Novel PRMT5 inhibitors and their use

Abstract

The present invention describes novel molecules having 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: [Case 1] TIFF2025533677000129.tif105150

Description

[Technical Field]

[0001] This application belongs to the field of drug synthesis and specifically relates to a PRMT5 inhibitor and its use. [Background technology]

[0002] Epigenetic changes are important mediators that promote and maintain the malignant phenotype of tumors. DNA methylation, histone acetylation and methylation, non-coding RNA, and changes in post-translational modifications are all epigenetic drivers of cancer development, independent of changes in DNA sequence. Arginine methylation is an important class of post-translational modifications that influence cell proliferation and growth, apoptosis, angiogenesis, and metastasis by regulating transcription and post-transcriptional RNA processing. There are three types of methylarginine: 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 and transfers methyl from S-adenosylmethionine (AdoMet) to histone and non-histone arginine side chains. The human genome has nine annotated PRMT genes, which are classified into Type I (PRMT1, 2, 3, 4, 6, 8), Type II (PRMT5 and PRMT9), and Type III enzymes (PRMT7) based on the type of methylarginine they produce. PRMT5 is primarily a Type II enzyme that catalyzes the symmetric dimethylation of arginine. PRMT5 was first identified in a two-hybrid assay that detects proteins that interact with Janus tyrosine kinase (Jak2).

[0003] PRMT5 is a versatile transcriptional repressor that forms complexes with other transcription factors such as BRG1, Hbrm, Blimp1, and Snail. PRMT5 is involved in various different cell biological processes through the methylation of various cytoplasmic and nuclear substrates, including histone H4 residues Arg3 (H4R3) and H3 residues Arg8 (H3R8). While H4R3 methylation is associated with transcriptional repression, H3R8 methylation is involved in both transcriptional activation and repression. In addition to the direct induction of repressive histone labeling by PRMT5, the enzyme's role in gene silencing is mediated by the formation of multiple repressive protein complexes involving NuRD components, HDACs, MDB proteins, and DNA methyltransferases. PRMT5 influences its substrate specificity through the interactions of numerous binding proteins. The central component of this protein complex is MEP50, which is required for PRMT5's enzymatic activity. PRMT5 has been found to be able to methylate proteins involved in RNA splicing, such as SmD3, and can be used to track the chemical activity of PRMT5 in cell biology.

[0004] PRMT5 plays a crucial role in tumorigenesis. Studies have shown that PRMT5 expression is upregulated in various tumors, including lymphoma, lung cancer, breast cancer, and colorectal cancer. Furthermore, PRMT5 expression was elevated in samples from mantle cell lymphoma (MCL) patients, and knockdown of PRMT5 suppressed MCL cell proliferation, suggesting that PRMT5 plays a vital role in MCL. PRMT5 overexpression promotes cell hyperplasia, and knockdown of PRMT5 can suppress the proliferation of melanoma, breast cancer, and lung cancer cell lines. Therefore, PRMT5 may be a potential target for cancer treatment.

[0005] Deficiency in methylthioadenosine phosphorylase (MTAP) leads to a selective dependence of cells on PRMT5 and its binding protein WDR77. MTAP is often deficient because it is located close to the tumor suppressor gene CDKN2A. In cells lacking MTAP, intracellular methylthioadenosine (MTA, a metabolite cleaved by MTAP) levels increase. MTA has a structure similar to S-adenosylmethionine (SAM), and when its concentration increases, MTA acts as a substance that essentially selectively inhibits the binding of SAM to PRMT5, thereby inhibiting the methyltransferase activity of PRMT5.

[0006] The major structural difference between MTAP-deficient cancer cells and MTAP wild-type cancer cells lies in the accumulation of MTA concentration, resulting in the PRMT5-MTA complex produced in MTAP-deficient cancer cells. Inhibitors developed against the PRMT5-MTA complex can selectively target MTAP-deficient cancer cells with minimal impact on normal cells, significantly improving the therapeutic index.

[0007] Thus, the identification and development of small molecules that inhibit PRMT5 activity is expected to be used as a therapeutic approach for various diseases and conditions associated with PRMT5, such as cancer. [Overview of the Initiative]

[0008] To solve the technical problems of the present invention, the present invention provides a group of novel compounds having good inhibitory activity against PRMT5.

[0009] Specifically, the present invention provides compounds represented by formula (I), pharmaceutically acceptable salts, esters, prodrugs, stereoisomers, or isotopic derivatives thereof. [ka] In the formula, W is C, In the formula, X3 is N or CR X3 And X4 is N or CR X4where X5 is N or CR X5 where X6 is N or CR X6 and wherein when X3 is CR X3 R X3 is 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 alkylthiol, hydroxy C1-C6 alkyl, or 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 substituted by 0 to 4 substituents selected from the group consisting of C3-C 10 cycloalkyl, C6-C 10 cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 aryl, 5-10 member heteroaryl, wherein when X4 is CR X4 R X4 [[ID=$$]]is hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen, -OR a , -SRa ,-P(O)R a R b -CN, -S(O)2R a ,-S(O)R a -SF5, -NR a R b These are halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, In the formula, X5 is CR X5 If R X5 is 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, a halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or 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 substituted by 0 to 4 substituents selected from the group consisting of C3-C 10 cycloalkyl, C6-C 10 cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 aryl, 5-10 member heteroaryl, and wherein, when X6 is CR X6 , R X6 is 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 , a halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), NRa 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 substituted by 0 to 4 substituents selected from the group consisting of C3-C 10 cycloalkyl, C6-C 10 cycloalkenyl, 4- to 10-membered heterocycloalkyl, 6- to 10-membered heterocycloalkenyl, C6-C 10 aryl, 5- to 10-membered heteroaryl, and where the A ring may optionally be fused with a 5- to 6-membered saturated or unsaturated ring containing 0 to 3 heteroatoms selected from O, N, S in the chemical bond between X3 and X4; where the A ring may optionally be fused with a 5- to 6-membered saturated or unsaturated ring containing 0 to 3 heteroatoms selected from O, N, S in the chemical bond between X4 and X5; where the A ring may optionally be fused with a 5- to 6-membered saturated or unsaturated ring containing 0 to 3 heteroatoms selected from O, N, S in the chemical bond between X5 and X6; where the A ring may also be 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 bThe molecules may be optionally substituted with 0, 1, 2, or 3 substituents selected from the group consisting of halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, and hydroxy C1-C6 alkyl, or with 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b Substituted by 0 to 4 substituents selected from the group consisting of; In the formula, R' is deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3-6 member saturated or unsaturated aliphatic heteromonocyclic, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b -CN, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 C1-C6 alkyl, C3-C alkyl substituted with 0 to 3 arbitrary substituents selected from the group consisting of the above. 10Cycloalkyl, 4-10 member heterocycloalkyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, Preferably, R is -CHR 2 R 3 or -CDR 2 R 3 And, In the formula, R 2 , R 3 These are hydrogen, deuterium, and -OR, respectively, independently. a , halogen, -CN, -C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, -C3-C 10 Cycloalkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b 4-10 member heterocycloalkyl, C6-C, substituted with 0-3 arbitrary substituents selected from the group consisting of 10 It is an aryl, 5-10 membered heteroaryl, In the formula, M1 is CR a R b , NR a , O, S or Se, In the formula, R L , R L’ Each is independently hydrogen, deuterium, and C1-C6 alkyl, or R L , RL’ These atoms, along with the atoms connected to them, form a 3-6 membered ring; In the formula, n and o are independently 0, 1, or 2.

[0010] In the formula, X1 is N or CR X1 And, In the formula, X2 is N or CR X2 And, In the formula, Y1 is CR Y1 R Y1’ , NR Y1 , O, S, Se, In the formula, Y2 is CR Y2 R Y2’ , NR Y2 , O, S, Se, In the formula, Y3 is CR Y3 R Y3’ , NR Y3 , O, S, Se, In the formula, R X1 , R X2 Each of these is independently 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, In the formula, R Y1 , R Y1’ , R Y2 , R Y2’ , R Y3 , R Y3’These do not exist independently of each other: 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, During the ceremony, [ka] It is either a single bond or a double bond. In the formula, R a , R b Each of these is independently hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, halogenated (C1-C6 alkyl), or R a , R b These atoms link together to form a 3-14 member saturated or unsaturated ring, which may optionally contain 0-2 heteroatoms selected from O, S, and N.

[0011] Furthermore, the present invention provides a compound having the structure of formula (II) below, a pharmaceutically acceptable salt, ester, prodrug, stereoisomer or isotopic derivative thereof. [ka] In the formula, W is C, In the formula, X3 is N or CR X3 And X4 is N or CR X4 And X5 is N or CR X5 And X6 is N or CR X6 And, In the formula, X3 is CR X3 If R X3 is 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 These are halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, In the formula, X4 is CR X4 If R X4 is 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 These are halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, In the formula, X5 is CR X5 If R X5 is 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 bThese are halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, In the formula, X6 is CR X6 If R X6 is 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 These are halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, hydroxy C1-C6 alkyl, or deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), NRa R b -CN, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, In the formula, R' is deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3-6 member saturated or unsaturated aliphatic heteromonocyclic, -OR a , oxo, hydroxy C1-C6 alkyl, NR a R b -CN, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 C1-C6 alkyl, C3-C alkyl substituted with 0 to 3 arbitrary substituents selected from the group consisting of the above. 10 Cycloalkyl, 4-10 member heterocycloalkyl, C6-C 10 It is an aryl, 5-10 membered heteroaryl, Preferably, R is -CHR 2 R 3 or -CDR 2 R 3 And, In the formula, R 2 , R 3 These are hydrogen, deuterium, and -OR, respectively, independently. a , halogen, -CN, -C1-C6 alkyl, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, hydroxy C1-C6 alkyl, -C3-C 10 Cycloalkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b 4-10 member heterocycloalkyl, C6-C, substituted with 0-3 arbitrary substituents selected from the group consisting of 10 It is an aryl, 5-10 membered heteroaryl, In the formula, M1 is CR a R b , NR a , O, S or Se, In the formula, R L , R L’ Each is independently hydrogen, deuterium, and C1-C6 alkyl, or R L , R L’ These atoms, together with the atoms they are linked to, form a 3-6 membered ring; In the formula, n and o are independently 0, 1, or 2.

[0012] In the formula, X1 is N or CR X1 And, In the formula, X2 is N or CR X2 And, In the formula, Y1 is CR Y1 R Y1’ , NR Y1 , O, S, Se, In the formula, Y2 is CR Y2 R Y2’ , NR Y2 , O, S, Se, In the formula, Y3 is CR Y3 R Y3’ , NR Y3 , O, S, Se, In the formula, R X1 , R X2 Each of these is independently 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, In the formula, R Y1 , R Y1’ , R Y2 , R Y2’ , R Y3 , R Y3’ These do not exist independently of each other: 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, During the ceremony, [ka] It is either a single bond or a double bond. In the formula, R a , R b Each of these is independently hydrogen, deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, halogenated (C1-C6 alkyl), or R a , R b These atoms link together to form a 3-14 member saturated or unsaturated ring, which may optionally contain 0-2 heteroatoms selected from O, S, and N.

[0013] In the preferred technical concept of the present invention, in the formula, [ka] It is a double bond.

[0014] In the preferred technical concept of the present invention, in the formula, X1 is CR X1 Or N, where R X1 These include hydrogen, deuterium, halogen, -CN, C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, and halogenated C1-C6 alkyl.

[0015] In the preferred technical concept of the present invention, X1 is CH, CF, or N in the formula.

[0016] In the preferred technical concept of the present invention, X2 is CH or CD in the formula.

[0017] In the preferred technical concept of the present invention, X2 is CH in the formula.

[0018] In the preferred technical concept of the present invention, X3 is CH, CD, or N in the formula.

[0019] In the preferred technical concept of the present invention, X3 is CH in the formula.

[0020] In the preferred technical concept of the present invention, in the formula, X4 is CR X4 Or N, where R X4 Hydrogen, deuterium, C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthiol, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, halogen, SF5, -S(O)2R a ,-P(O)R a R b , or cyano, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 4 substituents selected from the group consisting of 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 member heterocycloalkyl, 6-10 member heterocycloalkenyl, C6-C 10 It is an aryl, a 5-10 membered heteroaryl.

[0021] In the preferred technical concept of the present invention, in the formula, X4 is CR X4 And here, R X4 These are C1-C6 alkoxy, C1-C6 alkylthiol, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, halogenated C1-C6 alkyl, and -SF5.

[0022] In the preferred technical concept of the present invention, in the formula, X4 is CR X4 And here, R X4 These are halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthiol, halogenated C1-C6 alkyl, and -SF5.

[0023] In the preferred technical concept of the present invention, in the formula, X5 is CR X5 Or N, where R X5 These are hydrogen, deuterium, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogen, SF5, or cyano.

[0024] In the preferred technical concept of the present invention, X5 is CH in the formula.

[0025] In the preferred technical concept of the present invention, X6 is CH, CD, or N in the formula.

[0026] In the preferred technical concept of the present invention, X6 is CH in the formula.

[0027] In the preferred technical concept of the present invention, the chemical bond between Y1 and Y2 in the formula is a double bond.

[0028] In the preferred technical concept of the present invention, Y1 is CH, CD, or CCH3.

[0029] In the preferred technical concept of the present invention, Y2 is N in the formula.

[0030] In the preferred technical concept of the present invention, Y3 is CH, CD, CCH3, or CCH2OH.

[0031] In the preferred technical concept of the present invention, in the formula, R' is -CHR 2 R 3 or -CDR 2 R 3 And here, R 2 , R 3 Each of these is independently hydrogen, deuterium, and C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 3 substituents selected from the group consisting of 10 Cycloalkyl, 4-10 member heterocycloalkyl, C6-C 10 It is an aryl, a 5-10 membered heteroaryl.

[0032] In the preferred technical concept of the present invention, R' is -CHR 2 R 3 or -CDR 2 R 3 And here, R 2 R consists of hydrogen, deuterium, and C1-C6 alkyl. 3is hydrogen, deuterium, C1-C6 alkyl, or 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, C1-C6 alkyl halogens, C1-C6 alkoxy halogens, -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 Alternatively, -CONR a R b C3-C substituted with 0 to 3 substituents selected from the group consisting of 10 Cycloalkyl, 4-10 member heterocycloalkyl, C6-C 10 It is an aryl, a 5-10 membered heteroaryl.

[0033] In the preferred technical concept of the present invention, R' is deuterated, halogen, C1-C6 alkyl, hydroxy C1-C6 alkyl, -OR a -CN, NR a R b C3-C3 atoms are substituted with 0 to 3 substituents selected from the group consisting of halogenated C1-C6 alkyl and halogenated C1-C6 alkoxy atoms. 10 It is a cycloalkyl group.

[0034] In the preferred technical concept of the present invention, R' is C1-C 6ア The compounds are methyl (preferably ethyl) or deuterated C1-C6 alkyl (preferably deuterated methyl or deuterated ethyl) or C3-C6 cycloalkyl (preferably cyclopropyl).

[0035] In a preferred technical concept of the present invention, in the formula, M1 is O or S.

[0036] In a preferred technical concept of the present invention, in the formula, o is 1 or 2.

[0037] In a preferred technical concept of the present invention, in the formula, o is 1.

[0038] In a preferred technical concept of the present invention, in the formula, n is 0 or 1.

[0039] In a preferred technical concept of the present invention, in the formula, n is 0.

[0040] In a preferred technical concept of the present invention, in the formula, n is 1.

[0041] In a preferred technical concept of the present invention, in the formula, R L 、R L’ are each independently hydrogen or C1-C6 alkyl.

[0042] [[ID=3-- The present invention specifically provides the following compounds:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0043] Preferably, the pharmaceutical composition of the present invention may further contain a second active substance, the second active substance being an antitumor agent, the antitumor agent comprising one or more of a chemotherapeutic agent, a targeted anticancer agent, or an antibody anticancer agent.

[0044] The present invention also provides a method for treating a disease, preferably a tumor, by inhibiting the action of the compound of the present invention, its pharmaceutically acceptable salt, ester, prodrug, stereoisomer or isotopic derivative, or PRMT5. [Modes for carrying out the invention]

[0045] Definition: Unless otherwise specified, the term "alkyl" refers to a linear (i.e., unbranched), branched, or cyclic hydrocarbon group, or a combination thereof, either by itself or as part of another substituent, which may be saturated, monovalent, or polyunsaturated, and may contain divalent or polyvalent groups, and a certain number of carbon atoms (i.e., C1-C1). 10 A saturated hydrocarbon group has 1 to 10 carbon atoms. Examples of saturated hydrocarbon groups include, but are not limited to, radicals such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, cyclohexylmethyl, and cyclopropylmethyl (e.g., n-pentyl, n-hexyl, n-heptyl, n-octyl) and other congeners and isomers. An unsaturated alkyl group is an alkyl group having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, clotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher congeners and isomers. Alkyls limited to hydrocarbon groups are called "homoalkyls". The alkyl group described above is optionally substituted with one or more halogen atoms.

[0046] The term "alkyl halide" refers to an alkyl group in which one or more hydrogen atoms are substituted with halogen atoms, as defined above.

