Novel PRMT5 inhibitors and their applications

Abstract

The present invention describes novel molecules with protein arginine methyltransferase 5 inhibitory activity, as well as methods for synthesizing and using the compounds. Specifically, the present invention describes a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, or solvate thereof, as well as methods for synthesizing and using the compounds.

Description

Technical Field

[0001] The present application relates to the field of drug synthesis, and specifically to a PRMT5 inhibitor and its application. Background Art

[0002] Epigenetic alterations are key mediators driving and maintaining the malignant phenotype of tumors. Changes in DNA methylation, histone acetylation and methylation, noncoding RNAs, and post-translational modifications are all epigenetic drivers of cancer development, independent of DNA sequence changes. Arginine methylation is an important post-translational modification that influences cell growth and proliferation, apoptosis, angiogenesis, and metastasis by regulating transcription and post-transcriptional RNA processing. Three types of methylarginine exist: ω-NG, N'G-asymmetric dimethylarginine (ADMA) and ω-NG, N'G-symmetric dimethylarginine (SDMA). This modification is catalyzed by the protein arginine methyltransferase (PRMT) family, transferring a methyl group from S-adenosylmethionine (AdoMet) to arginine side chains on histones and non-histone proteins. Nine PRMT genes have been annotated in the human genome and are categorized as type I (PRMT1, 2, 3, 4, 6, and 8), type II (PRMT5 and PRMT9), and type III enzymes (PRMT7) based on the type of methylarginine produced. PRMT5 is primarily a type II enzyme that catalyzes the symmetric dimethylation of arginine. PRMT5 was first discovered in a two-hybrid assay to detect proteins that interact with Janus tyrosine kinase (Jak2).

[0003] PRMT5 is a universal transcriptional repressor that forms a complex with other transcription factors, including BRG1 and Hbrm, Blimp1, and Snail. PRMT5 participates in diverse cellular processes by methylating a variety of cytoplasmic and nuclear substrates, including histone H4 residue Arg3 (H4R3) and H3 residue Arg8 (H3R8). H4R3 methylation is associated with transcriptional repression, while H3R8 methylation is considered to be involved in both transcriptional activation and repression. In addition to directly inducing repressive histone marks, PRMT5's role in gene silencing is mediated by the formation of a multi-repressor protein complex, including NuRD components, HDACs, MDB proteins, and DNA methyltransferases. PRMT5 influences its substrate specificity through interactions with several binding proteins. A core component of this protein complex is MEP50, which is essential for PRMT5's enzymatic activity. Studies have shown that PRMT5 can methylate proteins involved in RNA splicing, such as SmD3, which can be used to track the chemical activity of PRMT5 in cellular biology. PRMT5 plays a crucial role in tumorigenesis. Studies have found that PRMT5 expression is upregulated in a variety of tumors, including lymphoma, lung cancer, breast cancer, and colorectal cancer. Furthermore, PRMT5 expression is elevated in samples from patients with mantle cell lymphoma (MCL), and PRMT5 knockout inhibits MCL cell proliferation, suggesting a key role for PRMT5 in MCL. PRMT5 overexpression promotes cell proliferation, while PRMT5 knockout inhibits cell proliferation in melanoma, breast cancer, and lung cancer cell lines. Therefore, PRMT5 is a potential target for cancer therapy.

[0004] Loss of methylthioadenosine phosphorylase (MTAP) confers a selective reliance on PRMT5 and its binding protein, WDR77. MTAP is frequently lost due to its proximity to the commonly deleted tumor suppressor gene CDKN2A. Cells harboring MTAP deletion have increased levels of intracellular methylthioadenosine (MTA), a metabolite cleaved by MTAP. MTA shares a similar structure to S-adenosylmethionine (SAM). As concentrations increase, MTA acts as an intrinsic, selective inhibitor, inhibiting the binding of SAM to PRMT5 and, consequently, the methyltransferase activity of PRMT5.

[0005] The most significant structural difference between MTAP-deficient and MTAP-wild-type cancer cells lies in the accumulation of MTA in MTAP-deficient cancer cells, which results in the formation of a PRMT5-MTA complex. Inhibitors developed against the PRMT5-MTA complex can selectively target MTAP-deficient cancer cells while minimizing the effect on normal cells, significantly improving the therapeutic index.

[0006] Therefore, identifying and developing small molecules that inhibit PRMT5 activity will be useful as therapeutic approaches for treating various PRMT5-associated diseases or disorders, such as cancer. Summary of the Invention

[0007] To solve the technical problem of the present invention, the present invention provides a class of compounds with novel structures that have excellent inhibitory activity against PRMT5.

[0008] Specifically, the present invention provides a compound represented by formula (I), and its pharmaceutically acceptable salts, esters, prodrugs, stereoisomers or isotopic derivatives,

[0009]

[0010] Where W represents C;

[0011] Where X3 represents N or CR X3 ; X4 represents N or CR X4 ; X5 represents N or CR X5 ; X6 represents N or CR X6 ;

[0012] Among them, when X3 represents CR X3 When R X3 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR bor -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0013] Among them, when X4 represents CR X4 When R X4 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0014] Among them, when X5 represents CR X5 When R X5 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a Rb 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0015] Among them, when X6 represents CR X6 When R X6 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b, -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0016] Wherein, the ring A may be optionally fused with a 5-6 membered saturated or unsaturated ring at the chemical bond between X3 and X4, and the ring may contain 0-3 heteroatoms selected from O, N, and S;

[0017] Wherein, the ring A may be optionally fused with a 5-6 membered saturated or unsaturated ring at the chemical bond between X4 and X5, and the ring may contain 0-3 heteroatoms selected from O, N, and S;

[0018] Wherein, the ring A may be optionally fused with a 5-6 membered saturated or unsaturated ring at the chemical bond between X5 and X6, and the ring may contain 0-3 heteroatoms selected from O, N, and S;

[0019] Wherein, the ring A can be arbitrarily replaced by 0, 1, 2, or 3 groups selected from deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3Ra 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0020] Wherein, R' represents 0-3 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3-6 membered saturated or unsaturated aliphatic monoheterocyclic group, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Substituted C1-C6 alkyl, C3-C 10 Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0021] Preferably, R' represents -CHR 2 R 3 or -CDR 2 R 3 ;

[0022] Among them, R 2 、R 3 Each independently represents hydrogen, deuterium, -OR a , halogen, -CN, -C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, -C3-C 10Cycloalkyl or 0-3 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0023] Among them, M 1 Indicates CR a R b NR a , O, S or S e ;

[0024] Among them, R L 、R L’ Each independently represents hydrogen, deuterium, C1-C6 alkyl, or R L 、R L’ Together with the atoms to which they are attached, they form a 3-6 membered ring;

[0025] Here, n and o each independently represent 0, 1 or 2.

[0026] Where X1 represents N or CR X1 ;

[0027] Where X2 represents N or CR X2 ;

[0028] Among them, Y1 represents CR Y1 R Y1’ NR Y1 , O, S, Se;

[0029] Among them, Y2 represents CR Y2 R Y2’ NR Y2 , O, S, Se;

[0030] Among them, Y3 represents CR Y3 R Y3’ NR Y3 , O, S, Se;

[0031] Among them, R X1 、R X2 Each independently represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -OR a 、-SR a 、-S(O)2R a 、-S(O)R a 、-CN、-OC(O)R a 、-OCONR a R b , halogen, -OSO3R a 、-NR a R b , -SF5;

[0032] Among them, R Y1 、R Y1’ 、R Y2 、R Y2’ 、R Y3 、R Y3’ Each independently represents absence, hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a 、-SR a 、-S(O)2R a 、-S(O)R a 、-CN、-OC(O)R a 、-OCONR a R b , halogen, -OSO3R a 、-NR a R b , -SF5;

[0033] in, Indicates a single bond or a double bond;

[0034] Among them, R a 、R b Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, halo(C1-C6 alkyl), or R a 、R bTogether with the atoms to which they are attached, they form a 3-14 membered saturated or unsaturated ring, which may optionally contain 0-2 heteroatoms selected from O, S, and N.

[0035] In addition, the present invention provides a compound, a pharmaceutically acceptable salt, ester, prodrug, stereoisomer or isotopic derivative thereof, which has the following structure:

[0036]

[0037] Where W represents C;

[0038] Where X3 represents N or CR X3 ; X4 represents N or CR X4 ; X5 represents N or CR X5 ; X6 represents N or CR X6 ;

[0039] Among them, when X3 represents CR X3 When R X3 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0040] Among them, when X4 represents CR X4 When R X4 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0041] Among them, when X5 represents CR X5 When R X5 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR aR b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0042] Among them, when X6 represents CR X6 When R X6 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、-CN、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, or 0-4 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0043] Wherein, R' represents 0-3 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3-6 membered saturated or unsaturated aliphatic monoheterocyclic group, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Substituted C1-C6 alkyl, C3-C 10 Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0044] Preferably, R' represents -CHR 2 R 3 or -CDR 2 R 3 ;

[0045] Among them, R 2 、R 3 Each independently represents hydrogen, deuterium, -OR a , halogen, -CN, -C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, -C3-C 10Cycloalkyl or 0-3 substituents selected from the following: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0046] Among them, M 1 Indicates CR a R b NR a , O, S or Se;

[0047] Among them, R L 、R L’ Each independently represents hydrogen, deuterium, C1-C6 alkyl, or R L 、R L’ Together with the atoms to which they are attached, they form a 3-6 membered ring;

[0048] Here, n and o each independently represent 0, 1 or 2.

[0049] Where X1 represents N or CR X1 ;

[0050] Where X2 represents N or CR X2 ;

[0051] Among them, Y1 represents CR Y1 R Y1’ NR Y1 , O, S, Se;

[0052] Among them, Y2 represents CR Y2 R Y2’ NR Y2 , O, S, Se;

[0053] Among them, Y3 represents CR Y3 R Y3’ NR Y3 , O, S, Se;

[0054] Among them, R X1 、R X2 Each independently represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -OR a 、-SR a 、-S(O)2R a 、-S(O)R a 、-CN、-OC(O)R a 、-OCONR a R b , halogen, -OSO3R a 、-NR a R b , -SF5;

[0055] Among them, R Y1 、R Y1’ 、R Y2 、R Y2’ 、R Y3 、R Y3’ Each independently represents absence, hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a 、-SR a 、-S(O)2R a 、-S(O)R a 、-CN、-OC(O)R a 、-OCONR a R b , halogen, -OSO3R a 、-NR a R b , -SF5;

[0056] in, Indicates a single bond or a double bond;

[0057] Among them, R a 、R b Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, halo(C1-C6 alkyl), or R a 、R bTogether with the atoms to which they are attached, they form a 3-14 membered saturated or unsaturated ring, which may optionally contain 0-2 heteroatoms selected from O, S, and N.

[0058] In the preferred technical solution of the present invention, Indicates a double bond.

[0059] In the preferred technical solution of the present invention, wherein X1 represents CR X1 or N, where R X1 represents hydrogen, deuterium, halogen, -CN, C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, or halogenated C1-C6 alkyl.

[0060] In a preferred technical solution of the present invention, X1 represents CH, CF or N.

[0061] In the preferred technical solution of the present invention, wherein X2 represents CH or CD

[0062] In a preferred technical solution of the present invention, X2 is represented by CH.

[0063] In a preferred technical solution of the present invention, X3 represents CH, CD or N.

[0064] In a preferred technical solution of the present invention, X3 represents CH.

[0065] In the preferred technical solution of the present invention, wherein X4 represents CR X4 or N, where R X4 represents hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, halogen, SF5, -S(O)2R a 、-P(O)R a R b , or cyano or is selected from 0-4 of the following substituents: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -S(O)2R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl.

[0066] In the preferred technical solution of the present invention, wherein X4 represents CR X4 ; Among them, R X4 It represents a C1-C6 alkoxy group, a C1-C6 alkylthio group, a halogenated C1-C6 alkoxy group, a halogenated C1-C6 alkylthio group, a halogenated C1-C6 alkyl group, or -SF5.

[0067] In the preferred technical solution of the present invention, wherein X4 represents CR X4 ; Among them, R X4 It represents a halogenated C1-C6 alkoxy group, a halogenated C1-C6 alkylthio group, a halogenated C1-C6 alkyl group, or -SF5.

[0068] In the preferred technical solution of the present invention, X5 represents CR X5 or N, where R X5 represents hydrogen, deuterium, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogen, SF5 or cyano.

[0069] In a preferred technical solution of the present invention, X5 represents CH.

[0070] In a preferred technical solution of the present invention, X6 represents CH, CD or N.

[0071] In a preferred technical solution of the present invention, X6 represents CH.

[0072] In a preferred technical solution of the present invention, the chemical bond between Y1 and Y2 is a double bond.

[0073] In a preferred technical solution of the present invention, Y1 represents CH, CD or CCH3.

[0074] In a preferred technical solution of the present invention, Y2 represents N.

[0075] In a preferred technical solution of the present invention, Y3 represents CH, CD, CCH3 or CCH2OH.

[0076] In the preferred technical solution of the present invention,

[0077] R' represents -CHR 2 R 3 or -CDR 2 R 3 , where R 2 、R 3 Each independently represents hydrogen, deuterium, C1-C6 alkyl, or 0-3 selected from halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-N R aCOR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl.

[0078] In the preferred technical solution of the present invention, wherein R' represents -CHR 2 R 3 or -CDR 2 R 3 , where R 2 represents hydrogen, deuterium, C1-C6 alkyl; R 3 represents hydrogen, deuterium, C1-C6 alkyl, or 0-3 selected from halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b , -CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-S(O)2R a 、-S(O)R a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a、-OC(O)NR a R b 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl.

[0079] In the preferred technical solution of the present invention, wherein R' represents 0-3 selected from deuterated, halogen, C1-C6 alkyl, hydroxy C1-C6 alkyl, -OR a 、-CN、NR a R b , C3-C substituted by halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy 10 Cycloalkyl.

[0080] In a preferred technical solution of the present invention, R' represents a C1-C6 alkyl group (preferably a methyl group or an ethyl group) or a deuterated C1-C6 alkyl group (preferably a deuterated methyl group or a deuterated ethyl group) or a C3-C6 cycloalkyl group (preferably a cyclopropyl group).

[0081] In a preferred technical solution of the present invention, M1 is O or S.

[0082] In a preferred technical solution of the present invention, o is 1 or 2.

[0083] In the preferred technical solution of the present invention, o is 1.

[0084] In a preferred technical solution of the present invention, n is 0 or 1.

[0085] In a preferred technical solution of the present invention, n is 0.

[0086] In a preferred technical solution of the present invention, n is 1.

[0087] In the preferred technical solution of the present invention, R L 、R L’ Each independently represents hydrogen or a C1-C6 alkyl group.

[0088] Specifically, the present invention provides the following compounds:

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] Unless otherwise indicated, the compounds of the present invention may be interpreted to include, in addition to the specific structures of the compounds, pharmaceutically acceptable salts of the compounds, their stereoisomers, isotope isomers (e.g., deuterated compounds), solvates, hydrates, prodrugs, and metabolites. In other words, pharmaceutically acceptable salts of the compounds, their stereoisomers, isotope isomers, solvates, hydrates, prodrugs, and metabolites also fall within the scope of protection of the compounds.

[0101] Preferably, the pharmaceutical composition of the present invention may further include a second active substance, wherein the second active substance is an anti-tumor drug, and the anti-tumor drug includes one or more of a chemotherapy drug, a targeted tumor treatment drug or a tumor treatment antibody drug.

[0102] In addition, the present invention also provides a compound of the present invention, a pharmaceutically acceptable salt, ester, prodrug, stereoisomer or isotope derivative thereof, for treating a disease by inhibiting the action of PRMT5, preferably the disease is a tumor. Definition:

[0103] Unless otherwise indicated, the term "alkyl" by itself or as part of another substituent refers to a straight chain (i.e., unbranched) or branched chain, or cyclic hydrocarbon radical, or combinations thereof, which may be saturated, mono- or polyunsaturated, and may include divalent or polyvalent groups, having the specified number of carbon atoms (i.e., C1-C 10Refers to one to ten carbon atoms). Examples of saturated hydrocarbon groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, cyclohexylmethyl, cyclopropylmethyl, and homologs and isomers such as n-pentyl, n-hexyl, n-heptyl, and n-octyl. Unsaturated alkyl groups are alkyl groups having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. Alkyl groups that are limited to hydrocarbon groups are referred to as "homoalkyl". The alkyl group is optionally substituted with one or more halogen atoms.

[0104] The term "haloalkyl" refers to an alkyl group as defined above wherein one or more hydrogen atoms are replaced by a halogen atom.

[0105] The term "alkylene" by itself or as part of another substituent refers to a divalent radical derived from an alkyl group, for example, but not limited to, -CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2C≡CCH2-, -CH2CH2CH(CH2CH2CH3)CH2-. Alkyl (or alkylene) groups typically have from 1 to 24 carbon atoms, with groups having 10 or fewer carbon atoms being preferred. "Lower alkyl" or "lower alkylene" refers to shorter chain alkyl or alkylene groups, typically having eight or fewer carbon atoms. The alkylene group is optionally substituted with one or more halogen atoms.

[0106] The term "alkynyl" refers to a carbon chain containing at least one carbon-carbon triple bond, which may be linear or branched, or a combination thereof. Examples of alkynyl groups include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl, and the like. The alkynyl group may be optionally substituted with one or more halogen atoms.

[0107] The term "cycloalkyl" refers to a monocyclic or bicyclic saturated carbocyclic ring, each having 3 to 10 carbon atoms. A "fused analog" of a cycloalkyl refers to a monocyclic ring fused to an aryl or heteroaryl group, wherein the point of attachment is on the non-aromatic portion. Examples of cycloalkyls and fused analogs thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydronaphthyl, decahydronaphthyl, and dihydroindanyl. The cycloalkyl group is optionally substituted with one or more halogen atoms. Furthermore, the term "cycloalkyl" as used herein includes bridged ring systems and spirocyclic ring systems.

[0108] The term "alkoxy" refers to a straight or branched chain alkoxy group having the indicated number of carbon atoms. 1-6 The alkoxy group includes, for example, methoxy, ethoxy, propoxy, isopropoxy and the like.

[0109] The term "heteroalkyl," by itself or in combination with another term, means, unless otherwise stated, a stable linear or branched chain, or cyclic hydrocarbon radical consisting of at least one carbon atom and at least one heteroatom selected from O, N, P, Si, S, or combinations thereof, wherein the nitrogen, phosphorus, or sulfur atom may be optionally oxidized and the nitrogen atom may be optionally quaternized. The heteroatoms O, N, P, S, and Si may be placed at any position within the heteroalkyl radical or at the position at which the alkyl radical is attached to the remainder of the molecule. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive. For example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. Similarly, the term "heteroalkylene," by itself or in combination with other terms, refers to a divalent radical derived from a heteroalkyl group, such as, but not limited to, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. For heteroalkylene, the heteroatom can be at either or both ends of the chain (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Additionally, for alkylene and heteroalkylene linking groups, the direction in which the linking group formula is written does not indicate the orientation of the linking group. For example, the formula -C(O)OR'- refers to both -C(O)OR'- and -R'OC(O)-. As described above, heteroalkyl groups as used herein include those groups that are attached to the rest of the molecule through a heteroatom, such as -C(O)R', -C(O)NR', ​​-NR'R", -OR', -SR', and / or -S02R'. Where "heteroalkyl" is mentioned followed by a specific heteroalkyl group such as -NR'R", it is understood that the terms heteroalkyl and -NR'R" are not redundant and are not mutually exclusive. Rather, these specific heteroalkyl groups are cited for clarity. Thus, the term "heteroalkyl" should not be construed herein to exclude specific heteroalkyl groups such as -NR'R".

