparative liquid chromatograph or flash chromatography instrument, with detection at 214 nM and 254 nM or using preparative liquid chromatography-mass spectrometry. Gradient elution is performed using the mobile phases of water / acetonitrile containing 0.1% hydrochloric acid, water / acetonitrile, water / acetonitrile containing 0.1% ammonium bicarbonate, water / acetonitrile containing 0.1% formic acid, 0.1% ammonia / acetonitrile, water / acetonitrile containing 0.1% trifluoroacetic acid, or other suitable solvent systems.

[0104] The structures of intermediates and compounds were characterized by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS). NMR spectrometers used were Bruker Ascend 400, Varian 400, ZKNJ BIXI-1 300 MHz, Bruker Avance III 400 MHz, or Bruker AVANCE Neo 400 MHz. Solvents used were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, or other deuterated solvents as noted. Spectral data are reported in terms of chemical shift δ (peak splitting number, coupling constant J (Hz), and number of hydrogen atoms). Tetramethylsilane was used as an internal standard for chemical shifts, and its chemical shift was set to zero (δ, 0 ppm). Abbreviations are as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), and br (broad).

[0105] Representative liquid chromatography-mass spectrometry (LCMS) methods for the structural characterization of intermediates and compounds are as follows:

[0106] Method 1: Agilent LC1260 system coupled to a 6120 single quadrupole mass spectrometer

[0107] Column: Waters CORTECS C-18, 2.7 μm, 4.6 x 30 mm. Solvent A: 0.05% formic acid in water, Solvent B: 0.05% formic acid in acetonitrile, 5% acetonitrile to 95% acetonitrile over 1 minute, hold for 1 minute, for a total of 2.5 minutes; Flow rate: 1.8 mL / min; Column temperature: 40°C.

[0108] Column: XSelect CSH C18, 3.5 μm, 4.6 x 50 mm. Solvent A: 0.05% ammonia in water, Solvent B: 0.05% ammonia in acetonitrile, 5% acetonitrile to 95% acetonitrile over 1 minute, hold for 1 minute, for a total of 2.5 minutes; Flow rate: 1.8 mL / min; Column temperature: 40°C.

[0109] Method 2: performed on an Agilent LC / MSD 1200 system coupled with a quadrupole mass spectrometer.

[0110] Column: ODS2000 (50×4.6 mm, 5 μm) (ES (+) or (-) ionization mode), temperature 30°C; flow rate 1.5 mL / min.

[0111] Example Synthesis General Method:

[0112] General Method: Synthesis of Intermediate A2

[0113] Synthesis route:

[0114] Step 1: Methyl 2,5-difluoro-4-nitrobenzoate (2)

[0115] 2,5-Difluoro-4-nitrobenzoic acid (1) (50 g, 246.18 mmol, 1 eq) was dissolved in methanol (500 mL) and thionyl chloride (43.93 g, 369.28 mmol, 26.79 mL, 1.5 eq) was added at 0°C. The reaction mixture was reacted at 40°C for 16 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to dryness under reduced pressure. After dilution with 300 mL of water, the mixture was extracted three times with 1 L of ethyl acetate. The organic phase was washed with saturated brine (400 mL), dried over anhydrous magnesium sulfate, and the filtrate was filtered and concentrated to dryness under reduced pressure. The crude product was slurried with petroleum ether at 25°C for 60 minutes to obtain methyl 2,5-difluoro-4-nitrobenzoate (2) (103 g, 474.38 mmol, 96.35% yield) as a white solid. HNMR: ES23714-64-P1A, 1H NMR (400MHz, CHLOROFORM-d) δppm 7.89 (td, J=9.51, 5.63Hz, 2H) 4.00 (s, 3H). 19F NMR(376MHz,CHLOROFORM-d)δppm-110.27(s,1F)-121.56(m,1F)

[0116] Step 2: Methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3)

[0117] Methyl 2,5-difluoro-4-nitrobenzoate (2) (80 g, 368.45 mmol, 1 eq) and 2-methyl-1H-imidazole (36.30 g, 442.14 mmol, 1.2 eq) were dissolved in dimethyl sulfoxide (1.2 L). The reaction mixture was reacted at 50 °C for 16 hours. LCMS showed that the reaction was complete. The reaction mixture was diluted with 4 L of water and extracted with 4.5 L of ethyl acetate. The organic phase was washed with saturated brine (3 L), dried over anhydrous magnesium sulfate, and the filtrate was filtered and concentrated to dryness under reduced pressure. The crude product was slurried with methyl tert-butyl methyl paraffin at 25 °C for 60 minutes (25.5 g). (2). The mother liquor was purified by column chromatography (silica, 50% tetrahydrofuran in petroleum ether) to give a yellow liquid which was then slurried at 25 degrees Celsius for 60 minutes to give a yellow solid (8.23 g). Yellow solid 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoic acid methyl ester (3) (25.5 g, 91.32 mmol, 24.79% yield) 2,5-difluoro-4-nitrobenzoic acid methyl ester (2) (raw material recovery) (20.34 g, 93.68 mmol, 25.43% yield). Yellow solid 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoic acid methyl ester (3) (8.2 g, 29.07 mmol, 7.89% yield, 99% purity) H NMR: ES23714-67-P1A1, 1H NMR (400 MHz, DMSO-d6) δ ppm 8.39 (d, J = 9.90 Hz, 1H) 8.08-8.20 (m, 1H) 7.23 (d, J = 1.32 Hz, 1H) 6.92 (s, 1H) 3.91 (s, 3H) 2.12 (s, 3H). 19F NMR (376 MHz, DMSO-d6) δ ppm -105.45 (br s, 1F)

[0118] Step 3: Methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4)

[0119] Methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) (13.9 g, 49.78 mmol, 1 eq) was dissolved in tetrahydrofuran (300 ml) and palladium hydroxide on carbon (2.8 g, 49.78 mmol, 20% purity, 1 eq) was added under a hydrogen atmosphere. The reaction system was replaced with hydrogen three times. The reaction solution was heated to 50 degrees Celsius under a hydrogen (1 eq) (50 psi) atmosphere for 32 hours. LCMS showed that the reaction was complete. The reaction solution was filtered through celite and the filter cake was washed four times with 300 ml of ethyl acetate. The filtrate was concentrated to dryness under reduced pressure to obtain a gray solid. The crude product was used directly in the next step. Methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4) (12 g, 48.15 mmol, 96.72% yield). 1H NMR (400 MHz, DMSO-d6) Shift 7.46 (d, J = 7.63 Hz, 1H), 7.07 (d, J = 1.38 Hz, 1H), 6.93 (d, J = 1.25 Hz, 1H), 6.59 (d, J = 13.51 Hz, 1H), 6.15 (br s, 2H), 3.68-3.77 (m, 3H), 2.01-2.12 (m, 3H). 19F NMR (376 MHz, DMSO-d6) Shift -109.31-108.47 (m, 1F).

[0120] Step 4: 7-Fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (5)

[0121] Methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4) (12 g, 48.15 mmol, 1 eq) was added to 1-methyl-2-pyrrolidone at 25°C, followed by 1,1-carbonyldiimidazole (19.52 g, 120.37 mmol, 2.5 eq). The reaction mixture was heated to 115°C for 16 hours. LCMS indicated the reaction was complete. The two batches of reaction mixture were combined. 600 mL of ethyl acetate and 600 mL of water were added to the reaction mixture and the mixture was slurried at 25°C for 16 hours. The slurry was filtered under reduced pressure, and the filter cake was washed with 100 mL of ethyl acetate. The solid was concentrated under reduced pressure to give methyl 7-fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (24.1 g, 87.56 mmol, 86.60% yield) as a gray solid. The crude product was used directly in the next reaction. 1H NMR (400 MHz, DMSO-d6) Shift 11.71 (br s, 1H), 8.40 (d, J = 6.38 Hz, 1H), 7.76 (s, 1H), 7.08 (d, J = 11.38 Hz, 1H), 3.88 (s, 3H), 2.89 (s, 3H). 19F NMR (376 MHz, DMSO-d6) Shift -111.43--110.58 (m, 1F)

[0122] Step 5: Methyl 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6)

[0123] Methyl 7-fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (12.0 g, 43.60 mmol, 1.0 eq), 2,4-dimethoxybenzylamine (10.9 g, 65.19 mmol, 9.82 ml, 1.50 eq), and 1,8-diazabicyclo[5.4.0]undec-7-ene (19.92 g, 130.80 mmol, 19.72 ml, 3.0 eq) were added to acetonitrile (240 ml). Benzotriazole-1-oxo-tris(dimethylaminophosphine)hexafluorophosphonium salt (25.07 g, 56.68 mmol, 1.3 eq) was added portionwise at 15-20°C. The reaction solution was slightly exothermic, became homogeneous, and solids precipitated. The reaction mixture was reacted at 15-20 degrees Celsius for 16 hours under nitrogen protection. LCMS showed that the starting material was completely consumed and the target compound was detected. The reaction suspension was filtered under reduced pressure and the filter cake was washed with 100 ml of acetonitrile. The solid was collected and dried under reduced pressure to obtain an off-white solid 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6) (15.3 g, 36.05 mmol, 82.68% yield). LCMS ES15882-1146-P1A: (ESI) m / z = 425.3 [M+1] +; RT = 1.721 min. 1H NMR (400 MHz, DMSO-d6) Shift 8.49(d,J=7.00Hz,1H),8.45(t,J=5.57Hz,1H),7.95(s,1H),7.23(d,J=12.51Hz,1H),7.18(d,J=8.38Hz,1H),6.58(d,J=2 .38Hz,1H),6.47(dd,J=2.38,8.38Hz,1H),4.66(d,J=5.25Hz,2H),3.88(s,3H),3.82(s,3H),3.73(s,3H),2.93(s,3H).19F NMR(376.5MHz,DMSO-d6)Shift-113.02.

[0124] Step 6: 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (7)

[0125] 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6) (16.3 g, 38.40 mmol, 1.0 eq) was added to dichloromethane (50 mL), followed by the addition of trifluoroacetic acid (250 mL). The reaction mixture was heated to 50°C for 16 hours. LCMS showed that the starting material was completely consumed and the target compound was detected. The reaction mixture was concentrated to dryness under reduced pressure to give a purple solid 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (7) (28.3 g, crude). The crude product was used directly in the next reaction. LCMS ES15882-1150-P1A: (ESI) m / z = 275.3 [M+1] +; RT = 0.607 min

[0126] Step 7: 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A2)

[0127] Methyl 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (7) (crude product from the previous step) (28.3 g, 38.70 mmol, 1 eq) was added to tetrahydrofuran (80 mL) and methanol (80 mL). Sodium hydroxide (7.74 g, 193.48 mmol, 5 eq) was dissolved in water (80 mL) and added to the reaction mixture. The reaction mixture was heated to 50°C and reacted for 4 hours. The desired compound was detected by LCMS. The reaction mixture was cooled to 20°C and concentrated under reduced pressure to remove the organic solvent. The residue was diluted with 10:1 water to methanol (300 mL) and filtered through celite. The filter cake was washed three times with 10:1 water to methanol (300 mL). All filtrates were combined and concentrated under reduced pressure to remove the methanol. The pH of the residue was adjusted to 5-6 with acetic acid. The resulting slurry was stirred at 15-20°C for 12 hours. The resulting solid was filtered under reduced pressure and washed with water. The collected solid was freeze-dried to yield a white solid. 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A2) (9.9 g, 37.55 mmol, 97.05% yield, 98.71% purity). LCMS ES15882-1154-P1C: (ESI) m / z = 261.1 [M+1]+; RT = 0.422 min. 1H NMR (400MHz, DMSO-d6)Shift 13.15(br s,1H),8.53(d,J=7.04Hz,1H),7.85(s,1H),7.68(s,2H),7.16(d,J=12.10Hz,1H),2.94(s,3H)

[0128] General Method: Synthesis of Intermediate A3

[0129] Synthesis route:

[0130] Step 1: 1-(5-Bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2)

[0131] 1-Bromo-2-chloro-5-fluoro-4-nitrobenzene (1) (3 g, 11 mmol, 1 eq) was added to a solution of 2-methyl-1H-imidazole (1.2 g, 14 mmol, 1.2 eq) in acetonitrile (50 mL). Potassium carbonate (4 g, 29 mmol, 2.5 eq) was then added to the reaction mixture. The mixture was heated to 80°C and stirred for 16 hours. The reaction mixture was detected to be completely reacted with the desired product. The acetonitrile was removed by concentration under reduced pressure, water (40 mL) was added, and the mixture was extracted with ethyl acetate (3 x 50 mL). The mixture was dried over anhydrous magnesium sulfate, and the filtrate was concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (silica, 35% tetrahydrofuran in petroleum ether) to give 1-(5-bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2) (4 g) as a white solid. H NMR: 1H NMR (400MHz, DMSO-d6) δ8.55 (s, 1H), 8.32 (s, 1H), 7.22 (d, J = 1.32Hz, 1H), 6.91 (d, J = 1.32Hz, 1H), 2.08-2.23 (m, 3H).