[0047] The term "alkylene" refers to a divalent group derived from alkyl, either by itself or as part of another substituent, such as, but not limited to, -CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2C≡CCH2-, and -CH2CH2CH(CH2CH2CH3)CH2-. Alkyl (or alkylene) typically has 1 to 24 carbon atoms, and groups having 10 or fewer carbon atoms are preferred in this invention. "Lower alkyl" or "lower alkylene" means a short-chain alkyl or alkylene, typically having 8 or fewer carbon atoms. The above alkylenes are optionally substituted with one or more halogen atoms.

[0048] The term "alkynyl" refers to a carbon chain containing at least one carbon-carbon triple bond, which may be linear, branched, or a combination thereof. Examples of alkynyls include ethynyl, propynyl, 3-methyl-1-pentynyl, and 2-heptynyl. The above alkynyls can be optionally substituted with one or more halogen atoms.

[0049] The term "cycloalkyl" refers to a monocyclic or bicyclic saturated carbon ring, each having 3 to 10 carbon atoms. A "condensation analogue" of a cycloalkyl is a monocyclic ring condensed with an aryl or heteroaryl atom, with the linkage position located in the non-aromatic moiety. Examples of "cycloalkyls" and their condensation analogues include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydronaphthalene, decahydronaphthalene, and dihydroindenyl. The above cycloalkyls are optionally substituted with one or more halogen atoms. Furthermore, in this invention, the term "cycloalkyl" includes bridging rings and spirocyclic rings.

[0050] The term "alkoxy" refers to a linear or branched alkoxy group having the indicated number of carbon atoms. 1-6 Examples of alkoxys include methoxy, ethoxy, propoxy, and isopropoxy.

[0051] Unless otherwise specified, the term “heteroalkyl” means, by itself or in combination with other terms, a stable linear or branched chain, or cyclic hydrocarbon group, or a combination thereof, comprising at least one carbon atom and at least one heteroatom selected from O, N, P, Si, and S, wherein the nitrogen, phosphorus, or sulfur atom may optionally be oxidized, and the nitrogen atom may optionally be quaternized. The heteroatoms O, N, P, S, and Si can be located at any position within the heteroalkyl or at a position where the alkyl is bonded to the rest of the molecule. For example, this includes, but is 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. Heteroatoms may be consecutive in groups of two or three. For example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. Similarly, the term “heteroalkylene,” either by itself or in combination with other terms, refers to a divalent group derived from a heteroalkyl, such as, but not limited to, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. In the case of heteroalkylenes, the heteroatom may be at either or both ends of the chain (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Furthermore, for linking groups of alkylenes and heteroalkylenes, the orientation of the linking group is not indicated by the direction in which the molecular formula of the linking group is written. For example, the molecular formula -C(O)OR'- represents -C(O)OR'- and -R'OC(O)-. As stated above, heteroalkyls as used herein include groups linked to the rest of the molecule via a heteroatom, such as -C(O)R', -C(O)NR', ​​-NR'R'', -OR', -SR', and / or -SO2R'.When referring to "heteroalkyl" and subsequently referring to specific heteroalkyls such as -NR'R'', the terms "heteroalkyl" and -NR'R'' are understood not to be overlapping or mutually exclusive. Instead, for clarity, these specific heteroalkyls are cited. Thus, the term "heteroalkyl" should not be construed herein to exclude specific heteroalkyls such as -NR'R''.

[0052] The term "cycloalkoxy" means a cycloalkyl as defined above, such as cyclopropoxy, bonded to an oxygen atom.

[0053] The term "halogenated alkoxy" means an alkoxy as defined above in which one or more hydrogen atoms are halogenated.

[0054] The term "aryl" means a monocyclic or bicyclic aromatic group containing only carbon atoms. The "condensed analog" of aryl means a condensation product of aryl with a monocyclic cycloalkyl or a monocyclic heterocycle where the linkage position is in the aromatic moiety. Examples of aryl and its condensed analogs include phenyl, naphthyl, indaryl, indenyl, tetrahydronaphthalene, 2,3-dihydrobenzofuran, dihydrobenzopyran, 1,4-benzodioxane, etc.

[0055] The term "heteroaryl" refers to a monocyclic or bicyclic aromatic ring containing at least one heteroatom selected from N, O, and S. A "condensation analogue" of a heteroaryl refers to a heteroaryl condensed with a monocyclic cycloalkyl or monocyclic heterocycle, where the linkage position is in the aromatic moiety. Examples of heteroaryls include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thiophene, pyrimidyl, pyridadinyl, pyrazinyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuran, benzothiophenyl, flu(2,3-b)pyridyl, quinolyl, indolyl, isoquinolyl, etc.

[0056] "Substituted or unsubstituted": Defined alkyl, aryl, and heteroaryl elements are either unsubstituted or substituted with at least one substituent selected from the group consisting of substituents.The above substituents include halogen atoms, alkyls having 1 to 6 carbon atoms, alkoxys having 1 to 6 carbon atoms, alkyl halides having 1 to 6 carbon atoms, alkoxy halides having 1 to 6 carbon atoms, -CN, alkynyls having 2 to 6 carbon atoms, alkanoyls having 1 to 6 carbon atoms, cycloalkyls having 3 to 7 cyclo atoms, heteroaryls, aryls, aralkoxys having 7 to 10 carbon atoms, arylcarbonyls, aminocarbonyls, alkenyls having 2 to 5 carbon atoms, alkylthiols having 1 to 6 carbon atoms, aminosulfinyls, aminosulfonyls, hydroxyl, -SF5, hydroxyalkyls having 1 to 4 carbon atoms, nitros, aminos, carboxyls, alkoxycarbonyls having 2 to 5 carbon atoms, alkoxyalkyls having 1 to 4 carbon atoms, alkylsulfonyls having 1 to 4 carbon atoms, alkanoylaminos having 1 to 4 carbon atoms, alkanoyl(alkyl)aminos having 1 to 6 carbon atoms, alkanoyls, and alkyls, all of which have 1 to 6 carbon atoms. Selected from the group consisting of alkanoylaminoalkyl having 1 carbon atoms, alkanoyl and alkanoyl(alkyl)aminoalkyl having 1 to 6 carbon atoms in each alkyl, alkylsulfonylamino having 1 to 4 carbon atoms, monoalkylaminocarbonyl or dialkylaminocarbonyl having 1 to 6 carbon atoms, monoalkylaminosulfinyl or dialkylaminosulfinyl having 1 to 6 carbon atoms, monoalkylaminosulfonyl or dialkylaminosulfonyl having 1 to 6 carbon atoms, aminoalkyl having 1 to 4 carbon atoms, monoalkylamino or dialkylamino having 1 to 6 carbon atoms, monoalkylaminoalkyl or dialkylaminoalkyl having 1 to 6 carbon atoms in each alkyl, aralkyl having 7 to 10 carbon atoms, heteroaralkyl having 1 to 4 carbon atoms in the alkyl, heteroarylalkoxy having 1 to 4 carbon atoms in the alkoxy, and alkylsulfonamide having 1 to 4 carbon atoms.

[0057] As used herein, the terms “heterocyclic,” “heterocyclic formula,” “heterocycloalkyl,” or “heterocyclo” mean a saturated, partially saturated, or unsaturated group (excluding aromatic) having a monocyclic or fused ring (including bridging ring systems and spiro-ring systems, having 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from nitrogen, sulfur, or oxygen), wherein one or more rings in the fused ring system may be cycloalkyl, aryl, or heteroaryl, provided that the linkage passes through a non-aromatic ring. In one embodiment, the nitrogen and / or sulfur atoms of the heterocyclic group are optionally oxidized to provide N-oxide, sulfinyl, and sulfonyl moieties. Examples of “heterocyclo” and its condensed analogues include pyrrolidinyl, piperidine, piperazine, imidazolidinyl, 2,3-dihydrofluoro(2,3-b)pyridine, benzoxazinyl, tetrahydroquinolyl, tetrahydroisoquinolyl, and dihydroindolyl. This term also includes non-aromatic partially unsaturated monocycles such as 2- or 4-pyridones linked via a nitrogen atom, and N-substituted-(1H,3H)-pyrimidine-2,4-dione (N-substituted uracil).

[0058] As used herein, the terms “substituted heterocyclic,” “substituted heterocycloalkyl,” or “substituted heterocyclo” mean a heterocyclic group substituted with 1 to 5 (e.g., 1 to 3) substituents, the substituents being the same as those defined for substituted cycloalkyls.

[0059] Unless otherwise specified, the terms "halogenated" or "halogen" refer to a fluorine, chlorine, bromine, or iodine atom, either by itself or as part of other substituents. Furthermore, the term "alkyl halide" includes monoalkyl halides and polyalkyl halides. For example, the term "(C1-C6) alkyl halide" includes, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 4-chlorobutyl, and 3-bromopropyl.

[0060] A "prodrug" is a substance that is converted into a parent drug in the body. Prodrugs are often used because, depending on the condition, they are easier to administer than the parent drug. For example, a prodrug can be orally bioavailable, while the parent drug cannot. In pharmaceutical compositions, prodrugs may have higher solubility than the parent drug. Examples of prodrugs, though not limited to, include any compound of formula (I) that is administered in ester form (prodrug) to facilitate transport across cell membranes where water solubility is unfavorable, and then, once inside the cell where water solubility is beneficial, the ester is metabolically hydrolyzed to a carboxylic acid, which is the active substance. Another example of a prodrug is a short peptide (polyamino acid) bound to an acid group, which is metabolized to release the active portion.

[0061] Optical isomers - diastereomers - geometric isomers - tautomers: Since the compounds of formula (I) contain one or more chiral centers, they can be used as racemates and racemic mixtures, single enantiomers, diastereomer mixtures, and single diastereomers. The present invention encompasses all such isomers of the compounds of formula (I).

[0062] Some of the compounds described herein contain alkenyl (olefin) double bonds, which, unless otherwise specified, include both E and Z geometric isomers.

[0063] Some of the compounds of the present invention constitute one or more ring systems, and therefore cis and trans isomers may exist. The present invention is intended to encompass all of these cis and trans isomers.

[0064] Among the compounds described herein, some have different hydrogen atom bonding sites, and these are known as tautomers. Examples of such compounds include keto and its enol derivatives, known as keto-enol tautomers. Individual tautomers and mixtures thereof are included in the compounds of the present invention.

[0065] The compounds of the present invention can be separated into diastereoisomeric pairs of enantiomers by multi-step crystallization from a suitable solvent such as methanol, ethyl acetate, or a mixture thereof. The pair of enantiomers thus obtained can be separated into separate stereoisomers by conventional methods, such as using optically active amines or acids as resolution reagents or by using a chiral HPLC column.

[0066] Alternatively, any enantiomer of the compound of the present invention can be obtained by stereosynthesis using optically pure starting materials or reagents of known configurations.

[0067] Stable isotope-labeled analogs: One or more protons in the compound of the present invention can be substituted with deuterium atoms, making it possible to provide deuterium-substituted analogs with improved pharmacological activity.

[0068] Salts and dosage forms When used herein, references to the compounds of the present invention should be understood to include pharmaceutically acceptable salts as well.

[0069] use The compounds provided in this invention can be used to treat diseases related to PRMT5.

[0070] The compounds of the present invention can be prepared according to the following reaction formula: Method A: [ka] Method A-SFC [ka] Here, R L , R L’ , R ’ , A ring, W, n, o, M1, X1, X2, X3, X4, X5, X6, Y1, Y2, Y3, [ka] This is as defined in claim 1.

[0071] Method A: Compound AP can be obtained by an amino acid condensation reaction between carboxylic acid A-1 and amine A-2, with HATU or PyBrOP as the coupling agent, DIPEA or TEA as the base, and DMF or DMAc as the solvent. If the amine used is a racemic mixture, it is resected by chiral SFC, and the stereochemistry of the resulting isomers is randomly assigned to either R or S.

[0072] AnalyticalHPLC Instrument: Agilent 1260; Column size: Agilent Poroshell HPH-C18 (3.0 × 50 mm, 2.7 μm); Two-component 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; Time: 12 min; Detector: DAD detector; Wavelength: 254 / 220 nm.

[0073] Preparative HPLC-MS HPLC instrument: Waters 2489; Column size: Ultimate μ XB-C18 (130A, 5μm, 30mm × 150mm); Vanilla-based 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 detector, Wavelength: 254 / 220 nm.

[0074] Mass spectrometer: Agilent G6125B.

[0075] The compounds of the present invention can be prepared by chemical synthesis, and examples thereof are shown below. Please note that the order of the steps described may be changed, the reagents, solvents, and reaction conditions mentioned in particular may be substituted, and reactive sites may be protected and deprotected as needed.

[0076] The following abbreviations have the following meanings: "ACN" refers to acetonitrile; "EA" refers to ethyl acetate; "CDI" refers to N,N'-carbonyldiimidazole; "DBU" refers to 1,8-diazabicyclo[5.4.0] refers to undeca-7-ene; "DIBAL-H" refers to diisobutylaluminum hydride; "DIEA" refers to diisopropylethylamine; "DMAP" refers to N,N-dimethyl-4-aminopyridine; "DME" refers to 1,2-dimethoxyethane; "DMF" refers to N,N-dimethylformamide; "DMA" and "DMAc" refer to N,N-dimethylformamide; "DMPE" refers to 1,2-bis(dimethylphosphino)ethane; "DMSO" refers to dimethyl sulfoxide; "DPPB" refers to 1,4-bis(diphenylphosphino)butane; "dppe" refers to 1,2-bis(diphenylphosphino) "dppf" refers to ethane; "dppm" refers to 1,1'-bis(diphenylphosphino)ferrocene; "DIAD" refers to diisopropyl azodicarboxylic acid; "EDCI" refers to 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; "HATU" refers to 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate; "HMPA" refers to hexamethyl phosphate triamide; "IPA" refers to isopropanol; "LDA" refers to lithium diisopropylamide; "LHMDS" refers to lithium "Bis(trimethylsilyl)amide" refers to bis(trimethylsilyl)amide; "LAH" refers to lithium aluminum hydride; "NCS" refers to N-chlorosuccinimide; "NaHMDS" refers to sodium bis(trimethylsilyl)amide; "PyBOP" refers to hexafluorophosphate (benzotriazole-1-yloxy)tripyrrolidinophosphonium; "PyBrOP" refers to bromotripyrrolidinophosphonium = hexafluorophosphate; "TDA-I" refers to tris[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"°C" refers to room temperature; "PMB" refers to the p-methoxybenzyl group; "Tosmic" refers to p-toluenesulfonylmethyl isocyanide; "(Boc)2O" refers to di-tert-butyl dicarbonate; "PE" refers to petroleum ether; and "o / n" indicates that the reaction takes place overnight.

[0077] The following preparations and examples illustrate the present invention, but do not limit it in any way.

[0078] The features and advantages of the subject matter of the present invention will be better understood in light of the detailed description of the selected embodiments. As to be understood, the disclosed and claimed subject matter can be modified in various ways without departing from the scope of the claims. Therefore, the description should be considered illustrative and not restrictive in nature. The entire scope of the subject matter of the present invention is described in the claims.

[0079] The present invention can be more readily understood by referring to the following embodiments, but these embodiments are intended to illustrate the present invention only and do not limit its scope.

[0080] intermediate The following intermediate raw materials were purchased from various suppliers: [Table 1] [Table 2] Intermediate 12: (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine [ka] Step 1: Synthesis of tert-butyl(S)-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (S)-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (200 mg, 0.99 mmol) was dissolved in dichloromethane (2 mL), and 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 using liquid chromatography-mass spectrometry (LC-MS). 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.

[0081] LCMS(ESI)m / z:304[M+H] + Step 2: Synthesis of tert-butyl(S)-methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate At 0°C, tert-butyl(S)-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (300 mg, 0.99 mmol) was dissolved in DMF (5.00 mL), followed by the addition of sodium hydride (48 mg, 2.00 mmol). The mixture was stirred at 0°C for 1 hour. Iodomethane (211 mg, 1.49 mmol) was added. The mixture was stirred at room temperature for 2 hours. The reaction was monitored using liquid chromatography-mass spectrometry (LC-MS). The residue was purified by flash column chromatography (20 g silica gel column, 15% ethyl acetate / petroleum ether) to obtain tert-butyl(S)-methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamate (200.0 mg, 64% yield) as a yellow liquid.

[0082] LCMS(ESI)m / z:318[M+H] + Step 3: Synthesis of (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine 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 1,4-dioxane solution of hydrogen chloride (5.00 mL). The mixture was stirred at room temperature for 0.5 hours. The reaction was monitored using liquid chromatography-mass spectrometry (LC-MS). The mixture was concentrated to obtain (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (130.0 mg, 95% yield) as a yellow liquid.

[0083] LCMS(ESI)m / z:218[M+H] + The following intermediates were prepared by adopting the preparation method and procedure of intermediate 12, replacing only the corresponding raw material intermediate: [Table 3] [Table 4] Intermediate 22(S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carbonitrile [ka] Step 1: tert-butyl(S)-(6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate The 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-methylpyrrolide (4 mL) was reacted in a microwave reactor at 135°C for 4 hours. After the reaction was complete, the system was cooled to room temperature, poured into water, extracted three times with ethyl acetate (30 mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (ethyl acetate:petroleum ether = 40:60) to obtain tert-butyl(S)-(6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (66 mg, 23.93% yield) as a colorless oil.