[0110] The term "cycloalkoxy" refers to a cycloalkyl group as defined above bound to an oxygen atom, such as cyclopropyloxy.

[0111] The term "haloalkoxy" refers to an alkoxy group as defined above in which one or more hydrogen atoms are replaced by a halo.

[0112] The term "aryl" refers to a monocyclic or bicyclic aromatic group containing only carbon atoms. A "fused analog" of an aryl group refers to an aryl group fused to a monocyclic cycloalkyl group or a monocyclic heterocyclic group, wherein the point of attachment is on the aryl portion. Examples of aryl groups and fused ring analogs thereof include phenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrochromenyl, 1,4-benzodioxanyl, and the like.

[0113] The term "heteroaryl" refers to a monocyclic or bicyclic aromatic group containing at least one heteroatom selected from N, O, and S. A "fused analog" of a heteroaryl group refers to a heteroaryl group fused to a monocyclic cycloalkyl group or a monocyclic heterocyclyl group, wherein the point of attachment is located on the aromatic portion. Examples of heteroaryl groups include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridinyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzothienyl, furo(2,3-b)pyridinyl, quinolinyl, indolyl, isoquinolinyl, and the like.

[0114] "Substituted or unsubstituted": the alkyl, aryl and heteroaryl groups are defined as being unsubstituted or substituted with at least one substituent selected from the group consisting of halogen atoms, alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, haloalkyl groups having 1 to 6 carbon atoms, haloalkoxy groups having 1 to 6 carbon atoms, -CN, alkynyl groups having 2 to 6 carbon atoms, alkanoyl groups having 1 to 6 carbon atoms, cycloalkyl groups having 3 to 7 ring atoms, heteroaryl groups, aryl groups, aralkyloxy groups having 7-10 carbon atoms, arylcarbonyl groups, aminocarbonyl groups, alkyl groups having 2 to 6 carbon atoms, alkynyl groups having 1 to 6 carbon atoms, alkanoyl groups having 1 to 6 carbon atoms, cycloalkyl groups having 3 to 7 ring atoms, heteroaryl groups, aryl groups, aralkyloxy groups having 7-10 carbon atoms, arylcarbonyl groups, aminocarbonyl groups, alkyl groups having 2 to 6 carbon atoms, alkyl groups having 1 to 6 carbon atoms, alkyl groups having 1 to 6 carbon atoms, alkyl groups having 3 to 7 ring atoms, heteroaryl groups, aryl groups, aralkyloxy groups having 7-10 carbon atoms, arylcarbonyl groups, aralkyloxy groups having 7-10 carbon atoms, aralkyl groups having 1 to ... an alkenyl group having 1 to 5 carbon atoms, an alkylthio group having 1 to 6 carbon atoms, an aminosulfinyl group, an aminosulfonyl group, a hydroxyl group, -SF5, a hydroxyalkyl group having 1 to 4 carbon atoms, a nitro group, an amino group, a carboxyl group, an alkoxycarbonyl group having 2 to 5 carbon atoms, an alkoxyalkyl group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1-4 carbon atoms, an alkanoylamino group having 1 to 4 carbon atoms, an alkanoyl(alkyl)amino group having 1 to 6 carbon atoms, an alkanoylaminoalkyl group having 1 to 6 carbon atoms in both the alkanoyl and alkyl moieties, an alkanoyl(alkyl)aminoalkyl group having 1 to 6 carbon atoms in both the alkanoyl and alkyl moieties, an alkylsulfonylamino group having 1 to 4 carbon atoms, a monoalkylaminocarbonyl group or a dialkylaminocarbonyl group having 1 to 6 carbon atoms, a monoalkylaminosulfinyl group or a dialkylaminosulfinyl group having 1 to 6 carbon atoms, a monoalkylaminosulfonyl group or a dialkylaminosulfonyl group having 1 to 6 carbon atoms dialkylaminosulfonyl, aminoalkyl having 1 to 4 carbon atoms, mono- or dialkylamino having 1 to 6 carbon atoms, mono- or dialkylaminoalkyl having 1 to 6 carbon atoms in each alkyl moiety, aralkyl having 7 to 10 carbon atoms, heteroaralkyl having 1 to 4 carbon atoms in the alkyl moiety, heteroarylalkoxy having from 1 to 4 carbon atoms in the alkoxy moiety, and alkylsulfonamide having 1 to 4 carbon atoms.

[0115] As used herein, the term "heterocycle" or "heterocyclic" or "heterocycloalkyl" or "heterocyclyl" refers to a saturated, partially saturated or unsaturated group (but not aromatic) having a single ring or a fused ring (including bridged ring systems and spiro ring systems) with 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from nitrogen, sulfur or oxygen in the ring. In a fused ring system, one or more rings can be cycloalkyl, aryl or heteroaryl, as long as the point of attachment is through the non-aromatic ring. In one embodiment, the nitrogen atom and / or sulfur atom of the heterocyclic group is optionally oxidized. , to provide N-oxide, sulfinyl and sulfonyl moieties. Examples of "heterocyclyl" and its fused analogs include pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, 2,3-dihydrofuryl (2,3-b) pyridinyl, benzoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, dihydroindolinyl, etc. The term also includes non-aromatic partially unsaturated monocyclic rings, such as 2- or 4-pyridones or N-substituted-(1H,3H)-pyrimidine-2,4-diones (N-substituted uracils) attached through a nitrogen atom.

[0116] As used herein, the term "substituted heterocyclic" or "substituted heterocycloalkyl" or "substituted heterocyclyl" refers to a heterocyclic group substituted with 1 to 5 (e.g., 1 to 3) substituents, the substituents being the same as those defined for substituted cycloalkyl.

[0117] Unless otherwise indicated, the term "halogenated" or "halogen" by itself or as part of another substituent refers to a fluorine, chlorine, bromine, or iodine atom. Additionally, the term "haloalkyl" is intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C6)alkyl" includes, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0118] "Prodrug" refers to a substance that is converted into the parent drug in vivo. In some cases, prodrugs are often used because they are easier to administer than the parent drug. For example, a prodrug may be bioavailable orally while the parent drug cannot. In a pharmaceutical composition, a prodrug may also have a higher solubility than the parent drug. Examples of prodrugs, but not limited to, may be any of the compounds of Formula I administered in the form of an ester (prodrug) to facilitate transcellular transport, where water solubility in the cell membrane is detrimental to migration, and once in the cell where water solubility is beneficial, the ester is subsequently metabolically hydrolyzed to the active substance, carboxylic acid. Another example of a prodrug may be a short peptide (polyamino acid) bonded to an acid group, wherein the peptide is metabolized to release the active portion.

[0119] Optical isomers - diastereomers - geometric isomers - tautomers:

[0120] The compounds of formula (I) contain one or more asymmetric centers and can occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The present invention is intended to encompass all such isomeric forms of the compounds of formula (I).

[0121] Some of the compounds described herein contain olefinic double bonds, and unless specified otherwise, are intended to include both E and Z geometric isomers.

[0122] Some of the compounds of the present invention may contain one or more than one ring system and may therefore exist as cis- and trans-isomers. The present invention is intended to include all such cis- and trans-isomers.

[0123] Some compounds described herein may have different sites of attachment to hydrogen atoms, known as tautomers. Examples of such are the ketone and its enol form, known as keto-enol tautomers. Individual tautomers as well as mixtures thereof are encompassed within the compounds of the present invention.

[0124] The compounds of the present invention can be separated into diastereomeric pairs of enantiomers, for example by fractional crystallization from a suitable solvent such as methanol or ethyl acetate or a mixture thereof. A pair of enantiomers thus obtained can be separated into individual stereoisomers by conventional methods, for example using an optically active amine or acid as a resolving agent or in a chiral HPLC column.

[0125] Alternatively, any enantiomer of a compound of the invention may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration.

[0126] Stable isotope-labeled analogs: One or more protons in the compounds of the present invention may be replaced with deuterium atoms to provide deuterated analogs with improved pharmacological activity.

[0127] Salt and dosage form

[0128] It will be understood that, as used herein, references to the compounds of the present invention also include the pharmaceutically acceptable salts.

[0129] application

[0130] The compounds of the present invention can be used to treat PRMT5-related diseases.

[0131] The compounds of the present invention can be prepared by the following reaction formula:

[0132] Method A:

[0133]

[0134] Method A-SFC

[0135]

[0136] Among them, R L 、R L’ ,R',A ring,W,n,o,M1,X1,X2,X3,X4,X5,X6,Y1,Y2,Y3, As defined in claim 1.

[0137] Method A: Compound AP can be prepared by the amine-acid condensation reaction of carboxylic acid A-1 and amine A-2. The condensing agent can be HATU or PyBrOP, the base can be DIPEA or TEA, and the solvent can be DMF or DMAc. If the amine used is a racemic form, it will be resolved by chiral SFC, and the stereochemistry of the resulting isomers will be randomly assigned to R or S.

[0138] Analytical HPLC

[0139] Equipment: Agilent 1260; Column dimensions: Agilent Poroshell HPH-C18 (3.0 × 50 mm, 2.7 μm); Binary solvent system: Mobile phase A: Water (0.1% v / v ammonium bicarbonate), Mobile phase B: Acetonitrile; Flow rate: 1 mL / min; Gradient: 10% B to 90% B; Flow duration: 12 min; Detector: DAD; Wavelength: 254 / 220 nm;

[0140] Preparative HPLC-MS

[0141] HPLC equipment: Waters 2489; Column specifications: Ultimate μXB-C18 (130A, 5 μm, 30 mm × 150 mm); Binary solvent system: Mobile phase A: Water (0.1% v / v ammonium bicarbonate), Mobile phase B: Acetonitrile; Flow rate: 60–100 mL / min; Gradient: 10% B to 90% B; Detector: DAD; Wavelength: 254 / 220 nm;

[0142] Mass spectrometer: Agilent G6125B.

[0143] The compounds of the present invention can be prepared by chemical synthesis, examples of which are shown below. It should be understood that the order of the steps in the process can be changed, those specifically mentioned reagents, solvents and reaction conditions can be replaced, and if necessary, reactive sites can be protected and deprotected.

[0144] The following abbreviations have the following meanings: ACN means acetonitrile; EA means ethyl acetate; CDI means N,N'-carbonyldiimidazole; DBU means 1,8-diazabicyclo[5.4.0]undec-7-ene; DIBAL-H means diisobutylaluminum hydride; DIEA means diisopropylethylamine; DMAP means N,N-dimethylaminopyridine; DME means 1,2-dimethoxyethane; DMF means N,N-dimethylformamide; DMA and DMAc mean N,N-dimethylformamide; DMPE means 1,2-bis(dimethylformamide) phosphino)ethane; DMSO denotes dimethyl sulfoxide; DPPB refers to 1,4-bis(diphenylphosphino)butane; dppe denotes 1,2-bis(diphenylphosphino)ethane; dppf denotes 1,1'-bis(diphenylphosphino)ferrocene; dppm denotes 1,1'-bis(diphenylphosphino)methane; DIAD denotes diisopropyl azodicarboxylate; EDCI denotes 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; HATU denotes 2-(7-aza-1H-benzotriazol-1-yl)-1,1, 3,3-Tetramethyluronium hexafluorophosphate; HMPA is hexamethylphosphoramide; IPA is isopropyl alcohol; LDA is lithium diisopropylamide; LHMDS is lithium bis(trimethylsilyl)amide; LAH is lithium aluminum hydride; NCS is N-chlorosuccinimide; NaHMDS is sodium bis(trimethylsilyl)amide; PyBOP is benzotriazol-1-yl-oxytripyrrolidinylphosphonium benzotriazole hexafluorophosphate; PyBrOP is tripyrrolidinylphosphonium bromide hexafluorophosphate; TDA-I is tris(trimethylsilyl)phosphonium benzotriazole hexafluorophosphate. (2-(2-methoxyethoxy)ethyl)amine; DCM refers to dichloromethane; TEA refers to triethylamine, TFA refers to trifluoroacetic acid; THF refers to tetrahydrofuran; NCS refers to N-chlorosuccinimide; NMM refers to N-methylmorpholine; NMP refers to N-methylpyrrolidone; PPh3 refers to triphenylphosphine, rt refers to room temperature; PMB refers to p-methoxybenzyl; Tosmic refers to p-toluenesulfonylmethyl isocyanide; (Boc)2O refers to di-tert-butyl dicarbonate; PE refers to petroleum ether; o / n refers to overnight reaction.

[0145] The following preparations and examples illustrate the present invention but do not limit it in any way.

[0146] The features and advantages of the subject matter of the present invention will become more apparent from the detailed description of selected embodiments. As will be appreciated, the disclosed and claimed subject matter is capable of modification in various respects, all without departing from the scope of the claims. Therefore, the description is to be regarded as illustrative in nature, not restrictive. The full scope of the subject matter of the present invention is set forth in the claims.

[0147] The present invention can be more easily understood by referring to the following examples, which are intended only to illustrate the present invention and are not intended to limit the scope of the present invention.

[0148] intermediates

[0149] The following intermediate raw materials were purchased from different suppliers:

[0150]

[0151]

[0152]

[0153] Intermediate 12: (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine

[0154]

[0155] Step 1: Synthesis of tert-butyl (S)-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate

[0156] (S)-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (200 mg, 0.99 mmol) was dissolved in dichloromethane (2 ml) and then di-tert-butyl dicarbonate (325 mg, 1.49 mmol) and triethylamine (200 mg, 1.98 mmol) were added. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The residue was purified by flash column chromatography (20 g silica gel column, 20% ethyl acetate / petroleum ether) to obtain tert-butyl (S)-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (300.0 mg, 99% yield) as a yellow liquid.

[0157] LCMS (ESI) m / z: 304 [M+H] +

[0158] Step 2: Synthesis of tert-butyl (S)-methyl (6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate

[0159] At 0°C, (S)-tert-butyl (6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (300 mg, 0.99 mmol) was dissolved in DMF (5.00 ml), and sodium hydride (48 mg, 2.00 mmol) was then added. The mixture was stirred at 0°C for 1 hour. Iodomethane (211 mg, 1.49 mmol) was then added. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The residue was purified by flash column chromatography (20 g silica gel column, 15% ethyl acetate / petroleum ether) to give (S)-methyl (6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (200.0 mg, 64% yield) as a yellow liquid.

[0160] LCMS (ESI) m / z: 318 [M+H] +

[0161] Step 3: Synthesis of (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine

[0162] At room temperature, tert-butyl (S)-methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (200.0 mg, 0.63 mmol) was dissolved in a solution of hydrogen chloride in 1,4-dioxane (5.00 ml). The mixture was stirred at room temperature for 0.5 hours. The reaction was monitored by LCMS. The mixture was concentrated to give (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (130.0 mg, 95% yield) as a yellow liquid.

[0163] LCMS (ESI) m / z: 218 [M+H] +

[0164] The following intermediates were prepared by using the preparation method and steps of intermediate 12, with only the corresponding raw material intermediates being replaced:

[0165]

[0166]

[0167]

[0168] Intermediate 22 (S)-3-(Methylamino)-2,3-dihydrobenzofuran-6-carbonitrile

[0169]

[0170] Step 1: (S)-tert-Butyl (6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0171] A reaction system of tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (328 mg, 1 mmol), cuprous cyanide (352 mg, 4 mmol), and N-methylpyrrolidone (4 ml) was reacted in a microwave reactor at 135°C for 4 hours. After completion of the reaction, the system was cooled to room temperature, poured into water, and extracted three times with ethyl acetate (30 ml). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 40:60) to afford tert-butyl (S)-(6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (66 mg, 23.93% yield) as a colorless oil.

[0172] LCMS(ESI):275[M+H] +

[0173] Step 2: (S)-3-(Methylamino)-2,3-dihydrobenzofuran-6-carbonitrile

[0174] Tert-butyl (S)-(6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (66 mg, 0.24 mmol) was added to a solution of hydrogen chloride in dioxane (3 mL, 4 mol / L), and the mixture was stirred for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure to afford crude (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carbonitrile (40 mg) as a white solid. The crude product was used directly in the next step without purification.

[0175] LCMS(ESI):175[M+H] +

[0176] Synthesis of Intermediate 23 (S)-6-methoxy-N-methyl-2,3-dihydrobenzofuran-3-amine

[0177]

[0178] A reaction system of tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (328 mg, 1 mmol), CuI (352 mg, 4 mmol), and a methanolic solution of sodium methoxide (180 mg, 40 mmol) in N,N-dimethylformamide (4 mL) was reacted at 120°C for four hours. After completion of the reaction, the system was cooled to room temperature and extracted three times with 10 mL of ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain (S)-6-methoxy-N-methyl-2,3-dihydrobenzofuran-3-amine (150 mg, 0.8 mmol, 80% yield) as a colorless oil.

[0179] LCMS(ESI):180[M+H] +

[0180] Intermediate 24 (S)-dimethyl (3-(methylamino)-2,3-dihydrobenzofuran-6-yl) phosphine oxide

[0181]

[0182] Step 1: A reaction system of tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (200 mg, 0.61 mmol), dimethylphosphine (57 mg, 0.73 mmol), potassium carbonate (101 mg, 0.73 mmol), palladium acetate (10 mg, 0.06 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (35 mg, 0.06 mmol), and 5 mL of dimethylformamide was reacted in a microwave reactor at 150°C for 20 minutes. After completion of the reaction, the system was cooled to room temperature and extracted with ethyl acetate (30 mL) (3 times, 10 mL each). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Tert-butyl (S)-(6-(dimethylphosphoryl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, 0.245 mmol, 40.35% yield) was obtained as a white solid.

[0183] LCMS(ESI):326[M+H] +

[0184] Step 2: tert-Butyl (S)-(6-(dimethylphosphino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, 0.25 mmol) was added to a 4M hydrochloric acid solution in dioxane (3 mL) and stirred for 2 hours. After the reaction, the resulting mixture was concentrated under reduced pressure. This afforded (S)-dimethyl(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)phosphine oxide (25 mg, 0.11 mmol, 45.14% yield) as a white solid.