[0132] Step 2: 4-Bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3)

[0133] 1-(5-Bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2) (3.5 g, 11 mmol, 1 eq) was dissolved in a mixture of water (8 mL), ethanol (16 mL) and tetrahydrofuran (16 mL). Ammonium chloride (8.9 g, 166 mmol, 15 eq) was added to the reaction solution. The temperature was raised to 70°C and iron powder (2.5 g, 44 mmol, 4 eq) was added to the reaction solution. The reaction was stirred at 90°C for 2 hours. LC-MS analysis showed that the reaction of the starting material was complete and the target product was produced. The reaction solution was filtered through celite and washed with ethyl acetate (40 mL x 3). The solvent was removed under reduced pressure to obtain 4-bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3) (3.1 g, 10.8 mmol, 98% yield) as a black solid. H NMR: 1H NMR (400MHz, DMSO-d6) δ7.41(s,1H),7.12(s,1H),7.07(s,1H),6.99(s,1H),5.45(s,2H),2.13(s,3H).

[0134] Step 3: 8-Bromo-7-chloro-1-methyl-5-hydrogen-imidazo[1,5-a]quinoxaline-4-one (4)

[0135] 4-Bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3) (3 g, 10.5 mmol, 1 eq) and 1,1-carbonyldiimidazole (2.6 g, 15.7 mmol, 1.5 eq) were dissolved in 1,2-dichlorobenzene (30 mL) in sequence. The reaction mixture was stirred at 130 °C for 16 h. LC-MS confirmed the complete reaction of the starting materials and the formation of the target product. The reaction mixture was stirred in a solution of ethyl acetate and water (2 / 1 15 mL) for 30 min and filtered to obtain a filter cake. The filter cake was concentrated under vacuum to obtain a black solid 8-bromo-7-chloro-1-methyl-5-hydrogen-imidazo[1,5-a]quinoxalin-4-one (4) (2.2 g, 7 mmol, 67% yield). H NMR: 1H NMR (400MHz, DMSO-d6) δ8.04 (s, 1H), 7.33 (d, J = 3.30Hz, 2H), 2.83 (s, 3H)

[0136] Step 4: 7-Chloro-1-methyl-4-oxo-5H-imidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (5)

[0137] 8-Bromo-7-chloro-1-methyl-5-hydrogen-imidazo[1,5-a]quinoxaline-4-one (4) (500 mg, 1.6 mmol, 1 eq) was dissolved in ethanol (5 mL). 1,8-diazabicyclo[5.4.0]undecane-7-ene (365 mg, 2.4 mmol, 362 μL, 1.5 eq) was added. The nitrogen atmosphere was replaced three times. Tributylphosphine tetrafluoroborate (46 mg, 160 μM, 0.1 eq), molybdenum hexacarbonyl (232 mg, 880 μM, 118 μL, 0.55 eq) and palladium acetate (36 mg, 160 μM, 0.1 eq) were added. The reaction was stirred at 90°C for two hours. LC-MS analysis showed that the reaction of the starting material was complete and the target product was produced. The ethanol was removed from the reaction solution under reduced pressure, water (10 mL) was added, and the mixture was extracted with 30 mL of ethyl acetate (10 mL * 3). The residue was dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 30% tetrahydrofuran in petroleum ether) gave methyl 7-chloro-1-methyl-4-oxo-5H-imidazole[1,5-a]quinoxaline-8-carboxylate (5) (500 mg) as a yellow solid.

[0138] Step 5: 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6)

[0139] Methyl 7-chloro-1-methyl-4-oxo-5H-imidazole[1,5-a]quinoxaline-8-carboxylate (5) (480 mg, 1.6 mmol, 1 eq) was dissolved in acetonitrile (5 mL) and added with 2,4-dimethoxybenzylamine (341 mg, 2 mmol, 306 μL, 1.3 eq), benzotriazole-1-oxo-tris(dimethylaminophosphonium) hexafluorophosphonium salt (1 g, 2.4 mmol, 1.5 eq) and 1,8-diazabicyclo[5.4.0]undecane-7-ene (1.2 g, 7.9 mmol, 1.2 mL, 5 eq). The reaction was stirred at room temperature for 16 hours. LC-MS analysis showed that the reaction of the starting material was complete and the target product was produced. The acetonitrile was removed from the reaction solution under reduced pressure, and water (8 mL) was added. The solution was extracted with 30 mL of ethyl acetate (10 mL*3). The solution was dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 30% tetrahydrofuran in petroleum ether) afforded methyl 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) as a yellow solid (500 mg, 1.1 mmol, 70% yield).

[0140] Step 6: 7-Chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3)

[0141] Methyl 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (500 mg, 1.1 mmol, 1 eq) was dissolved in water (5 mL) and ethanol (5 mL), and sodium hydroxide (132 mg, 3.3 mmol, 3 eq) was added. The reaction mixture was stirred at 50°C for 5 hours. LCMS analysis showed that the reaction of the starting material was complete and the target product was produced. The reaction mixture was added with 6 M hydrochloric acid (0.5 mL) and concentrated to dryness under reduced pressure. The crude product was used directly in the next step to obtain 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3) (350 mg, 820 μmol, 74.6% yield) as a white solid.

[0142] General Method: Synthesis of Intermediate A4

[0143] Synthesis route:

[0144] Step 1: Methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate

[0145] Methyl 3-fluoro-4-nitrobenzoate (1) (2.00 g, 10.04 mmol, 1 eq) was added to acetonitrile (40 eq) and potassium carbonate (4.16 g, 30.13 mmol, 3 eq) and 2,4-dimethyl-1H-imidazole (2) (965 mg, 10.04 mmol, 1 eq). The reaction mixture was reacted at 85°C for 16 hours. LC-MS showed that the starting material was completely consumed and the target product was produced. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was diluted with dichloromethane (80 eq) and filtered. The filtrate was dried over magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate (3) (2.6 g, 9.45 mmol, 94.05% yield) as a yellow solid. The crude product was used directly in the next step without further purification.

[0146] Step 2: Methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate

[0147] Methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate (3) (2.5 g, 9.08 mmol, 1 eq) was dissolved in ethanol (20 eq), tetrahydrofuran (20 eq), and water (10 eq). Iron powder (5.07 g, 90.82 mmol, 10 eq) and ammonium chloride (2.43 g, 45.41 mmol, 5 eq) were added at room temperature. The reaction mixture was reacted at 90°C for 16 hours. LC-MS showed that the starting material was completely consumed and the desired product was produced. The reaction mixture was filtered and concentrated to dryness under reduced pressure. The residue was diluted with water (50 equivalents) and extracted with ethyl acetate (40 equivalents * 3). The combined organic phases were dried over magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate (4) (1.9 g, 6.13 mmol, 67.49% yield, 79.13% purity) as a yellow solid. The crude product was used directly in the next step without further purification. LCMS: ES19974-375-P1C2, (ESI) m / z = 246.1 [M+1] +; RT = 0.61 min, purity: 79.13%

[0148] Step 3: 1,3-Dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester

[0149] Methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate (4) (500 mg, 2.04 mmol, 1 eq) and 1,1-carbonyldiimidazole (495 mg, 3.06 mmol, 1.5 eq) were dissolved in 1,2-dichlorobenzene (10 eq). The reaction mixture was reacted at 120°C under nitrogen for 16 h. LC-MS showed that the starting material was completely consumed and the target product was produced. The reaction mixture was filtered, the filter cake was slurried with water, and dried under reduced pressure to obtain brown solid methyl 1,3-dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (460 mg, 1.62 mmol, 79.48% yield, 95.55% purity). The crude product was used directly in the next step without further purification.

[0150] LCMS: ES19974-392-P1B1, (ESI) m / z=272.0[M+1]+; RT=0.61min, purity:95.55%

[0151] Step 4: Methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate

[0152] Methyl dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (700 mg, 2.58 mmol, 1 eq) was dissolved in acetonitrile (25 eq) and benzotriazole-1-oxo-tris(dimethylaminophosphonium) hexafluorophosphonium salt (1.83 g, 4.13 mmol, 1.6 eq) and 1,8-diazabicyclo[5.4.0]undecane-7-ene (1.96 g, 12.90 mmol, 1.94 eq, 5 eq) were added. The reaction mixture was reacted at room temperature for 0.5 h. (2,4-Dimethoxyphenyl)methylamine (647.2 mg, 3.87 mmol, 583.1 μL, 1.5 eq) was added to the reaction mixture, and the reaction mixture was reacted at 50°C for 15.5 h. LC-MS showed that the starting material was completely consumed and the target product was produced. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to obtain a brown solid of methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (900 mg, 2.08 mmol, 80.57% yield, 97.13% purity). The crude product was used directly in the next step without further purification. LCMS: ES19974-397-P1B1, (ESI) m / z = 421.1 [M+1] +; RT = 0.78 min, purity: 97.13%

[0153] Step 5: 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0154] Methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (850 mg, 2.02 mmol, 1 eq) was dissolved in methanol (10 eq), tetrahydrofuran (10 eq) and water (5 eq) and lithium hydroxide (424.2 mg, 10.11 mmol, 5 eq) was added. The reaction mixture was reacted at 50°C for 16 hours. LC-MS showed that the starting material was completely consumed and the target product was produced. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was adjusted to pH 6-7 with acetic acid. The residue was filtered and the filter cake was dried under reduced pressure to obtain a brown solid 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A4) (800 mg, 1.97 mmol, 97.37% yield). The crude product was used directly in the next step without further purification.

