Heterocyclic compound, preparation method therefor and pharmaceutical use thereof

Abstract

The present invention relates to a heterocyclic compound, a preparation method therefor and a pharmaceutical use thereof. Specifically, the present invention relates to a compound as shown in general formula (I), a preparation method therefor, a pharmaceutical composition comprising the compound, and a use thereof as a PRMT5 inhibitor. The compound and the pharmaceutical composition comprising the compound can be used for treating and / or preventing PRMT5 activity-related diseases, such as non-small cell lung cancer, tumor of mesothelial tissue, neurofibrosarcoma, pancreatic cancer, and solid tumors. The definition of each group in general formula (I) is the same as that in the description.

Description

Heterocyclic compounds, preparation methods and medical uses thereof Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to bicyclic heterocyclic compounds, methods for preparing the same, and pharmaceutical compositions containing the same, as well as their use as PRMT5 inhibitors in the treatment and / or prevention of diseases associated with PRMT5 activity. Background Art

[0002] Epigenetic regulation of gene expression is a crucial biological factor in protein production and cell differentiation, playing a significant role in the pathogenesis of many human diseases. Post-translational modifications (PTMs) are key to proteome diversity. Modifications at one or more sites on a protein can determine its conformation, subcellular localization, interactions with other proteins, stability, and activity. These PTMs are mediated by a variety of enzymes, including phosphorylation, acetylation, ubiquitination, methylation, and hydroxylation. They are also negatively regulated by enzymes such as phosphatases, deubiquitinating enzymes, deacetylases, and demethylases. Arginine methylation, among others, plays a crucial role in cellular processes, including cell signaling, gene transcription, RNA processing, and DNA recombination and repair.

[0003] Protein arginine methyltransferases (PRMTs) catalyze the methylation of specific arginine residues by transferring a methyl group from S-adenosylmethionine (SAM) to the guanidine nitrogen of arginine. PRMTs can be divided into three categories based on the specific mode of arginine methylation: type I (PRMT1, 2, 3, 4, 6, and 8) catalyze asymmetric dimethylation, type II (PRMT5 and PRMT9) catalyze symmetric dimethylation, and type III (PRMT7) catalyzes monomethylation. PRMT5 plays an essential role in cell growth and development and homeostatic hematopoiesis, participating in the survival and renewal of stem cells in the neural, muscle, hematopoietic, and reproductive systems. PRMT5 knockout in mice is embryonic lethal, while conditional PRMT5 deletion results in the absence of hematopoietic progenitor cells and lethal bone marrow atrophy. Furthermore, PRMT5-deficient hematopoietic stem and progenitor cells exhibit severe cytokine signaling impairment and upregulation of p53. PRMT5 interacts with the cofactor MEP50, enhancing its binding to SAM and substrates, as well as its methylation activity.

[0004] The methylation function of PRMT5 is closely related to tumorigenesis and progression through the regulation of gene expression, mRNA splicing, DNA damage response, cytokine signaling pathways, and tumor immunity. Numerous studies have confirmed that PRMT5 is overexpressed in different types and aggressive cancers, such as glioma, leukemia, B-cell and T-cell lymphoma, metastatic melanoma, breast cancer, prostate cancer, and colon cancer.

[0005] PRMT5 exerts important biological functions by epigenetically regulating the expression of target genes or directly methylating key signaling molecules through the methylation of various proteins (including histones and non-histones). PRMT5 methylates the terminal arginine residues of histones to activate or repress the expression of related genes. PRMT5 modification of histones often leads to the silencing of tumor suppressor genes such as p53, ST7, NM23, and Rb, thereby promoting the development and progression of tumors.

[0006] Overexpression of PRMT5 in tumor cells can promote tumor progression by inhibiting the expression of histone-dependent oncogenic miRNAs. For example, in B-cell lymphoma, PRMT5 inhibits miR-33b / 96, which upregulates cyclin D1 and c-Myc; in lung cancer, PRMT5 inhibits miR-99, which upregulates FGFR3; and in AML, PRMT5 inhibits miR-29b, which upregulates FLT3. PRMT5 methylates H4R3 and H3R8, promoting FGFR3 and eIF4E expression in colorectal cancer and AR expression in prostate cancer. PRMT5-methylated H3R2 participates in transcriptional activation, for example, by recruiting WDR5 and MLL coactivators, leading to trimethylation of H3K4, initiating FOXP1 expression to maintain breast cancer stem cell activity, and activating the transcription of redox-related genes. PRMT5 can also activate or repress the expression of related genes in a histone-independent manner. STRAP recruits PRMT5 in response to DNA damage, methylating p53 to alter its nuclear distribution and the expression of its target genes p21 and PUMA. PRMT5 methylation of p53 promotes lymphomagenesis. PRMT5 can also directly methylate E2F-1 and NF-KB / P65, inducing the expression of their target genes.