[0185] LCMS(ESI):226[M+H] +

[0186] Intermediate 25 (S)-N-methyl-6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-amine

[0187]

[0188] Step 1: A reaction system of tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (328 mg, 1 mmol), sodium methanesulfinate (122 mg, 1.2 mmol), L-proline (12 mg, 0.1 mmol), potassium carbonate (166 mg, 1.2 mmol), cuprous iodide (19 mg, 0.1 mmol), and N,N-dimethylformamide (5 mL) was reacted at 140°C in a microwave reactor for 2 hours. After completion of the reaction, the system was cooled to room temperature and extracted three times with 10 mL of ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (S)-tert-Butyl methyl(6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-yl)carbamate (60 mg, 0.18 mmol, 18.32% yield) was obtained as a colorless oil.

[0189] LCMS(ESI):328[M+H] +

[0190] Step 2: Add tert-butyl (S)-methyl(6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-yl)carbamate (60 mg, 0.24 mmol) to a 4 M solution of hydrogen chloride in 1,4-dioxane (3 mL) and stir for 2 hours. After the reaction, the resulting mixture was concentrated under reduced pressure to obtain (S)-N-methyl-6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-amine (30 mg, 0.13 mmol, 72.03% yield) as a white solid.

[0191] LCMS(ESI):228[M+H] +

[0192] Synthesis of Intermediate 101 (S)-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine

[0193]

[0194] Step 1: Preparation of tert-butyl (S)-(6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0195] Tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (300 mg, 0.914 mmol) and sodium cyclopropanesulfinate (118 mg, 0.914 mmol) were dissolved in DMSO (10 ml), and copper iodide (35 mg, 0.182 mmol) and (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (52 mg, 0.365 mmol) were added, and the reaction was stirred at 25°C for 10 hours. When LCMS showed the reaction was complete, water (10 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The mixture was purified by column chromatography (0-10% ethyl acetate / petroleum ether) to give (S)-tert-butyl (6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (150 mg, yield: 46.4%).

[0196] LCMS (ESI) m / z: 354.1 [M+H] +

[0197] Step 2: Synthesis of (S)-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine: Compound (S)-tert-butyl (6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (150 mg, 0.424 mmol) was dissolved in dichloromethane (3 ml) at room temperature, and trifluoroacetic acid (1 ml) was added. The reaction was stirred at room temperature for 16 hours, and the mixture was concentrated under reduced pressure to give crude S-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine (150 mg, crude product), which was used directly in the next step without purification.

[0198] LCMS (ESI) m / z: 254.1 [M+H] +

[0199] Synthesis of Intermediate 26 (S)-N-methyl-6-(1-trifluoromethyl)pyrazol-4-yl)-2,3-dihydrobenzofuran-3-amine

[0200]

[0201] Step 1: Synthesis of tert-butyl-(S)-methyl(6-(1-trifluoromethyl)-1H-pyrazol-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate

[0202] Under N2 atmosphere, tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (300 mg, 0.91 mmol) was dissolved in 1,4-dioxane and water (4:1, 10 ml), and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolin-2-yl)-1-trifluoromethylpyrazole (239.0 mg, 1.37 mmol), potassium carbonate (377 mg, 2.73 mmol), and 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride (67.0 mg, 0.09 mmol) were added. The reaction was carried out at 100°C for 16 hours. The system was poured into water (30 ml) and extracted with ethyl acetate (20 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was dried to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to give tert-butyl tert-butyl-(S)-methyl(6-(1-trifluoromethyl)-1H-pyrazol-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate (240.0 mg, 68.8% yield) as a white solid.

[0203] LCMS (ESI) m / z: 384.1 [M+H] +

[0204] Step 2: Synthesis of (S)-N-methyl-6-(1-trifluoromethyl)pyrazol-4-yl)-2,3-dihydrobenzofuran-3-amine

[0205] Tert-butyl (S)-methyl (6-(1-trifluoromethyl)-1H-pyrazol-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate (240 mg, 0.626 mmol) was dissolved in 4M hydrochloric acid and ethyl acetate (3 ml) at room temperature. After stirring at room temperature for 1 hour, the reaction solution was concentrated under reduced pressure to give the crude product (S)-N-methyl-6-(1-trifluoromethyl)pyrazol-4-yl)-2,3-dihydrobenzofuran-3-amine (25.0 mg, crude product).

[0206] LCMS (ESI) m / z: 283.1 [M+H] +

[0207] The following intermediates were prepared by using the same method and steps as for intermediate 26, replacing only the corresponding starting materials:

[0208]

[0209]

[0210]

[0211]

[0212] Intermediate 41 6-(Trifluoromethyl)benzo[b]thiophen-3(2H)-one

[0213]

[0214] Step 1: Synthesis of 2-fluoro-4-(trifluoromethyl)benzoic acid

[0215] To a solution of methyl 2-fluoro-4-(trifluoromethyl)benzoate (30.00 g, 135.05 mmol) in tetrahydrofuran:water (300 ml:30 ml) was added lithium hydroxide (9.70 g, 405.15 mmol) at room temperature, and the mixture was stirred at 50°C for 2 hours. The reaction was poured into water (100 ml), the pH was adjusted to 4 with formic acid solution, and then extracted with ethyl acetate (100 ml x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate = 30%) to provide 2-fluoro-4-(trifluoromethyl)benzoic acid (22.00 g, yield: 78%).

[0216] LCMS (ESI) m / z: 209.1 [M+H] +

[0217] Step 2: Synthesis of 2-((2-ethoxy-2-oxoethyl)thio)-4-(trifluoromethyl)benzoic acid

[0218] To a solution of ethyl 2-sulfanyl acetate (12.70 g, 105.72 mmol) in N,N-dimethylformamide (200.00 ml) at 0°C was added sodium hydrogen hydride (8.46 g, 211.43 mmol), and the mixture was stirred at room temperature for 0.5 hour. 2-Fluoro-4-(trifluoromethyl)benzoic acid (22.00 g, 105.71 mmol) was then added to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction solution was slowly poured into water (100 ml), extracted with ethyl acetate (200 ml), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate = 10%) to provide 2-((2-ethoxy-2-oxoethyl)thio)-4-(trifluoromethyl)benzoic acid (13.00 g, 40% yield).

[0219] LCMS (ESI) m / z: 309.2 [M+H] +

[0220] Step 3: Synthesis of 2-((carboxymethyl)thio)-4-(trifluoromethyl)benzoic acid

[0221] A mixture of 2-[(ethoxycarbonyl)methylthio]-4-(trifluoromethyl)benzoic acid (13.00 g, 42.17 mmol) and potassium hydroxide (4.72 g, 84.34 mmol) in methanol (130.00 mL) was stirred at room temperature for 2 hours and then concentrated to give 2-((carboxymethyl)thio)-4-(trifluoromethyl)benzoic acid (7.00 g, crude).

[0222] LCMS (ESI) m / z: 281.2 [M+H] +

[0223] Step 4: Synthesis of 6-(trifluoromethyl)benzo[b]thiophene-3-yl acetate

[0224] To a solution of 2-(carboxymethylthio)-4-(trifluoromethyl)benzoic acid (7.00 g, 24.98 mmol) in acetic anhydride (70.00 ml) was added sodium acetate trihydrate (10.19 g, 74.94 mmol) at room temperature. The mixture was stirred at 140°C for 30 minutes. The mixture was poured into water, extracted with ethyl acetate (100 ml), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash column chromatography (petroleum ether:ethyl acetate = 50%) to give 6-(trifluoromethyl)benzo[b]thiophen-3-yl acetate (5.00 g, 77% yield).

[0225] LCMS (ESI) m / z: 261.2 [M+H]+

[0226] Step 5: Synthesis of 6-(trifluoromethyl)benzo[b]thiophene-3(2H)-one

[0227] Aqueous hydrochloric acid (10.00 mL, 1 mol / L) was added to a solution of 6-(trifluoromethyl)benzo[b]thiophen-3-yl acetate (1.00 g, 3.84 mmol) in 1,4-dioxane (5.00 mL) at room temperature. The mixture was stirred at 100°C for 2 hours. The reaction mixture was cooled and poured into water, and the aqueous layer was extracted with ethyl acetate (2 x 200 mL). The combined organic layers were washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Purification by flash column chromatography (petroleum ether:ethyl acetate = 70%) afforded 6-(trifluoromethyl)benzo[b]thiophen-3(2H)-one (0.50 g, 59% yield) as a black solid.

[0228] LCMS (ESI) m / z: 219.2 [M+H] +

[0229] Intermediate 42 Synthesis of 1-methyl-7-trifluoromethylisochromen-4-one

[0230]

[0231] Step 1: Synthesis of 2-(1-allyloxy)ethyl)-1-bromo-4-trifluoromethylbenzene

[0232] At room temperature, 1-(2-bromo-5-trifluoromethylphenyl)ethane-1-ol (10 g, 37.16 mmol) was dissolved in tetrahydrofuran (100 ml), and potassium hydroxide (4.2 g, 74.33 mmol), tetrabutylammonium hydrogen sulfate (2.6 g, 7.433 mmol) and 3-bromopropylene (5.4 g, 44.60 mmol) were added. The reaction was carried out at 25°C for 4 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure and poured into water (100 ml), and extracted with ethyl acetate (100 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product (12 g, crude product), which was used directly in the next step without purification.

[0233] LCMS (ESI) m / z: 309.2 [M+H] +

[0234] Step 2: Synthesis of 1-methyl-4-methylene-7-trifluoromethylisochromane

[0235] To a solution of 2-(1-allyloxyethyl)-1-bromo-4-trifluoromethylbenzene (12 g, 38.82 mmol) in DMF (100 mL) were added cesium carbonate (15.5 g, 46.59 mmol), tris(o-methylphenyl)phosphine (5.1 g, 19.41 mmol), and palladium acetate (0.89 g, 3.882 mmol). Under a nitrogen atmosphere, the mixture was stirred at 90°C for 16 hours. The reaction solution was poured into 400 mL of water and extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous NaSO, and concentrated under reduced pressure to obtain the crude product. 1-Methyl-4-methylene-7-trifluoromethylisochromane (5.5 g, 62.1% yield) was obtained by column chromatography (petroleum ether:ethyl acetate = 10:1).

[0236] LCMS (ESI) m / z: 229.1 [M+H] +

[0237] Step 3: Synthesis of 4-hydroxymethyl-1-methyl-7-trifluoromethylisochromen-4-ol

[0238] 1-Methyl-4-methylene-7-trifluoromethylisochromene (5.5 g, 24.10 mmol) was dissolved in a mixed solvent of acetone (100 ml) and water (20 ml) at room temperature. N-methylmorpholine-N-oxide (9.2 g, 76.68 mmol) and potassium osmate (0.91 g, 2.410 mmol) were added. The reaction was stirred at 25°C under a nitrogen atmosphere for 16 hours. After completion of the reaction, solid sodium sulfite (5 g) was added to the reaction solution, which was stirred for 10 minutes. The solution was then concentrated under reduced pressure to remove a certain amount of acetone, poured into water (200 ml), and extracted with ethyl acetate (100 ml x 3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain crude 6-fluoro-4-hydroxymethyl-1-methylisochromen-4-ol (6.0 g, yield: 94.9%).

[0239] LCMS (ESI) m / z: 263.0 [M+H] +

[0240] Step 4: Synthesis of 1-methyl-7-trifluoromethylisochromen-4-one

[0241] At room temperature, 4-hydroxymethyl-1-methyl-7-trifluoromethylisochromen-4-ol (6.0 g, 22.88 mmol) was dissolved in a mixed solvent of tetrahydrofuran (100 ml) and water (3.5 ml), and sodium periodate (15.0 g, 68.64 mmol) was added. The mixture was stirred at 25° C. under a N2 atmosphere for 4 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate, filtered, washed, and the filtrate was combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product 6-fluoro-1-methylisochromen-4-one (5 g, crude product).

[0242] LCMS (ESI) m / z: 230.2 [M+H] +

[0243] The following intermediates were prepared by using the same method and steps as for intermediate 42, except that the corresponding raw materials were replaced:

[0244]

[0245]

[0246]

[0247] The following ketones were purchased from different suppliers:

[0248]

[0249]

[0250] Intermediate 55 N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride

[0251]

[0252] Step 1: Synthesis of 6-(trifluoromethyl)isochroman-4-ol

[0253] 4-(Hydroxymethyl)-6-(trifluoromethyl)isochroman-4-one (1.1 g, 5.09 mmol) was dissolved in THF (10 mL) at room temperature and cooled on ice. Sodium borohydride (251 mg, 6.62 mmol) was then added dropwise, followed by 2 drops of methanol. The reaction was allowed to proceed overnight at room temperature. After completion, the reaction was quenched with a 1M aqueous solution of hydrogen chloride (2 mL) and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over sodium sulfate, and filtered to obtain a crude product, which was then separated and purified by column chromatography to afford 6-(trifluoromethyl)isochroman-4-ol (1.16 g).

[0254] Step 2: Synthesis of tert-butyl(tert-butyloxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate

[0255] 6-(Trifluoromethyl)isochroman-4-ol (100.0 mg, 0.459 mmol) was dissolved in THF (5.0 mL) at room temperature. Bis(tert-butyloxycarbonyl)amine (110 mg, 0.505 mmol) and triphenylphosphine (132 mg, 0.505 mmol) were added. The reaction system was stirred at 0°C for 5 minutes. DIAD (102 mg, 0.505 mmol) was added dropwise. The reaction was stirred in an ice bath for 1 hour and then allowed to return to room temperature overnight. After completion of the reaction, the mixture was added to water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by column chromatography (pure petroleum ether) to afford tert-butyl(tert-butyloxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate (90.0 mg).

[0256] Step 3: Synthesis of tert-butyl (6-(trifluoromethyl)isochroman-4-yl)carbamate

[0257] Tert-butyl(tert-butoxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate (60.0 mg, 0.14 mmol) was dissolved in acetonitrile (2.0 ml), and lithium bromide (37.6 mg, 0.432 mmol) was added. The reaction system was reacted at 60°C for 20 hours. After completion of the reaction, saturated sodium bicarbonate (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to provide tert-butyl (6-(trifluoromethyl)isochroman-4-yl)carbamate (30 mg).

[0258] Step 4: Synthesis of tert-butyl N-methyl-6-(trifluoromethyl)isochroman-4-yl)carbamate

[0259] Dissolve tert-butyl (6-(trifluoromethyl)isochroman-4-yl)carbamate (360.0 mg, 1.14 mmol) in 5.0 mL of DMF and stir on ice for 2 minutes. Then, add sodium hydride (90.8 mg, 2.27 mmol) in one portion. Stir at this temperature for 1 hour, then add iodomethane (487 mg, 3.41 mmol). Stir overnight at room temperature. After completion of the reaction, add saturated sodium bicarbonate (15 mL), extract with ethyl acetate (15 mL x 3), dry the organic phase over sodium sulfate, filter, and spin-dry the filtrate to yield crude tert-butyl N-methyl-6-(trifluoromethyl)isochroman-4-yl)carbamate (400.0 mg).

[0260] Step 5: Synthesis of N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride

[0261] Dissolve tert-butyl N-methyl-6-(trifluoromethyl)isochroman-4-ylcarbamate (400 mg, 1.21 mmol) in 2 mL of dichloromethane. Add 4M hydrogen chloride in dioxane (2 mL) and stir overnight at room temperature. After completion, the reaction mixture was spin-dried to yield N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride (390 mg).

[0262] The following intermediates were prepared by using the same method and steps as for intermediate 55, replacing only the corresponding starting materials:

[0263]

[0264]

[0265] Intermediate 60 (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride

[0266]

[0267] Step 1: Synthesis of (S)-7-(trifluoromethyl)pyran-4-ol

[0268] 7-(Trifluoromethyl)pyran-4-one (500.0 mg, 2.31 mmol) was dissolved in DCM (10 mL) at room temperature. (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanediamine (p-isopropylbenzene) chloride (147.0 mg, 0.23 mmol) was added to the reaction mixture under ice-cooling. Formic acid (372.0 mg, 8.10 mmol) was then added dropwise, followed by triethylamine (701.0 mg, 6.94 mmol). The reaction was allowed to proceed overnight at room temperature. After completion of the reaction, saturated aqueous sodium bicarbonate (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over sodium sulfate, and filtered to obtain the crude product, which was then purified by column chromatography (petroleum ether:ethyl acetate = 0-40%) to afford (S)-7-(trifluoromethyl)pyran-4-ol (370 mg, 73% yield).

[0269] Step 2: Synthesis of tert-butyl (R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate

[0270] Under a nitrogen atmosphere, (S)-7-(Trifluoromethyl)pyran-4-ol (370.0 mg, 1.70 mmol) was dissolved in THF (5.0 ml). Bis(tert-butyloxycarbonyl)amine (405 mg, 1.88 mmol) and triphenylphosphine (489 mg, 1.88 mmol) were added. The reaction system was stirred at 0°C for 5 minutes. DIAD (377.0 mg, 1.88 mmol) was added dropwise. The reaction solution was stirred in an ice bath for 1 hour and then stirred at room temperature overnight. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (10 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (petroleum ether:ethyl acetate = 0-5%) to give tert-butyl (R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate (145.0 mg, yield: 20%).

[0271] 1 H NMR(400MHz, CDCl3)δ7.25-7.22(m,1H),7.12-7.09(m,1H),7.07(d,1H),5.59-5.54(m,1H) ,4.46-4.41(m,1H),4.18-4.10(m,1H),2.67-2.60(m,1H),2.16-2.10(m,1H),1.46(s,18H).

[0272] Step 3: Synthesis of tert-butyl (R)-(7-(trifluoromethyl)pyran-4-yl)carbamate

[0273] Tert-butyl (R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate (150.0 mg, 0.36 mmol) was dissolved in acetonitrile (5.0 ml), and lithium bromide (94.0 mg, 1.08 mmol) was added. The reaction system was reacted at 60°C for 20 hours. After completion of the reaction, saturated sodium bicarbonate (10 ml) was added, and the mixture was extracted with ethyl acetate (20 ml). The organic phase was dried over sodium sulfate, filtered, and evaporated to dryness to obtain crude tert-butyl (R)-(7-(trifluoromethyl)pyran-4-yl)carbamate (83.0 mg, crude product).

[0274] Step 4: Synthesis of tert-butyl (R)-methyl (7-(trifluoromethyl)pyran-4-yl)carbamate

[0275] Tert-butyl (R)-(7-(trifluoromethyl)pyran-4-yl)carbamate (83.0 mg, 0.26 mmol) was dissolved in 2.0 mL of DMF and stirred on ice for 2 minutes. Sodium hydride (26.2 mg, 0.66 mmol) was added all at once and stirred at this temperature for 1 hour. Methyl iodide (74.4 mg, 0.52 mmol) was then added and stirred at room temperature overnight. After completion of the reaction, saturated sodium bicarbonate (10 mL) was added and the mixture was extracted with ethyl acetate (15 mL). The organic phase was dried over sodium sulfate, filtered, and the filtrate was evaporated to dryness to yield crude tert-butyl (R)-methyl (7-(trifluoromethyl)pyran-4-yl)carbamate (100.0 mg, crude product).