[0155] General Method: Synthesis of Intermediate A6

[0156] Synthesis route:

[0157] Step 1: N-(4-Bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3)

[0158] To a solution of 4-bromo-2-fluoroaniline (5.78 g, 30.4 mmol, 1 eq) and 4-methyl-1H-pyrazole-5-carboxylic acid (4.60 g, 36.5 mmol, 1.2 eq) in Py (120 mL) was added POCl₃ (4.66 g, 30.4 mmol, 2.82 mL, 1 eq) and stirred at 0°C for 1 h. LC-MS (ET63399-4-R1A1) showed complete consumption of the starting material and a major peak with the desired m / z was detected. The reaction mixture was quenched with ice water (200 mL) and then extracted with ethyl acetate (60 mL x 6). The combined organic layers were washed with brine (30.0 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was triturated with ethyl acetate (50.0 mL) to give N-(4-bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3) (7.09 g, 23.8 mmol, 78.2% yield) as a white solid. 1 H NMR(400MHz, DMSO-d6)δ13.26(br s,1H)9.52(s,1H)7.92(t,J=8.52Hz,1H)7.70(s,1H)7.62(dd,J=10.31,1.85Hz,1H)7.41(br d,J=8.70Hz,1H)2.25(s,3H).LC-MS;[MH] + 298.0

[0159] Step 2: 8-Bromo-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (4)

[0160] To a solution of N-(4-bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3) (7.09 g, 23.8 mmol, 1 eq) in DMA (70 mL) was added NaH (1.43 g, 35.7 mmol, 60% purity, 1.5 eq). The mixture was stirred at 120 ° C for 16 hours. LC-MS showed complete consumption of the starting material and one major peak with the desired m / z. The reaction mixture was quenched with saturated ammonium chloride (300 mL), and the precipitate was collected and washed with water (50 mL), then concentrated under reduced pressure to give 8-bromo-3-methylpyrazolo[1,5-a]quinoxaline-4(5H)-one (4) (6.50 g, crude) as a white solid. 1 [MH] + 278.0

[0161] Step 3: 8-Bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5)

[0162] To a solution of 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (4) (1.00 g, 3.60 mmol, 1 eq) in MeCN (10.0 mL) was added PMBNH2 (1.23 g, 8.99 mmol, 1.16 mL, 2.5 eq), BOP (3.18 g, 7.19 mmol, 2 eq) and DBU (2.74 g, 18.0 mmol, 2.71 mL, 5 eq). The mixture was stirred at 50 ° C for 16 hours. LC-MS showed complete consumption of the starting material and a major peak with the desired m / z. The reaction mixture was diluted with saturated ammonium chloride (10.0 mL) and ethanol (5.00 mL). The precipitate was collected and washed with water (10.0 mL), then concentrated under reduced pressure to give 8-bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5) (1.09 g, crude) as a yellow solid. 1H NMR(400MHz, DMSO-d6)δ8.20(d,J=1.88Hz,1H)7.95(s,1H),7.44-7.48(m,2H)7.38(d,J=8.63Hz,2H)7.28(br s,1H)6.87(d,J=8.75Hz,2H)4.70(d,J=5.88Hz,2H)3.71(s,3H)2.53(s,3H).LC-MS; [MH] + 399.0

[0163] Step 4: 8-Bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6)

[0164] A solution of 8-bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5) (500 mg, 1.26 mmol, 1 equiv) in TFA (5 mL) was stirred at 60 °C for 12 hours. LC-MS showed complete consumption of the starting material and one major peak with the desired m / z. The reaction mixture was concentrated under reduced pressure to afford compound 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6) (400 mg, crude) as a yellow solid. LC-MS: [MH] + 277.0

[0165] Step 5: Ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7)

[0166] To a solution of 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6) (400 mg, 1.08 mmol, 1 eq) in EtOH (4 mL) was added Mo(CO)6 (143 mg, 541 umol, 72.9 uL, 0.5 eq), DBU (659 mg, 4.33 mmol, 652 uL, 4 eq), (t-Bu)3PBF4 (94.2 mg, 325 umol, 0.3 eq) and Pd(OAc)2 (36.5 mg, 162 umol, 0.15 eq). The mixture was stirred at 90 ° C for 12 hours. LC-MS (ET63399-28-R1A1) showed that the starting material was completely consumed and the desired mass was detected. The reaction mixture was diluted with saturated ammonium chloride (10 mL) and ethanol (10 mL). The precipitate was collected and washed with water (10.0 mL), then concentrated under reduced pressure to give ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7) (250 mg, 925 umol, 85.4% yield) as a yellow solid. LC-MS; [MH] + 271.1

[0167] Step 6: 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A6)

[0168] To a solution of ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7) (250 mg, 925 umol, 1 eq) in EtOH (3 mL) and H 2 O (1 mL) was added LiOH · H 2 O (116 mg, 2.77 mmol, 3 eq). The mixture was stirred at 25 ° C for 12 hours. LC-MS (ET63399-32-R1A1) showed that the starting material was completely consumed and the desired mass was detected. The reaction mixture was concentrated, diluted with water (10 mL), and washed with DCM (10.0 mL × 3) to remove impurities. The aqueous phase was adjusted to pH = 5 with 1 M HCl, and the precipitate was collected and purified by preparative HPLC (column: Phenomenex Luna C18 75*30mm*3um; mobile phase: [water (FA)-ACN]; B%: 1%-35%, 8 minutes, UV 220nm and 254nm) to give 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A6) (60.0 mg, 248umol, 26.8% yield) as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ12.94(br s,1H)8.67(d,J=1.96Hz,1H)7.96(s,1H)7.89(dd,J=8.44,1.96Hz,1H)7.52(d,J=8.44Hz,1H)7.19(br s,2H)2.49(br s,3H).LC-MS,[MH]+243.1

[0169] General Method: Synthesis of Intermediate A7

[0170] Synthesis route:

[0171] Step 1: Methyl 2-fluoro-5-(4-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3)

[0172] A mixture of 2,5-difluoro-4-nitrobenzoic acid methyl ester (20.0 g, 92.1 mmol, 1 equivalent) and 5-methyl-1H-imidazole (7.56 g, 92.1 mmol, 1 equivalent) in DMSO (200 mL) was stirred at 50 ° C for 12 hours. LCMS showed that the remaining starting material was detected and the required mass was detected. The residue was diluted with H o (400 mL) and extracted with EtOAc (200 mL × 3). The combined organic layer was washed with brine (200 mL), dried over Na sO , filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO , petroleum ether / ethyl acetate = 20 / 1 to 0 / 1) to obtain compound 2-fluoro-5- (4-methyl -1H- imidazole -1- bases) -4-nitrobenzoic acid methyl ester (9.30 g, 33.3 mmol, 34.2% yield) as a yellow solid. LC-MS (ESI) m / z = 280.1 [M+H] +

[0173] Step 2: Methyl 4-amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoate (4)

[0174] To a solution of 2-fluoro-5-(4-methyl-1H-imidazol-1-yl)-4-nitrobenzoic acid methyl ester (9.30 g, 33.3 mmol, 1 equivalent) and NH Cl (26.7 g, 499 mmol, 15 equivalents) in EtOH (90.0 mL) / THF (90.0 mL) / H O (45.0 mL) was added Fe powder (7.44 grams, 133 mmol, 4 equivalents). The mixture was stirred at 80 ° C for 2 hours. LCMS showed that the starting material had been consumed and the desired mass was detected. The reaction mixture was filtered, the filtrate was diluted with water (300 mL) and extracted with EtOAc (200 mL × 3). The combined organic layers were washed with brine (100 mL), dried over Na 2 SO 4 , and concentrated under reduced pressure to afford methyl 4-amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoate (8.00 g, 32.1 mmol, 95.2% yield) as a yellow solid. LC-MS (ESI) m / z = 250.0 [M+H] +

[0175] Step 3: 7-Fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (5)

[0176] 4-Amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoic acid methyl ester (500 mg, 2.01 mmol, 1 eq) and CDI (487 mg, 2.41 mmol, 1.2 eq) were placed in 1,2-dichlorobenzene (20.0 mL) in a microwave tube. The sealed tube was heated at 150 ° C for 3 hours under microwave. 11 parallel reactions were performed, and LCMS showed that the starting material had been consumed and the required mass was detected. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to obtain a residue. The crude product was ground with MTBE (20.0 mL) at 25 ° C for 30 minutes to obtain 7-fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6.00 g, 21.80 mmol, 85.0% yield) as a yellow solid. LC-MS (ESI) m / z = 276.0 [M + H] +

[0177] Step 4: 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6) To a solution of 7-fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester 5 (6.00 g, 21.8 mmol, 1 eq) in MeCN (120 mL) was added BOP (19.2 g, 43.6 mmol, 2 eq), DBU (16.5 g, 109 mmol, 16.4 mL, 5 eq) and PMBNH2 (7.48 g, 54.5 mmol, 7.05 mL, 2.5 eq). The mixture was stirred at 70 ° C for 16 hours. LCMS showed complete consumption of the starting material and the desired mass was detected. The reaction mixture was diluted with saturated ammonium chloride (20.0 mL). The filter cake was washed with water (10.0 mL) and concentrated under reduced pressure to give methyl 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 6 (5.00 g, 12.6 mmol, 50.0% yield) as a yellow solid. LC-MS (ESI) m / z = 395.1 [M+H] +

[0178] Step 5: 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (6)

[0179] A solution of 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester 6 (2.00 g, 5.07 mmol, 1 equivalent) in TFA (20.0 mL) was stirred at 75 ° C for 16 hours. LCMS showed that the starting material had been consumed and the required mass was detected. The reaction mixture was concentrated under reduced pressure to give 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester 7 (1.39 g, crude product) as a yellow solid. LC-MS (ESI) m / z = 275.0 [M + H] +

[0180] Step 6: 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A7)

[0181] To a solution of 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester 7 (1.39 g, 5.07 mmol, 1 equivalent) in EtOH (15 mL) / H2O (5.00 mL) was added LiOH (638 mg, 15.2 mmol, 3 equivalents). The mixture was stirred at 25 ° C for 12 hours. LCMS showed that the starting material had been consumed and a main peak with the desired mass was detected. The reaction mixture was diluted with H2O (20.0 mL) and extracted with (DCM 10.0 mL × 2) to remove impurities. The aqueous layer was adjusted to pH = 6 with 2M HCl, the precipitate was collected and dried under reduced pressure to give 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (intermediate A7) (1.30 g, 5.00 mmol, 98.5% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ13.15(br s,1H),9.12(s,1H),8.56(br d,J=6.4Hz,1H),7.35(br s,2H),7.13(br d,J=12.1Hz,1H),2.62(s,3H).LC-MS(ESI)m / z=261.0[M+H] +

[0182] General Method: Synthesis of Intermediate B1

[0183] Synthesis route:

[0184] Step 1: N-methoxy-N-methylpyrazolo[1,5-a]pyridine-2-carboxamide (1)

[0185] To a solution of pyrazolo[1,5-a]pyridine-2-carboxylic acid (2.0 g, 12.3 mmol) and HATU (7.0 g, 18.5 mmol) in DMF (100 mL) was added Et3N (6.2 g, 61.7 mmol) and methoxy(methyl)amine (1.9 g, 30.83 mmol) at 25°C. The mixture was then stirred at 25°C for 12 hours. The reaction was quenched with water (100 mL) and then extracted with EA (100 mL × 3). The organic solution was washed with brine (100 mL). The organic phase was dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with MeOH in DCM from 0% to 10% in 20 minutes to give N-methoxy-N-methylpyrazolo[1,5-a]pyridine-2-carboxamide (2.01 g, 79% yield) as a yellow solid. LC-MS: Rt=1.049min, (ESI)m / z.[M+H] + 206.1; C10H11N3O2.