[0007] PRMT5, as part of the spliceosome, is responsible for pre-mRNA splicing in the spliceosome and can affect mRNA splicing, transport, and degradation through methylation. Loss of Myc or PRMT5 leads to aberrant splicing (exon skipping or intron retention) of genes associated with cell cycle arrest or apoptosis. PRMT5 drives Myc-mediated lymphomagenesis by methylating the splicing factor SRSF1. The splicing factor E2F-1 is also an important substrate of PRMT5. PRMT5-mediated splicing regulates TIP60 / KAT5 to promote homologous recombination and genomic integrity.

[0008] PRMT5 also plays a crucial role in tumor-driven growth factor signaling pathways. In lung and colon cancer, PRMT5 promotes tumor development by transcriptionally activating FGFR genes. PRMT5 can also directly arginine methylate growth factors such as EGFR, PDGFR, and TFG-β. These signaling pathways are crucial for cancer cell proliferation, differentiation, and survival. Furthermore, PRMT5 regulates the assembly of DDR complexes and the expression of related genes through post-transcriptional modifications.

[0009] Loss of tumor suppressor genes is a key driver of tumorigenesis. Loss of tumor suppressor genes often leads to co-deletion of genes adjacent to these tumor suppressor genes. Loss of the tumor suppressor gene CDKN2A at chromosome 9p21 occurs in 15% of human tumors and results in co-deletion of MTAP, a key enzyme in the methionine and adenine salvage pathways. Loss of MTAP leads to elevated levels of MTA, which is structurally similar to the methyl donor SAM, a substrate for the type II methyltransferase PRMT5. Elevated MTA competes with SAM for PRMT5 binding, rendering the methyltransferase hypomorphic and susceptible to further inhibition by PRMT5. Multi-gene panel silencing screens across various tumor cell lines have revealed a strong correlation between MTAP loss and PRMT5 dependency. However, PRMT5 is a known essential gene. Conditional knockout of PRMT5 and siRNA interference studies have shown that PRMT5 inhibition in normal tissues may lead to adverse effects such as pancytopenia, infertility, skeletal muscle atrophy, and cardiac hypertrophy. Therefore, new strategies are needed to exploit this metabolic vulnerability and preferentially target PRMT5 in MTAP-deficient tumors while retaining PRMT5 activity in normal tissues. Targeting PRMT5 with MTA-synergistic small molecule inhibitors can preferentially target the MTA-bound state of PRMT5. In MTAP-deficient tumor cells, the MTA-bound state of PRMT5 is more enriched, providing a therapeutic window superior to that of normal cells. Summary of the Invention

[0010] After intensive research, the inventors designed and synthesized a series of bicyclic heterocyclic compounds and screened them for PRMT5 activity. The results showed that these compounds are potent and selective inhibitors of the PRMT5 / MTA complex, with selective inhibitory effects on MTAP-deficient cells, and can be developed as drugs for the treatment and / or prevention of diseases associated with PRMT5 activity.

[0011] Therefore, the object of the present invention is to provide a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof,

[0012] in:

[0013] is a single bond or a double bond;

[0014] X 1 CR 5 , CR 5 R 6 、N、NR 5 or C=O;

[0015] X 2 CR 7 , CR 7 R 8 、N、NR 7 or C=O;

[0016] X 3 、X 4 are each independently selected from CH2, O, S or NH;

[0017] X 5 、X 6 are each independently selected from a bond, CH2, O, S or NH;

[0018] Ring A is selected from saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclyl, phenyl or heteroaryl;

[0019] Each R 1 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a , -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p-NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a wherein said alkyl, alkenyl, alkynyl, heteroalkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted by one or more R 1a replace;

[0020] Each R 2 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, heteroalkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein said alkyl, heteroalkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally replaced by one or more R 2a replace

[0021] Each R 3 each independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;

[0022] Each R 4 Each is independently selected from hydrogen, halogen, hydroxy, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0023] Each R 1a are independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, heteroalkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a , -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a ;