[0276] LCMS (ESI) m / z: 276.1 [M+H]+

[0277] Step 5: Synthesis of (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride

[0278] Tert-butyl (R)-methyl (7-(trifluoromethyl)pyran-4-yl)carbamate (100 mg, 0.3 mmol) was dissolved in 2 mL of dichloromethane. A 4M solution of hydrogen chloride in dioxane (2 mL) was added, and the reaction was stirred at room temperature overnight. After completion of the reaction, the reaction mixture was directly spin-dried to obtain the crude product (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride (90 mg, crude product).

[0279] Using the method and steps used for intermediate 60, the following intermediate was prepared:

[0280]

[0281]

[0282] Intermediate 100 N,1,1-trimethyl-7-trifluoromethylisochromen-4-amine

[0283]

[0284] Step 1: Preparation of 1,1-dimethyl-7-trifluoromethylisochromen-4-ol

[0285] 1,1-Dimethyl-7-trifluoromethylisochromen-4-one (300 mg, 1.229 mmol) was dissolved in MeOH (3 mL), and sodium borohydride (70 mg, 1.844 mmol) was added. The mixture was stirred at 25° C. for 10 hours. When LCMS showed the reaction was complete, aqueous NH 4 Cl (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (0-10% ethyl acetate / petroleum ether) to give 1,1-dimethyl-7-trifluoromethylisochromen-4-ol (120 mg, yield: 39.4%).

[0286] LCMS (ESI) m / z: 247.1 [M+H] +

[0287] Step 2: Preparation of 1,1-dimethyl-7-trifluoromethylisochromene-4-methanesulfonate

[0288] 1,1-Dimethyl-7-trifluoromethylisochromen-4-ol (120 mg, 0.487 mmol) and methanesulfonyl chloride (112 mg, 0.974 mmol) were dissolved in DCM (10 mL), and triethylamine (148 mg, 1.461 mmol) was added. The mixture was stirred at 25°C for 10 hours. When LCMS indicated the reaction was complete, water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude 1,1-dimethyl-7-trifluoromethylisochromen-4-methanesulfonate (130 mg, yield: 13.3%).

[0289] LCMS (ESI) m / z: 326.1 [M+H] +

[0290] Step 3: Preparation of N,1,1-trimethyl-7-trifluoromethylisochromen-4-amine

[0291] Compound 1,1-dimethyl-7-trifluoromethylisochromene-4-methanesulfonate (130 mg, 0.398 mmol) was dissolved in DMF (3 ml) at room temperature, and a methylamine alcohol solution (0.3 ml, 30%) and DIEA (155 mg, 1.194 mmol) were added. The reaction was stirred at room temperature for 16 hours. When LCMS showed the reaction was complete, water (10 ml) was added and the mixture was extracted with ethyl acetate (20 ml x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (50-100% ethyl acetate / petroleum ether) to give N,1,1-trimethyl-7-trifluoromethylisochromene-4-amine (52 mg, yield: 50.4%).

[0292] LCMS (ESI) m / z: 260.1 [M+H] +

[0293] The following intermediates were prepared by using the preparation method and steps of intermediate 100, with only the corresponding raw material intermediates being replaced:

[0294]

[0295]

[0296] Intermediate 69 Synthesis of 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine

[0297]

[0298] Step 1: Synthesis of (5-bromo-2-(cyclopropylimino)methyl)phenol

[0299] To a solution of 4-bromo-2-hydroxybenzaldehyde (2.0 g, 9.95 mmol) in dichloromethane (40 ml) was added cyclopropylamine (1.13 g, 19.9 mmol) and anhydrous magnesium sulfate (4.79 g, 39.8 mmol) at room temperature. The mixture was stirred at 25°C for 20 hours. After the reaction was complete, the reaction solution was concentrated, ethyl acetate was added, and the filter cake was washed with ethyl acetate. The resulting filtrate was concentrated under reduced pressure to give a crude product (5-bromo-2-(cyclopropylimino)methyl)phenol (1.7 g, crude product) as a yellow solid, which was used directly in the next step without purification.

[0300] LCMS (ESI) m / z: 241.1 [M+H] +

[0301] Step 2: Synthesis of 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine

[0302] To a solution of trimethylsulfoxide iodide (3.89 g, 17.70 mmol) in THF (5 mL) at room temperature, potassium tert-butoxide (1.99 g, 17.70 mmol) was slowly added. The mixture was stirred at room temperature for 0.5 hours. (5-Bromo-2-(cyclopropylimino)methyl)phenol (1.7 g, 7.08 mmol) was then dissolved in THF and slowly added dropwise to the mixture. The resulting suspension was stirred at room temperature for 1 hour and then at 50°C for 3 hours. The reaction mixture was cooled to room temperature, and 1 equivalent of potassium tert-butoxide (0.79 g, 7.08 mmol) was added, followed by stirring at room temperature for 12 hours. The reaction mixture was filtered, and the filtrate was diluted with water and extracted with ethyl acetate. The organic layer was concentrated and the residue was purified by column chromatography with petroleum ether / ethyl acetate (3:1) to give 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine (1.0 g, 3.93 mmol) as a yellow oil.

[0303] LCMS (ESI) m / z: 255.1 [M+H] +

[0304] The intermediate amines in the following table can be prepared by using the synthetic steps described for intermediate 69, replacing only the corresponding starting materials, aldehydes or amines.

[0305]

[0306] Intermediate 71 Synthesis of 3-(cyclopropylamino)-2,3-dihydrobenzofuran-6-carbonitrile

[0307]

[0308] At room temperature, 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine (100 mg, 0.41 mmol) was placed in a 50 ml single-necked bottle, and Pd2(dba)3 (18.8 mg, 0.02 mmol), S-Phos (9.8 mg, 0.02 mmol), zinc cyanide (96.5 mg, 0.82 mmol) were added, and then N,N-dimethylacetamide (1.0 ml) was added. The reaction was heated to 110 ° C and stirred for 1 hour. Water (10 ml) was added and extracted with ethyl acetate (20 ml × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried to obtain a crude product. The crude product was mixed and purified by column chromatography with ethyl acetate: petroleum ether (0-50%) to give 3-(cyclopropylamino)-2,3-dihydrobenzofuran-6-carbonitrile (55 mg, yield: 67%).

[0309] 1H NMR(400MHz, CDCl3)δ7.44-7.42(dd,1H),7.2-7.19(dd,1H),7.07(d,1H),4.65-4.61(m,1H),4 .58-4.55(m,1H),4.47-4.46(m,1H),2.25-2.19(m,1H),0.52-0.49(m,2H),0.43-0.40(m,2H).

[0310] Intermediate 7 2(S)-6-(Fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine

[0311]

[0312] Step 1: Synthesis of tert-butyl (S)-(6-(hydroxymethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0313] A mixture of (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamic acid tert-butyl ester (400 mg, 1.22 mmol), 2,2-dibutyl-2-stannoxan-1-ol (391 mg, 1.22 mmol), and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (99 mg, 0.12 mmol) in dioxane (5 ml) was reacted at 100°C for 12 hours. After completion of the reaction, the reaction solution was poured into water (10 ml) and extracted with ethyl acetate (10 ml x 2). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether=40:60) to give tert-butyl (S)-(6-(hydroxymethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (110 mg, yield: 29%).

[0314] LCMS(ESI):280[M+H] +

[0315] Step 2: Synthesis of tert-butyl (S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0316] To a mixture of N-[(3S)-6-(hydroxymethyl)(2,3-dihydrobenzo[b]furan-3-yl)](tert-butoxy)-N-methylformamide (100 mg, 0.36 mmol) in dichloromethane (1.00 ml) was added [bis(2-methoxyethyl)amine]sulfur trifluoride (396 mg, 1.79 mmol) at -78° C. The mixture was stirred at -78° C. for 1 hour. After the reaction was completed, the reaction was quenched with ice water (5 ml), extracted with ethyl acetate (5 ml x 2), and the organic layers were combined, washed with saturated brine (5 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate: petroleum ether = 20:80) to give (S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamic acid tert-butyl ester (80 mg, yield: 70%).

[0317] LCMS(ESI):282[M+H] +

[0318] Step 3: Synthesis of (S)-6-(fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine

[0319] A mixture of (S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamic acid tert-butyl ester (80 mg, 0.36 mmol) and dioxane hydrochloride (1 ml) was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was directly spin-dried to give (S)-6-(fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine (100 mg, crude product).

[0320] LCMS(ESI):182[M+H] +

[0321] Intermediate 73: Synthesis of (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxamide

[0322]

[0323] Step 1: Synthesis of (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid

[0324] Methyl (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate (1 g, 3.25 mmol) was dissolved in methanol (10.00 ml) and tetrahydrofuran (10.00 ml) at room temperature. Sodium hydroxide (260 mg, 6.5 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to obtain (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid (600 mg, crude) as a white solid.

[0325] LCMS(ESI):294[M+H] +

[0326] Step 2: Synthesis of tert-butyl (S)-(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0327] (S)-3-((tert-Butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid (600 mg, 2.04 mmol) was dissolved in N,N-dimethylformamide (5.00 ml) at room temperature. 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.62 g, 4.38 mmol), N,N-diisopropylethylamine (1.38 g, 10.68 mmol), and ammonium chloride (360 mg, 6.78 mmol) were added. The mixture was reacted at room temperature for half an hour. After completion of the reaction, the mixture was diluted with ethyl acetate (30 ml) and water (30 ml), and the aqueous layer was extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by flash column chromatography (ethyl acetate / petroleum ether=17%) to give (S)-tert-butyl(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (240 mg, yield: 40%) as a yellow liquid.

[0328] LCMS(ESI):293[M+H]+

[0329] Step 3: Synthesis of (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxamide

[0330] Tert-butyl (S)-(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (240 mg, 0.82 mmol) was dissolved in dichloromethane (2.00 ml) and trifluoroacetic acid (0.40 ml) at room temperature. The mixture was stirred at room temperature for half an hour. After completion of the reaction, the reaction solution was concentrated to obtain (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxamide (100 mg, yield: 66.66%) as a black solid.

[0331] Example 74: (S)-N 3 -Synthesis of methyl-2,3-dihydrobenzofuran-3,6-diamine

[0332]

[0333] Step 1: Synthesis of tert-butyl (S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate

[0334] To a solution of tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (120 mg, 0.35 mmol) in 1,4-dioxane (5.00 ml) was added benzophenone imine (190 mg, 1.06 mmol), potassium tert-butoxide (80 mg, 0.71 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (40 mg, 0.07 mmol) and palladium acetate (8 mg, 0.03 mmol) at room temperature. The reaction was continued at 00°C for 2 hours. After the reaction, the reaction solution was poured into water (2 ml), extracted with ethyl acetate (2 ml × 2), and the organic layers were combined, washed with saturated brine (2 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by flash column chromatography (petroleum ether:ethyl acetate = 10%) to give tert-butyl (S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (90 mg, yield: 59%) as a white solid.

[0335] LCMS(ESI):429[M+H] +

[0336] Step 2: (S)-N 3 -Synthesis of methyl-2,3-dihydrobenzofuran-3,6-diamine

[0337] At room temperature, tert-butyl (S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (90 mg, 0.21 mmol) was dissolved in a hydrochloric acid methanol solution (5.00 ml), and the mixture was stirred at room temperature for 0.5 hours. After the reaction was completed, the reaction solution was poured into a dichloromethane solution to precipitate a solid to obtain (S)-N 3 -methyl-2,3-dihydrobenzofuran-3,6-diamine (60 mg, yield: 90%).

[0338] LCMS(ESI):165[M+H] +

[0339] Intermediate 75: Synthesis of (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide

[0340]

[0341] Step 1: Synthesis of tert-butyl (S)-methyl (6-((trifluoromethyl)sulfonamido)-2,3-dihydrobenzofuran-3-yl)carbamate

[0342] Tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (200 mg, 0.61 mmol), trifluoromethylsulfonamide (136 mg, 0.91 mmol), cuprous iodide (1.16 g, 6.09 mmol), and potassium phosphate (259 mg, 1.22 mmol) were sequentially dissolved in dimethylformamide (9 mL). The mixture was stirred at 90°C for 2 hours. After the reaction, the system was cooled to room temperature and extracted with ethyl acetate three times (10 mL each). The organic phases were combined, washed with saturated brine (10 mL x 3), and dried over anhydrous sodium sulfate. The reaction mixture was concentrated in vacuo to yield tert-butyl (S)-methyl(6-((trifluoromethyl)sulfonamido)-2,3-dihydrobenzofuran-3-yl)carbamate (800 mg, 51.85% yield) as a yellow liquid.

[0343] LCMS (ESI): 397.2 [M+H] +

[0344] Step 2: Synthesis of (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide

[0345] Dissolve (S)-tert-butyl methyl(6-((trifluoromethyl)sulfonamido)-2,3-dihydrobenzofuran-3-yl)carbamate (80 mg, 3.246 mmol) in a 4.0 M solution of hydrogen chloride in 1,4-dioxane (5 ml). Stir the mixture at room temperature for 2 hours. After the reaction, cool the system to room temperature, and concentrate the reaction mixture in vacuo to obtain (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide (60 mg, 1.18 mmol, 82.46% yield).

[0346] Intermediate 76 4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester

[0347]

[0348] Step 1: Synthesis of imidazo[1,5-a]imidazo[1,5-d]1,4-diazapiperazine-5,10-dione

[0349] Imidazole-5-carboxylic acid (100 g, 892.14 mmol) was added to thionyl chloride (500 ml), stirred at 80° C. for 12 hours, and the mixture was concentrated. The concentrated mixture was washed with toluene (500 ml×2) and filtered to obtain a solid product. The residual solvent was removed by vacuum oil pump to obtain imidazo[1,5-a]imidazo[1,5-d]1,4-diazapiperazine-5,10-dione (70 g, 0.37 mol, yield 42%) as a yellow solid.

[0350] LCMS (ESI) m / z: 189 [M+H] +

[0351] Step 2: Synthesis of N-(4-bromo-2-fluorophenyl)imidazol-5-ylcarboxamide

[0352] To a mixture of 4-bromo-2-fluorophenylamine (60.60 g, 318.91 mmol) in tetrahydrofuran (600 ml) was slowly added dropwise sodium bistrimethylsilylamide (318.91 ml, 637.82 mmol, 2 mol / L) at 0°C over a period of 1 hour. Imidazo[1,5-a]imidazo[1,5-d]1,4-diazapiperazine-5,10-dione (60 g, 318.91 mmol) was then added. The mixture was stirred at room temperature for 12 hours, poured into water, filtered, and the crude product was purified by silica gel chromatography (petroleum ether:ethyl acetate = 40%) to give N-(4-bromo-2-fluorophenyl)imidazol-5-ylformamide (60 g, 0.21 mol, 66% yield) as a white solid.

[0353] LCMS (ESI) m / z: 284.2 [M+H]+

[0354] Step 3: Synthesis of 8-bromo-10-hydroimidazo[1,5-a]quinoxaline-4-ol

[0355] N-(4-Bromo-2-fluorophenyl)imidazol-5-ylformamide (60 g, 211.20 mmol) and sodium hydride (16.88 g, 422.40 mmol, 60% content) were stirred in dimethylacetamide (600 ml) at 140° C. for 12 hours. The mixture was poured into water and filtered to give 8-bromo-10-hydroimidazo[1,5-a]quinoxaline-4-ol (50 g, 90% yield) as a yellow solid.

[0356] LCMS (ESI) m / z: 264.2 [M+H] +

[0357] Step 4: Synthesis of 8-bromo-4-chloro-10-hydroimidazo[1,5-a]quinoxaline

[0358] Phosphorus oxychloride (500 ml) was added to a mixture of 8-bromo-10-hydroimidazo[1,5-a]quinoxaline-4-ol (50 g, 189.34 mmol) and N,N-diisopropylethylamine (48.85 g, 378.67 mmol), and the mixture was stirred at 90° C. for 2 hours, followed by concentration. The residue was dissolved in acetonitrile and slowly added dropwise to ice water to precipitate a solid, which was filtered to give 8-bromo-4-chloro-10-hydroimidazo[1,5-a]quinoxaline (50 g, yield 93%).

[0359] LCMS (ESI) m / z: 282.2 [M+H] + .

[0360] Step 5: Synthesis of (8-bromo(10-hydroimidazo[1,5-a]quinoxalin-4-yl))[(4-methoxyphenyl)methyl]amine

[0361] 8-Bromo-4-chloro-10-hydroimidazo[1,5-a]quinoxaline (50 g, 176.98 mmol) and 4-methoxybenzylamine (29.13 g, 212.38 mmol) were added to dimethyl sulfoxide (500 ml), and N,N-diisopropylethylamine (45.66 g, 353.96 mmol) was added. The mixture was stirred at 80° C. for 2 hours, then poured into water and filtered to give (8-bromo(10-hydroimidazo[1,5-a]quinoxaline-4-yl))[(4-methoxyphenyl)methyl]amine (50 g, yield 74%) as a yellow oil.

[0362] LCMS (ESI) m / z: 383.2 [M+H] +

[0363] Step 6: Synthesis of methyl 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate

[0364] (8-Bromo(10-hydroimidazo[1,5-a]quinoxaline-4-yl))[(4-methoxyphenyl)methyl]amine (50 g, 130.47 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (10.83 g, 13.05 mmol), and potassium acetate (25.57 g, 260.93 mmol) were added to a solution of dimethylformamide (50 ml) and methanol (250 ml). The system was reacted at 100° C. under a carbon monoxide atmosphere (4 MPa) for 12 hours, then poured into water and filtered. The mixture was purified by silica gel chromatography (petroleum ether:ethyl acetate = 80%) to give methyl 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (30 g, 63% yield) as a yellow solid.

[0365] LCMS (ESI) m / z: 363.2 [M+H] +

[0366] Step 7: Synthesis of 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0367] Methyl 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (30 g, 82.87 mmol) was added to a mixed solution of methanol, water, and tetrahydrofuran (1:1:1, 150 ml), and potassium hydroxide (92.40 g, 165.0 mmol) was added. The mixture was reacted at 60° C. for 12 hours. The organic solvent was removed by vacuum concentration, and the mixture was poured into water and adjusted to pH 7-8 with HCl (2M). The mixture was then extracted with ethyl acetate (100 ml×3) and concentrated in vacuo to give 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (25 g, yield 86%).

[0368] LCMS (ESI) m / z: 349.2 [M+H] +

[0369] Step 8: 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0370] 4-((4-Methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid (100.0 mg, 0.28 mmol) was dissolved in trifluoroacetic acid (2 mL) at room temperature. The mixture was stirred at 100°C for 2 hours. The reaction was monitored by LCMS. The mixture was concentrated to afford 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (60.0 mg, 94% yield) as a white solid.

[0371] LCMS (ESI) m / z: 229.2 [M+H] +

[0372] The following intermediate carboxylic acids can be prepared by using the synthetic steps described for intermediate 76, substituting only the corresponding starting materials:

[0373]

[0374]

[0375] Intermediate 84 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0376]

[0377] Step 1: Synthesis of methyl 3-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate

[0378] Methyl 3-fluoro-4-nitrobenzoate (20.0 g, 100 mmol) was dissolved in acetonitrile (100 ml), and then 2-methyl-1H-imidazole (8.2 g, 100 mmol) and potassium carbonate (27.6 g, 200 mmol) were added. The mixture was stirred at 100° C. for 12 hours. The reaction was monitored by liquid chromatography-mass spectrometry. The reaction solution was poured into 300 ml of water, and ethyl acetate (200 ml × 3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and chromatographed on silica gel (petroleum ether:ethyl acetate = 10%) to give methyl 3-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (25.0 g, 95% yield) as a yellow solid.