[0186] Step 2: Pyrazolo[1,5-a]pyridine-2-carbaldehyde (3)

[0187] To a solution of N-methoxy-N-methylpyrazolo[1,5-a]pyridine-2-carboxamide (1.5 g, 7.31 mmol) in THF (20 mL) was added LiAlH4 (439 mg, 10.96 mmol) at -60 ° C. The mixture was then stirred at -60 ° C for 2 hours. The reaction was quenched with water (40 mL) and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with MeOH in DCM from 0% to 10% in 20 minutes to give pyrazolo[1,5-a]pyridine-2-carboxaldehyde (450 mg, 42% yield) as a yellow oil. LC-MS: Rt = 1.071 min, (ESI) m / z. [M + H] + 147.1; C8H6N2O

[0188] Step 3: N-(Pyrazolo[1,5-a]pyridin-2-ylmethyl)-1-(pyrimidin-2-yl)ethan-1-amine (Intermediate B1)

[0189] To a solution of pyrazolo[1,5-a]pyridine-2-carboxaldehyde (100 mg, 0.68 mmol) in MeOH (5 mL) was added 1-(pyrimidin-2-yl)ethylamine (126 mg, 1.03 mmol) and NaBH3CN (86 mg, 1.37 mmol) and reacted at 25°C. The mixture was then stirred at 25°C for 2 hours. The reaction was quenched with water (1 mL) and the solvent was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting from 0% to 5% with a solution of MeOH in DCM in 20 minutes to give N-(pyrazolo[1,5-a]pyridin-2-ylmethyl)-1-(pyrimidin-2-yl)ethylamine (Intermediate B1) (40 mg, 23% yield) as a yellow oil. LC-MS: Rt=0.521min, (ESI)m / z.[M+H]+254.2; C14H15N5

[0190] General Method: Synthesis of Intermediate B2

[0191] Synthesis route:

[0192] Step 1: 2-Dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3)

[0193] A mixture of 5-(trifluoromethyl)pyridin-2-amine (1) (30 g, 185 mmol, 1 eq), chlorobenzene (450 mL) and 1,1,3-trichloro-2-propanone (45 g, 277 mmol, 1.5 eq) was reacted at 135°C for 4 hours. LCMS confirmed the formation of the desired product. The reaction mixture was adjusted to pH 8 with sodium carbonate and extracted with ethyl acetate (500 mL x 3). The combined organic phases were dried over magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (silica, 15% ethyl acetate in petroleum ether) to afford 2-dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3) (30 g, 111 mmol, 60% yield) as a yellow solid. H NMR: ES19506-784-P1A, 1H NMR (400MHz, DMSO-d6) δ9.24 (s, 1H), 8.27 (s, 1H), 7.79 (d, J = 9.68Hz, 1H), 7.65 (s, 1H), 7.56 (dd, J = 1.65, 9.57Hz, 1H).

[0194] Step 2: 6-(Trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (4)

[0195] 2-Dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3) (30 g, 111 mmol, 1 eq), water (600 ml) and calcium carbonate (33 g, 334 mmol, 3 eq) were heated to 100°C and reacted for 2 hours. LCMS detected the formation of the target product. Celite and ethyl acetate (600 ml) were added to the reaction solution, stirred at room temperature for 30 minutes, filtered, and extracted with ethyl acetate (600 ml*2). The combined organic phases were dried over magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain a brown solid 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (4) (35 g). The crude product was used directly in the next step without further purification. H NMR: ES19506-789-P1A1, 1H NMR (400MHz, CHLOROFORM-d) δ 10.09-10.29 (m, 1H), 8.59 (s, 1H), 8.27 (s, 1H), 7.82 (br d, J=9.46Hz, 1H), 7.44 (br d,J=9.02Hz,1H).

[0196] Step 3: 1-Methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B2)

[0197] To a solution of 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (4) (100 mg, 467 umol, 1 eq) in DCM (2.00 mL) was added KOAc (91.7 mg, 934 umol, 2 eq) and 1-methyl-1H-pyrazol-4-amine (45.4 mg, 467 umol, 1 eq) at -5°C, and the reaction mixture was stirred at -5°C for 1 hour. NaBH(OAc)3 (198 mg, 934 umol, 2 eq) was then added and stirred at -5°C for another 3 hours. LCMS (ET63219-45-P1A1) showed complete consumption of Cpd.4 and several new peaks were observed on the LCMS. The reaction mixture was diluted with saturated aqueous Na2CO3 (3.00 mL) and extracted with dichloromethane (2.00 mL x 4). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (ethyl acetate / methanol=8 / 1) to give 1-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B2) (80.0 mg, 271 umol, 58.0% yield) as a yellow solid. 1H NMR(400 MHz, CDCl3) δ8.47(s,1H),7.69(d,J=9.5Hz,1H),7.64(s,1H),7.35(d,J=1.5 ,9.5Hz,1H),7.30(s,2H),6.97(s,1H),4.39(s,2H),3.81(s,3H); LC-MS,[MH] + 217.0.

[0198] General Method: Synthesis of Intermediate B3

[0199] Synthesis route:

[0200] Step 1: 1,3-Dimethyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B3)

[0201] 6-(Trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (4) (10 g, 46 mmol, 1 eq) and 1,3-dimethylpyrazol-4-amine (6 g, 56 mmol, 1.2 eq) were dissolved in dichloromethane (150 ml), and acetic acid (3 g, 56 mmol, 3 ml, 1.2 eq) was added. The reaction solution was reacted at 25 degrees Celsius for 1 hour. Sodium triacetoxyborohydride (25 g, 117 mmol, 2.5 eq) was added, and the reaction solution was reacted at 25 degrees Celsius for 3 hours. LCMS detected the production of the target product. The reaction solution was quenched with 200 ml of sodium bicarbonate and extracted with ethyl acetate (150 ml*2). The combined organic phases were washed with 400 ml of saturated brine, dried over magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 35% ethyl acetate:ethanol (3:1) in petroleum ether) gave 1,3-dimethyl-N-(6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B3) (14 g, 45 mmol, 97% yield) as a brown solid. H NMR:ES19506-795-P1A,1H NMR (400MHz, DMSO-d6) δ9.19(s,1H),7.92(s,1H),7.67(d,J=9.46Hz,1H),7.41(dd,J=1.76,9.46Hz,1H),6.93(s,1H),4.56(br s,1H),4.17(br d, J=3.96Hz, 2H), 3.52-3.61 (m, 3H), 2.04 (s, 3H). LCMS: ES19506-795-P1B, (ESI) m / z=310.3(M+1)+, RT=0.64min.

[0202] General Method: Synthesis of Intermediate B5

[0203] Synthesis route:

[0204] Step 1: Pyrazolo[1,5-a]pyridine-2-carboxylic acid (2)

[0205] To a solution of pyrazolo[1,5-a]pyridine-2-carboxylic acid (10.0 g, 61.67 mmol, 1 equivalent) and HATU (28.14 g, 74.01 mmol, 1.2 equivalents), DIEA (31.88 g, 246.69 mmol, 4 equivalents) in DCM (500 mL) was added N, O-dimethylhydroxylamine hydrochloride (12.03 grams, 123.35 mmol, 2 equivalents). The mixture was stirred at 25 ° C for 16 hours. LCMS (ET63565-18-P1A) showed that the starting material was completely consumed and the product was detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1 to 0 / 1) to obtain compound pyrazolo[1,5-a]pyridine-2-carboxylic acid (2) (12.1 g, 58.9 mmol, 95.6% yield) as a white solid. 1 H NMR(400MHz,CHLOROFORM-d)δ8.49(d,J=7.1Hz,1H),7.57(d,J=8.9Hz,1H),7.15(dd,J=7 .3,8.4Hz,1H),7.00(s,1H),6.85(t,J=6.9Hz,1H),3.79(s,3H),3.49(s,3H); LC-MS,[MH] + 206.22.

[0206] Step 2: 1-(Pyrazolo[1,5-a]pyridin-2-yl)ethan-1-one (3)

[0207] Under nitrogen atmosphere, MeLi (11.70 mL, 1.2 eq) was added to a solution of pyrazolo[1,5-a]pyridine-2-carboxylic acid (2) (2 g, 9.75 mmol, 1 eq) in THF (20.0 mL) at -60 ° C, and the mixture was stirred at 25 ° C for 16 hours. LCMS (ET63565-13-P1A1) showed that the starting material was completely consumed and the product was detected. The residue was purified by column chromatography (petroleum ether / ethyl acetate=50 / 1 to 3 / 1) to give 1-(pyrazolo[1,5-a]pyridin-2-yl)ethane-1-one (3) (277 mg, 1.73 mmol, 17.7% yield) as a white solid. LC-MS, [MH] + 161.1.

[0208] Step 3: N-ethyl-1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-amine (Intermediate B5)

[0209] A mixture of 1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-one (3) (0.1 g, 624.33 umol, 1 eq) and ethylamine (112.58 mg, 2.50 mmol, 163.39 uL, 4 eq) in DCM (2 mL) was stirred at -60 ° C for 0.5 h, and then NaBH 4 (264.64 mg, 1.25 mmol, 2 eq) was added to the mixture at 0 ° C and stirred at 25 ° C under N 2 atmosphere for 16 hours. LCMS (ET63565-9-P1A2) showed that the starting material was completely consumed and the product was detected. The reaction mixture was quenched with H 2 O (5 mL) and diluted with DCM (5 mL×3). The combined organic layers were concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (DCM:MeOH=10:1) to give N-ethyl-1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-amine (Intermediate B5) (50 mg, 264 umol, 42.3% yield) as a white solid. 1 H NMR(400MHz,CHLOROFORM-d)δ8.39(br d,J=6.9Hz,1H),7.46(br d,J=8.5Hz,1H),7.12-7.03(m,1H),6.70(br t,J=6.8Hz,1H),6.44(s,1H),4.13(br d,J=6.8Hz,1H),3.32-3.21(m,1H),2.77-2.56(m,2H),1.53(d,J=6.6Hz,4H),1.17-1.14(m,3H):LC-MS,[MH] + 190.1.