[0024] Each R 2a independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, heteroalkyl, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a ;

[0025] Each L is independently -O-, -NH-, alkylene, wherein the alkylene is optionally substituted with one or more groups selected from hydroxy, hydroxyalkyl and heteroaryl;

[0026] R 5 、R 6 Each is independently selected from hydrogen, deuterium, halogen, hydroxy, alkyl, hydroxyalkyl, aminoalkyl, haloalkyl, cyano, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NRa R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R b The alkyl, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0027] R 7 、R 8 Each is independently selected from hydrogen, deuterium, halogen, hydroxy, alkyl, hydroxyalkyl, aminoalkyl, haloalkyl, cyano, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NR a R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R bThe alkyl, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0028] Or, R 5 or R 6 With R 7 or R 8 and the atoms to which they are attached together form a saturated or partially saturated cycloalkyl, a saturated or partially saturated heterocyclyl, an aryl or a heteroaryl, wherein the cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more groups selected from deuterated, halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, deuterated alkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0029] R a and R b Each is independently selected from hydrogen, halogen, hydroxy, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or,

[0030] R a and R b together with the nitrogen atom to which it is attached, form a nitrogen-containing heterocyclic group, which is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl;

[0031] m1 is 0, 1, 2, 3, or 4;

[0032] m2 is 0, 1, 2, or 3;

[0033] m3 is 0, 1, 2 or 3;

[0034] m4 is 0, 1, or 2;

[0035] t is 0, 1, or 2;

[0036] p is an integer from 1 to 6.

[0037] In a preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0038] is a double bond;

[0039] X 1 CR 5 or NR 5 ;

[0040] X 2 CR 7 or NR 7 ;

[0041] R 5 、R 7 As defined in general formula (I).

[0042] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0043] is a double bond;

[0044] X 1 CR 5 or NR 5 ;

[0045] X 2 CR 7 or NR 7 ;

[0046] R 5 Selected from hydrogen, deuterium, halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Haloalkyl, cyano, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; said C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10Aryl and 5-6 membered heteroaryl are optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;

[0047] R 7 Selected from hydrogen, deuterium, halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Haloalkyl, cyano, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; said C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl are optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0048] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0049] is a double bond;

[0050] X 1 CR 5 or NR 5 ;

[0051] X 2 CR 7 or NR 7 ;

[0052] R 5 With R 7 and the atoms to which they are attached together form a saturated or partially saturated C5-C6 cycloalkyl, a saturated or partially saturated 5- to 6-membered heterocyclyl, a phenyl or a 5- to 6-membered heteroaryl, wherein the C5-C6 cycloalkyl, the 5- to 6-membered heterocyclyl, the phenyl or the 5- to 6-membered heteroaryl is optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, deuterated alkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0053] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0054] is a double bond;

[0055] X 1 CR 5 or NR 5 ;

[0056] X 2 CR 7 or NR 7 ;

[0057] R 5 With R 7 and the atoms to which it is attached together form a saturated or partially saturated C5-C6 cycloalkyl, a saturated or partially saturated 5- to 6-membered heterocyclic group, a phenyl group or a 5- to 6-membered heteroaryl group, preferably a cyclopentyl group, a cyclohexyl group, a dihydrofuranyl group, a pyrazolyl group; which is optionally selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Halogenated alkoxy, C 1-6 Cyanoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5- to 6-membered heterocyclic group, C 6-10 The alkyl group is substituted by one or more groups of aryl or 5- to 10-membered heteroaryl.

[0058] In another preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0059] is a double bond;

[0060] X 1 CR 5 or NR 5 ;

[0061] X 2 CR 7 or NR 7 ;

[0062] R 5 With R 7 and the atoms to which it is attached together form a saturated or partially saturated C5-C6 cycloalkyl, a saturated or partially saturated 5- to 6-membered heterocyclic group, a phenyl group or a 5- to 6-membered heteroaryl group, preferably a cyclopentyl group, a cyclohexyl group, a dihydrofuranyl group, a pyrazolyl group; which is optionally replaced by a C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 substituted with cyanoalkyl.