[0379] LCMS (ESI) m / z: 262 [M+H] +

[0380] Step 2: Synthesis of methyl 4-amino-3-(2-methyl-1H-imidazol-1-yl)benzoate

[0381] At room temperature, methyl 3-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (25 g, 95.8 mmol) was dissolved in methanol (300 ml), and then Raney nickel (2 g) was added. The mixture was stirred at room temperature for 12 hours under a hydrogen atmosphere. The reaction was monitored by LCMS. The residue was filtered to obtain a filtrate, which was concentrated and purified by silica gel chromatography (petroleum ether:ethyl acetate=20%) to give methyl 4-amino-3-(2-methyl-1H-imidazol-1-yl)benzoate (20.5 g, 93% yield) as a yellow solid.

[0382] LCMS (ESI) m / z: 232 [M+H] +

[0383] Step 3: Synthesis of 1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester

[0384] Methyl 4-amino-3-(2-methyl-1H-imidazol-1-yl)benzoate (2.0 g, 8.7 mmol) was dissolved in o-dichlorobenzene (40 ml) at room temperature, followed by the addition of carbonyldiimidazole (2.8 g, 17.4 mmol). The mixture was stirred at 180°C for 12 hours. The reaction was monitored by liquid chromatography-mass spectrometry. The mixture was filtered to obtain a filter cake, which was then slurried with ethyl acetate (5 ml) to afford methyl 1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (920.0 mg, 41% yield) as a black solid.

[0385] LCMS (ESI) m / z: 258 [M+H] +

[0386] Step 4: Synthesis of methyl 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate

[0387] Methyl 1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (100.0 mg, 0.39 mmol) was dissolved in phosphorus oxychloride (5 ml) at room temperature, and the mixture was stirred at 120°C for 4 hours. The reaction was monitored by liquid chromatography-mass spectrometry. The reaction solution was concentrated under reduced pressure, diluted with a small amount of acetonitrile, poured into water (5 ml), filtered, and the filter cake was washed with water and dried in vacuo to give methyl 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (60.0 mg, 56% yield) as a black solid.

[0388] LCMS (ESI) m / z: 276 [M+H] +

[0389] Step 5: Synthesis of methyl 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate

[0390] Methyl 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (60 mg, 0.22 mmol) was dissolved in dimethyl sulfoxide (2 ml) at room temperature, followed by the addition of (4-methoxyphenyl)methanamine (60 mg, 0.44 mmol) and N,N-diisopropylethylamine (67 mg, 0.52 mmol). The mixture was stirred at 100°C for 1 hour. The reaction was monitored by liquid chromatography-mass spectrometry. The reaction solution was poured into 10 ml of water and extracted with ethyl acetate (10 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel chromatography (petroleum ether:ethyl acetate = 40%) to give methyl 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (60 mg, 73% yield) as a black solid.

[0391] LCMS (ESI) m / z: 377 [M+H] +

[0392] Step 6: Synthesis of 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0393] At room temperature, methyl 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (60 mg, 0.16 mmol) was dissolved in methanol (1 ml), tetrahydrofuran (1 ml) and water (1 ml), and potassium hydroxide (26 mg, 0.44 mmol) was added. The mixture was stirred at room temperature for 1 hour. The reaction was monitored by liquid chromatography-mass spectrometry. The reaction solution was poured into 10 ml of water, acidified by the addition of formic acid, and extracted with ethyl acetate (10 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, 86% yield) as a white solid.

[0394] LCMS (ESI) m / z: 363 [M+H] +

[0395] Step 7: Synthesis of 4-amino-1-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0396] A solution of 4-{[(4-methoxyphenyl)methyl]amino}-1-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (400 mg, 1.10 mmol) in trifluoroacetic acid (5.00 ml) was stirred at 90° C. for 2 hours. The reaction solution was concentrated in vacuo to give 4-amino-1-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (250 mg, 94% yield).

[0397] LCMS (ESI) m / z: 243 [M+H] +

[0398] Intermediate 85 :4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0399]

[0400] Step 1: Synthesis of methyl 4-((tert-butoxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate

[0401] A solution of methyl 4-amino-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate (1.20 g, 4.34 mmol), di-tert-butyl dicarbonate (1.89 g, 8.67 mmol) and triethylamine (1.30 g, 13.01 mmol) in dichloromethane (10 ml) was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was poured into water (5 ml) and extracted with ethyl acetate (5 ml). The organic layer was dried over anhydrous sodium sulfate and filtered, then concentrated and purified by column chromatography (petroleum ether: ethyl acetate = 70:30) to give methyl 4-((tert-butoxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate (1.00 g, 61% yield) as a white solid.

[0402] LCMS(ESI):376[M+H] +

[0403] Step 2: Synthesis of methyl 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylate

[0404] Under nitrogen atmosphere, methyl 4-((tert-butyloxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate (0.20 g, 0.53 mmol), potassium ferricyanide (0.05 g, 0.16 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (0.09 g, 0.21 mmol), methanesulfonic acid (2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1 A mixture of 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (0.10 g, 51%) was stirred at 100° C. for 2 h. The mixture was concentrated and purified by column chromatography (petroleum ether:ethyl acetate=70:30) to give methyl 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylate (0.10 g, 51%) as a white solid.

[0405] LCMS(ESI):368[M+H] +

[0406] Step 3: Synthesis of methyl 4-amino-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylate

[0407] Trifluoroacetic acid (0.10 ml) was added to a solution of methyl 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylate (0.20 g, 0.54 mmol) in dichloromethane (0.50 ml), and the reaction was stirred at room temperature for 2 hours and then concentrated to give methyl 4-amino-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylate (0.05 g, crude product).

[0408] LCMS(ESI):268[M+H] +

[0409] Step 4: Synthesis of 4-amino-7-cyanoimidazolo[1,5-a]quinoxaline-8-carboxylic acid

[0410] A mixture of methyl 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate (0.20 g, 0.75 mmol), potassium hydroxide (0.07 g, 1.50 mmol), tetrahydrofuran / methanol / water (1 ml / 1 ml / 1 ml) was stirred at room temperature for 2 hours. After the reaction was completed, the system was concentrated to give a crude product of 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylic acid (210 mg, crude product). The crude product was used directly in the next step without purification.

[0411] LCMS(ESI):254[M+H] +

[0412] Intermediate 86 4-Amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0413]

[0414] Step 1: Synthesis of methyl 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate

[0415] A mixture of methyl 7-chloro-4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (200 mg, 0.5 mmol), potassium carbonate (300 mg, 2 mmol), dichloro[1,1'-bis(tert-butylphosphino)ferrocenepalladium(II)] (0.08 g, 0.1 mmol), and trimethylboroxane (0.01 g, 0.10 mmol) was stirred in dioxane (2.00 ml) at 100° C. for 12 hours, then poured into water, extracted with ethyl acetate (5 ml), dried over anhydrous sodium sulfate, filtered, and purified by column chromatography (petroleum ether:ethyl acetate=30%) to give methyl 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (100 mg, 53%).

[0416] LCMS(ESI):268[M+H] +

[0417] Step 2: Synthesis of 4-((4-methoxybenzyl)amino)-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0418] Methyl 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (500 mg, 1.86 mmol) was dissolved in methanol:tetrahydrofuran:saturated potassium hydroxide aqueous solution (1 ml:1 ml:1 ml). The reaction solution was reacted at 60° C. for 12 hours. After completion, the reaction solution was spin-dried and the pH was adjusted to 3 with formic acid. The solution was filtered to give 4-((4-methoxybenzyl)amino)-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (52 mg, 12% yield).

[0419] LCMS(ESI):363[M+H] +

[0420] Step 3: Synthesis of 4-amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0421] 4-((4-Methoxybenzyl)amino)-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (500 mg, 1.86 mmol) was dissolved in trifluoroacetic acid solution (5 ml), and the reaction solution was stirred at 100° C. for 12 hours. After the reaction was completed, the mixture was spin-dried to give 4-amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (520 mg, crude product).

[0422] LCMS(ESI):363[M+H] +

[0423] Using the method used in step 3 of intermediate 86, the following intermediate acid was prepared:

[0424]

[0425]

[0426] Intermediate 93 4-Amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid

[0427]

[0428] Step 1: Synthesis of 6-bromo-2-chloro-3-hydrazinoquinoxaline

[0429] 6-Bromo-2,3-dichloroquinoxaline (278.0 mg, 1.0 mmol) was placed in a 25 mL single-necked flask. Hydrazine hydrate (156 mg, 2.5 mmol, 80% wt) was added and the reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was filtered and the filter cake was washed with water (10 mL x 2) and ethyl acetate (5 mL x 2) to yield 6-bromo-2-chloro-3-hydrazinoquinoxaline (150 mg).

[0430] LCMS (ESI) m / z: 273.0 / 275.0 [M+H] +

[0431] Step 2: Synthesis of 8-bromo-4-chloro-[1,2,4]triazolo[4,3-a]quinoxaline

[0432] 6-Bromo-2-chloro-3-hydrazinoquinoxaline (116.0 mg, 0.5 mmol) was placed in a 25 mL single-necked vial at room temperature. Triethyl orthoformate (4.0 mL) was added. After the addition was complete, the reaction was heated to 100°C and allowed to react for 1 hour. After LCMS indicated completion of the reaction, the reaction mixture was cooled to room temperature and filtered. The filter cake was washed with methanol (3.0 mL x 3) and dried to yield 8-bromo-4-chloro-[1,2,4]triazolo[4,3-a]quinoxaline (110 mg).

[0433] 1 H NMR (400MHz, CDCl3) δ10.21(s,1H),8.84(d,1H),7.98(d,1H),7.90(dd,1H).

[0434] Step 3: Synthesis of 8-bromo-N-(4-methoxybenzyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4-amine

[0435] 8-Bromo-4-chloro-[1,2,4]triazolo[4,3-a]quinoxaline (1.70 g, 6.00 mmol) was dissolved in DMSO (15.0 ml), and p-methoxybenzylamine (1.23 g, 9.00 mmol) and DIEA (2.32 g, 18.00 mmol) were added. The reaction system was reacted at 90°C for 4 hours. After the reaction was completed, water (60 ml) was added and the mixture was extracted with ethyl acetate (40 ml x 3). The organic phase was dried over sodium sulfate, filtered, and evaporated to dryness to give crude 8-bromo-N-(4-methoxybenzyl)-[1,2,4]triazolo[4,3-a]quinoxalin-4-amine (2.20 g).

[0436] LCMS (ESI) m / z: 384.1 / 386.1 [M+H] +

[0437] Step 4: Synthesis of methyl 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate

[0438] To a solution of 8-bromo-N-(4-methoxybenzyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4-amine (2.2 g, 5.74 mmol) in MeOH (30 mL) and DMF (30 mL) were added potassium acetate (1.7 g, 17.2 mmol) and Pd(dppf)Cl2 (420 mg, 0.57 mmol). The mixture was stirred at 100°C under a CO atmosphere for 12 h. After removing the methanol by vortexing, 100 mL of water was added and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined and dried over anhydrous Na2SO4. The crude product was slurried in ethyl acetate (20 mL) to give methyl 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate (1.1 g, 53% yield).

[0439] LCMS (ESI) m / z: 364.2 [M+H] +

[0440] Step 5: Synthesis of methyl 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate.

[0441] Methyl 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate (300.0 mg, 0.78 mmol) was placed in a 25 mL single-necked flask. TFA (5.0 mL) was added and the reaction system was heated to 80°C and stirred at this temperature for 16 hours. After completion of the reaction, the reaction mixture was evaporated to dryness to obtain crude methyl 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate (190 mg).

[0442] LCMS (ESI) m / z: 244.3 [M+H] +

[0443] Step 6: Synthesis of 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid.

[0444] To the crude product from the previous step, methyl 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate (190 mg, 0.78 mmol), THF (5.0 ml) and methanol (5.0 ml) were added. The pH was adjusted to 7 with 3M aqueous potassium hydroxide solution. Lithium hydroxide (65.0 mg, 1.56 mmol) was then added and the reaction was stirred at 50°C for 16 hours. After the reaction was complete, the methanol and THF were removed by vortexing. The pH was adjusted to 6.5 with 1M dilute hydrochloric acid solution, filtered, and the filter cake was washed with water (5.0 ml x 3). The filter cake was dried and dehydrated to afford crude 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid (120 mg).

[0445] LCMS (ESI) m / z: 230.1 [M+H] +

[0446] Intermediate 94 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid

[0447]

[0448] Step 1: Synthesis of 6-bromo-N-(4-methoxybenzyl)-4-methylquinazolin-2-amine

[0449] At room temperature, the compound 6-bromo-2-chloro-4-methylquinazoline (0.5 g, 1.942 mmol) was dissolved in DMSO (15 ml), and DIEA (768 mg, 5.825 mmol) and 4-methoxybenzylamine (600 mg, 3.883 mmol) were added. The reaction was stirred at room temperature for 16 hours, and water (50 ml) was added. The mixture was extracted with ethyl acetate (20 ml×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The product was purified by column chromatography (0-10% methanol / dichloromethane) to give 6-bromo-N-(4-methoxybenzyl)-4-methylquinazoline-2-amine (500 mg, yield: 72%).

[0450] LCMS (ESI) m / z: 359.2 [M+H] +

[0451] Step 2: Synthesis of 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazolin-5-amine

[0452] At room temperature, compound 6-bromo-N-(4-methoxybenzyl)-4-methylquinazolin-2-amine (0.5 g, 1.396 mmol) was dissolved in DMSO (20 ml), and glycine (211 mg, 2.792 mmol), tert-butyl hydroperoxide (719 mg, 5.583 mmol), tetrabutylammonium iodide (104 mg, 0.2792 mmol) and acetic acid (251 mg, 4.1 87 mmol), under nitrogen protection, the reaction was stirred at 90 ° C for 16 hours, water (60 ml) was added, and the mixture was extracted with ethyl acetate (30 ml × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (0-10% methanol / dichloromethane) to give 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazolin-5-amine (300 mg, yield: 56%).

[0453] LCMS (ESI) m / z: 384.2 [M+H] +

[0454] Step 3: Synthesis of methyl 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylate

[0455] To a solution of 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazolin-5-amine (300 mg, 0.785 mmol) in MeOH (10 mL) and DMF (10 mL) was added potassium acetate (230 mg, 2.36 mmol) and Pd(dppf)Cl2 (58 mg, 0.0785 mmol) at room temperature. The mixture was stirred at 100°C under a CO atmosphere for 12 hours. After removing the methanol by vortexing, water (60 mL) was added and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification by column chromatography (0-10% methanol / dichloromethane) afforded methyl 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylate (200 mg, yield: 56%).

[0456] LCMS (ESI) m / z: 363.2 [M+H] +

[0457] Step 4: Synthesis of methyl 5-aminoimidazo[1,5-c]quinazoline-9-carboxylate

[0458] At room temperature, the compound 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylic acid methyl ester (200 mg, 0.5519 mmol) was placed in a 20 ml round-bottom flask, trifluoroacetic acid (2 ml) was added and the mixture was stirred at 78°C for 3 hours. After the trifluoroacetic acid was removed by vortexing, water (5 ml) was added and the mixture was filtered. The filter cake was washed with water to obtain 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid methyl ester (200 mg, crude product).

[0459] LCMS (ESI) m / z: 243.2 [M+H] +

[0460] Step 5: Synthesis of 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid

[0461] At room temperature, the compound 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylic acid methyl ester (200 mg, 0.8257 mmol) was dissolved in a mixed solvent of methanol / tetrahydrofuran / water (9 ml, 4:4:1), and lithium hydroxide (40 mg, 1.651 mmol) was added. The mixture was stirred at 78°C for 3 hours. After removing the trifluoroacetic acid by vortexing, water (5 ml) was added and the mixture was filtered. The filter cake was washed with water to obtain 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid (200 mg, crude product).

[0462] LCMS (ESI) m / z: 243.2 [M+H] +

[0463] Intermediate 95 Synthesis of (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazo[1,5-a]quinoxaline-8-carboxamide

[0464]

[0465] Step 1: Synthesis of 6-bromo-2-chloro-3-hydrazinoquinoxaline

[0466] A mixture of 6-bromo-2,3-dichloroquinoxaline (6.00 g, 21.73 mmol) and hydrazine hydrate (1.15 g, 35.98 mmol) in ethanol (50.00 ml) was stirred at 0°C for 1 hour and then at room temperature for 2 hours. The mixture was filtered and washed with ethanol (100 ml) to give 6-bromo-2-chloro-3-hydrazinoquinoxaline (5.00 g, 18.31 mmol).

[0467] LCMS(ESI):273[M+H] +

[0468] Step 2: Synthesis of 8-bromo-4-chlorotetrazo[1,5-a]quinoxaline

[0469] A mixture of 6-bromo-2-chloro-3-hydrazinylquinoxaline (5.00 g, 18.31 mmol) and sodium nitrite (2.52 g, 36.56 mmol) in aqueous hydrochloric acid (0.2 M, 50.00 ml) was stirred at room temperature for 2 hours. The mixture was extracted with ethyl acetate (100 ml x 3), and the organic phase was concentrated in vacuo and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 70%) to give 8-bromo-4-chlorotetrazo[1,5-a]quinoxaline (4.00 g, 18.28 mmol).

[0470] LCMS(ESI):284[M+H] +

[0471] Step 3: Synthesis of 8-bromo-N-(4-methoxybenzyl)tetrazo[1,5-a]quinoxaline-4-amine

[0472] A mixture of 8-bromo-4-chlorotetrazolo[1,5-a]quinoxaline (4.00 g, 14.13 mmol), p-methoxybenzylamine (3.87 g, 28.26 mmol), and N,N-diisopropylethylamine (7.30 g, 56.52 mmol) in dichloromethane (50 ml) was stirred at room temperature for 2 hours, extracted with ethyl acetate (100 ml×3), and the organic phase was concentrated in vacuo and purified by silica gel column chromatography (petroleum ether:ethyl acetate=30%) to give 8-bromo-N-(4-methoxybenzyl)tetrazo[1,5-a]quinoxalin-4-amine (3.00 g, 7.81 mmol).

[0473] LCMS(ESI):385[M+H] +

[0474] Step 4: Synthesis of methyl 4-((4-methoxybenzyl)amino)tetrazo[1,5-a]quinoxaline-8-carboxylate

[0475] 8-Bromo-N-(4-methoxybenzyl)tetrazo[1,5-a]quinoxaline-4-amine (3.00 g, 7.81 mmol), potassium acetate (1.53 g, 15.61 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (0.63 g, 0.77 mmol) were dissolved in a mixed solution of N,N-dimethylformamide and methanol (1:1, 30 ml), and the mixture was reacted in an autoclave under carbon monoxide (4 mPa) at 100°C overnight. The mixture was filtered and extracted with water (100 ml) and ethyl acetate (100 ml×3). The organic phase was concentrated in vacuo to give methyl 4-((4-methoxybenzyl)amino)tetrazo[1,5-a]quinoxaline-8-carboxylate (3.00 g, 8.24 mmol).