[0210] General Method: Synthesis of Intermediate B6

[0211] Synthesis route:

[0212] Step 1: 2-(Dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3)

[0213] A mixture of 4-(trifluoromethyl)benzene-1,2-diamine (5 g, 28.3 mmol, 1 eq) and 2,2-dichloroacetic acid (7.32 g, 56.7 mmol, 4.66 mL, 2 eq) in HCl (125 mL) (4 M) was stirred at 100 ° C for 10 minutes. 16 hours. LCMS (ET60224-68-P1A) showed that Cpd.1 was consumed and the desired mass was detected. The reaction mixture was filtered and the filter cake was washed with water. The combined filtrate was extracted with DCM (20 ml * 3). The combined organic layer was washed with brine (100 mL), dried over MgSO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 5 / 1) to give compound 2-(dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3) (4.4 g, 16.3 mmol, yield 57.6%) as a yellow oil. 1 H NMR (400MHz, CHLOROFORM-d) δ8.01 (s, 1H), 7.79 (d, J = 8.6Hz, 1H), 7.66 (d, J = 8.2Hz, 1H), 7.26 (s, 1H); LCMS: [M+H] + 268.9

[0214] Step 2: 6-(Trifluoromethyl)-1H-benzo[d]imidazole-2-carbaldehyde (4)

[0215] To a suspension of 2-(dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3) (1 g, 3.72 mmol, 1 eq) in H 2 O (20 mL) was added CaCO 3 (1.12 g, 11.1 mmol, 3 eq). The mixture was stirred at 100 ° C for 8 hours. LCMS (ET60224-74-P1B) showed that Cpd.3 was consumed and the desired mass was detected. The reaction mixture was diluted with H 2 O (50 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layers were concentrated under reduced pressure to give 6-(trifluoromethyl)-1H-benzo[d]imidazole-2-carbaldehyde (4) (310 mg, 1.45 mmol, 38.9% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ14.28-13.59(m,1H),10.02(s,1H),8.40-8.30(m,2H),8.23-8.05(m,3H),8.00(dd,J=5.4,8.5Hz,1H),7.89(br s,1H),7.83-7.61(m,3H),7.34(br d,J=7.9Hz,1H),7.27(d,J=7.6Hz,1H); LCMS:[M+H]+ 215.2

[0216] Step 3: 2-Methyl-N-((6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)methyl)propan-1-amine (Intermediate B6)

[0217] To a solution of 6-(trifluoromethyl)-1H-benzo[d]imidazole-2-carbaldehyde (4) (0.31 g, 1.45 mmol, 1 eq) and 2-methylpropan-1-amine (105 mg, 1.45 mmol, 143 uL, 1 eq) in DCM (6.2 mL) was added KOAc (170 mg, 1.74 mmol, 1.2 eq) at 25°C. The mixture was stirred at 25°C for 0.5 h, and then NaBH(OAc)3 (398 mg, 1.88 mmol, 1.3 eq) was added to the mixture at 25°C, and the mixture was stirred at 25°C for 15.5 h. LCMS (ET60224-77-P1A) showed that Cpd.4 was consumed and the desired mass was detected. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (2 mL×3). The combined organic layers were washed with brine (10 mL), dried over MgSO , filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (petroleum ether / ethyl acetate=0 / 1) to give 2-methyl-N-((6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)methyl)propan-1-amine (Intermediate B6) (100 mg, 368 μmol, 25.4% yield, 100% purity) as a colorless oil. 1 H NMR(400MHz,CHLOROFORM-d)7.80(s,1H),7.56(d,J=8.4Hz,1H),7.43(d,J=8.5Hz,1H),4 .07(s,2H),2.45(d,J=6.8Hz,2H),1.82-1.68(m,1H),0.89(d,J=6.6Hz,6H); LCMS:[M+H] + 272.0

[0218] General Method: Synthesis of Intermediate B7

[0219] Step: 2-Methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-1-amine (Intermediate B7)

[0220] To a solution of 2-methylpropan-1-amine (68.3 mg, 933 μmol, 92.8 μL, 1 eq) and 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (0.2 g, 933 μmol, 1 eq) in DCM (4 mL) was added KOAc (109 mg, 1.12 mmol, 1.2 eq) at 25°C. The mixture was stirred at 25°C for 0.5 h. NaBH(OAc) (257 mg, 1.21 mmol, 1.3 eq) was then added to the mixture at 25°C, and the mixture was stirred at 25°C for 15.5 h. LCMS (ET60224-75-P1A) indicated that Cpd.4 was consumed and the desired mass was detected. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (2 mL x 3) to remove impurities. The aqueous layer was basified with saturated Na2CO3 to pH = 8 and then extracted with DCM (10 mL * 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to afford the intermediate 2-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-1-amine (Intermediate B7) (108 mg, 398 umol, 42.6% yield, 100% purity) as a colorless oil. 1 H NMR(400MHz,CHLOROFORM-d)δ8.49(s,1H),7.70-7.62(m,2H),7.33(br d,J=9.2Hz,1H),4.01(s,2H),2.54(d,J=6.7Hz,2H),1.84(quind,J=6.6,13.3Hz,1H),0.97(d,J=6.6Hz,6H); LC-MS,[MH] + 272.0.

[0221] Synthesis of Example 4

[0222] Step 7: (E)-4-(((dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1)

[0223] To a solution of 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (intermediate A6) (50.0 mg, 206 umol, 1 equiv) in DCM (2 mL) was added HCl / dioxane (4 M). The mixture was stirred at 25 ° C for 0.5 hours. The reaction mixture was then concentrated, evaporated with hexane (10 mL×3), and cooled to 0 ° C. Oxalyl chloride (157 mg, 1.24 mmol, 108 uL, 6 equiv) and DMF (3.02 mg, 41.3 umol, 3.18 uL, 0.2 equiv) were added dropwise at 0 ° C. The mixture was stirred at 0 ° C for 2 hours. A sample was taken in MeOH. LC-MS showed that the desired mass of methyl ester was detected. The mixture was concentrated under reduced pressure to give a residue. The residue was triturated with n-hexane (5 mL) and then concentrated under reduced pressure to give (E)-4-(((dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1) (50.0 mg, 158 umol, 76.7% yield) as a yellow solid. LC-MS, [MH] + 243.1

[0224] Step 8: (E)-4-(((dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2)

[0225] To a solution of (E)-4-(((dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1) (50.0 mg, 158 umol, 1 eq) and DIEA (81.9 mg, 633 umol, 110 uL, 4 eq) in THF (2 mL) was added Int.6 (44.7 mg, 158 umol, 1 eq) at 0° C. The mixture was stirred at 0° C. for 2 hours. LC-MS showed complete consumption of the starting material and the desired mass was detected. The mixture was concentrated under reduced pressure to afford (E)-4-(((dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2) (61.0 mg, 109 umol, 68.6% yield) as a yellow solid. LC-MS, [MH]+ 562.2.

[0226] Step 9: 4-amino-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Example 4)

[0227] A solution of (E)-4-(((dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2) (60.0 mg, 107 umol, 1 eq) in NH / MeOH (2 mL) was stirred at 70 ° C for 2 hours. LC-MS showed complete consumption of the starting material and the desired mass was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150*40mm*10um; mobile phase: [water(NH4HCO3)-ACN]; B%: 25%-65%, 8 minutes, UV 220&254nm) to give 4-amino-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Example 4) (19.2 mg, 37.9 umol, 35.48% yield, 100% purity) as a white solid. 1 H NMR(400MHz, DMSO-d6)δ8.84(br s,1H)8.79(br d,J=4.63Hz,2H)8.28(br s,1H)8.13(br d,J=6.75Hz,1H)7.80-8.01(m,1H)7.49-7.72(m,3H)7.29-7.47(m,1H)6.99(br s,2H)5.43(br s,1H)4.92(br d,J=17.01Hz,1H)4.52(br d,J=16.51Hz,1H)2.50(br s,3H)1.62(br d,J=7.00Hz,3H).LC-MS,[MH] + 507.2

[0228] Synthesis of Example 15

[0229] Step 1: 4-(2,4-dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3)

[0230] 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (h) (100 mg, 246.0 μmol, 1 eq) was dissolved in N,N-dimethylformamide (2.5 ml), and N-(5-(trifluoromethyl)pyridin-2-ylmethyl)ethanamine (60.3 mg, 295.2 μmol, 1.2 eq), N,N,N,N-tetramethylchloroformamidine hexafluorophosphate (138 mg, 492.1 μmol, 2 eq) and N-methylimidazole (101 mg, 1.23 mmol, 98.1 μL, 5 eq) were added. The reaction mixture was reacted at 50°C for 16 hours. LCMS showed that the starting material was completely consumed and the target product was produced. The reaction mixture was concentrated to dryness under reduced pressure and purified by column chromatography ( 4 grams Silica gel column, 0-2.3% methanol / dichloromethane system, flow rate 20 ml / min) to give a yellow oil 4-(2,4-dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3) (150 mg, 233.45 μmol, 94.88% yield, 92.23% purity). LCMS: ES19974-409-P1B1, (ESI) m / z = 593.2 [M+1] +; RT = 0.82 min, purity: 92.23%

[0231] Step 2: 4-amino-N-ethyl-1,3-dimethyl-N-((5-trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 15)

[0232] 4-(2,4-Dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3) (140 mg, 236.2 μmol, 1 eq) was dissolved in dichloromethane (0.5 ml) and trifluoroacetic acid (2.5 ml) was added. The reaction solution was reacted at 50°C for 16 hours. LC-MS detected that the starting material was completely consumed and the target product was generated. The reaction solution was concentrated to dryness under reduced pressure, 10% sodium carbonate (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over magnesium sulfate, filtered, concentrated to dryness, and purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 150 x 30 mm x 5 μm; mobile phase: [water (ammonia)-acetonitrile]; B% gradient: 30%-50% over 9 minutes) to give 4-amino-N-ethyl-1,3-dimethyl-N-((5-trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 15) (30 mg, 67.81 μmol, 28.70% yield) as a white solid. LCMS: ES19974-414-P1B1, (ESI) m / z=443.1[M+1]+; RT=2.623min, purity: 100.0%. HNMR: ES19974-414-P1A,1H NMR (400MHz, DMSO-d6)Shift=8.97(br s,1H),8.20(br d,J=6.9Hz,1H),7.97(br s,1H),7.61(br d,J=8.1Hz,1H),7.39(br s,2H),6.72(br s,2H),4.82(br s,2H),3.45(q,J=6.9Hz,2H),3.10-2.60(m,3H),2.56(br s,3H),1.14(t,J=7.0Hz,3H)

[0233] Synthesis of Example 86

[0234] Step 1: 7-Chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methylpyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3)

[0235] 7-Chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3) (145 mg, 339 μmol, 1 eq) was dissolved in 1-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-4-amine (Intermediate B2) (100 mg, 339 μmol, 1 eq) in acetonitrile (2 mL). N,N,N,N-tetramethylchloroformamidine hexafluorophosphate (285 mg, 1 mmol, 3 eq) and N-methylimidazole (139 mg, 1.7 mmol, 135 μL, 5 eq) were then added sequentially. The reaction was stirred at 50°C for 16 hours. LC-MS analysis confirmed the complete reaction of the starting material and the formation of the desired product. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (silica, 10% methanol in dichloromethane) to afford 7-chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methylpyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3) as a yellow oil (130 mg, 185 μmol, 54.5% yield).

[0236] Step 2: 4-amino-7-chloro-1-methyl-N-(1-methyl-1H-pyrazol-4-yl)-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl])methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 86)

[0237] 7-Chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methylpyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3) (120 mg, 170 μmol, 1 eq) was added to a mixture of trifluoroacetic acid (0.4 mL) and dichloromethane (1 mL) and dissolved. The reaction was stirred at 50°C for 16 hours. LC-MS showed that the starting material was completely reacted and the desired product was produced. The reaction mixture was concentrated to dryness under reduced pressure. Purification by reverse phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water and acetonitrile as eluent) gave 4-amino-7-chloro-1-methyl-N-(1-methyl-1H-pyrazol-4-yl)-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl])methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 86) as a white product (30 mg, 53 μmol, 31% yield, 98% purity). H NMR: 1H NMR(400MHz,DMSO-d6)δ9.15-9.31(m,1H),8.33(s,1H),8.07-8.16(m,1H),7.92-8.03(m,1H),7.76-7.86(m,1H),7.58-7.75(m,2H),7.41-7 .55(m,3H),7.18-7.33(m,1H),4.71-5.17(m,2H),3.53-3.83(m,3H),2.80-2.97(m,3H).LCMS:(ESI)m / z=554.2(M+1)+,RT=1.544min,purity of 98.7%.