[0063] In another preferred embodiment, the compound of formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

[0064] in:

[0065] R 5 Selected from hydrogen, deuterium, halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Haloalkyl, cyano, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; said C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl are optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;

[0066] R 7 Selected from hydrogen, deuterium, halogen, hydroxyl, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Haloalkyl, cyano, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl; said C 1-6 Alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, C 6-10Aryl and 5-6 membered heteroaryl are optionally substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

[0067] Ring A, X 3 、X 4 、X 5 、X 6 、R 1 、R 2 、R 3 、R 4 , m1, m2, m3, m4, and t are as defined in the general formula (I).

[0068] In another preferred embodiment, the compound of formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

[0069] in:

[0070] R 5 selected from hydrogen;

[0071] R 7 Selected from C 1-6 alkyl;

[0072] Ring A, X 3 、X 4 、X 5 、X 6 、R 1 、R 2 、R 3 、R 4 , m1, m2, m3, m4, and t are as defined in the general formula (I).

[0073] In another preferred embodiment, the compound of formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

[0074] in:

[0075] Ring E is selected from saturated or partially saturated 5-6 membered cycloalkyl, saturated or partially saturated 5-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl;

[0076] Each R 9 are each independently selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl;

[0077] s is 0, 1, 2, or 3;

[0078] Ring A, X 3 、X 4 、X 5 、X 6 、R 1 、R 2 、R 3 、R 4 , m1, m2, m3, m4, and t are as defined in the general formula (I).

[0079] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, ring A is selected from phenyl or 5- to 6-membered heteroaryl.

[0080] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring A is selected from phenyl, pyridyl, furyl, pyrazolyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolyl, and imidazolyl.

[0081] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, ring A is selected from phenyl and pyridyl.

[0082] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein: Selected from

[0083] in:

[0084] X 3 Selected from CH2, O or NH, preferably CH2 and O;

[0085] X 4 selected from CH2;

[0086] X 5 Selected from CH2, O, S or NH, preferably O;

[0087] X 6 Selected from CH2 or a bond, preferably a bond;

[0088] t is 0, 1, or 2, preferably 1;

[0089] Among them, R 1 、R 2 、R 3 , m1, m2, and m3 are as defined in the general formula (I).

[0090] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein: Selected from

[0091] Among them, R 1 、R 2 、R 3 , m1, m2, and m3 are as defined in the general formula (I).

[0092] In another preferred embodiment, the compound represented by the general formula (III) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring E is selected from R 10a and R 10b Each independently selected from hydrogen, halogen, amino, hydroxyl, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl;

[0093] R 10c Selected from halogen, amino, hydroxy, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 1-6 Cyanoalkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl.

[0094] In another preferred embodiment, the compound represented by the general formula (III) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring E is selected from R 10a Selected from C 1-6 Alkyl or C 1-6 Deuterated alkyl.

[0095] In another preferred embodiment, the compound represented by the general formula (III) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring E is selected from R 10b Selected from hydrogen or C 1-6 alkyl.

[0096] In another preferred embodiment, the compound represented by the general formula (III) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein ring E is selected from R 10a Selected from hydrogen or C 1-6 Alkyl; R 10c Selected from hydrogen, C 1-6 Alkyl, C 1-6 Cyanoalkyl.

[0097] In another preferred embodiment, the compound of general formula (I) according to the present invention, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, is a compound of general formula (IVA) or general formula (IVB), or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof,

[0098] in:

[0099] X 3 Selected from O or CH2, preferably O;

[0100] R 10a Selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl; preferably C 1-6 Alkyl, C 1-6 deuterated alkyl;

[0101] R 10b Selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl; preferably C 1-6 alkyl;

[0102] R 10c Selected from hydrogen, C 1-6 Alkyl; preferably hydrogen;

[0103] R 1 、R 3 、R 4 , m3, and m4 are as defined in the general formula (I).

[0104] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, each R 1 are each independently selected from halogen, C1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -NHC(=O)R a , where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl are optionally substituted with one or more R 1a replace;

[0105] Each R 1a independently selected from halogen, cyano, C 1-6 alkyl, 4 to 8 membered heteroalkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-(CH2) p -NR a R b ;

[0106] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl;

[0107] R b Selected from hydrogen, C 1-6 alkyl;

[0108] m1 is 0, 1, 2, 3, or 4;

[0109] p is an integer from 1 to 4.