[0084] LCMS(ESI):275[M+H] + Step 2: (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carbonitri 66 mg, 0.24 mmol of tert-butyl(S)-(6-cyano-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate was added to a dioxane solution of hydrogen chloride (3 mL, 4 mol / L), and the mixture was stirred for 2 hours. After the reaction was complete, the resulting mixture was concentrated under reduced pressure to obtain a white solid crude product of (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carbonitrile (40 mg). The crude product was used directly in the next step without purification.

[0085] LCMS(ESI):175[M+H] + Synthesis of the intermediate 23(S)-6-methoxy-N-methyl-2,3-dihydrobenzofuran-3-amine [ka] The 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 methanol solution of sodium methoxide (180 mg, 40 mmol) in N,N-dimethylformamide (4 mL) was reacted at 120 °C for 4 hours. After the reaction was complete, the system was cooled to room temperature and extracted three times in 10 mL portions with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. (S)-6-methoxy-N-methyl-2,3-dihydrobenzofuran-3-amine (150 mg, 0.8 mmol, yield 80%) was obtained as a colorless oil.

[0086] LCMS(ESI):180[M+H] + Intermediate 24(S)-dimethyl(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)phosphine oxide [ka] Step 1: A reaction system consisting of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (200 mg, 0.61 mmol), dimethylphosphine oxide (57 mg, 0.73 mmol), potassium carbonate (101 mg, 0.73 mmol), palladium acetate (10 mg, 0.06 mmol), 4,5-bis(diphenylphosphin)-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 the reaction was complete, the system was cooled to room temperature and extracted three times with ethyl acetate (30 mL) in 10 mL increments. The organic phases were combined, washed with saturated brine, dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. tert-butyl(S)-(6-(dimethylphosphinyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, 0.245 mmol, 40.35% yield) was obtained as a white solid.

[0087] LCMS(ESI):326[M+H] + Step 2: tert-butyl(S)-(6-(dimethylphosphinyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, 0.25 mmol) was added to 4 M dioxane hydrochloride solution (3 mL) and stirred for 2 hours. After the reaction was complete, the resulting mixture was concentrated under reduced pressure. (S)-dimethyl(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)phosphine oxide (25 mg, 0.11 mmol, 45.14% yield) was obtained as a white solid.

[0088] LCMS(ESI):226[M+H] + Intermediate 25(S)-N-methyl-6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-amine [ka] Step 1: The reaction system of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (328 mg, 1 mmol), sodium methanesulfonate (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 in a microwave reactor at 140 °C for 2 hours. After the reaction was complete, the system was cooled to room temperature and extracted three times in 10 mL portions with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. tert-butyl(S)-methyl(6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-yl)carbamate (60 mg, 0.18 mmol, 18.32% yield) was obtained as a colorless oil.

[0089] LCMS(ESI):328[M+H] + Step 2: tert-butyl(S)-methyl(6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-yl)carbamate (60 mg, 0.24 mmol) was added to a 1,4-dioxane solution in 4 M hydrogen chloride (3 mL) and stirred for 2 hours. After the reaction was complete, the resulting mixture was concentrated under reduced pressure. (S)-N-methyl-6-(methylsulfonyl)-2,3-dihydrobenzofuran-3-amine (30 mg, 0.13 mmol, 72.03% yield) was obtained as a white solid.

[0090] LCMS(ESI):228[M+H] + Synthesis of intermediate 101(S)-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine [ka] Step 1: Preparation of tert-butyl(S)-(6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate 300 mg, 0.914 mmol of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate and 118 mg, 0.914 mmol of cyclopropane sulfinate were dissolved in 10 mL of DMSO. 35 mg, 0.182 mmol of cuprous iodide and 52 mg, 0.365 mmol of (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine were added, and the mixture was stirred at 25°C for 10 hours. After confirming the completion of the reaction by LC-MS, water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried on anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The filtrate was purified by column chromatography (0-10% ethyl acetate / petroleum ether) to obtain tert-butyl(S)-(6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (150 mg, yield 46.4%).

[0091] LCMS(ESI)m / z:354.1[M+H] + Step 2: Synthesis of (S)-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine At room temperature, the compound tert-butyl(S)-(6-(cyclopropylsulfonyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (150 mg, 0.424 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added. The reaction was stirred at room temperature for 16 hours and then concentrated under reduced pressure to obtain the crude product S-6-cyclopropylsulfonyl-N-methyl-2,3-dihydrobenzofuran-3-amine (150 mg, crude product), which was used directly in the next step without purification.

[0092] LCMS(ESI)m / z:254.1[M+H] + Synthesis of the intermediate 26(S)-N-methyl-6-(1-trifluoromethyl)pyrazole-4-yl)-2,3-dihydrobenzofuran-3-amine [ka] Step 1: Synthesis of tert-butyl-(S)-methyl(6-(1-trifluoromethyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate tert-butyl Under an 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), to which 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolidine-2-yl)-1-trifluoromethylpyrazole (239.0 mg, 1.37 mmol) and potassium carbonate were added. (377 mg, 2.73 mmol) and 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (67.0 mg, 0.09 mmol) were added, and the mixture was reacted at 100°C for 16 hours. The system was poured into water (30 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, anhydrous sodium sulfate was removed by filtration, and the filtrate was spin-dried to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain tert-butyl-(S)-methyl(6-(1-trifluoromethyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate tert-butyl (240.0 mg, yield 68.8%) as a white solid.

[0093] LCMS(ESI)m / z:384.1[M+H] + Step 2: Synthesis of (S)-N-methyl-6-(1-trifluoromethyl)pyrazole-4-yl)-2,3-dihydrobenzofuran-3-amine tert-butyl-(S)-methyl(6-(1-trifluoromethyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzofuran-3-yl)carbamate tert-butyl (240 mg, 0.626 mmol) was dissolved in 4M ethyl acetate hydrochloride (3 mL) at room temperature. After stirring at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure to obtain the crude product (S)-N-methyl-6-(1-trifluoromethyl)pyrazole-4-yl)-2,3-dihydrobenzofuran-3-amine (25.0 mg, crude product).

[0094] LCMS(ESI)m / z:283.1[M+H] + The following intermediates were prepared by employing the method and procedure of intermediate 26, replacing only the corresponding raw materials: [Table 5] [Table 6] [Table 7] [Table 8] Intermediate 41: 6-(trifluoromethyl)benzo[b]thiophene-3(2H)-one [ka] Step 1: Synthesis of 2-fluoro-4-(trifluoromethyl)benzoic acid To a tetrahydrofuran:water (300 mL:30 mL) solution of methyl 2-fluoro-4-(trifluoromethyl)benzoate (30.00 g, 135.05 mmol), lithium hydroxide (9.70 g, 405.15 mmol) was added at room temperature, and the mixture was stirred at 50°C for 2 hours. The reaction solution was poured into water (100 mL), the pH was adjusted to 4 with aqueous formic acid, and then extracted with ethyl acetate (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by rapid column chromatography on silica gel (petroleum ether:ethyl acetate = 30%) to obtain 2-fluoro-4-(trifluoromethyl)benzoic acid (22.00 g, yield: 78%).

[0095] LCMS(ESI)m / z:209.1[M+H] + Step 2: Synthesis of 2-((2-ethoxy-2-oxoethyl)thio)-4-(trifluoromethyl)benzoic acid Sodium hydrogen (8.46 g, 211.43 mmol) was added at 0°C to a solution of 2-(ethylsulfanyl)acetic acid (12.70 g, 105.72 mmol) in N,N-dimethylformamide (200.00 mL), and the mixture was stirred at room temperature for 0.5 hours. Next, 2-fluoro-4-(trifluoromethyl)benzoic acid (22.00 g, 105.71 mmol) was added to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was slowly poured into water (100 mL), extracted with ethyl acetate (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by rapid column chromatography on silica gel (petroleum ether:ethyl acetate = 10%) to obtain 2-((2-ethoxy-2-oxoethyl)thio)-4-(trifluoromethyl)benzoic acid (13.00 g, yield: 40%).

[0096] LCMS(ESI)m / z:309.2[M+H] + Step 3: Synthesis of 2-((carboxymethyl)thio)-4-(trifluoromethyl)benzoic acid 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 obtain 2-((carboxymethyl)thio)-4-(trifluoromethyl)benzoic acid (7.00 g, crude product).

[0097] LCMS(ESI)m / z:281.2[M+H] + Step 4: Synthesis of 6-(trifluoromethyl)benzo[b]thiophene-3-ylacetate To a solution of 2-(carboxymethylthio)-4-(trifluoromethyl)benzoic acid (7.00 g, 24.98 mmol) in acetic anhydride (70.00 mL), sodium acetate trihydrate (10.19 g, 74.94 mmol) was added 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 high-performance column chromatography (petroleum ether:ethyl acetate = 50%) to obtain 6-(trifluoromethyl)benzo[b]thiophene-3-yl acetate (5.00 g, yield: 77%).

[0098] LCMS(ESI)m / z:261.2[M+H] + Step 5: Synthesis of 6-(trifluoromethyl)benzo[b]thiophene-3(2H)-one A 10.00 mL, 1 mol / L aqueous hydrochloric acid solution was added at room temperature to a 5.00 mL solution of 1,4-dioxane in 1.00 g, 3.84 mmol of 6-(trifluoromethyl)benzo[b]thiophene-3-yl acetate. The mixture was stirred at 100 °C for 2 hours. The reaction mixture was cooled, poured into water, and the aqueous layer was extracted with ethyl acetate (2 × 200 mL). The organic layers were combined, washed with saturated brine (300 mL), dried on anhydrous sodium sulfate, and concentrated to obtain the crude product. Purification by high-performance column chromatography (petroleum ether:ethyl acetate = 70%) yielded 6-(trifluoromethyl)benzo[b]thiophene-3(2H)-one (0.50 g, yield: 59%) as a black solid.

[0099] LCMS(ESI)m / z:219.2[M+H] + Intermediate 42: Synthesis of 1-methyl-7-trifluoromethylisochromen-4-one [ka] Step 1: Synthesis of 2-(1-allyloxy)ethyl)-1-bromo-4-trifluoromethylbenzene At room temperature, 1-(2-bromo-5-trifluoromethylphenyl)ethane-1-ol (10 g, 37.16 mmol) was dissolved in tetrahydrofuran (100 mL), potassium hydroxide (4.2 g, 74.33 mmol), tetrabutylammonium bisulfate (2.6 g, 7.433 mmol), and 3-bromopropene (5.4 g, 44.60 mmol) were added, and the mixture was reacted at 25°C for 4 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, poured into water (100 mL), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product (12 g, crude product), which was used directly in the next step without purification.

[0100] LCMS(ESI)m / z:309.2[M+H] + Step 2: Synthesis of 1-methyl-4-methylene-7-trifluoromethylisochromane To a solution of 2-(1-allyloxy)ethyl)-1-bromo-4-trifluoromethylbenzene (12 g, 38.82 mmol) in DMF (100 mL), cesium carbonate (15.5 g, 46.59 mmol), tri(o-tolyl)phosphine (5.1 g, 19.41 mmol), and palladium acetate (0.89 g, 3.882 mmol) were added. The mixture was stirred at 90°C for 16 hours under an N2 atmosphere. The reaction solution was poured into 400 mL of water and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude product. This was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain 1-methyl-4-methylene-7-trifluoromethylisochromane (5.5 g, yield: 62.1%).

[0101] LCMS(ESI)m / z:229.1[M+H] + Step 3: Synthesis of 4-hydroxymethyl-1-methyl-7-trifluoromethylisochromen-4-ol 1-Methyl-4-methylene-7-trifluoromethylisochroman (5.5 g, 24.10 mmol) was dissolved in a solvent mixture of acetone (100 mL) and water (20 mL), and N-methylmorpholine-N-oxide (9.2 g, 76.68 mmol) and potassium osmite (0.91 g, 2.410 mmol) were added at room temperature. The reaction was stirred at 25°C for 16 hours under an N2 atmosphere. After the reaction was complete, sodium sulfite solid (5 g) was added to the reaction mixture, stirred for 10 minutes, concentrated under reduced pressure to remove a certain amount of acetone, poured into water (200 mL), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude product 6-fluoro-4-hydroxymethyl-1-methylisochroman-4-ol (6.0 g, yield: 94.9%).

[0102] LCMS(ESI)m / z:263.0[M+H] + Step 4: Synthesis of 1-methyl-7-trifluoromethylisochromen-4-one At room temperature, 6.0 g of 4-hydroxymethyl-1-methyl-7-trifluoromethylisochromen-4-ol (22.88 mmol) was dissolved in a mixed solvent of 100 mL of tetrahydrofuran and 3.5 mL of water. Sodium periodate (15.0 g of 68.64 mmol) was added, and the mixture was stirred at 25°C for 4 hours under an N2 atmosphere. After the reaction was complete, the reaction solution was diluted with ethyl acetate, filtered, washed, and the filtrates were combined. After drying over anhydrous sodium sulfate, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product 6-fluoro-1-methylisochromen-4-one (5 g, crude product).

[0103] LCMS(ESI)m / z:230.2[M+H] + The following intermediates were prepared by employing the method and procedure of intermediate 42, replacing only the corresponding raw materials: [Table 9] The following ketones were purchased from different suppliers: [Table 10] Intermediate 55: N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride [ka] Step 1: Synthesis of 6-(trifluoromethyl)isochroman-4-ol At room temperature, 4-(hydroxymethyl)-6-(trifluoromethyl)isochroman-4-one (1.1 g, 5.09 mmol) was first dissolved in THF (10 mL) in an ice bath, sodium borohydride (251 mg, 6.62 mmol) was added, followed by the addition of 2 drops of methanol. After the addition, the reaction was left at room temperature overnight. After the reaction was complete, the mixture was quenched with 1 M aqueous hydrogen chloride (2 mL), extracted with ethyl acetate (10 mL x 2), the organic phases were combined, dried on sodium sulfate, and filtered to obtain the crude product. This was purified by column chromatography to obtain 6-(trifluoromethyl)isochroman-4-ol (1.16 g).

[0104] Step 2: Synthesis of tert-butyl(tert-butoxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate At room temperature, first, 6-(trifluoromethyl)isochroman-4-ol (100.0 mg, 0.459 mmol) was dissolved in THF (5.0 mL), bis(tert-butoxycarbonyl)amine (110 mg, 0.505 mmol) and triphenylphosphine (132 mg, 0.505 mmol) were added, and the reaction system was stirred at 0°C for 5 minutes. Then, DIAD (102 mg, 0.505 mmol) was added dropwise, the mixture was stirred in an ice bath for 1 hour, and then allowed to react overnight at room temperature. After the reaction was complete, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried on anhydrous sodium sulfate, filtered, and the resulting filtrate was concentrated under reduced pressure to obtain the crude product. This was purified by column chromatography (pure petroleum ether) to obtain tert-butyl(tert-butoxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate (90.0 mg).

[0105] Step 3: Synthesis of tert-butyl(6-(trifluoromethyl)isochroman-4-yl)carbamate 60.0 mg, 0.14 mmol of tert-butyl (tert-butoxycarbonyl)(6-(trifluoromethyl)isochroman-4-yl)carbamate was dissolved in 2.0 mL of acetonitrile, and lithium bromide (37.6 mg, 0.432 mmol) was added. The reaction system was allowed to react at 60°C for 20 hours. After the reaction was complete, 10 mL of saturated sodium bicarbonate was added, and the mixture was extracted with 20 mL of ethyl acetate. The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain 30 mg of tert-butyl (6-(trifluoromethyl)isochroman-4-yl)carbamate.

[0106] Step 4: Synthesis of tert-butyl-N-methyl-(6-(trifluoromethyl)isochroman-4-yl)carbamate 360.0 mg, 1.14 mmol of tert-butyl(6-(trifluoromethyl)isochroman-4-yl)carbamate was dissolved in 5.0 mL of DMF and stirred in an ice bath for 2 minutes. 90.8 mg, 2.27 mmol of sodium hydride was added all at once, and the mixture was stirred at this temperature for 1 hour. Subsequently, 487 mg, 3.41 mmol of iodomethane was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, 15 mL of saturated sodium bicarbonate was added, and the mixture was extracted with ethyl acetate (15 mL x 3). The organic phase was dried over sodium sulfate, filtered, and the filtrate was spin-dried to obtain the crude product tert-butyl-N-methyl-(6-(trifluoromethyl)isochroman-4-yl)carbamate (400.0 mg).

[0107] Step 5: Synthesis of N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride 400 mg, 1.21 mmol of tert-butyl-N-methyl-(6-(trifluoromethyl)isochroman-4-yl)carbamate was dissolved in 2 mL of dichloromethane, and 2 mL of 4 M hydrogen chloride dioxane solution was added. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction system was spin-dried to obtain N-methyl-6-(trifluoromethyl)isochroman-4-amine hydrochloride (390 mg).

[0108] The following intermediates were prepared by employing the method and procedure of intermediate 55, replacing only the corresponding raw materials: [Table 11] Intermediate 60 (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride [ka] Step 1: Synthesis of (S)-7-(trifluoromethyl)pyran-4-ol 7-(trifluoromethyl)pyran-4-one (500.0 mg, 2.31 mmol) was first dissolved in DCM (10 mL) in an ice bath, (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanediamine (p-isopropylbenzene) chloride (147.0 mg, 0.23 mmol) was added, formic acid (372.0 mg, 8.10 mmol) was added dropwise, and finally triethylamine (701.0 mg, 6.94 mmol) was added dropwise, and the mixture was reacted overnight at room temperature. After the reaction was complete, saturated sodium bicarbonate aqueous solution (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, filtered, and the crude product was obtained. This was separated and purified by column chromatography (petroleum ether:ethyl acetate = 0-40%) to obtain (S)-7-(trifluoromethyl)pyran-4-ol (370 mg, yield: 73%).