[0238] Synthesis of Example 143

[0239] Synthesis method:

[0240] Step 1: 2-Methyl-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3)

[0241] 5-(Trifluoromethyl)picolinaldehyde (2) (300 mg, 1.71 mmol, 1.0 eq), 2-methylpyridin-3-amine (1) (216 mg, 2.00 mmol, 1.17 eq) and acetic acid (123 mg, 2.05 mmol, 117.25 μL, 1.2 eq) were dissolved in dichloromethane (10 ml) and stirred at room temperature for 4 hours. Sodium triacetoxyborohydride (1.09 g, 5.15 mmol, 3.01 eq) was added and stirred at room temperature for 12 hours. LCMS showed that the starting material was completely consumed and the product was generated. The reaction solution was diluted with water and dichloromethane, and the pH was adjusted to about 12 with 2M aqueous potassium carbonate solution. The layers were separated, and the aqueous phase was extracted with dichloromethane (10 ml * 3). The combined organic phases were dried over sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography ( 12 grams Silica gel column, 0-25% ethyl acetate / petroleum ether system, flow rate, 30 ml / min) to give 2-methyl-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (205 mg, 44.7% yield) as a yellow syrup. LCMS ES15882-1182-P1A: (ESI) m / z = 268.1 [M+1] +; RT = 0.65 min

[0242] Step 2: 4-amino-7-fluoro-1-methyl-N-(2-methylpyridin-3-yl)-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 143)

[0243] 2-Methyl-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (60 mg, 224.51 μmol, 1.0 eq), 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (60 mg, 230.57 μmol, 1.03 eq) and N-ethyl-N-isopropyl-2-propanamine (119 mg, 925.11 μmol, 161.13 μL, 4.12 eq) were dissolved in 1-methylpyrrolidin-2-one (1.0 mL). 2-Chloro-1,3-dimethyl-4,5-dihydroimidazol-1-yl chloride (DMC) (57.82 mg, 342.02 μmol, 1.52 eq) was added and the reaction mixture was reacted at 80°C for 16 hours. The target product was detected by LCMS. The reaction solution was purified by reverse-phase preparative liquid chromatography (formic acid conditions; Boston Prime C18 column, 150*30 mm*5 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 15%-35% B, 10 minutes) to give 4-amino-7-fluoro-1-methyl-N-(2-methylpyridin-3-yl)-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 143) (8 mg, 6.29% yield) as a yellow solid. LCMS ES15882-1205-P1B1: (ESI) m / z=510.1[M+1]+; RT=0.996min.1H NMR(400MHz, DMSO-d6)Shift 8.84-8.98(m,1H),8.17-8.27(m,2H),7.72-7.87(m,4H),7.50(s,2H),7.13(dd,J=4.64,7.91Hz,1H) ,7.01(d,J=11.29Hz,1H),5.44(d,J=15.81Hz,1H),5.01(d,J=15.81Hz,1H),2.82(s,3H),2.37(s,3H)

[0244] 19F NMR(376.5MHz,DMSO-d6)Shift-60.71,-60.85,-116.51,-119.18

[0245] Synthesis of Example 145

[0246] Synthesis method:

[0247] Step 1: 2-Methoxy-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3)

[0248] 5-(Trifluoromethyl)picolinaldehyde (2) (300 mg, 1.71 mmol, 1.0 eq), 2-methoxypyridin-3-amine (1) (248 mg, 2.00 mmol, 1.17 eq) and acetic acid (123 mg, 2.05 mmol, 1.2 eq) were dissolved in dichloromethane (10 ml) and stirred at room temperature for 4 hours. Sodium triacetoxyborohydride (1.09 g, 5.15 mmol, 3.01 eq) was added and stirred at room temperature for 12 hours. LCMS showed that the starting material was completely consumed and the product was generated. The reaction solution was diluted with water and dichloromethane, and the pH was adjusted to about 12 with 2M potassium carbonate aqueous solution. The layers were separated, and the aqueous phase was extracted with dichloromethane (10 ml * 3). The combined organic phases were dried over sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography ( 12 grams Silica gel column, 0-15% ethyl acetate / petroleum ether system, flow rate, 30 mL / min) to give 2-methoxy-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (430 mg, 1.52 mmol, 88.61% yield) as a yellow syrup.

[0249] LCMS ES15882-1189-P1A: (ESI)m / z=284.0[M+1]+; RT=0.82min

[0250] Step 2: 4-amino-7-fluoro-N-(2-methoxypyridin-3-yl)-1-methyl-N-(5-trifluoromethyl)pyridin-2-methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 145)

[0251] 2-Methoxy-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (64 mg, 226 μmol, 1.0 eq.), 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (61 mg, 234 μmol, 1.04 eq.), and N-ethyl-N-isopropyl-2-propanamine (119 mg, 919 μmol, 4.07 eq.) were dissolved in 1-methylpyrrolidin-2-one (1.0 ml), and 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-yl chloride (DMC) (58 mg, 344 μmol, 1.52 eq.) was added. The reaction mixture was reacted at 80°C for 16 hours. The target product was detected by LCMS. The reaction solution was purified by reverse phase preparative liquid chromatography (formic acid conditions; Boston Prime C18 column, 150*30 mm*5 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 23%-43% B over 10 minutes) to afford 4-amino-7-fluoro-N-(2-methoxypyridin-3-yl)-1-methyl-N-(5-trifluoromethyl)pyridin-2-methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 145) as a yellow solid (12 mg, 8.79% yield, 87% purity). LCMS ES15882-1206-P1B1: (ESI) m / z=526.1 [M+1]+; RT=1.140 min

[0252] 1H NMR(400MHz,DMSO-d6)Shift 8.91(s,1H),8.23-8.30(m,1H),8.20(s,0.2H,HCOOH),7.91-7.95(m,1H),7.86(d,J=6.53Hz,1H),7.77-7.83(m,2H),7.72-7.76(m,1H),7.50(br s,2H),7.03(d,J=11.29Hz,1H),6.89(dd,J=5.02,7.53Hz,1H),5.34(d,J=16.06Hz,1H),5.04(d,J=15.81Hz,1H),3.74(s,3H),2.80(s,3H)

[0253] 19F NMR(376.5MHz,DMSO-d6)Shift-60.72,-117.24

[0254] Synthesis of Example 188

[0255] Synthesis method:

[0256] Step 1: 1,3-Dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazol-4-amine (2)

[0257] 5-(Trifluoromethyl)picolinaldehyde (1) (300 mg, 1.71 mmol, 1 eq) was dissolved in dichloromethane (8 mL). 1,3-Dimethyl-1H-pyrazol-4-amine (247 mg, 2.22 mmol, 1.3 eq) and glacial acetic acid (134 mg, 2.23 mmol, 127.74 μL, 1.3 eq) were added. The reaction was stirred at 15-20°C for 4 hours. Sodium acetate borohydride (1.09 g, 5.15 mmol, 3.01 eq) was then added, and the mixture was stirred at 15-20°C for 12 hours. LC-MS analysis indicated that the reaction of the starting material was complete and the target product was produced. The reaction solution was added with 10 ml of water and 10 ml of dichloromethane, basified to pH 12 with a 2 mol / L aqueous solution of potassium carbonate, separated, extracted with dichloromethane (10 ml * 3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (12 g silica gel column, eluent 0-40% (ethyl acetate:ethanol=3:1) / petroleum ether, flow rate @ 30 ml / min), and concentrated under reduced pressure to give 1,3-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazol-4-amine (3) (311 mg, 1.15 mmol, 67.17% yield) as a yellow solid.

[0258] LCMS ES15882-1166-P1A: (ESI)m / z=228.2[M+1]+; RT=0.601min

[0259] Step 2: 4-amino-N-(1,3-dimethyl-1H-pyrazol-4-yl)-1,7-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 188)

[0260] To a reaction mixture of 4-amino-1,7-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (c) (42 mg, 163.90 μmol, 1 eq), 1,3-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazol-4-amine (b) (45.18 mg, 167.17 μmol, 1.02 eq), and N,N-diisopropylethylamine (84.73 mg, 655.59 μmol, 114.19 μL, 4 eq) in 1-methyl-2-pyrrolidone (1 mL) was added 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-yl chloride (41.56 mg, 245.84 μmol, 1.5 eq). The reaction was stirred at 50°C for 3 hours. LC-MS confirmed the complete reaction of the starting material and the formation of the desired product. Purification by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm column length, using decreasingly polar mixtures of water (containing 0.05% formic acid) and acetonitrile (20%-40%) as eluents) afforded a yellow solid. The crude product was then purified by reverse phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water (containing 0.05% formic acid) and acetonitrile (18%-38%) as eluents) to afford 4-amino-N-(1,3-dimethyl-1H-pyrazol-4-yl)-1,7-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 188) as a white solid (10 mg, 19.67 μmol, 12.00% yield). LCMS: ES13685-1063-P1D, (ESI) m / z=509.1[M+1]+; RT=1.066min NMR: ES13685-1063-P1A, 1H NMR (400MHz, DMSO-d6) 8.95 (s, 1H), 8.25 (br d,J=8.60Hz,1H),8.15(s,1H),7.75(s,2H),7.56-7.72(m,2H),7.23(br d,J=4.84Hz,3H),5.14(s,2H),3.51-3.70(m,3H),2.79(s,3H),2.38-2.44(m,3H),1.83(s,3H),19F NMR(376MHz,DMSO-d6)-60.68(br s,3F)

[0261] Synthesis of Example 210

[0262] Synthesis method:

[0263] Step 1: 1,4-Dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazol-2-amine (2)

[0264] 2-Bromo-1,4-dimethyl-1H-imidazole (210 mg, 1.20 mmol, 1 eq) was dissolved in tetrahydrofuran (12 ml), and (5-(trifluoromethyl)pyridin-2-yl)methanamine (1) (211 mg, 1.20 mmol, 1 eq), sodium trimethylsilyl (oxide) (267 mg, 1.44 mmol, 1.2 eq) and bromo[dicyclohexyl[3-(1 The reaction was stirred at 80 degrees Celsius for 16 hours. LC-MS analysis showed that the reaction of the starting material was complete and the target product was produced. The mixture was cooled to room temperature, 10 ml of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 ml * 3). The organic phase was dried over anhydrous magnesium sulfate, and the filtrate was filtered and concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (4 g + 4 g silica gel column, eluent 0-40% ethyl acetate / petroleum ether, flow rate 20 ml / min) to obtain a yellow liquid 1,4-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazole-2-amine (2) (50 mg, 185.01 μmol, 15.42% yield).