[0110] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, each R 1 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-10 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl, wherein C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-10 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl are optionally substituted by one or more R 1a replace;

[0111] Each R 1a Independently selected from deuterated, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-O-(CH2) p -NR a R b 、-C(=O)NR a R b 、-C(=O)NR b -(CH2) p -、-(CH2) p -NR a R b ; the C 1-6 Alkyl, 4-6 membered heterocyclic group is optionally selected from amino, C 1-6 substituted with one or more groups of an alkyl group;

[0112] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 3-6 Heterocyclic group; said C 3-6 The heterocyclic group is selected from halogen, C 1-6 One or more groups are substituted on the alkyl group;

[0113] R b Selected from hydrogen, C 1-6 Alkyl; or

[0114] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group;

[0115] m1 is 0, 1, 2, 3, or 4;

[0116] p is an integer from 1 to 4.

[0117] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 5-6 membered heteroaryl group, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a is substituted by one or more groups;

[0118] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl;

[0119] R b Selected from hydrogen, C 1-6 alkyl.

[0120] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 5-6 membered heteroaryl or phenyl group, which is optionally substituted by one or more R 1a replace;

[0121] Each R 1aIndependently selected from deuterated, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-O-(CH2) p -NR a R b 、-C(=O)NR b -(CH2) p -NR a R b 、-(CH2) p -NR a R b The 4-6 membered heterocyclic group is optionally C 1-6 Alkyl substituted;

[0122] R a Selected from C 1-6 Alkyl, C 3-6 Heterocyclic group; said C 3-6 The heterocyclic group is selected from halogen, C 1-6 One or more groups are substituted on the alkyl group;

[0123] R b Selected from hydrogen, C 1-6 Alkyl; or

[0124] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group;

[0125] m1 is 1;

[0126] p is an integer of 1 to 4, preferably 1 or 2.

[0127] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 5-10 membered heteroaryl group, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C1-6 Halogenated alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-CH2-NR a R b 、-CH2-CH2-NR a R b is substituted by one or more groups;

[0128] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 1-6 Heteroalkyl, C 3-6 heterocyclic group;

[0129] R b Selected from hydrogen, C 1-6 Alkyl; or

[0130] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group.

[0131] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is pyrazolyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, thienyl, which is optionally substituted by one or more R 1a replace;

[0132] Each R 1a Independently selected from deuterated, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NRa R b 、-O-(CH2) p -NR a R b 、-C(=O)NR b -(CH2) p -NR a R b 、-(CH2) p -NR a R b The 4-6 membered heterocyclic group is optionally C 1-6 Alkyl substituted;

[0133] R a Selected from C 1-6 Alkyl, C 3-6 Heterocyclic group; said C 3-6 The heterocyclic group is selected from halogen, C 1-6 One or more groups are substituted on the alkyl group;

[0134] R b Selected from hydrogen, C 1-6 Alkyl; or

[0135] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group;

[0136] m1 is 1;

[0137] p is an integer of 1 to 4, preferably 1 or 2.

[0138] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 5-10 membered aryl group, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a、-C(=O)NR a R b 、-NHC(=O)R a 、-CH2-NR a R b 、-CH2-CH2-NR a R b is substituted by one or more groups;

[0139] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 1-6 Heteroalkyl, C 3-6 heterocyclic group;

[0140] R b Selected from hydrogen, C 1-6 Alkyl; or

[0141] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group.

[0142] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 9-10 membered heterocyclic group or a 9-10 membered heteroaryl group, preferably It is optionally C 1-6 Alkyl substitution;

[0143] m1 is 1.

[0144] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 5-6 membered heterocyclic group, preferably a dihydrofuranyl group, a tetrahydrofuranyl group, a piperidinyl group, which is optionally substituted by C 1- 6 alkyl substitution;

[0145] m1 is 1.

[0146] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is phenyl, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-(CH2) p -NR a R b is substituted by one or more groups;

[0147] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 1-6 Heteroalkyl, C 3-6 heterocyclic group;

[0148] R b Selected from hydrogen, C 1-6 Alkyl; or

[0149] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group;

[0150] p is 1 or 2.

[0151] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is pyrazolyl, which is optionally further selected from C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Halogenated alkyl, C3-6 Cycloalkyl, 4-6 membered heterocyclic group, -(CH2) p -OR a Substituents substituted;

[0152] R a Selected from C 1-6 alkyl;

[0153] p is an integer of 1-4, preferably an integer of 1-2, and more preferably 2.

[0154] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is thiazolyl, which is optionally further substituted with cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl and -C(=O)NR a R b substituted by a substituent;

[0155] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl;

[0156] R b Selected from hydrogen, C 1-6 alkyl.