[0476] LCMS(ESI):365[M+H] +

[0477] Step 5: Synthesis of methyl 4-aminotetrazo[1,5-a]quinoxaline-8-carboxylate

[0478] 4-((4-methoxybenzyl)amino)tetrazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (3.00 g, 8.24 mmol) was dissolved in dioxane hydrochloride solution (4 M, 50 ml) and stirred at room temperature for 2 hours. The mixture was extracted with water (100 ml) and ethyl acetate (100 ml×3). The organic phase was concentrated in vacuo and purified by silica gel column chromatography (petroleum ether:ethyl acetate=50%) to give 4-aminotetrazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (2.00 g, 8.16 mmol).

[0479] LCMS(ESI):245[M+H] +

[0480] Step 6: Synthesis of 4-aminotetrazo[1,5-a]quinoxaline-8-carboxylic acid

[0481] 4-Aminotetrazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (2.00 g, 8.16 mmol) and potassium hydroxide (0.91 g, 16.32 mmol) were dissolved in tetrahydrofuran (10 ml), water (10 ml) and methanol (10 ml), stirred at 60° C. for 2 hours, extracted with water (50 ml) and ethyl acetate (50 ml×3), and the organic phase was concentrated in vacuo to give 4-aminotetrazo[1,5-a]quinoxaline-8-carboxylic acid (1.00 g, 4.34 mmol).

[0482] LCMS(ESI):231[M+H] +

[0483] Intermediate 96 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid

[0484]

[0485] Step 1: Synthesis of N-(2,6-dichloropyridin-3-yl)-1H-imidazole-5-carboxamide

[0486] 2,6-Dichloropyridin-3-amine (163 mg, 1.0 mmol) was dissolved in tetrahydrofuran (5.0 mL) and cooled in an ice bath for 5 minutes. NaHMDS (0.125 mL, 2.0 M, 2.5 mmol) was then added dropwise to the reaction system. After stirring for 1 hour, 5H,10H-diimidazole[1.5-a:1',5'-d]pyrazine-5,10-dione (188.0 mg, 1.0 mmol) was added and stirred at room temperature overnight. The reaction was monitored by LCMS. Upon completion, the reaction solution was poured into water and the pH was adjusted to 7. Solid precipitated, which was collected by filtration and dried to yield crude N-(2,6-dichloropyridin-3-yl)-1H-imidazole-5-carboxamide (175 mg, crude product).

[0487] LCMS (ESI) m / z: 257.0 [M+H] +

[0488] Step 2: Synthesis of 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazin-6(5H)-one

[0489] The compound N-(2,6-dichloropyridin-3-yl)-1H-imidazole-5-carboxamide (256.0 mg, 1.0 mmol) was dissolved in N,N-dimethylacetamide (2.0 ml), and potassium carbonate (276 mg, 2.0 mmol) was added. The reaction was heated to 140°C and stirred for 2 hours. When LCMS showed that the reaction was complete, it was cooled to room temperature and poured into water. The reaction solution was stirred for 30 minutes. Solid precipitated and was filtered and the solid was collected to give crude 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-6(5H)-one (153.0 mg, crude product).

[0490] LCMS (ESI) m / z: 221.1 [M+H] +

[0491] Step 3: Synthesis of methyl-6-hydroxyimidazole[1,5-a]pyridine[3,2-e]pyrazine-2-carboxylate

[0492] To a solution of 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazin-6(5H)-one (2.0 g, 9.09 mmol) in MeOH (30.00 mL) and DMF (310.00 mL) were added potassium acetate (1.78 g, 18.18 mmol) and Pd(dppf)Cl2 (660 mg, 0.91 mmol). The mixture was stirred at 100°C under a CO atmosphere for 12 h. After removing the methanol by vortexing, the reaction solution was added to 100 mL of water. Solids precipitated and were filtered to obtain a crude solid. The crude product was slurried with ethyl acetate (50 mL x 2) to obtain methyl 6-hydroxyimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (1.5 g, 68% yield) as a brown solid.

[0493] LCMS (ESI) m / z: 245.2 [M+H] +

[0494] Step 4: Synthesis of methyl 6-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate

[0495] Methyl 6-hydroxyimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (200.0 mg, 0.82 mmol) was added to phosphorus oxychloride (2.0 mL), and N,N-diisopropylethylamine (528 mg, 4.09 mmol) was added dropwise. The reaction was heated to 90°C for 2.5 hours. Once LCMS indicated completion of the reaction, the phosphorus oxychloride was removed by vortexing. The reaction mixture was then added to ice water and stirred for 15 minutes. The filter cake was filtered and dried to yield crude methyl 6-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (200.0 mg).

[0496] LCMS (ESI) m / z: 263.0 [M+H] +

[0497] Step 5: Synthesis of methyl 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate

[0498] Methyl 6-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (200.0 mg, 0.76 mmol) was dissolved in DMSO (3.0 mL), and p-methoxybenzylamine (125.0 mg, 0.92 mmol) and N,N-diisopropylethylamine (295 mg, 2.29 mmol) were added. The reaction was heated to 90°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature and poured into ice water with stirring. The mixture was extracted with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 0-5%) to provide methyl 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (250 mg, 91% yield).

[0499] LCMS (ESI) m / z: 364.2 [M+H] +

[0500] Step 6: Synthesis of methyl 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate

[0501] Methyl 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (250 mg, 0.69 mmol) was dissolved in TFA (5.0 mL) and heated to 80°C for 16 hours. The reaction was allowed to complete, cooled, and the TFA was removed by vortexing to yield crude methyl 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (160 mg, crude product).

[0502] LCMS (ESI) m / z: 244.1 [M+H] +

[0503] Step 7: Synthesis of 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid

[0504] Methyl 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate (160.0 mg, 0.66 mmol) was dissolved in a mixture of THF / MeOH (1.2 mL / 1.2 mL). Lithium hydroxide solution (0.66 mL, 2.0 M, 1.32 mmol) was added and the reaction was heated to 50°C for 16 hours. After the reaction was complete, the solvent was removed by vortexing and the pH was adjusted to approximately 7 with 1N aqueous HCl. The mixture was stirred for 15 minutes, filtered, and the solid was collected to yield 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid (83 mg, 55% yield).

[0505] LCMS (ESI) m / z: 230.1 [M+H] +

[0506] Intermediate 97 Synthesis of (S)-4-amino-N,6-dimethyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0507]

[0508] Step 1: Synthesis of N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboxamide

[0509] At room temperature, 4-bromo-2-fluoro-6-methylaniline (15 g, 73.89 mmol) was added to a 500 ml round-bottom flask, followed by the addition of tetrahydrofuran (150 ml). Sodium bis(trimethylsilyl)amide (74 ml) was added at 0°C. The resulting mixture was stirred at 0°C for half an hour before the addition of 5H,10H-diimidazole[1,5-a:1',5'-d]pyrazine-5,10-dione (6.95 g, 36.95 mmol). The mixture was allowed to react for an additional hour at room temperature. The reaction mixture was poured into ice water and filtered. The residue was the product, N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboxamide (10 g, 90.9% yield), as a red solid.

[0510] LCMS (ESI) m / z: 298 [M+H] +

[0511] Step 2: Synthesis of 8-bromo-6-methylimidazo[1,5-a]quinoxaline-4-ol

[0512] At room temperature, N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboxamide (10 g, 33.67 mmol) was added to a 250 ml round-bottom flask. N,N-dimethylacetamide (100 ml) was then added, followed by potassium carbonate (13.94 g, 101.01 mmol). The resulting mixture was stirred at 140°C for 2 hours. The reaction mixture was poured into water, the pH adjusted to 3-4, and filtered. The residue was the product, 8-bromo-6-methylimidazo[1,5-a]quinoxalin-4-ol (8 g, 85.74% yield), as a black solid.

[0513] LCMS (ESI) m / z: 278 [M+H] +

[0514] Step 3: Synthesis of 8-bromo-4-chloro-6-methylimidazo[1,5-a]quinoxaline

[0515] 8-Bromo-6-methylimidazo[1,5-a]quinoxaline-4-ol (8 g, 28.88 mmol) was added to a 250 ml round-bottom flask at room temperature. Phosphorus oxychloride (100 ml) was then added to the flask, and the resulting mixture was stirred and reacted at 120° C. for 12 hours. The reaction solution was concentrated to dryness, poured into ice water, and filtered. The filter residue was the product, yielding 8-bromo-4-chloro-6-methylimidazo[1,5-a]quinoxaline (7.5 g, 88.23% yield) as a black solid.

[0516] LCMS (ESI) m / z: 296 [M+H] +

[0517] Step 4: Synthesis of 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine

[0518] 8-Bromo-4-chloro-6-methylimidazo[1,5-a]quinoxaline (7.5 g, 25.42 mmol) was added to a 250 ml round-bottom flask at room temperature. Dimethyl sulfoxide (75 ml) was then added to the flask, followed by (4-methoxyphenyl)methanamine (4.18 g, 30.51 mmol) and N,N-diisopropylethylamine (6.56 g, 50.85 mmol). The resulting mixture was stirred at 90°C for 2 hours. The reaction solution was concentrated to dryness, poured into ice water, and filtered. The residue was the product, yielding 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine (6 g, 60% yield) as a red solid.

[0519] LCMS (ESI) m / z: 397 [M+H] +

[0520] Step 5: Synthesis of methyl 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylate

[0521] At room temperature, 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine (6 g, 15.15 mmol) was added to a 250 ml autoclave. N,N-dimethylformamide (30 ml) and methanol (60 ml) were then added to the autoclave. Potassium acetate (2.97 g, 30.3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.24 g, 1.52 mmol) were then added. Carbon monoxide (4 MPa) was then introduced, and the resulting mixture was stirred at 100°C for 12 hours. The reaction solution was concentrated to dryness, poured into water, and filtered. The residue was the product, methyl 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylate (4 g, 70.54% yield), as a red solid.

[0522] LCMS (ESI) m / z: 377 [M+H] +

[0523] Step 6: Synthesis of 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0524] At room temperature, methyl 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylate (4 g, 10.61 mmol) was added to a 250 ml round-bottom flask. Then, tetrahydrofuran (40 ml) and methanol (40 ml) were poured into the flask, and potassium hydroxide (1.19 g, 21.22 mmol) was added. The resulting mixture was stirred and reacted at 60° C. for 1 hour. The reaction solution was concentrated and spin-dried, poured into water, and the pH was adjusted to around 3 with formic acid. Solids precipitated and were filtered. The filter residue was the product, yielding 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (2.9 g, 76.31% yield) as a red solid.

[0525] LCMS (ESI) m / z: 363 [M+H] +

[0526] Step 7: Synthesis of 4-amino-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0527] 4-((4-Methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (2.9 g, 7.99 mmol) was added to a 50 ml round-bottom flask at room temperature. Trifluoroacetic acid (30 ml) was then poured into the flask, and the resulting mixture was stirred at 100°C for 3 hours. The reaction solution was concentrated and vortexed to obtain 4-amino-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (1.5 g, yield: 51.28%) as a black solid.

[0528] LCMS (ESI) m / z: 243 [M+H] +

[0529] Intermediate 98 Synthesis of 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0530]

[0531] Step 1: Synthesis of methyl 3-(2-cyclopropyl-1H-imidazol-1-yl)-4-nitrobenzoate

[0532] A mixture of methyl 3-(2-bromoimidazolyl)-4-nitrobenzoate (1.00 g, 3.07 mmol), cyclopropylboronic acid (0.26 g, 3.07 mmol), 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (0.25 g, 0.31 mmol) and potassium carbonate (0.85 g, 6.13 mmol) in dioxane (10 ml) was stirred at 100°C for 2 hours. After completion of the reaction, the reaction solution was poured into water (10 ml) and extracted with ethyl acetate (10 ml x 2). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether=10:90) to give methyl 3-(2-cyclopropyl-1H-imidazol-1-yl)-4-nitrobenzoate (0.50 g, yield: 57%).

[0533] LCMS(ESI):288[M+H] +

[0534] Step 2: Synthesis of methyl 4-amino-3-(2-cyclopropyl-1H-imidazol-1-yl)benzoate

[0535] A mixture of methyl 3-(2-cyclopropyl-1H-imidazol-1-yl)-4-nitrobenzoate (1.00 g, 3.48 mmol), biboric acid (0.94 g, 10.44 mmol), and 4,4-bipyridine (0.27 g, 1.74 mmol) in dimethylformamide (10.00 ml) was stirred at room temperature for 2 hours and then concentrated. The reaction solution was poured into water (10 ml) and extracted with ethyl acetate (10 ml x 2). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 53%) to obtain methyl 4-amino-3-(2-cyclopropyl-1H-imidazol-1-yl)benzoate (0.60 g, yield: 67%).

[0536] LCMS(ESI):257[M+H] +

[0537] Step 3: Synthesis of 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester

[0538] A mixture of methyl 4-amino-3-(2-cyclopropyl-1H-imidazol-1-yl)benzoate (500 mg, 1.94 mmol) and carbonylimidazole (377 mg, 2.33 mmol) in chlorobenzene (5 ml) was stirred at 140° C. for 12 hours and then concentrated. The reaction solution was poured into water (10 ml) and extracted with ethyl acetate (10 ml×2). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 20%) to obtain methyl 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (500 mg, yield: 91%).

[0539] LCMS(ESI):283[M+H] +

[0540] Step 4: Synthesis of methyl 4-chloro-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylate

[0541] A solution of 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (500.00 mg, 1.77 mmol) in phosphorus oxychloride (5.00 ml) was stirred at 120° C. for 12 hours and then concentrated to give 4-chloro-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (300 mg, crude product).

[0542] LCMS(ESI):301[M+H]+

[0543] Step 5: Synthesis of methyl 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylate

[0544] A mixture of methyl 4-chloro-1-cyclopropyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate (300 mg, 0.99 mmol), p-methoxybenzylamine (417 mg, 2.98 mmol), and diisopropylethylamine (384 mg, 2.98 mmol) in dimethyl sulfoxide (3 ml, 0.00 mmol) was stirred at 120°C for 2 hours, then concentrated and the reaction solution was poured into water (10 ml), extracted with ethyl acetate (10 ml x 2), and the organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 42%) to give methyl 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylate (300 mg, yield: 75%).

[0545] LCMS(ESI):402[M+H] +

[0546] Step 6: Synthesis of 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid

[0547] Lithium hydroxide (89.45 mg, 2.24 mmol) was added to a solution of a mixture of 1-cyclopropyl-4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (300 mg, 0.75 mmol) in methanol / tetrahydrofuran (5.00 ml / 5 ml), and the reaction solution was stirred at 60° C. for 2 hours and then concentrated to give 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid (160 mg, yield: 55%).

[0548] LCMS(ESI):388[M+H] +

[0549] Step 7: Synthesis of 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid

[0550] A mixture of 1-cyclopropyl-4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (200 mg, 0.51 mmol) in trifluoroacetic acid (2.00 ml) was heated at 100° C. for 2 hours and then concentrated to give 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid (100 mg, crude).

[0551] LCMS(ESI):268[M+H] +

[0552] Example 1

[0553] Synthesis of 4-amino-N-(2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0554]

[0555] 4-Aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, 0.21 mmol) was dissolved in N,N-dimethylformamide (2.50 ml) and 4-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (125 mg, 0.32 mmol), N,N-diisopropylethylamine (85 mg, 0.65 mmol) and N-methyl-2,3-dihydrobenzofuran-3-amine (33 mg, 0.21 mmol) were added. The mixture was stirred at room temperature for half an hour. After the reaction, the reaction solution was concentrated, and the crude product was purified by high-performance liquid chromatography: column: XBridge BEH Shield RP18 5m, 30 mm × 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 36% B over 8 minutes; wavelength: 254 nm / 220 nm; to obtain 4-amino-N-(2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (2 mg, 0.01 mmol, 3% yield) as a white solid.

[0556] 1 H NMR(400MHz,DMSO-d6)δ9.19(s,1H),8.33(s,1H),7.92(s,1H),7.48-7.41(m,5H),7.30-7.26(m ,1H),7.00-6.99(m,1H),6.91-6.89(M,1H),6.35-5.76(m,1H),4.79-4.56(m,2H),2.68(s,3H).

[0557] LCMS (ESI): 360.30 [M+H] +

[0558] The following examples can be prepared by using the same synthetic steps as described in Example 1, replacing only the corresponding starting materials:

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567]

[0568]

[0569]

[0570]

[0571] Example 58 (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazo[1,5-a]quinoxaline-8-carboxamide

[0572]

[0573] Step 1: Synthesis of (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazo[1,5-a]quinoxalin-4-yl)amino)methylene)-N-methylmethanamine

[0574] 4-Aminotetrazo[1,5-a]quinoxaline-8-carboxylic acid (131 mg, 0.56 mmol) was dissolved in N,N-dimethylformamide (4.5 ml). 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (325 mg, 0.85 mmol), N,N-diisopropylethylamine (220 mg, 1.71 mmol), and (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (144 mg, 0.660 mmol) were added to the solution. The mixture was reacted at room temperature for half an hour. The mixture was diluted with ethyl acetate (30 ml) and water (30 ml), and the aqueous layer was extracted with ethyl acetate (2 x 20 ml). The organic layers were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to give (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazo[1,5-a]quinoxalin-4-yl)amino)methylene)-N-methylmethanamine (100 mg, 0.19 mmol, 33.33% yield) as a white solid.

[0575] LCMS(ESI):528[M+H] +

[0576] Step 2: Synthesis of (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazo[1,5-a]quinoxaline-8-carboxamide

[0577] (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazo[1,5-a]quinoxalin-4-yl)amino)methylene)-N-methylmethanamine (100 mg, 0.19 mmol) was dissolved in tetrahydrofuran solution (4 ml), an equal volume of water (4 ml) was added, and then lithium hydroxide (27 mg, 1.12 mmol) was added. The reaction solution was concentrated and the crude product was purified by high performance liquid chromatography: Chromatography column specifications: YMC TriartC 18 ExRs 5m, 30mm*150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 25% B to 50% B in 10 minutes; wavelength: UV 254 nm / 220 nm; retention time (min): 8.43 / 9.23, to obtain the product (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazolo[1,5-a]quinoxaline-8-carboxamide (3.58 mg, 0.01 mmol, yield 4.41%) as a white solid.

[0578] 1 H NMR(400MHz,DMSO-d6)δ12.10(s,1H),8.01-7.95(d,2H),7.74-7.71(d,2H),7.4 1-7.39(d,1H),7.33(s,1H),6.50-5.74(m,1H),4.91-4.73(m,2H),3.94(s,3H).