[0265] LCMS: ES13685-1155-P1C, (ESI) m / z=271.1[M+1]+; RT=0.926min

[0266] Step 2: 4-amino-N-(1,4-dimethyl-1H-imidazol-2-yl)-7-fluoro-1-methyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 210)

[0267] To a reaction mixture of 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (40 mg, 153.71 μmol, 1 eq), 1,4-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazol-2-amine (2) (41.54 mg, 153.71 μmol, 1 eq), and N,N-diisopropylethylamine (79.46 mg, 614.86 μmol, 107.09 μL, 4 eq) in 1-methyl-2-pyrrolidone (1 mL) was added 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-yl chloride (38.98 mg, 230.5 μmol, 1.5 eq). The reaction was stirred at 60°C for 16 hours. LC-MS analysis indicated the presence of a small amount of starting material and the desired product. Purification by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5-micron silica, 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water (containing 0.05% formic acid) and acetonitrile (10%-30%) as eluent) afforded a yellow solid. The crude product was further purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5-micron silica, 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water (containing 0.05% ammonia) and acetonitrile (28%-48%) as eluent) to afford 4-amino-N-(1,4-dimethyl-1H-imidazol-2-yl)-7-fluoro-1-methyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 210) as a white solid (3 mg, 5.56 μmol, 3.62% yield). LCMS: ES13685-1158-P1C1, (ESI)m / z=513.3[M+1]+; RT=0.759min

[0268] NMR: ES13685-1158-P1A, 1H NMR (400MHz, DMSO-d6) 8.91 (br s, 1H), 8.23 ​​(br d, J=8.13Hz, 1H), 7.97 (br d, J=6.75Hz, 1H), 7.89 (br s,1H),7.81(s,1H),7.34(br s,2H),7.13(br d,J=11.26Hz,1H),6.50(s,1H),5.17(br s,2H),3.23-3.28(m,3H),2.87(s,3H),1.96(br s,3H).NMR:ES13685-1158-P1A,19F NMR(376MHz,DMSO-d6)-60.85(s,3F),-118.24(br s,1F)

[0269] Synthesis of Example 213

[0270] Synthesis method:

[0271] Step 1: 1-Methyl-N-(1-trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3)

[0272] 1-Methylpyrazol-5-amine (133 mg, 1.37 mmol, 1.5 eq) and 1-trifluoromethylpyrazole-4-carboxaldehyde (1) (150 mg, 914.16 μmol, 1 eq) were dissolved in methanol (5 ml) and acetic acid (164 mg, 2.74 mmol, 156.99 μL, 3 eq) was added. The reaction solution was stirred at room temperature for half an hour. Sodium cyanoborohydride (172.34 mg, 2.74 mmol, 3 eq) was added and stirred at room temperature for 15.5 hours. LCMS showed that the reaction was complete and the target product was generated. The mixture was concentrated to dryness under reduced pressure, sodium bicarbonate (10 ml) was added, and the mixture was extracted with ethyl acetate (10 ml * 3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography ( 12g Silica gel column, 0-60% ethyl acetate:ethanol (3:1) / petroleum ether system, flow rate, 35 mL / min). 1-Methyl-N-(1-trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3) (220 mg, 897.21 μmol, 86.86% yield) was obtained as a colorless oil. LCMS: ES13683-1961-P1C, (ESI) m / z = 246.1 (M+1)+, RT = 0.786 min

[0273] Step 2: 4-amino-7-fluoro-1-methyl-1-methylpyrazol-5-yl)-N-(1-trifluoromethyl-1H-pyrazol-4-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 213)

[0274] 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (63.6 mg, 244.69 μmol, 1 eq) and 1-methyl-N-(1-trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3) (60 mg, 244.69 μmol, 1 eq) were dissolved in 1-methylpyrrolidin-2-one (1.5 ml), and N-ethyl-N-isopropyl-2-propanamine (158 mg, 1.22 mmol, 213.11 μl, 5 eq) and 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (DMC) (62.0 mg, 367.04 μmol, 1.5 eq) were added. The reaction mixture was reacted at 50°C for 16 hours. LCMS analysis showed that the starting material was completely consumed and the target product was produced. The reaction solution was diluted with acetonitrile (1 mL) and water (1 mL), filtered, and the filtrate was purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 150*30 mm*5 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 18%-38% over 10 minutes) to afford 4-amino-7-fluoro-1-methyl-1-methylpyrazol-5-yl)-N-(1-trifluoromethyl-1H-pyrazol-4-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 213) as a white solid (50 mg, 100.53 μmol, 41.08% yield, 98% purity). LCMS: ES13683-1966-P1C, (ESI) m / z=488.3[M+1]+, RT=0.767min HNMR: ES13683-1966-P1A, 1H NMR (400MHz, DMSO-d6)Shift 8.39(s,1H),8.17(s,0.2H),7.84-8.07(m,2H),7.80(s,1H),7.52(s,2H),7.23(br s,1H),7.06(br d,J=11.01Hz,1H),6.06(br s,1H),4.49-5.28(m,2H),3.48-3.74(m,3H),2.87(s,3H).FNMR:19F NMR(376MHz,DMSO-d6)Shift-59.21(s,3F),-118.10(br s,1F)

[0275] Synthesis of Example 220

[0276] Synthesis method:

[0277] Step 1: 4-Fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) 6-(Trifluoromethyl)imidazo[1,2-a]pyridine-2-carbaldehyde (1) (700 mg, 3.27 mmol, 1 eq) was dissolved in methanol (14 mL), 4-fluoro-1-methyl-1H-pyrazol-5-amine (2) (402.61 mg, 3.50 mmol, 1.07 eq) and glacial acetic acid (255.18 mg, 4.25 mmol, 243.26 μL, 1.3 eq) were added, and the reaction was stirred at 25°C for 1 hour. Sodium cyanoborohydride (616.24 mg, 9.81 mmol, 3 eq) was then added, and the mixture was stirred at 25°C for 15 hours. LC-MS showed that the reaction of the starting materials was complete and the target product was generated. The reaction solution was concentrated and dried by rotary evaporation. 14 mL of 2 mol / L sodium carbonate solution and 14 mL of ethyl acetate were added, the mixture was separated, and the mixture was extracted with ethyl acetate (14 mL*3). The organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (12 g + 4 g silica gel column, eluent 0-30% ethyl acetate: ethanol 3:1 / petroleum ether, flow rate 30 mL / min), and concentrated under reduced pressure to obtain a light yellow solid 4-fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) (970 mg, 3.10 mmol, 94.73% yield). LCMS: ES13685-1213-P1A, (ESI) m / z = 314.0 [M+1] +; RT = 1.537 min. NMR: ES13685-1213-R2A, 1H NMR (400MHz, CHLOROFORM-d) 7.11 (d, J = 4.38Hz, 1H), 3.57 (s, 3H), 3.24 (br s, 2H). HNMR:ES13685-1213-R2A,19F NMR(376MHz,CHLOROFORM-d)-185.48(s,1F)

[0278] Step 2: 4-amino-7-fluoro-N-(4-fluoro-1-methyl-1H-pyrazol-5-yl)-1-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 220)

[0279] To a reaction mixture of 4-amino-7-fluoro-methyl-imidazo[1,5-a]quinoxaline-8-carboxylic acid (500 mg, 1.92 mmol, 1 eq), 4-fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) (601.89 mg, 1.92 mol, 1 eq) and N,N-diisopropylethylamine (993.30 mg, 7.69 mol, 1.34 mL, 4 eq) dissolved in 1-methyl-2-pyrrolidinone (10 mL) was added 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-yl chloride (487.23 mg, 2.88 mmol, 1.5 eq). The reaction was stirred at 50°C for 16 hours. LC-MS analysis indicated 47% starting material remaining and 36% desired product. 2-Chloro-1,3-dimethyl-4,5-dihydroimidazolium-1-chloride (74.71 mg, 441.93 μmol, 0.23 equiv) was then added. The reaction was stirred at 50°C for 16 hours. LC-MS analysis indicated 49% starting material remaining and 38% desired product. Purification by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water (containing 0.05% formic acid) and acetonitrile (20%-40%) as eluents) afforded a pale yellow solid.NMR analysis revealed partial formation of a formate salt. The crude product was then purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using decreasingly polar mixtures of water (containing 0.05% ammonia) and acetonitrile (32%-52%) as eluents) to afford 4-amino-7-fluoro-N-(4-fluoro-2-methyl-pyrazol-3-yl)-1-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (Example 220) as a white solid (155 mg, 276.26 μmol, 14.38% yield). LCMS: ES13685-1217-P1D1, (ESI) m / z=556.1[M+1]+; RT=1.929minNMR: ES13685-1217-P1B,1H NMR(400MHz, DMSO-d6)9.28(br s,1H),8.10(br s,1H),7.92(br s,1H),7.82(s,1H),7.73(d,J=9.76Hz,1H),7.55(s,2H),7.47(br d,J=8.63Hz,1H),7.29(br s,1H),7.11(br d,J=10.38Hz,1H),5.19(br s,1H),5.04-5.14(m,1H),3.59(s,3H),2.86(br s,3H). NMR: ES13685-1217-P1B, 19F NMR (376MHz, DMSO-d6) -60.47 (s, 3F), -118.15 (s, 1F), -173.54 (s, 1F).

[0280] Synthesis of Example 287

[0281] Step 1: (E)-4-(((dimethylamino)methylene)amino)-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A7-1)

[0282] To a stirred solution of 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A7) (270 mg, 1.04 mmol, 1 eq) in DCM (3.00 mL) was added HCl / dioxane (4 M, 778 uL, 3 eq). The mixture was stirred at 25 ° C for 0.5 hours. The reaction mixture was then concentrated, evaporated and dried with toluene (10 mL x 3), the crude material was dissolved in DCM (3.00 mL) and cooled to 0 ° C, and oxalyl chloride (790 mg, 6.23 mmol, 544 uL, 6 eq added dropwise at 0 ° C) and DMF (75.8 mg, 1.04 mmol, 79.8 uL, 1 eq) were added. The mixture was stirred at 25 ° C for 12 hours. LCMS showed that the starting material had been consumed and a main peak with the desired mass was detected. The reaction mixture was concentrated, digested with n-hexane (10 mL x 3), and dried under reduced pressure to obtain (E)-4-(((dimethylamino)methylene)amino)-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A7-1) (340 mg, crude) as a yellow solid. LC-MS (ESI) m / z = 330.0 [M+H] +

[0283] Step 2: 4-amino-7-fluoro-N-(1-methoxypropan-2-yl)-3-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 287)

[0284] To a solution of 1-methoxy-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-2-amine (50.0 mg, 174 umol, 1 eq) and DIEA (89.9 mg, 696 umol, 121 uL, 4 eq) in THF (1.00 mL) was added Int.24-COCl (63.9 mg, 191 umol, 1.1 eq)) at 0 ° C. The mixture was stirred at 25 ° C for 3 hours. The reaction mixture was quenched by adding MeOH (1.00 mL) at 25 ° C and concentrated under reduced pressure to obtain a residue. The residue was dissolved in MeOH (1 mL) and NH3 / MeOH (7 M, 1 mL). The mixture was stirred at 70 ° C for 2 hours. LCMS showed that the starting material had been consumed and a major peak with the desired mass was detected. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30mm*10um; mobile phase: [water(NH4HCO3)-ACN]; B%: 30%-60%, 8 minutes, UV 220nm&254nm) to give 4-amino-7-fluoro-N-(1-methoxypropan-2-yl)-3-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 287) (58.1 mg, 109umol, 64.2% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ9.29-8.54(m,2H),8.18-7.78(m,2H),7.66(br s,1H),7.38(br d,J=7.9Hz,1H),7.13(br s,1H),6.72(br s,2H),4.95-4.39(m,2H),4.22-3.82(m,1H),3.52(br s,2H),3.31-3.10(m,3H),2.63(s,3H),1.21(br s,3H)LC-MS(ESI)m / z=530.2[M+H] +

[0285] The following compounds were synthesized based on the general method. The product structures are shown in Tables 1 and 2. The product characterizations are as follows:

[0286] Table 3

[0287] Biological test example 2: In vitro proliferation inhibition experiment of HCT116 and HCT116-MTAP-KO cells

[0288] Experimental Materials

[0289] The HCT116 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences. The MTAP gene was knocked out using CRISPR / Cas9 technology to obtain the HCT116-MTAP-KO cell line.