[0109] Step 2: Synthesis of tert-butyl(R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate Under a nitrogen atmosphere, (S)-7-(trifluoromethyl)pyran-4-ol (370.0 mg, 1.70 mmol) was first dissolved in THF (5.0 mL), bis(tert-butoxycarbonyl)amine (405 mg, 1.88 mmol) and triphenylphosphine (489 mg, 1.88 mmol) were added, and the reaction system was stirred at 0°C for 5 minutes. Then, DIAD (377.0 mg, 1.88 mmol) was added dropwise, and the reaction mixture was stirred in an ice bath for 1 hour, followed by continued stirring at room temperature overnight. After the reaction was complete, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried on anhydrous sodium sulfate, filtered, and the resulting filtrate was concentrated under reduced pressure to obtain the crude product. This was purified by column chromatography (petroleum ether:ethyl acetate = 0-5%) to obtain tert-butyl(R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate (145.0 mg, yield 20%).

[0110] 1H NMR (400 MHz, CDCl3) δ 7.25-7.22(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). Step 3: Synthesis of tert-butyl(R)-(7-(trifluoromethyl)pyran-4-yl)carbamate 150.0 mg, 0.36 mmol of tert-butyl(R)-(tert-butoxycarbonyl)(7-(trifluoromethyl)pyran-4-yl)carbamate was dissolved in 5.0 mL of acetonitrile, and lithium bromide (94.0 mg, 1.08 mmol) was added. The reaction system was allowed to react at 60°C for 20 hours. After the reaction was complete, 10 mL of saturated sodium bicarbonate was added, and the mixture was extracted with 20 mL of ethyl acetate. The organic phase was dried over sodium sulfate, filtered, and evaporated to obtain the crude product tert-butyl(R)-(7-(trifluoromethyl)pyran-4-yl)carbamate (83.0 mg, crude product).

[0111] Step 4: Synthesis of tert-butyl(R)-methyl-(7-(trifluoromethyl)pyran-4-yl)carbamate 83.0 mg, 0.26 mmol of tert-butyl(R)-(7-(trifluoromethyl)pyran-4-yl)carbamate was dissolved in 2.0 mL of DMF and stirred in an ice bath for 2 minutes. 26.2 mg, 0.66 mmol of sodium hydride was added all at once, and the mixture was stirred at this temperature for 1 hour. Subsequently, 74.4 mg, 0.52 mmol of iodomethane was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, 10 mL of saturated sodium bicarbonate 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 spin-dried to obtain the crude product tert-butyl(R)-methyl-(7-(trifluoromethyl)pyran-4-yl)carbamate (100.0 mg, crude product).

[0112] LCMS(ESI)m / z:276.1[M+H] + Step 5: Synthesis of (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride 100 mg, 0.3 mmol of tert-butyl(R)-methyl-(7-(trifluoromethyl)pyran-4-yl)carbamate was dissolved in 2 mL of dichloromethane, and 2 mL of 4 M hydrogen chloride dioxane solution was added. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction system was spin-dried to obtain the crude product (R)-N-methyl-7-(trifluoromethyl)pyran-4-amine hydrochloride (90 mg, crude product).

[0113] The following intermediates were prepared using the same method and procedure as for intermediate 60: [Table 12] [Table 13] Intermediate 100 N,1,1-trimethyl-7-trifluoromethylisochromen-4-amine [ka] Step 1: Preparation of 1,1-dimethyl-7-trifluoromethylisochromen-4-ol 1,1-dimethyl-7-trifluoromethylisochromen-4-one (300 mg, 1.229 mmol) was dissolved in MeOH (3 mL), sodium borohydride (70 mg, 1.844 mmol) was added, and the mixture was stirred at 25°C for 10 hours. After confirming the completion of the reaction by LC-MS, NH4Cl 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 filtrate was purified by column chromatography (0-10% ethyl acetate / petroleum ether) to obtain 1,1-dimethyl-7-trifluoromethylisochromen-4-ol (120 mg, yield: 39.4%).

[0114] LCMS(ESI)m / z:247.1[M+H] + Step 2: Preparation of 1,1-dimethyl-7-trifluoromethylisochromene-4-methanesulfonate 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), triethylamine (148 mg, 1.461 mmol) was added, and the mixture was stirred at 25°C for 10 hours. After confirming that the reaction was complete by LC-MS, 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 obtain the crude product 1,1-dimethyl-7-trifluoromethylisochromen-4-methanesulfonate (130 mg, yield: 13.3%).

[0115] LCMS(ESI)m / z:326.1[M+H] + Step 3: Preparation of N,1,1-trimethyl-7-trifluoromethylisochromen-4-amine Compound 1,1-dimethyl-7-trifluoromethylisochromen-4-methanesulfonate (130 mg, 0.398 mmol) was dissolved in DMF (3 mL) at room temperature, and methylamine solution (0.3 mL, 30%) and DIEA (155 mg, 1.194 mmol) were added. The mixture was stirred at room temperature and allowed to react for 16 hours. After confirming that the reaction was complete by LC-MS, 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. Purification was performed by column chromatography (50-100% ethyl acetate / petroleum ether) to obtain N,1,1--trimethyl-7-trifluoromethylisochromen-4-amine (52 mg, yield: 50.4%).

[0116] LCMS(ESI)m / z:260.1[M+H] + The following intermediates were prepared by adopting the preparation method and procedure of intermediate 100, replacing only the corresponding raw material intermediate: [Table 14] Synthesis of intermediate 69 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine [ka] Step 1: Synthesis of (5-bromo-2-(cyclopropylimino)methyl)phenol To a solution of 4-bromo-2-hydroxybenzaldehyde (2.0 g, 9.95 mmol) in dichloromethane (40 mL), cyclopropylamine (1.13 g, 19.9 mmol) and anhydrous magnesium sulfate (4.79 g, 39.8 mmol) were added 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 mixture was filtered. The filtered cake was washed with ethyl acetate, and the resulting filtrate was concentrated under reduced pressure to obtain a yellow solid crude product (5-bromo-2-(cyclopropylimino)methyl)phenol (1.7 g, crude product), which was used directly in the next step without purification.

[0117] LCMS(ESI)m / z:241.1[M+H] + Step 2: Synthesis of 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine Potassium tert-butoxide (1.99 g, 17.70 mmol) was slowly added at room temperature to a solution of trimethylsulfoxonium iodide (3.89 g, 17.70 mmol) in THF (5 mL), and the mixture was stirred at room temperature for 0.5 hours. Next, (5-bromo-2-(cyclopropylimino)methyl)phenol (1.7 g, 7.08 mmol) was dissolved in THF and slowly added dropwise to the mixture, and the resulting suspension was stirred at room temperature for 1 hour, then at 50°C for 3 hours. The reaction mixture was cooled to room temperature, 1 eq of potassium tert-butoxide (0.79 g, 7.08 mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was filtered, the filtrate was diluted with water, and extracted with ethyl acetate. After concentrating the organic layer, the residue was purified by column chromatography using petroleum ether / ethyl acetate (3:1) to obtain 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine (1.0 g, 3.93 mmol) as a yellow oil.

[0118] LCMS(ESI)m / z:255.1[M+H] + The intermediate amines in the table below can be prepared using the synthesis procedure described for intermediate 69, by substituting only the aldehyde or amine of the corresponding starting material. [Table 15] Intermediate 71: Synthesis of 3-(cyclopropylamino)-2,3-dihydrobenzofuran-6-carbonitrile [ka] At room temperature, 6-bromo-N-cyclopropyl-2,3-dihydrobenzofuran-3-amine (100 mg, 0.41 mmol) was first placed in a 50 mL single-port flask, and Pd2(dba)3 (18.8 mg, 0.02 mmol), S-Phos (9.8 mg, 0.02 mmol), and zinc cyanide (96.5 mg, 0.82 mmol) were added. Subsequently, N,N-dimethylacetamide (1.0 mL) was added, and the mixture was heated to 110 °C by microwave and stirred for 1 hour. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried on anhydrous sodium sulfate, filtered, and the organic phase was spin-dried to obtain the crude product. This was thoroughly mixed and purified by column chromatography using ethyl acetate:petroleum ether (0-50%) to obtain 3-(cyclopropylamino)-2,3-dihydrobenzofuran-6-carbonitrile (55 mg, yield: 67%).

[0119] 1 H NMR (400 MHz, CDCl3) δ 7.44-7.42(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). Intermediate 72: (S)-6-(fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine [ka] Step 1: Synthesis of tert-butyl(S)-(6-(hydroxymethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate A mixture of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (400 mg, 1.22 mmol), 2,2-dibutyl-2-stanoxycyclohexane-1-ol (391 mg, 1.22 mmol), and 1,1'-bis(diphenylphosphin)ferrocenepalladium dichloride (99 mg, 0.12 mmol) was reacted in dioxane (5 mL) at 100 °C for 12 hours. After the reaction was complete, the reaction mixture was poured into water (10 mL), 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 the crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 40:60), and eluted to obtain tert-butyl(S)-(6-(hydroxymethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (110 mg, yield: 29%).

[0120] LCMS(ESI):280[M+H] + Step 2: Synthesis of tert-butyl(S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate At -78°C, 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 mixed with [bis(2-methoxyethyl)amine]sulfur trifluoride (396 mg, 1.79 mmol). The mixture was stirred at -78°C for 1 hour. After the reaction was complete, the mixture was quenched with ice water (5 mL), extracted with ethyl acetate (5 mL x 2), the organic layers were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. This was purified by column chromatography (ethyl acetate:petroleum ether = 20:80) to obtain tert-butyl(S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, yield: 70).

[0121] LCMS(ESI):282[M+H] + Step 3: Synthesis of (S)-6-(fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine A mixture of tert-butyl(S)-(6-(fluoromethyl)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (80 mg, 0.36 mmol) and dioxane hydrochloride (1 mL) was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was spin-dried to obtain (S)-6-(fluoromethyl)-N-methyl-2,3-dihydrobenzofuran-3-amine (100 mg, crude product).

[0122] LCMS(ESI):182[M+H] + Intermediate 73: Synthesis of (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-formamide [ka] Step 1: Synthesis of (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid At room temperature, (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid methyl (1 g, 3.25 mmol) was dissolved in methanol (10.00 mL) and tetrahydrofuran (10.00 mL), and sodium hydroxide (260 mg, 6.5 mmol) was added, and the mixture was stirred for 1 hour. After the reaction was complete, the reaction mixture was concentrated to obtain (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylic acid (600 mg, crude) as a white solid.

[0123] LCMS(ESI):294[M+H] + Step 2: Synthesis of tert-butyl(S)-(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (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. To this solution, 2-(7-azabenzotriazolyl)-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 g, 6.78 mmol) were added, and the mixture was reacted at room temperature for 30 minutes. After the reaction was complete, the solution 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 on anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (ethyl acetate / petroleum ether = 17%) to obtain tert-butyl(S)-(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (240 mg, yield: 40%) as a yellow liquid.

[0124] LCMS(ESI):293[M+H] + Step 3: Synthesis of (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-formamide At room temperature, tert-butyl(S)-(6-carbamoyl-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (240 mg, 0.82 mmol) was dissolved in a solution of dichloromethane (2.00 mL) and trifluoroacetic acid (0.40 mL), and the mixture was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was concentrated to obtain (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-formamide as a black solid (100 mg, yield: 66.66%).

[0125] Example 74: (S)-N 3 Synthesis of methyl-2,3-dihydrobenzofuran-3,6-diamine [ka] Step 1: Synthesis of tert-butyl(S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate At room temperature, tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (120 mg, 0.35 mmol) was dissolved in 1,4-dioxane (5.00 mL) solution, and benzophenonimide (190 mg, 1.06 mmol), potassium tert-butoxide (80 mg, 0.71 mmol), 4,5-bis(diphenylphosphin)-9,9-dimethylxanthene (40 mg, 0.07 mmol), and palladium acetate (8 mg, 0.03 mmol) were added. The mixture was reacted at 100°C for 2 hours. After the reaction was complete, the reaction mixture was poured into water (2 mL), extracted with ethyl acetate (2 mL x 2), the organic layers were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. Purification by flash column chromatography (petroleum ether:ethyl acetate = 10%) yielded tert-butyl(S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate (90 mg, yield: 59%) as a white solid.

[0126] LCMS(ESI):429[M+H] + Step 2: (S)-N 3 Synthesis of methyl-2,3-dihydrobenzofuran-3,6-diamine 90 mg, 0.21 mmol of tert-butyl(S)-(6-((diphenylmethylene)amino)-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate was dissolved in methanol hydrochloride (5.00 mL) at room temperature, and the mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the reaction solution was poured into a dichloromethane solution to precipitate a solid, and (S)-N3-methyl-2,3-dihydrobenzofuran-3,6-diamine (60 mg, yield: 90%) was obtained as a white solid.

[0127] LCMS(ESI):165[M+H] + Intermediate 75: Synthesis of (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide [ka] Step 1: Synthesis of tert-butyl(S)-methyl(6-(trifluoromethyl)sulfonamide)-2,3-dihydrobenzofuran-3-yl)carbamate 200 mg, 0.61 mmol of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate, 136 mg, 0.91 mmol of trifluoromethylsulfonamide, 1.16 g, 6.09 mmol of cuprous iodide, and 259 mg, 1.22 mmol of potassium phosphate were sequentially dissolved in 9 mL of dimethylformamide solution, and the mixture was stirred at 90°C for 2 hours. After the reaction was complete, the system was cooled to room temperature and extracted three times with ethyl acetate in 10 mL increments. The organic phases were combined, washed with saturated brine (10 mL x 3), dried on anhydrous sodium sulfate, and the reaction mixture was concentrated under vacuum to obtain 800 mg, yield 51.85%, of tert-butyl(S)-methyl(6-(trifluoromethyl)sulfonamide)-2,3-dihydrobenzofuran-3-yl)carbamate as a yellow liquid.

[0128] LCMS(ESI):397.2[M+H] + Step 2: Synthesis of (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide tert-butyl(S)-methyl(6-(trifluoromethyl)sulfonamide)-2,3-dihydrobenzofuran-3-yl)carbamate (80 mg, 3.246 mmol) was dissolved in a 1,4-dioxane solution of hydrogen chloride (4.0 M, 5 mL). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the system was cooled to room temperature, and the reaction solution was concentrated under vacuum to obtain (S)-1,1,1-trifluoro-N-(3-(methylamino)-2,3-dihydrobenzofuran-6-yl)methanesulfonamide (60 mg, 1.18 mmol, yield 82.46%).

[0129] Intermediate 76: 4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylate methyl [ka] Step 1: Synthesis of imidazo[1,5-a]imidazo[1,5-d]1,4-diazapiperazine-5,10-zion Imidazole-5-carboxylic acid (100 g, 892.14 mmol) was added to dichlorosulfoxide (500 mL) and stirred at 80°C for 12 hours. The mixture was concentrated, washed with toluene (500 mL x 2), filtered to obtain a solid product, and the residual solvent was removed with a 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.

[0130] LCMS(ESI)m / z:189[M+H] + Step 2: Synthesis of N-(4-bromo-2-fluorophenyl)imidazole-5-ylformamide To a mixture of 4-bromo-2-fluorophenylamine (60.60 g, 318.91 mmol) in tetrahydrofuran (600 mL), sodium bis(trimethylsilyl)amide (318.91 mL, 637.82 mmol, 2 mol / liter) was slowly added dropwise at 0°C for 1 hour. Then, imidazo[1,5-a]imidazo[1,5-d]1,4-diazapiperazine-5,10-dione (60 g, 318.91 mmol) was added, and the mixture was stirred at room temperature for 12 hours. The mixture was poured into water, filtered, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 40%) to obtain N-(4-bromo-2-fluorophenyl)imidazole-5-ylformamide (60 g, 0.21 mol, yield 66%) as a white solid. LCMS(ESI)m / z:284.2[M+H] + Step 3: Synthesis of 8-bromo-10-hydroimidazo[1,5-a]quinoxaline-4-ol N-(4-bromo-2-fluorophenyl)imidazole-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 obtain 8-bromo-10-hydroimidazo[1,5-a]quinoxaline-4-ol (50 g, 90% yield) as a yellow solid.

[0131] LCMS(ESI)m / z:264.2[M+H] + Step 4: Synthesis of 8-bromo-4-chloro-10-hydroimidazo[1,5-a]quinoxaline 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) was mixed with phosphorus oxychloride (500 mL), stirred at 90°C for 2 hours, then concentrated. The residue was dissolved in acetonitrile, slowly added dropwise to ice water to precipitate as a solid, filtered, and 8-bromo-4-chloro-10-hydroimidazo[1,5-a]quinoxaline (50 g, 93% yield) was obtained.

[0132] LCMS(ESI)m / z:282.2[M+H] + .