[0579] LCMS (ESI) m / z: 430.10 [M+H] +

[0580] Example 59

[0581] Synthesis of 4-amino-N-cyclopropyl-N-(7-(5-(trifluoromethyl)pyridin-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridin-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0582]

[0583] N-cyclopropyl-7-(5-(trifluoromethyl)pyridin-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridin-4-amine (45 mg, 0.13 mmol) was dissolved in DMF (3.0 ml), and then 4-amino-7-fluoroimidazo[1,5-a]quinoxaline-8-carboxylic acid (33 mg, 0.13 mmol), PyBrop (79 mg, 0.17 mmol), and DIPEA (50.3 mg, 0.39 mmol) were added to the reaction solution. The reaction was stirred at room temperature overnight and water was added. (10 ml), extracted with ethyl acetate (20 ml), the organic phase was washed with water (10 ml × 2), washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, the filtrate was evaporated to dryness, and the crude product was separated and purified by column chromatography (methanol / dichloromethane 0-7%) to give compound 4-amino-N-cyclopropyl-N-(7-(5-(trifluoromethyl)pyridin-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridin-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (18.0 mg, yield: 25%).

[0584] 1H NMR (400MHz, DMSO-d6) δ9.31(s,1H),9.18(s,1H),9.02(s,1H),8.63(d,J=29.1Hz,2H),8.24(s,1H),7.93(s,1H),7.77( s,1H),7.56(s,2H),7.22(d,J=11.4Hz,1H),5.39(s,1H),4.46(d,J=53.9Hz,2H),3.05(s,1H),2.35(s,2H),0.70(s,4H).

[0585] LCMS (ESI) m / z: 564.30 [M+H] +

[0586] The following examples can be prepared by using the same synthetic steps as described in Example 59, replacing only the corresponding starting materials:

[0587]

[0588]

[0589]

[0590]

[0591] Example 75 Synthesis of (S)-4-amino-N-(methyl-d3)-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboxamide

[0592]

[0593] A mixture of (S)-N-(methyl-d3)-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (0.10 g, 0.45 mmol), 4-aminoimidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboxylic acid (0.10 g, 0.45 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (0.19 g, 0.68 mmol), and N-methylimidazole (0.11 g, 1.36 mmol) in N,N-dimethylformamide (1 mL) was stirred for 1 hour. After the reaction, the system was cooled to room temperature and extracted with ethyl acetate three times (10 mL each). The organic phases were combined, washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (HPLC) using the following column specifications: YMC Triart C18 ExRs5m, 30 mm*150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 25% B to 55% B in 8 minutes; wavelength: 254 nm / 220 nm; retention time (minimum): 7.22) to give (S)-4-amino-N-(methyl-d3)-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboxamide (11.94 mg, yield 6.09%).

[0594] 1 H NMR (400MHz, DMSO-d6) δ9.31-9.30(m,1H),8.70-8.65(m,1H),8.56-8.51(m,1H),7.99(d,J=4.0Hz, 1H),7.79–7.56(m,3H),7.36-7.34(m,1H),7.27-7.25(m,1H),6.44-6.02(m,1H),4.89-4.71(m,2H).

[0595] LCMS (ESI): 431.85 [M+H] +

[0596] The same method and steps as in Example 75 were used, except that the corresponding starting materials were replaced, to prepare the following compound:

[0597]

[0598] Example 77 Synthesis of 4-amino-N-(4S)-6-fluoro-1-methylisochroman-4-yl)-N-methylimidazo[1,5-a]quinoline-8-carboxamide

[0599]

[0600] At room temperature, (4S)-6-fluoro-N,1-dimethylisochromen-4-amine hydrochloride (100 mg, 0.5122 mmol) was dissolved in N,N-dimethylacetamide (4 ml), and 4-amino-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (140 mg, 0.6146 mmol), T3P (489 mg, 0.7683 mmol) and N,N-diisopropylethylamine (338 mg, 3.561 mmol) were added. After stirring at 25°C for 2 hours, the reaction solution was poured into water (40 ml) and extracted with ethyl acetate (20 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (dichloromethane:methanol=10:1) to give the product 4-amino-N-(4S)-6-fluoro-1-methylisochroman-4-yl)-N-methylimidazo[1,5-a]quinoline-8-carboxamide (70 mg, yield: 33.7%).

[0601] 1 H NMR(400MHz, DMSO-d6)δ9.27–9.11(m,1H),8.38(d,J=17.2Hz,1H),7.92(s,1H),7.60–6.98(m,7H), 5.62(s,1H),4.94–4.67(m,1H),4.38–3.79(m,2H),2.87–2.66(m,3H),1.49(dd,J=13.0,6.2Hz,3H).

[0602] 19 F NMR(376MHz,DMSO-d6)δ-115.37,-115.39.

[0603] LCMS (ESI) m / z: 406 [M+H] +

[0604] Example 78 (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide Example 111 (S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0605]

[0606] Step 1: 4-Aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (500 mg, 2.19 mmol) was dissolved in tetrahydrofuran (15 mL). The reaction mixture was cooled to -40°C, and n-butyllithium (5 mL, 15.34 mmol) was added and allowed to react for 10 minutes. Bromine (2.5 mL, 21.91 mmol) was then added. The mixture was stirred at -40°C for 10 minutes, then returned to room temperature and reacted for another 10 minutes. After the reaction, the reaction mixture was concentrated in vacuo, added with dichloromethane, and filtered to obtain a solid. The solid was then purified by silica gel column chromatography (formic acid:acetonitrile = 1:1) to obtain 4-amino-1-bromoimidazo[1,5-a]quinoxaline-8-carboxylic acid (30 mg, 4.46% yield) as a white solid.

[0607] LCMS (ESI) m / z: 307 [M+H] +

[0608] Step 2: 4-Amino-1-bromoimidazo[1,5-a]quinoxaline-8-carboxylic acid (10 mg, 0.03 mmol), 2-(7-azobenzotriazole)-tetramethyluronium hexafluorophosphate (19 mg, 0.05 mmol), and N,N-diisopropylethylamine (8 mg, 0.05 mmol) were dissolved in dimethylacetamide (1 ml), and (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (7 mg, 0.03 mmol) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction, the mixture was extracted with ethyl acetate (10 ml x 3), the organic phases were combined, washed with saturated brine (10 ml x 3) and dried over anhydrous sodium sulfate, and the combined organic phases were concentrated in vacuo to give Example 111 (S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (30 mg, 0.03 mmol, yield 80%) as a yellow liquid.

[0609] 1 H NMR(400MHz,DMSO-d6)δ9.09(s,1H),7.98(s,1H),7.65–7.55(m,2H),7.52(d,J=7.9Hz,3H ),7.32(d,J=7.7Hz,1H),7.25(s,1H),6.42(s,1H),4.79–4.72(m,2H),2.75–2.62(m,3H).

[0610] LCMS (ESI) m / z: 506 [M+H] +

[0611] Step 3: (S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (20 mg, 0.16 mmol), zinc powder (26 mg, 1.58 mmol), and deuterated formic acid (19 mg, 1.58 mmol) were dissolved in deuterated methanol (1 ml) and heavy water (1 ml). The mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was extracted with ethyl acetate (10 ml × 3), the organic phases were combined, washed with saturated brine (10 ml × 3), and dried over anhydrous sodium sulfate. The combined organic phases were concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography: Chromatography column specifications: Kinetex 5m EVO C 18 The reaction mixture was prepared by filtration using a 30 mm × 150 mm column; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B over 10 min; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide-1-d (3.34 mg, 0.16 mmol, 8.45% yield) was obtained as a white solid.

[0612] 1 H NMR (400MHz, DMSO-d6) δ8.34(s,1H),7.92(d,J=7.3Hz,1H),7.64-7.62(d,J=7.6Hz,1H),7 .49(s,4H),7.33-7.32(m,1H),7.26(s,1H),6.48(m,1H),4.73-4.72(m,2H),2.68(s,3H).

[0613] LCMS (ESI) m / z: 428 [M+H] +

[0614] The following examples can be prepared by using the same synthetic procedures as described in Example 78, replacing only the corresponding starting materials:

[0615]

[0616] Example 109 Synthesis of 4-amino-N-cyclopropyl-N-(4-(trifluoromethoxy)benzyl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0617]

[0618] 4-Amino-3-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (30 mg, 0.06 mmol), potassium acetate (17 mg, 0.18 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (4 mg, 0.01 mmol) were added to a mixed solvent of N,N-dimethylformamide (5 ml) and ethanol (5 ml), and the autoclave was filled with carbon monoxide (4 mPa) and stirred at 110°C overnight. After the reaction was completed, the mixture was diluted with ethyl acetate (10 ml) and water (5 ml), and the aqueous layer was extracted with ethyl acetate (2×10 ml). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by high pressure preparative chromatography (Column: Sunfire C 18 The product was purified by centrifugation at 4 °C (5 m, 30 mm × 150 mm; Mobile Phase A: Water (0.1% FA), Mobile Phase B: ACN; Flow rate: 60 ml / min mL / min; Gradient: 37% B to 64% B in 10 min; Wave Length: 254 nm / 220 nm nm; RT1 (min): 5.97) to give ethyl 4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylate (4.22 mg, 14%).

[0619] 1 H NMR(400MHz, DMSO-d6)δ9.36(s,1H),9.30-8.61(m,1H),8.46(s,1H),8.10-7.92(m,1H),7.70-7.64(m,1H),7.61–7.50(m,2H),7. 34-7.30(m,1H),7.26(s,1H),6.38-5.76(m,1H),4.84-4.70(m,2H),4.32-4.37(m,2H),2.68-2.66(m,3H),1.37(t,J=7.1Hz,3H).

[0620] LCMS (ESI): 500.10 [M+H] +

[0621] Example 80: Synthesis of (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0622]

[0623] Step 1: Synthesis of (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine

[0624] Tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (200 mg, 0.6 mmol) was dissolved in 1,4-dioxane (4 M hydrochloric acid, 2 mL) and stirred at room temperature. After completion of the reaction, the reaction solution was concentrated to obtain the crude product (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine (85 mg, 61% yield).

[0625] LCMS (ESI) m / z: 228 [M+H] +

[0626] Step 2: Synthesis of (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0627] Dissolve 4-amino-7-fluoroimidazo[1,5-a]quinoxaline-8-carboxylic acid (92 mg, 0.37 mmol) in dimethylacetamide (2 ml). Add 1-propylphosphonic acid cyclic anhydride (172 mg, 0.74 mmol), N,N-diisopropylethylamine (144 mg, 1.12 mmol), and (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine (85 mg, 0.37 mmol). Stir overnight at room temperature. After completion of the reaction, extract with ethyl acetate (30 ml) three times, 10 ml each time. Combine the organic phases, wash with saturated brine, and dry over anhydrous sodium sulfate. Concentrate the combined organic phases under reduced pressure. Column chromatography (dichloromethane:methanol=10%) gave (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (35 mg, 20% yield).

[0628] LCMS (ESI) m / z: 456 [M+H] +

[0629] Step 3: Synthesis of (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0630] (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (35 mg, 0.08 mmol) was dissolved in dioxane (1 ml), and methanesulfonic acid-2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl (2-amino-1,1'-biphenyl-2-yl (33 mg, 0.07 mmol), sodium tert-butoxide (11 mg, 0.12 mmol), methanesulfonic acid-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl (33 mg, 0.07 mmol), sodium tert-butoxide (11 mg, 0.12 mmol), methanesulfonic acid- 2-(Di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl (2-amino-1,1'-biphenyl-2-yl) palladium (65 mg, 0.07 mmol) and methanol (86 mg, 2.68 mmol) were stirred at room temperature for 10 hours under nitrogen protection. After the reaction was completed, it was extracted with ethyl acetate (30 ml) three times, 10 ml each time. The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. High pressure preparation (Column: XBridge BEH Shield RP18 5m,30mm×150mm;Mobile Phase A:Water (10mmol / L NH4HCO3),Mobile Phase B:ACN;Flow rate:60ml / min mL / min;Gradient:28%B to 50%B in 8min;Wave Length:254nm / 220nm nm;RT1(min):7.88) to obtain (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (14.25 mg, 44.60% yield).

[0631] 1 H NMR(400MHz, DMSO-d6)δ9.12(s,1H),8.46-8.29(m,1H),7.92(d,J=3.6Hz,1H),7.59(s,2H),7.28–7.20(m, 2H), 6.58–6.31(m,2H),,5.42–4.79(m,1H),4.74–4.53(m,2H),3.74(d,J=10.0Hz,3H),2.70–2.58(m,3H).

[0632] LCMS (ESI) m / z: 408.10 [M+H] +

[0633] Example 122 Synthesis of (S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carboxamido)-2,3-dihydrobenzofuran-6-carboxylic acid methyl ester

[0634]

[0635] Step 1: Synthesis of methyl (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate

[0636] At room temperature, tert-butyl (S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (100 mg, 0.31 mmol) was dissolved in methanol (5.00 ml) and N,N-dimethylformamide (10.00 ml). [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (56 mg, 0.07 mmol) and potassium acetate (68 mg, 0.69 mmol) were added. The mixture was stirred overnight at 100°C under an atmosphere of carbon monoxide (4 MPa). After completion of the reaction, the reaction solution was concentrated, poured into water, and filtered. The filter cake was slurried with petroleum ether to yield methyl (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate (70 mg, yield: 77.78%) as a red solid. LCMS (ESI): 308 [M+H]. +

[0637] Step 2: Synthesis of methyl (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxylate

[0638] (S)-methyl 3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate (70 mg, 0.23 mmol) was dissolved in a 1,4-dioxane solution of hydrogen chloride (2 mL, 4 M), and the mixture was reacted at room temperature for half an hour. After completion of the reaction, the reaction mixture was concentrated to provide methyl (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxylate (46 mg, 97% yield) as a black solid.

[0639] LCMS(ESI):208[M+H] +

[0640] Step 3: Synthesis of (S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carboxamido)-2,3-dihydrobenzofuran-6-carboxylic acid methyl ester

[0641] 4-Aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, 0.22 mmol) was dissolved in N,N-dimethylformamide (4.00 ml), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (128 mg, 0.34 mmol), N,N-diisopropylethylamine (87 mg, 0.67 mmol), and (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxylic acid methyl ester (46 mg, 0.22 mmol) were added. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, the mixture was diluted with ethyl acetate (10 ml) and water (5 ml), and the aqueous layer was extracted with ethyl acetate (2 x 10 ml). The organic layers were combined, washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by high performance liquid chromatography: column Sunfire C18 5 m, 30 mm × 150 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 5% B to 30% B in 8 minutes; wavelength: 254 nm / 221 nm; to give (S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carboxamido)-2,3-dihydrobenzofuran-6-carboxylic acid methyl ester (21 mg, yield: 1%) as a white solid.

[0642] 1 H NMR(400MHz,DMSO-d6)δ9.21(s,1H),8.35(s,1H),8.13(s,1H),8.01-7.92(s,1H),7.62-7.60(m,1H),7.56-7.54(m,1H),7.50- 7.49(m,2H),7.40-7.36(m,1H),6.39-5.66(s,1H),4.87-4.68(m,2H),3.85-3.84(m,3H),2.67-2.64(m,3H),0.98-0.79(m,3H)

[0643] LCMS (ESI): 418.20 [M+H] +

[0644] Example 112 Synthesis of (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0645]

[0646] Step 1: Synthesis of (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine

[0647] (S)-tert-Butyl (6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (100 mg, 0.30 mmol) was dissolved in a 4.0 M solution of hydrogen chloride in 1,4-dioxane (10 ml). The mixture was stirred at room temperature for 2 hours. After the reaction, the reaction solution was concentrated in vacuo to afford (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine (80 mg, 0.30 mmol, 95.46% yield) as a yellow liquid. LCMS (ESI): 228 [M+H] +

[0648] Step 2: Synthesis of (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0649] 4-Aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (80 mg, 0.35 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (200 mg, 0.53 mmol), and N,N-diisopropylethylamine (136 mg, 1.05 mmol) were dissolved in dimethylacetamide (3 ml), and (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine (80 mg, 0.35 mmol) was added. The mixture was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was extracted with ethyl acetate three times, each time with 10 ml. The organic phases were combined, washed with saturated brine (10 ml x 3) and dried over anhydrous sodium sulfate. The reaction solution was spin-dried, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 80:20) to give (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (70 mg, 0.35 mmol, yield 45.54%).

[0650] LCMS(ESI):438[M+H] +

[0651] Step 3: Synthesis of (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide

[0652] N-((3S)-6-bromo(2,3-dihydrobenzo[b]furan-3-yl))(4-amino(10-hydroimidazo[1,5-a]quinoxalin-8-yl))-N-methylformamide (30 mg, 0.07 mmol) was dissolved in tetrahydrofuran (2 ml), and [Ir(dF(CF3)ppy)2(dpy)]PF6 (8 mg, 0.007 mmol), nickel bromide glycol dimethyl ether complex (1 mg, 0.003 mmol), PPh3(CF2H)2 (17 mg, 0.14 mmol), and diphenylphenanthroline (2 mg, 0.007 mmol) were added in sequence. The mixture was reacted at room temperature at a wavelength of 465 nm overnight. After completion of the reaction, the mixture was extracted three times with 10 ml of ethyl acetate each time. The combined organic phases were washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by high-performance liquid chromatography: column specifications: Kinetex 5m EVO C18, 30 mm x 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboxamide (0.84 mg, 0.07 mmol, 2.96% yield) was obtained as a white solid.

[0653] 1 H NMR(400MHz,Methanol-d4)δ9.11(s,1H),8.29(s,1H),7.98(s,1H),7.57-7.54(m,3H ),7.17-7.16(s,1H),7.03(s,1H),4.73-4.62(s,2H),4.60-4.52(m,1H),2.78(s,3H).

[0654] LCMS(ESI):409[M+H] +

[0655] Example 115 Synthesis of (R)-4-amino-7-fluoro-N-methyl-N-(7-(1-methylpyrazol-5-yl)benzopyran-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0656]

[0657] At room temperature, the compound 4-amino-7-fluoroimidazo[1,5-a]quinoxaline-8-carboxylic acid (200 mg, 0.813 mmol) was dissolved in DMAc (3 ml), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (187 mg, 0.976 mmol) and 1-hydroxybenzotriazole (133 mg, 0.976 mmol) were added, followed by the addition of DIEA (421 mg, 3.264 mmol), and the reaction was stirred at room temperature for 0.5 hour. (R)-N-methyl-7-(1-methylpyrazol-5-yl)chromen-4-amine (1.0 g, 4.184 mmol) was then added, and the mixture was reacted at room temperature for 16 hours. When LCMS showed the reaction was complete, water (30 mL) was poured into the mixture, and the mixture was extracted with ethyl acetate (10×3 mL). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was separated and purified by column chromatography (0-5% methanol / dichloromethane) to give (R)-4-amino-7-fluoro-N-methyl-N-(7-(1-methylpyrazol-5-yl)chromen-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (45.0 mg, yield: 11.75%).