[0290] McCoy's 5A medium (Gibco, catalog number 16600082), fetal bovine serum (Gibco, catalog number 10099141C), penicillin-streptomycin double antibody (Gibco, catalog number 15140122), trypsin (Gibco, catalog number 25200056), CellTiter-Glo detection kit (Promega, catalog number G7572), 384-well clear flat-bottom black wall cell culture plates (Corning, catalog number 3764), ultra-micropipette reader (Tecan, catalog number D300e), multi-function microplate reader (Biotek, catalog number SynergyHTX)

[0291] Experimental methods

[0292] 1. Cell culture: HCT116 and HCT116-MTAP-KO cells were cultured in McCoy's 5A medium + 10% fetal bovine serum + 1% penicillin-streptomycin. The cells were kept in the logarithmic growth phase and had a cell viability greater than 95%.

[0293] 2. Preparation of compound concentration gradient: The test compound was added to a 384-well plate using an ultra-micropipette, starting from 30 μM (HCT116 cells) or 3 μM (HCT116-MTAP-KO cells), and diluted 3-fold with DMSO for a total of 9 concentrations in triplicate.

[0294] 3. Compound-treated cells: Add 40 μL of the trypsinized HCT116 or HCT116-MTAP-KO cell suspension to a 384-well plate containing the test compound, i.e., 100 cells per well, with a final DMSO concentration of 0.4%. Incubate the cell culture plate in a 37°C, 5% CO2 incubator for 6 days.

[0295] 4. Detection: Add 20 μL of CellTiter-Glo reagent to each well of the cell culture plate and incubate at room temperature with shaking for 30 minutes. Detect the luminescence signal at 578 nm using a multi-function microplate reader.

[0296] 5. Data Analysis:

[0297] GraphPad Prism 8.0 software was used to fit the data using a four-parameter inhibitor-response model to obtain the IC values ​​of the test compounds. 50 value (half maximal inhibitory concentration).

[0298] The biological activities of some compounds were determined by experimental methods. "A" represents IC 50 (nm)<100, “B” means 100 <IC 50 (nm)<1000, “C” means 1000 <IC 50 (nm)<10000, see Table 4:

[0299] Among them, the first column is the cell proliferation inhibition rate HCT116MTAP WT IC 50 (nm), the second column is the cell proliferation inhibition rate HCT116-MTAP null IC 50 (nm)

[0300] The corresponding structures of the tested products are shown in Table 1 and Table 2, and the activity test results are as follows:

[0301] Table 4

[0302] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound represented by the following formula I or II, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof: in, Ra is W is O or S; X1 and X2 are each independently selected from the following group: CR, N; X3 is N; The L1 is selected from the following group: chemical bond, -CHR-, -C(R)R-; Ring A is selected from the following group: a substituted or unsubstituted 7-12 membered bridged ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 7-12 membered spirocyclic ring (including a carbocyclic ring or a heterocyclic ring), a substituted or unsubstituted 8-12 membered fused bicyclic heterocyclic group (including a carbocyclic ring or a heterocyclic ring, preferably a pentahedral and hexacyclic ring), a substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl group (preferably a pentahedral and hexacyclic ring), or the ring A is a substituted or unsubstituted 3-7 membered carbocyclic ring or heterocyclic ring, a substituted or unsubstituted 5-6 membered aromatic ring or heteroaromatic ring; The E ring is selected from the following group: a substituted or unsubstituted 3-7 membered monocyclic heterocycle, a substituted or unsubstituted 7-12 membered bridged heterocycle, a substituted or unsubstituted 7-12 membered spiro heterocycle, a substituted or unsubstituted 8-12 membered fused polycyclic heterocyclyl (e.g., a fused bicyclic ring); R8 is selected from the group consisting of H, deuterium, halogen, cyano, C2-C6 alkynyl, -SF5, amino, nitro, hydroxyl, mercapto, aldehyde, carboxyl, substituted or unsubstituted or halogenated C1-C6 alkyl, substituted or unsubstituted or halogenated C1-C6 alkoxy, or R8 is R8' is selected from the following groups: H, deuterium, halogen, cyano, amino, nitro, hydroxyl, thiol, aldehyde, carboxyl, unsubstituted or halogenated C1-C6 alkyl, substituted or unsubstituted phenyl ring, substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated), substituted or unsubstituted 3-12 membered heterocycle (including saturated or partially unsaturated), or R8' is The L3 is selected from the following group: chemical bond, -O-, -CHR-, -C(R)R-, carbonyl, S, -NH-; Ring B is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including saturated or partially unsaturated rings), and a substituted or unsubstituted 3-7 membered heterocyclic ring (including saturated or partially unsaturated rings). R2 and R2' are each independently selected from the following group: R7, -L2R7; wherein, L2 is selected from the following group: -O-, -CHR-, -C(R)R-; wherein R7 is selected from the following groups: hydrogen or none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 aromatic ring, substituted or unsubstituted 5-12 membered (preferably 5-6 membered or 8-10 membered) heteroaromatic ring, substituted or unsubstituted C3-C 10 Carbocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings), substituted or unsubstituted 3-10 membered heterocycle (including saturated or partially unsaturated, including monocyclic, condensed, spirocyclic or bridged rings); n is 0, 1, 2 or 3; R3 is selected from the group consisting of H, deuterium, halogen, cyano, substituted or unsubstituted C1-C6 alkyl; R4 and R5 together with the ring atoms to which they are connected form a 5-12 membered saturated or unsaturated ring, and the ring may be substituted or unsubstituted; R is H, deuterium, halogen, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy, or substituted or unsubstituted C3-C6 cycloalkyl; Unless otherwise specified, in the above formulae, the substitution refers to the substitution of hydrogen atoms on the corresponding groups by one or more substituents selected from the following groups: deuterium, tritium, halogen, hydroxyl, carboxyl, mercapto, benzyl, C1-C 12 Alkoxycarbonyl, C1-C6 aldehyde, amino, C1-C6 amide, nitro, cyano, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-, unsubstituted or halogenated C1-C6 alkyl-O-C1-C6 alkyl-O-, unsubstituted or halogenated C1-C6 alkylene-OH, unsubstituted or halogenated C3-C8 cycloalkyl, C2-C 10 Alkenyl, unsubstituted or halogenated C1-C6 alkoxy, C1-C6 alkyl-amino, C6-C 10 Aryl, five-membered or six-membered heteroaryl, five-membered or six-membered non-aromatic heterocyclic group, -O-(C6-C 10 aryl), -O-(five-membered or six-membered heteroaryl), C1-C 12 Alkylaminocarbonyl, unsubstituted or halogenated C2-C 10 Acyl, sulfonyl (-SO2-OH), phosphoryl (-PO3-OH), unsubstituted or halogenated C1-C4 alkyl-S(O)2-, unsubstituted or halogenated C1-C4 alkyl-SO-, -SF5.

2. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The Ra is selected from the following group: Wherein, the R9 is selected from the following group: deuterium, tritium, halogen, hydroxyl, carboxyl, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or substituted C1-C6 alkyl-OH, -NH (unsubstituted or halogenated C1-C6 alkyl), -N (unsubstituted or halogenated C1-C6 alkyl) 2; m is selected from 0, 1, 2 or 3; preferably, the R9 is selected from the group consisting of deuterium, tritium, halogen, and unsubstituted or halogenated C1-C6 alkyl.

3. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The L1 is -CHR-, -C(R)R-; Ring A is selected from the following groups: substituted or unsubstituted 8-12 membered fused bicyclic heterocyclic group, substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl; R8 is selected from the following groups: H, halogen, cyano, amino, C2-C6 alkynyl, SF5, hydroxyl, thiol, aldehyde, carboxyl, unsubstituted or halogenated C1-C6 alkyl, The B ring is selected from the following group: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocycle, a substituted or unsubstituted 3-6 membered heterocycle; L3 is selected from the following group: a chemical bond, -O-, -CHR-, carbonyl, S, or -NH-.

4. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The R2 is selected from the following groups: R7, -L2R7; wherein the L2 is selected from the following groups: -O-, -CHR-, carbonyl, S, -NH-; wherein R7 is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 an aromatic ring, or a substituted or unsubstituted 5-12 membered heteroaromatic ring.

5. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The R2 is an ortho-substituted 5-membered or 6-membered heteroaromatic ring, as shown in the following formula: Among them, R 10 is a substituent located at the ortho position of the attachment site, which is selected from the group consisting of hydrogen, deuterium, halogen, halogenated or unhalogenated C1-C3 alkyl, halogenated or unhalogenated C1-C3 alkoxy; The D ring is selected from the following groups: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, preferably, the D ring is selected from the following groups:

6. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The L1 is -CH2-, -CH(CH3)-; the A ring is selected from the following group: wherein the C ring is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including saturated or partially unsaturated rings), and a substituted or unsubstituted 3-6 membered heterocyclic ring (including saturated or partially unsaturated rings); Or ring A is selected from the following group: R8 is a halogenated or non-halogenated C1-C6 alkyl group, or The B ring is selected from the following group: a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5-6 membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocycle, a substituted or unsubstituted 3-6 membered heterocycle; L3 is selected from the following group: a chemical bond, -O-, -CHR-, carbonyl, S, or -NH-.

7. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, characterized in that: R2 is selected from the following groups: R7, -(CHR)R7; wherein R7 is selected from the following groups: hydrogen or none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C 6-10 substituted or unsubstituted 5-12 membered heteroaromatic ring, substituted or unsubstituted C3-C8 carbocyclic ring (including saturated or partially unsaturated cases, including monocyclic, condensed, spirocyclic or bridged rings), substituted or unsubstituted 3-8 membered heterocyclic ring (including saturated or partially unsaturated cases, including monocyclic, condensed, spirocyclic or bridged rings).

8. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The compound has a structure shown in the following formula: Wherein, Q is O, NH, CH2, or a chemical bond (i.e., is a five-membered ring); R8 is defined as above; The R 8a and R 8b Each independently selected from the following group: H; or said R 8a and R 8b and the carbon atoms to which they are connected together form a 4-7 membered carbocyclic or heterocyclic ring; And when the R 8a and R 8b When each independently is H, the R 8a or R 8b may be optionally substituted with R8; when said R 8a and R 8b When the carbon atoms connected to them together form a 4-7 membered carbocyclic ring or heterocyclic ring, the R8 can be located on the carbocyclic ring or heterocyclic ring.

9. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein: The R3 is selected from the following group: H, deuterium, halogen, cyano, substituted or unsubstituted C1-C6 alkyl.

10. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, characterized in that: The R2 is a substituted or unsubstituted 5-7 membered heteroaromatic ring; and the A ring is selected from the following group: a substituted or unsubstituted 5-6 membered aromatic ring or heteroaromatic ring, a substituted or unsubstituted 7-10 membered fused bicyclic heteroaryl group; and the R8 is CF3.

11. The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, characterized in that: The compound is selected from the following group: Table 1 12. A compound selected from the group consisting of: Table 2 13. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a therapeutically effective amount of the compound according to any one of claims 1 to 12, one or more of its pharmaceutically acceptable salts, racemates, optical isomers, stereoisomers or tautomers, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, excipients and / or diluents.

14. Use of a compound according to any one of claims 1 to 12, a racemate, stereoisomer or pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing diseases associated with abnormal gene levels or abnormal expression of PRMT5 (such as corresponding nucleic acid mutations, deletions, or abnormal MTAP gene levels, or ectopic or fusion or overexpression of the methyltransferase).

15. The use according to claim 14, characterized in that The disease is selected from the group consisting of ovarian cancer, esophageal cancer, lung cancer, lymphoma, glioblastoma, colon cancer, melanoma, gastric cancer, pancreatic cancer or bladder cancer.

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