[0133] Step 5: Synthesis of (8-bromo(10-hydroimidazo[1,5-a]quinoxaline-4-yl))[(4-methoxyphenyl)methyl]amine 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), followed by the addition of N,N-diisopropylethylamine (45.66 g, 353.96 mmol). The mixture was stirred at 80°C for 2 hours, then poured into water and filtered to obtain (8-bromo(10-hydroimidazo[1,5-a]quinoxaline-4-yl))[(4-methoxyphenyl)methyl]amine (50 g, yield 74%) as a yellow oil.

[0134] LCMS(ESI)m / z:383.2[M+H] + Step 6: Synthesis of 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (8-bromo(10-hydroimidazo[1,5-a]quinoxaline-4-yl))[(4-methoxyphenyl)methyl]amine (50 g, 130.47 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (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 under a carbon monoxide atmosphere (4 MPa) at 100 °C for 12 hours. The system was then poured into water, filtered, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 80%) to obtain 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (30 g, yield 63%) as a yellow solid.

[0135] LCMS(ESI)m / z:363.2[M+H] + Step 7: Synthesis of 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl (30 g, 82.87 mmol) was added to a mixture 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, and then concentrated under vacuum. The organic solvent was removed, the mixture was poured into water, and the pH was adjusted to 7-8 with hydrochloric acid (2 M). The mixture was extracted with ethyl acetate (100 mL x 3) and concentrated under vacuum to obtain 4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (25 g, yield 86%).

[0136] LCMS(ESI)m / z:349.2[M+H] + Step 8: 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid 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 using liquid chromatography-mass spectrometry (LC-MS). The mixture was concentrated to obtain 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (60.0 mg, yield: 94%) as a white solid.

[0137] LCMS(ESI)m / z:229.2[M+H] + The intermediate carboxylic acids in the table below can be prepared using the synthesis procedure described for intermediate 76, by simply substituting the corresponding starting materials. [Table 16] Intermediate 84 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid [ka] Step 1: Synthesis of methyl 3-(2-methyl-1H-imidazole-1-yl)-4-nitrobenzoate Methyl 3-fluoro-4-nitrobenzoate (20.0 g, 100 mmol) was dissolved in acetonitrile (100 mL), and 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 LC-MS. The reaction solution was poured into 300 mL of water, and 200 mL x 3 of ethyl acetate were added for extraction. The organic phases were combined and dried on anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10%) to obtain methyl 3-(2-methyl-1H-imidazole-1-yl)-4-nitrobenzoate (25.0 g, yield 95%) as a yellow solid.

[0138] LCMS(ESI)m / z:262[M+H] + Step 2: Synthesis of methyl 4-amino-3-(2-methyl-1H-imidazole-1-yl)benzoate At room temperature, methyl 3-(2-methyl-1H-imidazole-1-yl)-4-nitrobenzoate (25 g, 95.8 mmol) was dissolved in methanol (300 mL), and Raney nickel (2 g) was added. The mixture was stirred under a hydrogen atmosphere at room temperature for 12 hours. The reaction was monitored using liquid chromatography-mass spectrometry (LC-MS). The residue was filtered to obtain a filtrate, which was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 20%) to obtain methyl 4-amino-3-(2-methyl-1H-imidazole-1-yl)benzoate (20.5 g, yield 93%) as a yellow solid.

[0139] LCMS(ESI)m / z:232[M+H] + Step 3: Synthesis of methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl At room temperature, methyl 4-amino-3-(2-methyl-1H-imidazole-1-yl)benzoate (2.0 g, 8.7 mmol) was dissolved in o-dichlorobenzene (40 mL), carbonyldiimidazole (2.8 g, 17.4 mmol) was added, and the mixture was stirred at 180 °C for 12 hours. The reaction was monitored by LC-MS, and the mixture was filtered to obtain a filtration cake. This was pulped with ethyl acetate (5 mL) to obtain methyl 1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (920.0 mg, yield: 41%) as a black solid.

[0140] LCMS(ESI)m / z:258[M+H] + Step 4: Synthesis of methyl 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate 1-Methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (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 LC-MS, the reaction solution was concentrated under reduced pressure, diluted with a small amount of acetonitrile, poured into water (5 mL), filtered, the filtered cake was washed with water, and dried under vacuum to obtain 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl (60.0 mg, yield: 56%) as a black solid.

[0141] LCMS(ESI)m / z:276[M+H] + Step 5: Synthesis of 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl 60 mg, 0.22 mmol of 4-chloro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl (60 mg, 0.44 mmol) was dissolved in dimethyl sulfoxide (2 mL) at room temperature, followed by the addition of (4-methoxyphenyl)methylamine (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 LC-MS. 10 mL of the reaction solution was poured into water, ethyl acetate was added for extraction (10 mL x 3), the organic phases were combined and dried over anhydrous sodium sulfate, the organic phase was concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 40%) to obtain 60 mg, yield: 73%, of 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl (60 mg, yield: 73%) as a black solid.

[0142] LCMS(ESI)m / z:377[M+H] + Step 6: Synthesis of 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid 60 mg, 0.16 mmol of 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl was dissolved in methanol (1 mL), tetrahydrofuran (1 mL), and water (1 mL) at room temperature, followed by the addition of potassium hydroxide (26 mg, 0.44 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was monitored by LC-MS. (10 mL) of the reaction solution was poured into water, formic acid was added to make it acidic, and 10 mL x 3 of ethyl acetate was added for extraction. The organic phases were combined and dried on anhydrous sodium sulfate, and the organic phase was concentrated to obtain 4-((4-methoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, yield 86%) as a white solid.

[0143] LCMS(ESI)m / z:363[M+H] + Step 7: Synthesis of 4-amino-1-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid 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 mixture was concentrated under vacuum to obtain 4-amino-1-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid (250 mg, yield 94%).

[0144] LCMS(ESI)m / z:243[M+H] + Intermediate 85: 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylic acid [ka] Step 1: Synthesis of 4-((tert-butoxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate methyl A solution of 4-amino-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (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 the reaction was complete, the solution was poured into water (5 mL), extracted with ethyl acetate (5 mL), the organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 70:30) to obtain methyl 4-((tert-butoxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (1.00 g, 61% yield) as a white solid.

[0145] LCMS(ESI):376[M+H] + Step 2: Synthesis of 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate methyl Under a nitrogen atmosphere, a mixture of methyl 4-((tert-butoxycarbonyl)amino)-7-chloroimidazo[1,5-a]quinoxaline-8 carboxylate (0.20 g, 0.53 mmol), potassium iron cyanide (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-1,1'-biphenyl-2-yl)palladium(II) (0.17 g, 0.21 mmol), and potassium acetate (0.01 g, 0.07 mmol) in a dioxane / aqueous solution (4.00 mL) was stirred at 100°C for 2 hours. Subsequently, the solution was concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 70:30) to obtain 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate methyl (0.10 g, 51%) as a white solid.

[0146] LCMS(ESI):368[M+H]+ Step 3: Synthesis of methyl 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate To a solution of 4-((tert-butoxycarbonyl)amino)-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate methyl (0.20 g, 0.54 mmol) in dichloromethane (0.50 mL), trifluoroacetic acid (0.10 mL) was added, the reaction mixture was stirred at room temperature for 2 hours, and then concentrated to obtain 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylate methyl (0.05 g, crude product).

[0147] LCMS(ESI):268[M+H] + Step 4: Synthesis of 4-amino-7-cyanoimidazo[1,5-a]quinoxaline-8-carboxylic acid 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), and tetrahydrofuran / methanol / water (1 mL / 1 mL / 1 mL) was stirred at room temperature for 2 hours. After the reaction was complete, the system was concentrated to obtain the crude product 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.

[0148] LCMS(ESI):254[M+H] + Intermediate 86 4-amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid [ka] Step 1: Synthesis of 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl A mixture of 7-chloro-4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (200 mg, 0.5 mmol), potassium carbonate (300 mg, 2 mmol), dichloro[1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) (0.08 g, 0.1 mmol), and trimethylboroxane (0.01 g, 0.10 mmol) l) was stirred in dioxane (2.00 mL) at 100°C for 12 hours, then poured into water, extracted with ethyl acetate (5 mL), dried on anhydrous sodium sulfate, filtered, and purified by column chromatography (petroleum ether:ethyl acetate = 30%) to obtain 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (100 mg, 53%).

[0149] LCMS(ESI):268[M+H] + Step 2: Synthesis of 4-((4-methoxybenzyl)amino)-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid 4-{[(4-methoxyphenyl)methyl]amino}-7-methyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl (500 mg, 1.86 mmol) was dissolved in methanol:tetrahydrofuran:saturated potassium hydroxide aqueous solution (1 mL:1 mL:1 mL), and the reaction mixture was allowed to react at 60°C for 12 hours. After the reaction was complete, it was spin-dried, the pH was adjusted to 3 with formic acid, and the mixture was filtered to obtain 4-((4-methoxybenzyl)amino)-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (52 mg, yield 12%).

[0150] LCMS(ESI):363[M+H] + Step 3: Synthesis of 4-amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid 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 mixture was stirred at 100°C for 12 hours. After the reaction was complete, 4-amino-7-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (520 mg, crude product) was obtained by spin-drying.

[0151] LCMS(ESI):363[M+H] + The following intermediate acids were prepared using the method used in step 3 of intermediate 86: [Table 17] Intermediate 93 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid [ka] Step 1: Synthesis of 6-bromo-2-chloro-3-hydrazinoquinoxaline 6-bromo-2,3-dichloroquinoxaline (278.0 mg, 1.0 mmol) was added to a 25 mL single-port flask at room temperature, and hydrazine hydrate (156 mg, 2.5 mmol, 80% wt) was added. The mixture was reacted 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 obtain 6-bromo-2-chloro-3-hydrazinoquinoxaline (150 mg).

[0152] LCMS(ESI)m / z:273.0 / 275.0[M+H] + Step 2: Synthesis of 8-bromo-4-chloro-[1,2,4]triazolo[4,3-a]quinoxaline At room temperature, 6-bromo-2-chloro-3-hydrazinoquinoxaline (116.0 mg, 0.5 mmol) was first placed in a 25 mL single-port flask, triethyl orthoformate (4.0 mL) was added, and the reaction was heated to 100 °C and allowed to proceed for 1 hour. After confirming that the reaction was complete by LC-MS, the mixture was cooled to room temperature, filtered, and the filtered cake was washed with methanol (3.0 mL x 3) and dried to obtain 8-bromo-4-chloro-[1,2,4]triazolo[4,3-a]quinoxaline (110 mg).

[0153] 1 H NMR (400 MHz, CDCl3) δ 10.21(s, 1H), 8.84(d, 1H), 7.98(d, 1H), 7.90(dd, 1H). Step 3: Synthesis of 8-bromo-N-(4-methoxybenzyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4-amine 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 carried out at 90°C for 4 hours. After the reaction was complete, 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 obtain the crude product 8-bromo-N-(4-methoxybenzyl)-[1,2,4]triazolo[4,3-a]quinoxaline-4-amine (2.20 g).

[0154] LCMS(ESI)m / z:384.1 / 386.1[M+H] + Step 4: Synthesis of 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate methyl 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), potassium acetate (1.7 g, 17.2 mmol) and Pd(dppf)Cl2 (420 mg, 0.57 mmol) were added. The mixture was stirred at 100°C for 12 hours under a CO atmosphere. After spin-off of methanol, 100 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, and spin-dried. The crude product was pulped with ethyl acetate (20 mL) to obtain 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate methyl (1.1 g, yield: 53%).

[0155] LCMS(ESI)m / z:364.2[M+H] + Step 5: Synthesis of methyl 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate.

[0156] Compound 4-((4-methoxybenzyl)amino)-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate methyl (300.0 mg, 0.78 mmol) was placed in a 25 mL single-port 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 the reaction was complete, the reaction mixture was spin-dried to obtain the crude product 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylate methyl (190 mg).

[0157] LCMS(ESI)m / z:244.3[M+H] + Step 6: Synthesis of 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid.

[0158] To the crude product 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid methyl (190 mg, 0.78 mmol) from the previous step, THF (5.0 mL) and methanol (5.0 mL) were added, and the pH was adjusted to 7 with 3 M potassium hydroxide aqueous solution. Then lithium hydroxide (65.0 mg, 1.56 mmol) was added, and the mixture was stirred at 50°C for 16 hours. After waiting for the reaction to complete, methanol and THF were spun off from the system, the pH was adjusted to 6.5 with 1 M dilute hydrochloric acid solution, the mixture was filtered, the filter cake was washed with water (5.0 mL x 3), the filter cake was dried, and the water was removed to obtain the crude product 4-amino-[1,2,4]triazolo[4,3-a]quinoxaline-8-carboxylic acid (120 mg).

[0159] LCMS(ESI)m / z:230.1[M+H] + Intermediate 94 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid [ka] Step 1: Synthesis of 6-bromo-N-(4-methoxybenzyl)-4-methylquinazoline-2-amine 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 mixture was stirred at room temperature for 16 hours. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried on anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The filtrate was purified by column chromatography (0-10% methanol / dichloromethane) to obtain 6-bromo-N-(4-methoxybenzyl)-4-methylquinazoline-2-amine (500 mg, yield: 72%).

[0160] LCMS(ESI)m / z:359.2[M+H] + Step 2: Synthesis of 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazoline-5-amine Compound 6-bromo-N-(4-methoxybenzyl)-4-methylquinazoline-2-amine (0.5 g, 1.396 mmol) was dissolved in DMSO (20 mL) at room temperature, 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, 4.187 mmol) were added. The mixture was stirred at 90°C for 16 hours under nitrogen protection. 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. The mixture was purified by column chromatography (0-10% methanol / dichloromethane) to obtain 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazoline-5-amine (300 mg, yield: 56%).

[0161] LCMS(ESI)m / z:384.2[M+H] + Step 3: Synthesis of 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylate methyl To a solution of 9-bromo-N-(4-methoxybenzyl)imidazo[1,5-c]quinazoline-5-amine (300 mg, 0.785 mmol) in MeOH (10 mL) and DMF (10 mL), potassium acetate (230 mg, 2.36 mmol) and Pd(dppf)Cl2 (58 mg, 0.0785 mmol) were added at room temperature. The mixture was stirred at 100°C for 12 hours under a CO atmosphere, and after the methanol was spun off, 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) was obtained to obtain 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylate methyl (200 mg, yield: 56%).

[0162] LCMS(ESI)m / z:363.2[M+H] + Step 4: Synthesis of methyl 5-aminoimidazo[1,5-c]quinazoline-9-carboxylate Compound 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylate methyl (200 mg, 0.5519 mmol) was placed in a 20 mL round-bottom flask at room temperature, trifluoroacetic acid (2 mL) was added, and the mixture was stirred at 78 °C for 3 hours to spin off the trifluoroacetic acid. After that, water (5 mL) was added, the mixture was filtered, and the filtered cake was washed with water to obtain 5-aminoimidazo[1,5-c]quinazoline-9-carboxylate methyl (200 mg, crude product).

[0163] LCMS(ESI)m / z:243.2[M+H] + Step 5: Synthesis of 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid Compound 5-((4-methoxybenzyl)amino)imidazo[1,5-c]quinazoline-9-carboxylic acid methyl (200 mg, 0.8257 mmol) was dissolved at room temperature in a methanol / tetrahydrofuran / water (9 mL, 4:4:1) mixed solvent. Lithium hydroxide (40 mg, 1.651 mmol) was added, and the mixture was stirred at 78°C for 3 hours. After spinning off the trifluoroacetic acid, water (5 mL) was added, the mixture was filtered, and the filtered cake was washed with water to obtain 5-aminoimidazo[1,5-c]quinazoline-9-carboxylic acid (200 mg, crude product).

[0164] LCMS(ESI)m / z:243.2[M+H]+ Intermediate 95 Synthesis of (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazolo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: Synthesis of 6-bromo-2-chloro-3-hydrazinoquinoxaline 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, followed by 2 hours at room temperature. The mixture was filtered and washed with ethanol (100 mL) to obtain 6-bromo-2-chloro-3-hydrazinoquinoxaline (5.00 g, 18.31 mmol).

[0165] LCMS(ESI):273[M+H] + Step 2: Synthesis of 8-bromo-4-chlorotetrazolo[1,5-a]quinoxaline A mixture of 6-bromo-2-chloro-3-hydrazinoquinoxaline (5.00 g, 18.31 mmol) and sodium nitrite (2.52 g, 36.56 mmol) in an aqueous hydrochloric acid solution (0.2 M, 50.00 mL) was stirred at room temperature for 2 hours. Extraction was performed with ethyl acetate (100 mL x 3), and the organic phase was concentrated under vacuum. 8-bromo-4-chlorotetrazolo[1,5-a]quinoxaline (4.00 g, 18.28 mmol) was obtained by silica gel column chromatography (petroleum ether:ethyl acetate = 70%).

[0166] LCMS(ESI):284[M+H] + Step 3: Synthesis of 8-bromo-N-(4-methoxybenzyl)tetrazolo[1,5-a]quinoxaline-4-amine 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. The mixture was extracted with ethyl acetate (100 mL x 3), the organic phase was concentrated under vacuum, and 8-bromo-N-(4-methoxybenzyl)tetrazolo[1,5-a]quinoxaline-4-amine (3.00 g, 7.81 mmol) was obtained by silica gel column chromatography (petroleum ether:ethyl acetate = 30%).