[0157] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is oxazolyl, which is optionally further substituted with C 1-6 Alkyl or C 1-6 Haloalkyl substitution.

[0158] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is pyrimidinyl, which is optionally further selected from C 1-6 Alkyl or C 1-6 Haloalkyl substitution.

[0159] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is pyridyl, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Heteroalkyl, C 1-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-(CH2) p -NR a R b is substituted by one or more groups;

[0160] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 1-6 Heteroalkyl, C 3-6 heterocyclic group;

[0161] R b Selected from hydrogen, C 1-6 Alkyl; or

[0162] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group;

[0163] p is 1 or 2. In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, according to the present invention, wherein R 1 Selected from halogen, C 1-6 Alkoxy, C 3-6 Cycloalkyl, -NHC(=O)R a ;

[0164] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl.

[0165] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 It is a halogen.

[0166] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 It is a cyano group.

[0167] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 1-6 Alkoxy.

[0168] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 1-6 Halogenated alkyl.

[0169] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 1-6 Halogenated alkoxy.

[0170] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 3-6 Cycloalkyl.

[0171] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 -NHC(=O)R a ; R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl.

[0172] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 4-10 membered heterocyclic group, preferably a 4-6 membered heterocyclic group, which is optionally further selected from halogen, C 1-6 Alkyl or C 1-6 Haloalkyl, -C(=O)R a 、-C(=O)NR a R b is substituted by one or more groups;

[0173] R a Selected from C 1-6 Alkyl, C 3-6 Cycloalkyl;

[0174] R b Selected from hydrogen.

[0175] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 is a 4-6 membered saturated or partially saturated heterocyclic group, which is optionally further selected from halogen, C 1-6 Alkyl, C1-6 Haloalkyl, -C(=O)R a 、-C(=O)NR a R b is substituted by one or more groups;

[0176] R a Selected from C 1-6 Alkyl, C 3-6 Cycloalkyl;

[0177] R b Selected from hydrogen.

[0178] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 2-6 Alkynyl, which is optionally further selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -(CH2) p -OR a 、-NR a R b 、-(CH 2)p -NR a R b is substituted by one or more groups;

[0179] R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 1-6 Heteroalkyl, C 3-6 heterocyclic group;

[0180] R b Selected from hydrogen, C 1-6 Alkyl; or

[0181] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group;

[0182] p is 1 or 2.

[0183] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 1 C 2-6 Alkynyl, preferably propynyl, which is optionally replaced by R 1a replace;

[0184] Each R 1a Independently selected from C 3-6 Cycloalkyl, -OR a 、-NR a R b ;

[0185] R a Selected from C 1-6 Alkyl, C 3-6 Heterocyclic group; said C 3-6 Heterocyclic group is C 1-6 Alkyl substitution;

[0186] R b Selected from hydrogen, C 1-6 Alkyl; or

[0187] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group;

[0188] m1 is 1.

[0189] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein: Selected from:

[0190] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, each R 2Each independently selected from hydrogen, halogen, hydroxy, amino, cyano, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 4 to 8 membered heterocyclyl, phenyl, 5 to 6 membered heteroaryl, wherein the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 4 to 8 membered heterocyclyl, phenyl, 5 to 6 membered heteroaryl are optionally substituted with one or more R 2a replace;

[0191] Each R 2a independently selected from hydrogen, halogen, hydroxy, amino, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, heteroalkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 3-6 Cycloalkyl, 4- to 8-membered heterocyclic group, phenyl, 5- to 6-membered heteroaryl, -LC 1-6 Haloalkyl, -(CH2) p -OR a 、-LC 3-6 Cycloalkyl, -L-4 to 8-membered heterocyclic group, -L-phenyl, -L-5 to 6-membered heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a Rb 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a ;

[0192] Each L is independently -O-, -NH-, C 1-6 alkylene;

[0193] m2 is 0, 1, or 2;

[0194] R a 、R b As defined in general formula (I);

[0195] Preferably, m2 is 0.

[0196] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 2 For hydrogen.

[0197] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein m2 is 0.

[0198] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein m1 is 1.