[0658] 1 H NMR (400MHz, DMSO-d6) δ9.17(d,J=22.9Hz,1H),8.42(d,J=6.6Hz,1H),7.94(d,J=8.6Hz,1H),7.61(d ,J=12.8Hz,2H),7.46(dd,J=6.2,1.9Hz,1H),7.27(td,J=11.3,7.6Hz,2H),7.14(dd,J=7.9,1.8Hz,1 H),6.98(dd,J=27.5,1.8Hz,1H),6.41(dd,J=9.7,1.9Hz,1H),6.10-5.04(m,1H),4.49-4.05(m,2H), 3.86(d,J=15.6Hz,3H),2.82-2.62(m,3H),2.32(dd,J=17.9,7.3Hz,1H),2.13(dd,J=7.1,3.6Hz,1H).

[0659] LCMS (ESI) m / z: 472.1 [M+H] +

[0660] The following examples can be prepared by using the same synthetic steps as described in Example 115, replacing only the corresponding starting materials:

[0661]

[0662]

[0663] Example 128 Synthesis of (S)-4-amino-3-(hydroxymethyl)-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0664]

[0665] Step 1: (S)-4-Amino-3-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (60 mg, 0.12 mmol) was dissolved in dimethylformamide (2 mL) and methanol (2 mL). Dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (3 mL, 0.004 mmol) and potassium acetate (23 mg, 0.24 mmol) were added. The mixture was placed in an autoclave and stirred at 100°C under 4 MPa of carbon monoxide for 16 hours. After completion of the reaction, the reaction mixture was poured into water and extracted with ethyl acetate three times (10 mL each time). The combined organic phases were washed with saturated brine (10 ml × 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo and purified by silica gel column chromatography (alkaline water:acetonitrile = 1:1) to give (S)-4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylic acid methyl ester (40 mg, yield: 69.53%) as a brown solid.

[0666] LCMS (ESI): 485.42 [M+H] +

[0667] Step 2: Dissolve (S)-methyl 4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylate (30 mg, 0.06 mmol) in methanol (2 mL) and add sodium borohydride (5 mg, 0.12 mmol). Stir the mixture at room temperature for 2 hours. After the reaction, extract with ethyl acetate three times (10 mL each). Combine the organic phases, wash with saturated brine (10 mL x 3), and dry over anhydrous sodium sulfate. Concentrate the combined organic phases in vacuo. The crude product was purified by high-performance liquid chromatography using a Kinetex 5m EVOC18 column (30 mm x 150 mm); mobile phase A: water (10 mmol / L ammonium bicarbonate); mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (S)-4-amino-3-(hydroxymethyl)-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (3.17 mg, 11.13% yield) was obtained as a white solid.

[0668] 1 H NMR (400MHz, DMSO-d6) δ9.11(s,1H),8.31(s,1H),7.64(d,J=7.8Hz,1H),7.59(s,2H),7.47(d,J=5.5Hz,2H),7.34(d,J=7. 8Hz,1H),7.27–7.23(m,1H),6.36(t,J=5.2Hz,1H),4.85(d,J=5.1Hz,3H),4.73(d,J=4.6Hz,1H),3.31(s,1H),2.67(S,3H).

[0669] LCMS (ESI): 457.41 [M+H] +

[0670] Example 132 Synthesis of 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}-4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboxamide

[0671]

[0672] A stirred solution of methyl 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}-4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboxylate (30 mg, 0.06 mmol) in methanol (0.50 ml) / tetrahydrofuran (0.50 ml) / aqueous ammonia (0.5 ml) was stirred at 60° C. for 2 hours. After the reaction was complete, the reaction solution was poured into water (5 ml), and the mixture was taken out with ethyl acetate and then dried to obtain a crude product, which was purified by high pressure preparative chromatography (column, C18 silica gel; mobile phase, water in acetonitrile, gradient from 10% to 50% in 10 minutes; detector, UV 254 nm) to give 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}-4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboxamide (1.06 mg, 4%) as a white solid.

[0673] 1 H NMR (400MHz, DMSO-d6) δ10.01(s,1H),9.32(s,1H),8.42(s,1H),8.21(s,1H),7.92(s,1H),7.78(s,1H),7.64(d,J=7.9Hz,1H), 7.54(d,J=8.2Hz,1H),7.48(d,J=11.1Hz,1H),7.34(d,J=7.8Hz,1H),7.27(s,1H),6.43(s,1H),4.73(s,2H),2.76–2.63(m,3H).

[0674] LCMS (ESI): 470.80 [M+H] +

[0675] Example 131 Synthesis of (S)-4-amino-3-(hydroxymethyl)-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboxamide

[0676]

[0677] To a stirred solution of 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}-4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboxamide (30.00 mg, 0.06 mmol) in dichloromethane (2.00 ml) were added pyridine (10.09 mg, 0.13 mmol) and trifluoroacetic anhydride (26.79 mg, 0.26 mmol) and the mixture was reacted at room temperature for 12 hours. After completion of the reaction, the reaction solution was poured into water (5 ml), and the mixture was taken with ethyl acetate and dried to obtain a crude product, which was then purified by high pressure preparative chromatography (column, C 18 Silica gel; mobile phase, water in acetonitrile, gradient 10% to 50% over 10 minutes; detector, UV 254 nm) to afford N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)](4-amino-3-cyano(10-hydroimidazo[1,5-a]quinoxalin-8-yl))-N-methylformamide (1.61 mg, 5%) as a white solid.

[0678] 1 H NMR (400MHz, DMSO-d6) δ9.46 (s, 1H), 8.48 (s, 1H), 7.64 (d, J = 8.7Hz, 3H), 7.33 (s, 1H), 7.26 ( s,1H),7.12(s,2H),6.40(s,1H),4.72(d,J=10.5Hz,1H),4.71(s,1H),2.67(d,J=2.8Hz,3H).

[0679] LCMS (ESI): 452.80 [M+H] +

[0680] Example 81 & 82 :

[0681] Example 81 (R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide & Example 82 (S)-Synthesis of 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide

[0682]

[0683] Compound 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide (90 mg) was subjected to SFC (column: DAICEL CHIRALCEL OJ (250mm×30mm,10um, conditions: CO2-i-PrOH (0.1% NH3·H2O), 50% / 50%, flow rate: 80ml / min) to give (R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide (34.61 mg) and (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide (28.15 mg).

[0684] Example 81 (R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide

[0685] LCMS (ESI) m / z: 465.3 [M+H] +

[0686] 1 H NMR(400MHz, DMSO-d6)δ9.18(s,1H),8.37(d,J=1.8Hz,1H),7.91(s,1H),7.59(dd,J=8.4,1.8Hz,1H),7.55–7.39(m,4H),7.18–7.09(m,2H),5 .86(dd,J=9.3,4.1Hz,1H),4.81(t,J=9.7Hz,1H),4.65(dd,J=10.2,4.2Hz,1H),2.90(p,J=3.2Hz,1H),0.45–0.26(m,2H),0.24–0.03(m,2H).

[0687] Example 82 (S)-4-Amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxamide

[0688] LCMS (ESI) m / z: 465.3 [M+H] +

[0689] 1H NMR(400MHz,DMSO-d6)δ9.24(d,J=3.6Hz,1H),8.40(d,J=3.5Hz,1H),8.03(s,1H),7.8 5(s,2H),7.62(dd,J=8.4,3.3Hz,1H),7.47(dd,J=8.2,3.3Hz,2H),7.13(dd,J=8.7,3.3 Hz,2H),5.86(dt,J=8.6,4.0Hz,1H),4.80(td,J=9.7,3.6Hz,1H),4.65(dt,J=9.2,4.1 Hz,1H),2.90(d,J=6.7Hz,1H),0.44–0.26(m,2H),0.13(ddd,J=45.3,10.6,4.8Hz,2H).

[0690] Using the procedures described in Example 81 and Example 82 and the corresponding chiral SFC separation conditions, the compounds shown in the following table were obtained:

[0691]

[0692]

[0693]

[0694]

[0695]

[0696] Biochemical evaluation

[0697] 1. Experimental study on the inhibitory activity of compounds on tumor cell proliferation

[0698] Test Example 1: Inhibitory activity of compounds on HCT-116 MTAP(- / -) deficient cell proliferation

[0699] Materials and cells: HCT-116 MTAP(- / -)-deficient cells were purchased from Kangyuan Bochuang (China); RPMI-1640 medium, fetal bovine serum, and penicillin-streptomycin were purchased from Thermo Fisher Scientific (USA); 384-well plates were purchased from PerkinElmer (USA); and the Cell-Titer Glo kit was purchased from Promega (USA).

[0700] Cell culture: HCT116 MTAP(- / -)-deficient cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C and 5% CO2. Cells in the logarithmic growth phase were used for experiments.

[0701] Cell proliferation inhibitory activity assay: The Cell-Titer Glo kit was used to detect the inhibitory activity of compounds against HCT-116 MTAP(- / -)-deficient cells. The cell concentration was adjusted, and 40 μL was inoculated into each well of a 384-well plate. The plate was incubated overnight at 37°C and 5% CO2. 40 nL of compound was added to each well to a final concentration of 0-10,000 nM (starting concentration 10,000 nM, 3-fold dilution, 10 points), with a DMSO content of 0.1%. The cell plates were incubated at 37°C and 5% CO2 for 8 days. 40 μL of Cell-Titer Glo reagent was added to detect cell viability. The test results are shown in Table 1.

[0702] Test Example 2: Experimental study on the inhibitory activity of compounds on HCT-116 wild-type cell proliferation

[0703] Materials and cells: HCT-116 wild-type cells were purchased from Kangyuan Bochuang (China); RPMI-1640 medium, fetal bovine serum, and penicillin-streptomycin were purchased from Thermo Fisher Scientific (USA); 384-well plates were purchased from PerkinElmer (USA); and the Cell-TiterGlo kit was purchased from Promega (USA).

[0704] Cell culture: HCT-116 wild-type cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C and 5% CO2. Cells in the logarithmic growth phase were used in experiments.

[0705] Cell proliferation activity assay: The Cell-Titer Glo kit was used to detect the inhibitory activity of the compound on HCT-116 wild-type cells. The cell concentration was adjusted, and 40 μL was inoculated into each well of a 384-well plate and cultured overnight at 37°C and 5% CO2. 40 nL of compound was added to each well to a final concentration of 0-10,000 nM (starting concentration 10,000 nM, 2-fold dilution, 10 points) with a DMSO content of 0.1%. The cell plates were incubated at 37°C and 5% CO2 for 8 days. 40 μL of Cell-Titer Glo reagent was added to detect cell activity. The test results are shown in Table 1.

[0706] Table 1 shows the inhibitory activity of the compounds in the examples on the proliferation of HCT116 MTAP(- / -) deficient cells and HCT116 wild-type cells.

[0707] Table 1

[0708]

[0709]

[0710]

[0711]

[0712]

[0713] 2. Pharmacokinetic Experiments in Mice

[0714] 1. Test compounds

[0715] The compounds used in this test are from the specific examples of the present invention, and the reference compound AMG473 is the compound of Example 473 of Amgen's patent WO 2022 / 169948A1.

[0716] 2. Experimental Animals

[0717] ICR male mice, N=3 / group; original source: Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd.

[0718] 3. Drug preparation and administration

[0719] Single oral (PO) administration in ICR mice: Weigh the compound and dissolve it in dimethyl sulfoxide. Add a certain volume of polyethylene glycol 400 and water for injection, and adjust the solution to a clear solution with a small amount of 1 mol / L hydrochloric acid solution. Administer the compound to three mice by gavage after an overnight fast at a dose of 10 mg / kg.

[0720] Single intravenous (IV) administration in ICR mice: Weigh the compound and dissolve it in dimethyl sulfoxide. Add a certain volume of polyethylene glycol 400 and water for injection, and adjust the solution to a clear solution with a small amount of 1 mol / L hydrochloric acid solution. Administer 3 mg / kg of the compound to the tail vein of 3 mice after an overnight fast.

[0721] 4. Sample Collection

[0722] Approximately 30 μL of blood was collected from the dorsal foot vein at each time point. Anticoagulated with dipotassium EDTA, placed on ice, and centrifuged within 1 hour to separate plasma (centrifugation conditions: 4000 g / min, 5 minutes, 4°C). Blood was collected at 0.0833 (intravenous injection), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. Samples were stored at -20°C.

[0723] To 30 μL of plasma sample (10 μL sample + 20 μL blank plasma sample), add 200 μL of ice-cold acetonitrile containing internal standard, vortex for 30 seconds, and centrifuge at 4000 g / min for 20 minutes. Transfer 100 μL of the supernatant to a 96-well plate, add 200 μL of ultrapure water, vortex for 30 seconds, and inject 5 μL or 10 μL into LC-MS / MS for analysis.

[0724] Table 2: Pharmacokinetic data

[0725]

[0726] "NA": does not exist.

[0727] By comparing the pharmacokinetic parameters, it can be found that compared with AMG473, at the same dose and the same administration method, the embodiments of the present invention have higher plasma exposure, which can reduce the effective dose of the compound and increase the safety window.

[0728] 3. hERG ion channel inhibition experiment

[0729] 1. Test compounds

[0730] The compounds used in this test are from the specific examples of the present invention, and the reference compound AMG473 is the compound of Example 473 of Amgen's patent WO 2022 / 169948A1.

[0731] 2. Cell Lines and Cell Culture

[0732] HEK293 cells stably expressing the hERG ion channel (Cat. No. K1236) were purchased from Invitrogen. The cell line was cultured in a medium containing 85% DMEM, 10% dialyzed fetal bovine serum, 0.1 mM non-essential amino acid solution, 100 U / mL penicillin-streptomycin solution, 25 mM HEPES, 5 μg / mL blasticidin, and 400 μg / mL geneticin. When the cell density reached 40% to 80% of the culture dish bottom area, the cells were trypsinized and passaged three times per week. Prior to the experiment, cells were cultured at a density of 5 × 10⁵ in 3.5 cm culture dishes and induced with 1 μg / mL doxycycline for 48 hours. The cells were then digested and plated on glass slides for subsequent manual patch clamp experiments.

[0733] 3. Experimental Procedure

[0734] 1) Place a small glass slide containing HEK293 cells in a culture dish into the perfusion chamber on the microscope stage. 2) Under an Olympus IX71 or IX73 inverted microscope, center the field of view of an appropriate cell. Using a ×10x objective, locate the tip of the glass electrode and position it in the center of the field of view. Then, use the micromanipulator to lower the electrode while adjusting the coarse focus knob to slowly approach the cell. 3) Once close to the cell, switch to a ×40x objective for observation and use the micromanipulator to fine-tune the electrode to gradually approach the cell surface. 4) Apply negative pressure to create a seal with a resistance greater than 1 GΩ between the electrode tip and the cell membrane. 5) Compensate for the transient capacitive current, Cfast, in voltage-clamp mode. Then, repeatedly apply brief negative pressure to rupture the membrane, ultimately establishing whole-cell recording mode. 6) With the membrane potential clamped at -60 mV, compensate for the slow capacitive current, Cslow, the cell membrane capacitance (Cm), and the input membrane resistance (Ra). 7) After cells stabilize, change the clamping voltage to -90 mV, set the sampling frequency to 20 kHz, and the filtering frequency to 10 kHz. Leakage current is measured with the clamping voltage set to -80 mV and a time duration of 500 ms. 8) hERG current measurement is as follows: apply a 4.8-second depolarizing command voltage to depolarize the membrane potential from -80 mV to +30 mV, followed by a 5.2-second repolarizing voltage to reduce the membrane potential to -50 mV to eliminate channel inactivation, allowing observation of the hERG tail current. The peak value of the tail current is the magnitude of the hERG current. 9) hERG currents used for testing test compounds are recorded continuously for 120 seconds prior to administration to assess the stability of hERG current generation in the test cells. Only stable cells within the acceptable range of the evaluation criteria are used for subsequent compound testing. 10) Testing for the inhibitory effect of test compounds on hERG current: Initially, the hERG current measured in extracellular fluid containing 0.1% DMSO is used as the baseline for the assay. After the hERG current remains stable for at least 5 minutes, the solution containing the compound to be tested is perfused around the cells in sequence from low concentration to high concentration. After each perfusion, wait for about 5 minutes to allow the compound to fully act on the cells and record the hERG current synchronously. After the recorded current stabilizes, record the last 5 hERG current values, and take the average value as the final current value at a specific concentration. After testing the compound, add 450nM dofetilide to the same cell to completely inhibit its current as a positive control for the cell. At the same time, the positive compound dofetilide is synchronously detected using the same patch clamp system before and after the end of the test drug experiment to ensure the reliability and sensitivity of the entire detection system. The above test steps will be repeated on two separate test cells (n=2).

[0735] 4. Data Analysis

[0736] Only data meeting the above criteria can be analyzed according to the following steps: Note: Data is output from PatchMaster software. 1) After perfusing the blank solvent or compound gradient solution, obtain the average of five consecutive stable current values ​​and use them as the "Tail Current Size Blank" and "Tail Current Size Compound" values, respectively. The percentage of current inhibition is calculated using the following formula.

[0737] 2) Dose-effect curves were fitted using Graphpad Prism 8.0 software, and IC50 values ​​were calculated. 3) The standard deviation of the three data sets was less than 15 (SD < 15). 4) A widely accepted criterion for evaluating the inhibitory potency of hERG channel assay compounds is as follows:

[0738] 1) Low inhibitory efficacy: IC 50 >10μM

[0739] 2) Moderate inhibitory potency: 1 μM <IC 50 <10 μM

[0740] 3) High inhibitory efficacy: IC 50 <1 μM

[0741] Data quality control standards

[0742] Only data that meet the following criteria can be analyzed: 1) initial sealing resistance greater than 1 GΩ; 2) series resistance less than 15 MΩ and the series resistance voltage error less than 5 mV; 3) leakage current at the detection voltage is less than 50% of the current value under that condition; 4) tail current greater than the platform current of the pre-pulse, and the initial tail current value is greater than 250 pA; 5) membrane rupture resistance Ra is less than 15 MΩ; 6) the tail current decay rate per minute is less than 2.5%.

[0743] 5.hERG IC 50 (μM) data are shown in Table 3:

[0744] Table 3: hERG IC 50 data

[0745] Example No. <![CDATA[hERG IC 50 (μM)]]> 4 5.29 29 11.4 16 2.3 107 4.5 104 1.51 130 1.36 41 8.16 AMG473 <0.37

[0746] By comparing hERG IC 50 The data showed that compared with AMG473, the embodiments of the present invention showed a lower risk of cardiac toxicity and side effects and a larger safety window.

[0747] Although the preferred embodiment has been described above, it will be apparent to those skilled in the art that modifications may be made without departing from the invention. Such modifications are considered to be possible variations within the scope of the invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound has the following structure:

2. A pharmaceutical composition comprising the compound according to claim 1 and a second active substance, wherein the second active substance is an anti-tumor drug.

3. The pharmaceutical composition according to claim 2, wherein the anti-tumor drug comprises one or more of a chemotherapy drug, a targeted tumor therapy drug or a tumor therapy antibody drug.

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