[0167] LCMS(ESI):385[M+H] + Step 4: Synthesis of 4-((4-methoxybenzyl)amino)tetrazolo[1,5-a]quinoxaline-8-carboxylate methyl 8-Bromo-N-(4-methoxybenzyl)tetrazolo[1,5-a]quinoxaline-4-amine (3.00 g, 7.81 mmol), potassium acetate (1.53 g, 15.61 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (0.63 g, 0.77 mmol) were dissolved in a mixed solution of N,N-dimethylformamide and methanol (1:1,30 mL). The mixture was reacted overnight at 100°C in carbon monoxide (4 mPa) using an autoclave, filtered, extracted with water (100 mL) and ethyl acetate (100 mL x 3), and the organic phase was concentrated under vacuum to obtain 4-((4-methoxybenzyl)amino)tetrazolo[1,5-a]quinoxaline-8-carboxylate methyl (3.00 g, 8.24 mmol).

[0168] LCMS(ESI):365[M+H] + Step 5: Synthesis of methyl 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylate 4-((4-methoxybenzyl)amino)tetrazolo[1,5-a]quinoxaline-8-carboxylate methyl (3.00 g, 8.24 mmol) was dissolved in a 4M dioxane hydrochloride solution (50 mL), stirred at room temperature for 2 hours, extracted with water (100 mL) and ethyl acetate (100 mL x 3), and the organic phase was concentrated under vacuum. 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylate methyl (2.00 g, 8.16 mmol) was obtained by silica gel column chromatography (petroleum ether:ethyl acetate = 50%).

[0169] LCMS(ESI):245[M+H] + Step 6: Synthesis of 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylic acid 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylic acid methyl (2.00 g, 8.16 mmol) and potassium hydroxide (0.91 g, 16.32 mmol) were dissolved in tetrahydrofuran-water-methanol (10 mL:10 mL:10 mL), stirred at 60°C for 2 hours, extracted with water (50 mL) and ethyl acetate (50 mL x 3), and the organic phase was concentrated under vacuum to obtain 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylic acid (1.00 g, 4.34 mmol).

[0170] LCMS(ESI):231[M+H] + Intermediate 96 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid [ka] Step 1: Synthesis of N-(2,6-dichloropyridine-3-yl)-1H-imidazole-5-carboamide Compound 2,6-dichloropyridine-3-amine (163 mg, 1.0 mmol) was dissolved in tetrahydrofuran (5.0 mL), cooled in an ice bath for 5 minutes, and then NaHMDS (0.25 mL, 2.0 M, 2.5 mmol) was added dropwise to the reaction system. After stirring for 1 hour, 5H,10H-diimidazo[1.5-a:1',5'-d]pyrazine-5,10-dione (188.0 mg, 1.0 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction was monitored by LC-MS, and after the reaction was complete, the reaction solution was poured into water to adjust the pH to 7, the solid was precipitated, filtered, the solid was collected, and dried to obtain the crude product N-(2,6-dichloropyridine-3-yl)-1H-imidazole-5-carboamide (175 mg, crude product).

[0171] LCMS(ESI)m / z:257.0[M+H] + Step 2: Synthesis of 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-6(5H)-one The compound N-(2,6-dichloropyridine-3-yl)-1H-imidazole-5-carbamide (256.0 mg, 1.0 mmol) was dissolved in N,N-dimethylacetamide (2.0 mL), potassium carbonate (276 mg, 2.0 mmol) was added, the mixture was heated to 140°C and stirred for 2 hours, and the reaction was confirmed to be complete by LC-MS. The reaction mixture was then cooled to room temperature, poured into water, and stirred for 30 minutes to precipitate the solid. The mixture was filtered, and the solid was recovered to obtain the crude product 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-6(5H)-one (153.0 mg, crude product).

[0172] LCMS(ESI)m / z:221.1[M+H] + Step 3: Synthesis of methyl-6-hydroxyimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl To a solution of 2-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-6(5H)-one (2.0 g, 9.09 mmol) in MeOH (30.00 mL) and DMF (310.00 mL), potassium acetate (1.78 g, 18.18 mmol) and Pd(dppf)Cl2 (660 mg, 0.91 mmol) were added. The mixture was stirred at 100°C for 12 hours under a CO atmosphere. After spinning off methanol, the reaction solution was added to 100 mL of water to precipitate the solid, and the crude product was filtered to obtain a solid. This was pulped with ethyl acetate (50 mL x 2) to obtain the product methyl 6-hydroxyimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (1.5 g, yield: 68%) as a brown solid.

[0173] LCMS(ESI)m / z:245.2[M+H] + Step 4: Synthesis of methyl 6-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate 6-hydroxyimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (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 mixture was heated to 90°C and reacted for 2.5 hours. After confirming that the reaction was complete by LC-MS, the phosphorus oxychloride was filtered off, the reaction mixture was added to ice water, stirred for 15 minutes, filtered, and a filter cake was obtained. The cake was dried to obtain 6-chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (200.0 mg, crude product).

[0174] LCMS(ESI)m / z:263.0[M+H] + Step 5: Synthesis of 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl 6-Chloroimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (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 mixture was heated to 90°C and reacted for 16 hours. After the reaction was complete, the mixture was cooled to room temperature, and the reaction system was added to ice water while stirring. Extraction was performed with ethyl acetate (20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purification by silica gel column chromatography (dichloromethane:methanol = 0-5%) was obtained to obtain 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (250 mg, yield: 91%).

[0175] LCMS(ESI)m / z:364.2[M+H] + Step 6: Synthesis of methyl 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate Compound 6-((4-methoxybenzyl)amino)imidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (250 mg, 0.69 mmol) was dissolved in TFA (5.0 mL), and the reaction mixture was heated to 80°C and reacted for 16 hours. After waiting for the reaction to complete, the mixture was cooled and the TFA in the system was spun off to obtain the crude product 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylate methyl (160 mg, crude product).

[0176] LCMS(ESI)m / z:244.1[M+H] + Step 7: Synthesis of 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid methyl (160.0 mg, 0.66 mmol) was dissolved in a THF / MeOH (1.2 mL / 1.2 mL) mixed solvent, and lithium hydroxide aqueous solution (0.66 mL, 2.0 M, 1.32 mmol) was added. The mixture was heated to 50°C and reacted for 16 hours. After waiting for the reaction to complete, the solvent in the reaction system was spun off, and the pH was adjusted to approximately 7 with 1N hydrochloric acid aqueous solution. The mixture was stirred for 15 minutes, and the solid was collected by filtration to obtain the product 6-aminoimidazo[1,5-a]pyrido[3,2-e]pyrazine-2-carboxylic acid (83 mg, yield: 55%).

[0177] LCMS(ESI)m / z:230.1[M+H] + Intermediate 97 (S)-4-amino-N,6-dimethyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: Synthesis of N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboamide At room temperature, 15 g of 4-bromo-2-fluoro-6-methylaniline (73.89 mmol) was added to a 500 mL round-bottom flask, and 150 mL of tetrahydrofuran was poured into the flask. 74 mL of bis(trimethylsilyl)amide sodium was added at 0°C, and the resulting mixture was stirred at 0°C for 30 minutes. Then, 6.95 g of 5H,10H-diimidazo[1,5-a:1',5'-d]pyrazine-5,10-dione (6.95 g, 36.95 mmol) was added, and the mixture was reacted at room temperature for a further 1 hour. The reaction solution was poured into ice water, filtered, and the residue was used as the product to obtain N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboamide (10 g, yield: 90.9%), a red solid.

[0178] LCMS(ESI)m / z:298[M+H] + Step 2: Synthesis of 8-bromo-6-methylimidazo[1,5-a]quinoxaline-4-ol First, N-(4-bromo-2-fluoro-6-methylphenyl)-1H-imidazole-5-carboamide (10 g, 33.67 mmol) was added to a 250 mL round-bottom flask at room temperature. Next, N,N-dimethylacetamide (100 mL) was poured into the flask, and potassium carbonate (13.94 g, 101.01 mmol) was added. The resulting mixture was stirred at 140 °C for 2 hours. The reaction solution was poured into water to adjust the pH to 3-4, filtered, and the filtrate was used as the product to obtain 8-bromo-6-methylimidazo[1,5-a]quinoxaline-4-ol (8 g, yield: 85.74%), a black solid.

[0179] LCMS(ESI)m / z:278[M+H] + Step 3: Synthesis of 8-bromo-4-chloro-6-methylimidazo[1,5-a]quinoxaline At room temperature, 8-bromo-6-methylimidazo[1,5-a]quinoxaline-4-ol (8 g, 28.88 mmol) was first added to a 250 mL round-bottom flask, then phosphorus oxychloride (100 mL) was poured into the flask, and the resulting mixture was stirred at 120 °C for 12 hours. The reaction mixture was concentrated, spin-dried, poured into ice water, filtered, and the filtration residue was used as the product to obtain 8-bromo-4-chloro-6-methylimidazo[1,5-a]quinoxaline (7.5 g, yield: 88.23%) as a black solid.

[0180] LCMS(ESI)m / z:296[M+H] + Step 4: Synthesis of 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine At room temperature, first, 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, then dimethyl sulfoxide (75 mL) was poured into the flask, and (4-methoxyphenyl)methylamine (4.18 g, 30.51 mmol) and N,N-diisopropylethylamine (6.56 g, 50.85 mmol) were added. The resulting mixture was stirred at 90°C for 2 hours. The reaction mixture was concentrated, spin-dried, poured into ice water, filtered, and the filtration residue was used as the product to obtain 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine (6 g, yield: 60%) as a red solid.

[0181] LCMS(ESI)m / z:397[M+H] + Step 5: Synthesis of 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl At room temperature, 8-bromo-N-(4-methoxybenzyl)-6-methylimidazo[1,5-a]quinoxaline-4-amine (6 g, 15.15 mmol) was first added to a 250 mL high-pressure reactor. Next, N,N-dimethylformamide (30 mL) and methanol (60 mL) were poured into the reactor, potassium acetate (2.97 g, 30.3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.24 g, 1.52 mmol) were added, carbon monoxide (4 MPa) was charged, and the resulting mixture was stirred at 100 °C for 12 hours. The reaction mixture was concentrated, spin-dried, poured into water, filtered, and the filtration residue product yielded 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylate methyl (6 g, yield: 60%) as a red solid.

[0182] LCMS(ESI)m / z:377[M+H] + Step 6: Synthesis of 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl (4 g, 10.61 mmol) was first added to a 250 mL round-bottom flask at room temperature, then tetrahydrofuran (40 mL) and methanol (40 mL) were poured into the flask, potassium hydroxide (1.19 g, 21.22 mmol) was added, the resulting mixture was stirred, and the reaction was carried out at 60°C for 1 hour. The reaction solution was concentrated, spin-dried, poured into water, and the pH was adjusted to around 3 with formic acid until solids precipitated, then filtered, and the filtration residue was used as the product to obtain 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (2.9 g, yield: 76.31%) as a red solid.

[0183] LCMS(ESI)m / z:363[M+H] + Step 7: Synthesis of 4-amino-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid At room temperature, 2.9 g of 4-((4-methoxybenzyl)amino)-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (7.99 mmol) was first added to a 50 mL round-bottom flask, and then 30 mL of trifluoroacetic acid was poured into the flask. The resulting mixture was stirred at 100°C for 3 hours. The reaction solution was concentrated and spin-dried to obtain 1.5 g of 4-amino-6-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (yield: 51.28%) as a black solid.

[0184] LCMS(ESI)m / z:243[M+H] + Intermediate 98 Synthesis of 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid [ka] Step 1: Synthesis of methyl 3-(2-cyclopropyl-1H-imidazole-1-yl)-4-nitrobenzoate In 10 mL of dioxane, 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) was stirred at 100°C for 2 hours. After the reaction was complete, the reaction mixture was poured into 10 mL of water, 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 the crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 10:90) elution to obtain methyl 3-(2-cyclopropyl-1H-imidazole-1-yl)-4-nitrobenzoate (0.50 g, yield: 57%).

[0185] LCMS(ESI):288[M+H] + Step 2: Synthesis of methyl 4-amino-3-(2-cyclopropyl-1H-imidazole-1-yl)benzoate A mixture of methyl 3-(2-cyclopropyl-1H-imidazole-1-yl)-4-nitrobenzoate (1.00 g, 3.48 mmol), bis(boronic 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, then concentrated. The reaction mixture was poured into water (10 mL), 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 by suction, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 53%) elution to obtain methyl 4-amino-3-(2-cyclopropyl-1H-imidazole-1-yl)benzoate (0.60 g, yield 67%).

[0186] LCMS(ESI):257[M+H] + Step 3: Synthesis of 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl A mixture of methyl 4-amino-3-(2-cyclopropyl-1H-imidazole-1-yl)benzoate (500 mg, 1.94 mmol) and carbonyldiimidazole (377 mg, 2.33 mmol) in chlorobenzene (5 mL) was stirred at 140°C for 12 hours, concentrated, and the reaction mixture 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 by suction, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 20%) elution to obtain methyl 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (500 mg, yield: 91%).

[0187] LCMS(ESI):283[M+H] + Step 4: Synthesis of 4-chloro-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylate methyl A solution of 1-cyclopropyl-4-oxo-5,10-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (500.00 mg, 1.77 mmol) in phosphorus oxychloride (5.00 mL) was stirred at 120°C for 12 hours, and then concentrated to obtain 4-chloro-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylate methyl (300 mg, crude product).

[0188] LCMS(ESI):301[M+H] + Step 5: Synthesis of 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylate methyl A solution of 4-chloro-1-cyclopropyl-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylate methyl (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. The reaction mixture was concentrated, poured into water (10 mL), 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 by suction, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (ethyl acetate:petroleum ether = 42%) elution to obtain 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylate methyl (300 mg, yield: 75%).

[0189] LCMS(ESI):402[M+H] + Step 6: Synthesis of 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid A methanol / tetrahydrofuran (5.00 mL / 5 mL) solution of 1-cyclopropyl-4-{[(4-methoxyphenyl)methyl]amino}-10-hydroimidazo[1,5-a]quinoxaline-8-carboxylic acid methyl mixture (300 mg, 0.75 mmol) was mixed with lithium hydroxide (89.45 mg, 2.24 mmol). The reaction mixture was stirred at 60°C for 2 hours, and then concentrated to obtain 1-cyclopropyl-4-((4-methoxybenzyl)amino)imidazo[1,5-a]quinoxaline-8-carboxylic acid (160 mg, yield: 55%).

[0190] LCMS(ESI):388[M+H] + Step 7: Synthesis of 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid A mixed solution 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 concentrated to obtain 4-amino-1-cyclopropylimidazo[1,5-a]quinoxaline-8-carboxylic acid (100 mg, crude product).

[0191] LCMS(ESI):268[M+H] + Example 1 Synthesis of 4-amino-N-(2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide [ka] 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, 0.21 mmol) was dissolved in N,N-dimethylformamide (2.50 mL), and 2-(7-azabenzotriazolyl)-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 to this mixture, and the mixture was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was concentrated and the crude product was purified by high-performance liquid chromatography (HPLC) (column: XBridge BEH Shield RP18 5m, 30mm × 150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 20%B to 36%B for 8 minutes; wavelength: 254 nm / 220 nm); 4-amino-N-(2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (2 mg, 0.01 mmol, yield 3%) was obtained as a white solid.

[0192] 1 H NMR (400 MHz, DMSO-d6) δ 9.19 (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). LCMS(ESI):360.30[M+H] + The examples in the table below can be prepared using the synthesis procedure described in Example 1, by simply substituting the corresponding aldehyde or amine of the starting material. [Table 18] [Table 19] [Table 20] [Table 21] [Table 22] [Table 23] [Table 24] [Table 25] [Table 26] [Table 27] [Table 28] [Table 29] Example 58 (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazolo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: Synthesis of (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazolo[1,5-a]quinoxaline-4-yl)amino)methylidene)-N-methylmethylamine 4-aminotetrazolo[1,5-a]quinoxaline-8-carboxylic acid (131 mg, 0.56 mmol) was dissolved in N,N-dimethylformamide (4.5 mL), and 2-(7-azabenzotriazolyl)-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 reacted in this solution at room temperature for 30 minutes. The solution was diluted with ethyl acetate (30 mL) and water (30 mL), and the aqueous layer was extracted with ethyl acetate (2 × 20 mL). The organic layers were combined, washed with saturated saline solution (30 mL), dried on anhydrous sodium sulfate, and concentrated to obtain (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazolo[1,5-a]quinoxaline-4-yl)amino)methylidene)-N-methylmethylamine (100 mg, 0.19 mmol, yield 33.33%) as a white solid.

[0193] LCMS(ESI):528[M+H] + Step 2: Synthesis of (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)tetrazolo[1,5-a]quinoxaline-8-carboamide (S)-N-((dimethylamino)((8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)tetrazolo[1,5-a]quinoxaline-4-yl)amino)methylidene)-N-methylmethylamine (100 mg, 0.19 mmol) was dissolved in tetrahydrofuran solution (4 mL), and an equal volume of water (4 mL) was added. Then, lithium hydroxide (27 mg, 1.12 mmol) was added. The reaction mixture was concentrated, and the crude product was purified by high-performance liquid chromatography (chromatography column: YMC Triart C18 ExRs 5 m, 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 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-carboamide (3.58 mg, 0.01 mmol, yield 4.41%) as a white solid.

[0194] 1 H NMR (400 MHz, DMSO-d6) δ 12.10 (s, 1H), 8.01-7.95 (d, 2H), 7.74-7.71 (d, 2H), 7.41-7.39 (d, 1H), 7.33(s,1H), 6.50-5.74(m,1H), 4.91-4.73(m,2H), 3.94(s,3H). LCMS(ESI)m / z:430.10[M+H] + Example 59 Synthesis of 4-amino-N-cyclopropyl-N-(7-(5-(trifluoromethyl)pyridine-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridine-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide [ka] Dissolve N-cyclopropyl-7-(5-(trifluoromethyl)pyridine-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridine-4-amine (45 mg, 0.13 mmol) in DMF (3.0 mL), then add 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) to the reaction mixture, stir overnight at room temperature, add water (10 mL), and ethyl acetate (2 Extraction was performed with 0 mL of water, the organic phase was washed with water (10 mL x 2), washed with saturated saline solution (10 mL), dried on anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The crude product was purified by column chromatography (methanol / dichloromethane 0-7%) to obtain compound 4-amino-N-cyclopropyl-N-(7-(5-(trifluoromethyl)pyridine-3-yl)-3,4-dihydro-2H-pyrano[3,2-b]pyridine-4-yl)imidazo[1,5-a]quinoxaline-8-carboxamide (18.0 mg, yield: 25%).

[0195] 1 H NMR (400 MHz, DMSO-d6) δ 9.31 (s, 1H), 9.18 (s, 1H), 9.02 (s, 1H), 8.63 (d, J = 29.1 Hz, 2H),8.24 (s, 1H), 7.93 (s, 1H), 7.77 (s, 1H), 7.56 (s, 2H), 7.22 (d, J = 11.4 Hz, 1H), 5.39 (s, 1H), 4.46 (d, J= 53.9 Hz, 2H), 3.05 (s, 1H), 2.35 (s, 2H), 0.70 (s, 4H). LCMS(ESI)m / z:564.30[M+H]+ The examples in the table below can be prepared using the synthesis procedure described in Example 59, by simply substituting the aldehyde or amine of the corresponding starting material. [Table 30] [Table 31] [Table 32] 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-2-carboamide [ka] (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), chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (0.19 g, 0.68 mmol), and N-methylimidazole (0.11 g, 1.36 mmol) were stirred in N,N-dimethylformamide (1 mL) for 1 hour. After the reaction was complete, the system was cooled to room temperature and extracted three times with ethyl acetate in 10 mL increments. The organic phases were combined, washed with saturated brine (10 mL x 3), dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. The crude product was subjected to high-performance liquid chromatography (chromatography column: YMC Triart C18 ExRs 5 m, 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 over 8 minutes; wavelength: 254 nm / 220 nm; retention time (min): 7.22) to obtain (S)-4-amino-N-(methyl-d3)-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]pyrido[3,4-e]pyrazine-8-carboamide (11.94 mg, yield 6.09%).

[0196] 1H NMR (400 MHz, DMSO-d6) δ 9.31-9.30 (m, 1H), 8.70-8.65 (m, 1H),8.56-8.51 (m, 1H), 7.99 (d, J = 4.0 Hz, 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). LCMS(ESI):431.85[M+H] + The following compounds were prepared using the same method and steps as in Example 75, but with the corresponding raw materials replaced: [Table 33] Example 77 Synthesis of 4-amino-N-(4S)-6-fluoro-1-methylisochroman-4-yl)-N-methylimidazo[1,5-a]quinoline-8-carboamide [ka] 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, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (40 mL), extracted with ethyl acetate (20 mL x 3), combined the organic phases, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the product 4-amino-N-(4S)-6-fluoro-1-methylisochroman-4-yl)-N-methylimidazo[1,5-a]quinoline-8-carboamide (70 mg, yield: 33.7%).

[0197] 1 H NMR (400 MHz, DMSO-d6) δ 9.27 - 9.11 (m, 1H), 8.38 (d, J = 17.2 Hz, 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, 6.2 Hz, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -115.37, -115.39. LCMS(ESI)m / z:406[M+H]+ Example 78 (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide-1-d Example 111 (S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (500 mg, 2.19 mmol) was dissolved in tetrahydrofuran (15 mL) solution, the reaction mixture was cooled to -40°C, n-butyllithium (5 mL, 15.34 mmol) was added over 10 minutes, then bromine (2.5 mL, 21.91 mmol) was added, the mixture was stirred at -40°C for 10 minutes, then transferred to room temperature and stirred for another 10 minutes. After the reaction was complete, the reaction mixture was concentrated under vacuum, dichloromethane was added, and the mixture was filtered to obtain a solid. This was 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, yield: 4.46%) as a white solid.

[0198] LCMS(ESI)m / z:307[M+H] + Step 2: Dissolve 4-amino-1-bromoimidazo[1,5-a]quinoxaline-8-carboxylic acid (10 mg, 0.03 mmol), 2-(7-azobenzotriazolyl)-tetramethyluronium hexafluorophosphate (19 mg, 0.05 mmol), and N,N-diisopropylethylamine (8 mg, 0.05 mmol) in dimethylacetamide (1 ml), add (S)-N-methyl-6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-amine (7 mg, 0.03 mmol), and stir the mixture at room temperature for 2 hours. After the reaction was complete, the organic phases were extracted with ethyl acetate (10 mL x 3), washed with saturated saline solution (10 mL x 3), dried over anhydrous sodium sulfate, and the combined organic phases were concentrated under vacuum to obtain Example 111(S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide (30 mg, 0.03 mmol, yield 80%) as a yellow liquid.

[0199] 1 H NMR (400 MHz, DMSO-d6) δ 9.09 (s, 1H), 7.98 (s, 1H), 7.65 - 7.55 (m, 2H), 7.52 (d, J = 7.9 Hz, 3H), 7.32 (d, J = 7.7 Hz, 1H), 7.25 (s, 1H), 6.42(s,1H),4.79 - 4.72 (m, 2H), 2.75 - 2.62 (m, 3H). LCMS(ESI)m / z:506[M+H] + Step 3: (S)-4-amino-1-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide (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 complete, the organic phases were extracted with ethyl acetate (10 mL x 3), washed with saturated brine (10 mL x 3), dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. The crude product was purified by high-performance liquid chromatography (column: Kinetex 5 m EVO C18, 30 mm × 150 mm; mobile phase A: water (10 mmol ammonium bicarbonate per liter), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (min): 9.35). (S)-4-amino-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide-1-d (3.34 mg, 0.16 mmol, yield 8.45%) was obtained as a white solid.

[0200] 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H),7.92 (d, J = 7.3 Hz, 1H), 7.64-7.62 (d, J = 7.6 Hz, 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). LCMS(ESI)m / z:428[M+H] + The examples in the table below can be prepared using the synthesis procedure described in Example 78, by simply substituting the aldehyde or amine of the corresponding starting material.

[0201] [Table 34] Example 109 Synthesis of 4-amino-N-cyclopropyl-N-(4-(trifluoromethoxy)benzyl)imidazo[1,5-a]quinoxaline-8-carbamide [ka] 4-amino-3-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide (30 mg, 0.06 mmol), potassium acetate (17 mg, 0.18 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (4 mg, 0.01 mmol) were added to a mixed solvent of N,N-dimethylformamide (5 mL) and ethanol (5 mL). The mixture was stirred overnight at 110 °C using an autoclave filled with carbon monoxide (4 mPa). After the reaction was complete, 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 on anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by high-pressure preparative chromatography (column: Sunfire C18 5m, 30mm × 150mm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 37% B to 64% B in 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 5.97) to obtain ethyl 4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylate (4.22 mg, 14%).

[0202] 1H NMR (400 MHz, DMSO-d6) δ 9.36 (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 = 7.1 Hz, 3H). LCMS(ESI): 500.10[M+H] + Example 80: Synthesis of (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: Synthesis of (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine 200 mg, 0.6 mmol of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate was dissolved in 1,4-dioxane hydrochloric acid (4 M, 2 mL) and stirred at room temperature. After the reaction was complete, the reaction mixture was concentrated to obtain the crude product (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine (85 mg, 61% yield).

[0203] LCMS(ESI)m / z:228[M+H] + Step 2: Synthesis of (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboamide 4-amino-7-fluoroimidazo[1,5-a]quinoxaline-8-acid (92 mg, 0.37 mmol) was dissolved in dimethylacetamide (2 mL), and 1-propyl 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) were stirred overnight at room temperature. After the reaction was complete, the mixture was extracted three times in 10 mL portions with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. Column chromatography (dichloromethane:methanol = 10%) yielded (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (35 mg, yield 20%).

[0204] LCMS(ESI)m / z:456[M+H] + Step 3: Synthesis of (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-7-fluoro-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (35 mg, 0.08 mmol) is dissolved in dioxane (1 mL) and [(2-di-tert-butylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methanesulfone Salt (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 complete, the mixture was extracted three times in 10 mL portions with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine, dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. (S)-4-amino-7-fluoro-N-(6-methoxy-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (14.25 mg, yield 44.60%) was obtained by high-pressure preparation (column: XBridge BEH ShieldRP185m, 30 mm × 150 mm; mobile phase A: water (10 mmol / LNH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient from 28% B to 50% B over 8 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 7.88).

[0205] 1H NMR (400 MHz, DMSO-d6) δ 9.12 (s, 1H),8.46-8.29 (m, 1H), 7.92 (d, J = 3.6 Hz, 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 = 10.0 Hz, 3H), 2.70 -2.58(m, 3H). LCMS(ESI)m / z:408.10[M+H] + Example 122 Synthesis of methyl(S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carbamide)-2,3-dihydrobenzofuran-6-carboxylate [ka] Step 1: Synthesis of (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate methyl 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, and the mixture was stirred overnight at 100°C with carbon monoxide (4 MPa). After the reaction was complete, the reaction mixture was concentrated, poured into water, filtered, and the filter cake was pulped with petroleum ether to obtain (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate methyl (70 mg, yield: 77.78%) as a red solid. LCMS(ESI):308[M+H] + Step 2: Synthesis of methyl (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxylate (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2,3-dihydrobenzofuran-6-carboxylate methyl (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 30 minutes. After the reaction was complete, the reaction solution was concentrated to obtain (S)-3-(methylamino)-2,3-dihydrobenzofuran-6-carboxylate methyl (46 mg, yield: 97%) as a black solid.

[0206] LCMS(ESI):208[M+H] + Step 3: Synthesis of (S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carboamide)-2,3-dihydrobenzofuran-6-carboxylate methyl 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (50 mg, 0.22 mmol) was dissolved in N,N-dimethylformamide (4.00 mL) solution. To this solution, 2-(7-azabenzotriazolyl)-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-carboxylate methyl (46 mg, 0.22 mmol) were added, and the mixture was stirred at room temperature for 3 hours. After the reaction was complete, 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 obtain the crude product, which was purified by high-performance liquid chromatography (Sunfire C18 column, 5 m, 30 mm × 150 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: from 5% B to 30% B over 8 minutes; wavelength: 254 nm / 221 nm); (S)-3-(4-amino-N-methylimidazo[1,5-a]quinoxaline-8-carbamide)-2,3-dihydrobenzofuran-6-carboxylate methyl (21 mg, yield: 1%) was obtained as a white solid.

[0207] 1 H NMR (400 MHz, 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) LCMS(ESI):418.20[M+H] + Example 112 Synthesis of (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide [ka] Step 1: Synthesis of (S)-6-bromo-N-methyl-2,3-dihydrobenzofuran-3-amine 100 mg, 0.30 mmol of tert-butyl(S)-(6-bromo-2,3-dihydrobenzofuran-3-yl)(methyl)carbamate was dissolved in a 1,4-dioxane solution of hydrogen chloride (4.0 M, 10 mL). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated under vacuum to obtain (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] + Step 2: Synthesis of (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide 4-aminoimidazo[1,5-a]quinoxaline-8-carboxylic acid (80 mg, 0.35 mmol), 2-(7-azabenzotriazolyl)-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 then (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 the reaction was complete, the mixture was extracted three times in 10 mL portions with ethyl acetate. The organic phases were combined, washed with saturated saline solution (10 mL x 3), dried on anhydrous sodium sulfate, the reaction mixture was spin-dried, and the crude product was obtained by column chromatography (petroleum ether:ethyl acetate = 80:20) to obtain (S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (70 mg, 0.35 mmol, 45.54% yield).

[0208] LCMS(ESI):438[M+H] + Step 3: Synthesis of (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide N-((3S)-6-bromo(2,3-dihydrobenzo[b]furan-3-yl))(4-amino(10-hydroimidazo[1,5-a]quinoxaline-8-yl))-N-methylcarboxamide (30 mg, 0.07 mmol) was dissolved in tetrahydrofuran solution (2 mL), and to this, [Ir(dF(CF3)ppy)2(dpy)]PF6 (8 mg, 0.007 mmol), nickel bromide ethylene glycol dimethyl ether complex (1 mg, 0.003 mmol), PPh3(CF2H)2 (17 mg, 0.14 mmol), and diphenyl o-phenanthroline (2 mg, 0.007 mmol) were added in order. The mixture was reacted overnight at room temperature at 465 nm. After the reaction was complete, the mixture was extracted three times in 10 mL portions with ethyl acetate. The organic phases were combined, washed with saturated saline solution (10 mL x 3), dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under reduced pressure. The crude product was purified by high-performance liquid chromatography (column: Kinetex 5 m EVO C18, 30 mm x 150 mm; mobile phase A: water (10 mmol ammonium bicarbonate per liter), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (min): 9.35). (S)-4-amino-N-(6-(difluoromethyl)-2,3-dihydrobenzofuran-3-yl)-N-methylimidazo[1,5-a]quinoxaline-8-carboamide (0.84 mg, 0.07 mmol, yield 2.96%) was obtained as a white solid.

[0209] 1 H NMR (400 MHz, Methanol-d4) δ 9.11 (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). LCMS(ESI):409[M+H] + Example 115 Synthesis of (R)-4-amino-7-fluoro-N-methyl-N-(7-(1-methylpyrazole-5-yl)benzopyran-4-yl)imidazo[1,5-a]quinoxaline-8-carboamide [ka] Compound 4-amino-7-fluoroimidazo[1,5-a]quinoxaline-8-carboxylic acid (200 mg, 0.813 mmol) was dissolved in DMAc (3 mL) at room temperature, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (187 mg, 0.976 mmol) and 1-hydroxybenzotriazole (133 mg, 0.976 mmol), followed by DIEA (421 mg, 3.264 mmol), were added, and the reaction mixture was stirred at room temperature for 0.5 hours. Next, (R)-N-methyl-7-(1-methylpyrazole-5-yl)benzopyran-4-amine (1.0 g, 4.184 mmol) was added, and the mixture was reacted at room temperature for 16 hours. After confirming the completion of the reaction by LC-MS, the mixture was poured into water (30 mL), extracted with ethyl acetate (10 × 3 mL), washed with saturated saline solution (30 mL), dried the organic phase over anhydrous sodium sulfate, filtered, and concentrated the filtrate under reduced pressure to obtain the crude product. This was separated and purified by column chromatography (0-5% methanol / dichloromethane) to obtain (R)-4-amino-7-fluoro-N-methyl-N-(7-(1-methylpyrazole-5-yl)benzopyran-4-yl)imidazo[1,5-a]quinoxaline-8-carboamide (45.0 mg, yield: 11.75%).

[0210] 1H NMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 22.9 Hz, 1H), 8.42 (d, J = 6.6 Hz, 1H), 7.94 (d, J = 8.6 Hz, 1H), 7.61 (d, J = 12.8 Hz, 2H), 7.46 (dd, J = 6.2, 1.9 Hz, 1H), 7.27 (td, J = 11.3, 7.6 Hz, 2H), 7.14 (dd, J = 7.9, 1.8 Hz, 1H), 6.98 (dd, J = 27.5, 1.8 Hz, 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 = 15.6 Hz, 3H), 2.82 -2.62 (m, 3H), 2.32 (dd, J = 17.9, 7.3 Hz, 1H), 2.13 (dd, J = 7.1, 3.6 Hz, 1H). LCMS(ESI)m / z:472.1[M+H] + The examples in the table below can be prepared using the synthesis procedure described in Example 115, by simply substituting the aldehyde or amine of the corresponding starting material. [Table 35] [Table 36] 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-carboamide [ka] Step 1: (S)-4-amino-3-bromo-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide (60 mg, 0.12 mmol) was dissolved in dimethylformamide (2 mL) with methanol (2 mL) added. The mixture was then placed in an autoclave with dichloro[1,1'-bis(diphenylphosphino)ferrocene], palladium (3 mL, 0.004 mmol), and potassium acetate (23 mg, 0.24 mmol), and stirred at 100 °C for 16 hours under a carbon monoxide atmosphere of 4 MPa. After the reaction was complete, the reaction mixture was poured into water and extracted three times with ethyl acetate in 10 mL portions. The organic phases were combined, washed with saturated saline solution (10 mL x 3), dried over anhydrous sodium sulfate, concentrated under vacuum, and purified by silica gel column chromatography (alkaline water:acetonitrile = 1:1) to obtain (S)-4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylate methyl (40 mg, yield: 69.53%) as a brown solid.

[0211] LCMS(ESI):485.42[M+H] + Step 2: (S)-4-amino-8-(methyl(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)carbamoyl)imidazo[1,5-a]quinoxaline-3-carboxylate methyl (30 mg, 0.06 mmol) was dissolved in methanol (2 mL) solution, and sodium borohydride (5 mg, 0.12 mmol) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, it was extracted three times in 10 mL portions with ethyl acetate. The organic phases were combined, washed with saturated brine (10 mL x 3), dried on anhydrous sodium sulfate, and the combined organic phases were concentrated under vacuum. The substance was purified by high-performance liquid chromatography (column: Kinetex 5 m EVO C18, 30 mm × 150 mm; mobile phase A: water (10 mmol ammonium bicarbonate per liter), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (min): 9.35). (S)-4-amino-3-(hydroxymethyl)-N-methyl-N-(6-(trifluoromethyl)-2,3-dihydrobenzofuran-3-yl)imidazo[1,5-a]quinoxaline-8-carboamide (3.17 mg, yield 11.13%) was obtained as a white solid.

[0212] 1 H NMR (400 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.31 (s, 1H), 7.64 (d, J = 7.8 Hz, 1H), 7.59 (s, 2H), 7.47 (d, J = 5.5 Hz, 2H), 7.34 (d, J = 7.8 Hz, 1H), 7.27 - 7.23 (m, 1H), 6.36 (t, J = 5.2 Hz, 1H), 4.85 (d, J = 5.1 Hz, 3H), 4.73 (d, J = 4.6 Hz, 1H), 3.31 (s, 1H),2.67(S,3H). LCMS(ESI):457.41[M+H] + 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-carbamide [ka] 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}-4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboxylate methyl (30 mg, 0.06 mmol) was stirred at 60°C for 2 hours in a methanol (0.50 mL) / tetrahydrofuran (0.50 mL) / ammonia (0.5 mL) solution. After the reaction was complete, the reaction mixture was poured into water (5 mL), extracted with ethyl acetate, and spin-dried to obtain the crude product. This was purified by high-pressure preparative chromatography (column: C18 silica gel; mobile phase: water in acetonitrile; gradient: 10% to 50% over 10 minutes; detector: UV 254 nm) to obtain 8-{N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)]-N-methylcarbamoyl}4-amino-10-hydroimidazo[1,5-a]quinoxaline-3-carboamide (1.06 mg, 4%) as a white solid.

[0213] 1 H NMR (400 MHz, DMSO-d6) δ 10.01 (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 = 7.9 Hz, 1H), 7.54 (d, J = 8.2 Hz, 1H), 7.48 (d, J = 11.1 Hz, 1H), 7.34 (d, J = 7.8 Hz, 1H), 7.27 (s, 1H), 6.43 (s, 1H), 4.73 (s, 2H), 2.76 - 2.63 (m, 3H). LCMS(ESI):470.80[M+H]+ 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-carboamide [ka] 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), pyridine (10.09 mg, 0.13 mmol) and trifluoroacetic anhydride (26.79 mg, 0.26 mmol) were added, and the mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction mixture was poured into water (5 mL), and the crude product was spin-dried on ethyl acetate. The crude product was purified by high-pressure preparative chromatography (column: C18 silica; mobile phase: water in acetonitrile; gradient: 10% to 50% over 10 minutes; detector: UV 254 nm) to obtain N-[(3S)-6-(trifluoromethyl)(2,3-dihydrobenzo[b]furan-3-yl)](4-amino-3-cyano(10-hydroimidazo[1,5-a]quinoxaline-8-yl))-N-methylformamide (1.61 mg, 5%) as a white solid.

[0214] 1 H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.48 (s, 1H), 7.64 (d, J = 8.7 Hz, 3H), 7.33 (s, 1H), 7.26 (s, 1H), 7.12 (s, 2H),6.40(s,1H), 4.72 (d, J = 10.5 Hz, 1H),4.71(s,1H), 2.67 (d, J = 2.8 Hz, 3H). LCMS(ESI):452.80[M+H] + Examples 81 & 82: Synthesis of Example 81(R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide and Example 82(S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide [ka] Compound 4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide (90 mg) was placed in SFC (column: DAIEL CHIRALCEL). The samples were purified using an OJ (250mm x 30mm, 10μm) under the conditions of CO2-i-PrOH (0.1% NH3-H2O), 50% / 50%, and a flow rate of 80mL / min, to obtain (R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide (34.61mg) and 82(S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide (28.15mg).

[0215] Example 81 (R)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carboamide LCMS(ESI)m / z:465.3[M+H]+ 1H NMR (400 MHz, DMSO-d6) δ 9.18 (s, 1H), 8.37 (d, J = 1.8 Hz, 1H), 7.91 (s, 1H), 7.59 (dd, J = 8.4, 1.8 Hz, 1H), 7.55 - 7.39 (m, 4H), 7.18 - 7.09 (m, 2H), 5.86 (dd, J = 9.3, 4.1 Hz, 1H), 4.81 (t, J = 9.7 Hz, 1H), 4.65 (dd, J = 10.2, 4.2Hz, 1H), 2.90 (p, J = 3.2 Hz, 1H), 0.45 - 0.26 (m, 2H), 0.24 - 0.03 (m, 2H). Example 82 ( S)-4-amino-N-(6-bromo-2,3-dihydrobenzofuran-3-yl)-N-cyclopropylimidazo[1,5-a]quinoxaline-8-carbamide LCMS(ESI)m / z:465.3[M+H]+ 1 H NMR (400 MHz, DMSO-d6) δ 9.24 (d, J = 3.6 Hz, 1H), 8.40 (d, J = 3.5 Hz, 1H), 8.03 (s, 1H), 7.85 (s, 2H), 7.62 (dd, J = 8.4, 3.3 Hz, 1H), 7.47 (dd, J = 8.2, 3.3 Hz, 2H), 7.13 (dd, J = 8.7, 3.3 Hz, 2H), 5.86(dt, J = 8.6, 4.0 Hz, 1H), 4.80 (td, J = 9.7, 3.6 Hz, 1H), 4.65 (dt, J = 9.2, 4.1 Hz, 1H), 2.90 (d, J = 6.7 Hz,1H), 0.44 - 0.26 (m, 2H), 0.13 (ddd, J = 45.3, 10.6, 4.8 Hz, 2H). Using the procedures described in Examples 81 and 82 and the corresponding chiral SFC resolution conditions, the compounds shown in the table below were obtained: [Table 37] [Table 38] [Table 39] [Table 40] [Table 41] [Table 42] Biochemical evaluation 1. Assay of compound inhibitory activity against tumor cell proliferation Experimental Example 1: Assay on the inhibitory activity of a compound on the proliferation of HCT-116 MTAP(- / -)-deficient cells. Materials and Cells: HCT-116 MTAP(- / -) deficient cells were purchased from Kang Yuan Bochuang (China); RPMI-1640 medium, fetal bovine serum, and penicillin-streptomycin were purchased from Thermo Fisher (USA); 384-well plates were purchased from PerkinElmer (USA); and Cell-Titer Glo kits were purchased from Promega (USA).

[0216] Cell culture: HCT116 MTAP(- / -) deficient cells were cultured in RPMI1640 medium containing 10% fetal bovine serum + 1% penicillin-streptomycin at 37°C and 5% CO2. Cells in the logarithmic growth phase should be used for experimental purposes only.

[0217] Cell proliferation inhibitory activity assay: The proliferation inhibitory activity of the compound against HCT-116 MTAP(- / -) deficient cells was detected using the Cell-Titer Glo kit. Cell concentrations were adjusted, and 40 μL / well was inoculated into a 384-well plate and incubated overnight at 37°C and 5% CO2. 40 nL of the compound was added to each well to achieve a final concentration of 0-10,000 nM (starting concentration 10,000 nM, 3-fold dilution, 10 spots). The DMSO content was 0.1%. The cell plates were incubated at 37°C and 5% CO2 for 8 days. Cell activity was measured by adding 40 μL of Cell-Titer Glo reagent. The assay results are shown in Table 1.

[0218] Experimental Example 2: Assay on the inhibitory activity of compounds on the proliferation of HCT-116 wild-type cells Materials and Cells: HCT-116 MTAP(- / -) wild-type cells were purchased from Kang Yuan Bochuang (China); RPMI-1640 medium, fetal bovine serum, and penicillin-streptomycin were purchased from Thermo Fisher (USA); 384-well plates were purchased from PerkinElmer (USA); and the Cell-Titer Glo kit was purchased from Promega (USA).

[0219] Cell culture: HCT116 wild-type cells were cultured in RPMI1640 medium containing 10% fetal bovine serum + 1% penicillin-streptomycin at 37°C and 5% CO2. Cells in the logarithmic growth phase should be used for experimental purposes only.

[0220] Cell proliferation activity assay: The growth inhibitory activity of compounds against HCT-116 wild-type cells was detected using the Cell-Titer Glo kit. Cell concentrations were adjusted, and 40 μL / well was inoculated into a 384-well plate and incubated overnight at 37°C and 5% CO2. 40 nL of the compound was added to each well to achieve a final concentration of 0-10,000 nM (starting concentration 10,000 nM, 2-fold dilution, 10 spots). The DMSO content was 0.1%. The cell plates were incubated at 37°C and 5% CO2 for 8 days. Cell activity was measured by adding 40 μL of Cell-Titer Glo reagent. The assay results are shown in Table 1.

[0221] Table 1 below shows the inhibitory activity of the compounds used in the examples against the proliferation of HCT116 MTAP(- / -)-deficient cells, as well as HCT116 wild-type cells.

[0222] Table 1 [Table 43] [Table 44] [Table 45] [Table 46] 2. Mouse pharmacokinetic experiments Test substance This test substance is derived from a specific example compound of the present invention, and the reference compound AMG473 is a compound of Embodiment 473 of Amgen's patent WO2022 / 169948A1.

[0223] Laboratory animals ICR mouse, male, N=3 / group; Origin: Zhejiang Vital River Laboratory Animal Technologies Co. Ltd. Preparation and administration of pharmaceuticals Single oral (PO) administration to ICR mice: Each compound was weighed, dissolved in dimethyl sulfoxide, a fixed amount of polyethylene glycol 400 and sterile water for injection were added, and a clarified solution was prepared with a small amount of 1 mol / L hydrochloric acid aqueous solution. Three mice were fasted overnight and then orally administered 10 mg / kg.

[0224] ICR mice were administered a single dose by intravenous injection (IV): Each compound was weighed, dissolved in dimethyl sulfoxide, a fixed amount of polyethylene glycol 400 and sterile water for injection were added, and a clarified solution was prepared with a small amount of 1 mol / L hydrochloric acid solution; three mice were fasted overnight and then administered a dose of 3 mg / kg via tail vein injection.

[0225] Sample collection Blood was collected from the dorsal pedis vein at approximately 30 μL / time, treated with dipotassium ethylenediaminetetraacetate for anticoagulation, placed on ice after collection, and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 4000 g / min, 5 minutes, 4°C). Blood collection times were 0.0833 (intravenous injection), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. Samples were stored in a refrigerator at -20°C.

[0226] For plasma samples, 30 μL (10 μL sample + 20 μL blank plasma sample) was mixed with 200 μL of ice-cold acetonitrile containing an internal standard, vortexed for 30 seconds, and then centrifuged at 4000 g / min for 20 minutes. 100 μL of the supernatant was transferred to a 96-well plate, 200 μL of ultrapure water was added, and vortexed for 30 seconds. 5 μL or 10 μL was then injected into an LC-MS / MS for analysis.

[0227] Table 2: Pharmacokinetic data [Table 47] "NA": Not applicable.

[0228] By comparing pharmacokinetic parameters, it can be seen that all embodiments of the present invention, at the same dose and administration method, result in higher plasma exposure compared to AMG473, a reduced starting dose of the compound, and a wider safety window.

[0229] 3. hERG ion channel inhibition assay Test substance This test substance is derived from a specific example compound of the present invention, and the reference compound AMG473 is a compound of Embodiment 473 of Amgen's patent WO2022 / 169948A1.

[0230] Cell lines and cell cultures The HEK293 cell line (K1236), which stably expresses the hERG ion channel, was 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 blastosidine, and 400 μg / mL Geneticin. When the cell density grew to 40-80% of the bottom area of ​​the culture dish, the cells were digested with trypsin and passaged three times a week. Prior to the experiment, cells were cultured in a 3.5 cm culture dish at a rate of 5 × 10⁶. 5 The cells were cultured at a specific density and induced with 1 μg / mL doxycycline for 48 hours. The cells were then digested, implanted onto slides, and subjected to subsequent manual patch-clamp experiments.

[0231] Experimental Procedure 1) Place a small slide containing HEK293 cells in a culture dish onto the perfusion tank of the microscope stand. 2) Position a suitable cell tune in the center of the field of view of an Olympus IX71 or IX73 inverted microscope, locate the tip of the glass electrode using a 10x objective lens, and position it in the center of the field of view. Next, move the electrode downward using a micromanipulator while coarsely adjusting the focus so that the electrode slowly approaches the cell. 3) Once close to the cell, switch the objective lens to 40x and observe, gradually moving the electrode closer to the cell surface through the fine adjustment gear of the micromanipulator. 4) Apply negative pressure to form a seal with a resistance of 1 GΩ or more between the electrode tip and the cell membrane. 5) Compensate for instantaneous capacitive current Cfast in voltage clamp mode. Then, break the membrane by repeatedly applying short bursts of negative pressure to obtain a whole-cell recording pattern. 6) Clamp the membrane potential to -60 mV to compensate for slow capacitive current Cslow, cell membrane capacitance (Cm), and input membrane resistance (Ra), respectively. 7) After the cells stabilized, the clamp voltage was changed to -90mV, the sampling frequency was set to 20kHz, and the filter frequency to 10kHz. Leakage current was detected with a clamp voltage of -80mV over a time range of 500ms. 8) hERG current was tested by depolarizing the membrane potential from -80mV to +30mV over 4.8 seconds using a depolarization command voltage, and then lowering the membrane potential to -50mV over 5.2 seconds using a repolarization voltage to deactivate the channel and allow observation of the hERG tail current. The peak of the tail current represents the magnitude of the hERG current. 9) The hERG current used for detecting the test substance was continuously recorded for 120 seconds before administration to evaluate the stability of the hERG current generated from the test cells. Only stable cells within the acceptable range of the evaluation criteria were allowed to proceed to the subsequent compound assay. 10) Testing of hERG current inhibition by the test substance: hERG current measured in extracellular fluid containing 0.1% DMSO was used as the baseline for the assay. After the hERG current stabilized for at least 5 minutes, the solution containing the test substance was sequentially perfused around the cells from low to high concentrations. After each perfusion, approximately 5 minutes were allowed for the compound to reach the cells, and the hERG current was recorded simultaneously. After the recorded current stabilized, the last five hERG current values ​​were recorded and averaged to obtain the final current value at a specific concentration.After testing the compounds, 450 nM dofetilide was added to the same cells to completely inhibit the current and serve as a positive control for the cells. The positive compound, dofetilide, was also assayed simultaneously with the test drug experiment using the same membrane clamp system before and after completion to confirm the reliability and sensitivity of the entire assay system. The above test procedure was repeated with two separate test cell groups (n=2).

[0232] Data Analysis Only data that meets the above conditions can be analyzed as follows: Note: Data is exported by PatchMaster software. 1) After perfusion with a blank solvent or compound gradient solution, stabilize the current values ​​five times in a row, and the average value is taken as the "tail current". ブランク "Tail current 化合物 The current suppression rate was calculated using the following formula. [ka] 2) Fit the dose-response curve and use Graphpad Prism 8.0 software for IC 50 The values ​​were calculated. 3) The range of standard deviations for the three datasets is less than 15 (SD<15); 4) The widely accepted criteria for evaluating the inhibitory efficacy of hERG channel assay compounds are as follows: 1) Low inhibitory effect: IC 50 >10μM 2) Moderate inhibitory effect: 1 μM <IC 50 <10μM 3) High inhibitory effect: IC 50 <1μM Data Quality Management Standards Subsequent analysis can only be performed if the following criteria are met: 1) Initial seal resistance is greater than 1 GΩ; 2) Series resistance is less than 15 MΩ and series resistance voltage error is less than 5 mV; 3) Leakage current at detection voltage is less than 50% of the current value in that state; 4) Tail current is greater than the plateau current before the pulse and the initial tail current value is greater than 250 pA; 5) Access resistance Ra is less than 15 MΩ; 6) Tail current decay rate per minute is less than 2.5%.

[0233] hERG IC 50 The (μM) data is shown in Table 3.

[0234] Table 3: hERG IC 50 data [Table 48] hERG IC 50 Comparing the data, it can be seen that the embodiment of the present invention has a lower risk of cardiotoxicity and a wider safety window compared to AMG473.

[0235] While preferred embodiments have been described above, it will be apparent to those skilled in the art that modifications are possible without departing from the present invention. Such modifications are considered to be variations that may fall within the scope of the present invention.

Claims

1. Compounds having the following structure, pharmaceutically acceptable salts thereof, or isotopic derivatives. 【Chemistry 1】

2. A pharmaceutical composition comprising the compound described in Claim 1, a pharmaceutically acceptable salt thereof, or an isotopic derivative thereof, The pharmaceutical composition further comprises a second active substance, wherein the second active substance is an antitumor drug.

3. The pharmaceutical composition according to claim 2, wherein the antitumor agent comprises one or more of a chemotherapeutic agent, a targeted anticancer agent, or an antibody anticancer agent.

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