[0199] In another preferred embodiment, the compound represented by general formula (I) or general formula (II) according to the present invention, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0200] R 5 、R 6 are independently selected from hydrogen, deuterium, halogen, hydroxyl, C l -C6 alkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkyl, cyano, -CH2-C 6-10 Aryl, -CH2-5-6 membered heteroaryl, -CH2-C 3-8 Cycloalkyl, -CH2-4-8 membered heterocyclic group, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NR a R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R b ; the C l -C6 alkyl, -CH2-C 6-10 Aryl, -CH2-5-6 membered heteroaryl, -CH2-C 3-8 Cycloalkyl, -CH2-4-8 membered heterocyclic group, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, C l -C6 alkyl, C l -C6 alkoxy, C l-C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 substituted by one or more groups of aryl or 5-6 membered heteroaryl;

[0201] R a and R b Each independently selected from hydrogen, halogen, hydroxyl, mercapto, C l -C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, the C l -C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, C l -C6 alkyl, C l -C6 alkoxy, C l -C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 aryl, or one or more 5- to 6-membered heteroaryl groups; or

[0202] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4- to 8-membered nitrogen-containing heterocyclic group, wherein the 4- to 8-membered nitrogen-containing heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C l -C6 alkyl, C l -C6 alkoxy, C l -C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 The aryl group or one or more 5- to 6-membered heteroaryl groups are substituted.

[0203] In another preferred embodiment, the compound represented by general formula (I) or general formula (II) according to the present invention, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein:

[0204] R 7 、R 8 are independently selected from hydrogen, deuterium, halogen, hydroxyl, C l -C6 alkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkyl, cyano, -CH2-C 6-10 Aryl, -CH2-5-6 membered heteroaryl, -CH2-C 3-8 Cycloalkyl, -CH2-4-8 membered heterocyclic group, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NR a R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R b ; the C l -C6 alkyl, -CH2-C 6-10 Aryl, -CH2-5-6 membered heteroaryl, -CH2-C 3-8 Cycloalkyl, -CH2-4-8 membered heterocyclic group, C3-C8 cycloalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, C l -C6 alkyl, C l -C6 alkoxy, C l-C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 substituted by one or more groups of aryl or 5-6 membered heteroaryl;

[0205] R a and R b Each independently selected from hydrogen, halogen, hydroxyl, mercapto, C l -C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl, the C l -C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 Aryl, 5-6 membered heteroaryl are optionally selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, C l -C6 alkyl, C l -C6 alkoxy, C l -C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 aryl, or one or more 5- to 6-membered heteroaryl groups; or

[0206] R a and R b Together with the nitrogen atom to which it is attached, it forms a 4- to 8-membered nitrogen-containing heterocyclic group, wherein the 4- to 8-membered nitrogen-containing heterocyclic group is optionally selected from halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, C l -C6 alkyl, C l -C6 alkoxy, C l -C6 haloalkyl, C l -C6 hydroxyalkyl, C l -C6 aminoalkyl, C l -C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C 3-8 Cycloalkyl, 4-8 membered heterocyclic group, C 6-10 The aryl group or one or more 5- to 6-membered heteroaryl groups are substituted.

[0207] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 3 For hydrogen.

[0208] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein m3 is 0.

[0209] In another preferred embodiment, the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomorph, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof according to the present invention, wherein R 4 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, m4 is 0 or 1.

[0210] In another preferred embodiment, according to the compound represented by general formula (I), general formula (II), general formula (III), general formula (IVA), general formula (IVB) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, wherein m4 is 0.

[0211] Typical compounds of the present invention include, but are not limited to:

[0212] Its tautomers, meso racemates, racemates, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts.

[0213] Another aspect of the present invention provides a method for preparing the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, comprising the following steps:

[0214] In the presence of a condensation agent and an alkaline agent, compound Ie is subjected to a condensation reaction with compound If to obtain a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof,

[0215] Among them, the condensation reagent is preferably N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, and the alkaline reagent is preferably N-methylimidazole;

[0216] in, Ring A, R 1 、R 2 、R 3 、R 4 、m1、m2、m3、m4、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 As defined in general formula (I).

[0217] Another aspect of the present invention provides a pharmaceutical composition comprising the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0218] The present invention further provides use of the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, in the preparation of a PRMT5 inhibitor.

[0219] The present invention further provides the use of the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition containing the same in the preparation of a drug for preventing and / or treating diseases related to PRMT5 activity.

[0220] The present invention further provides the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or a pharmaceutical composition containing the same, for use as a medicament.

[0221] The present invention further provides the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or pharmaceutical composition containing the same, which is used as a PRMT5 inhibitor.

[0222] The present invention further provides a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, for use in preventing and / or treating diseases associated with PRMT5 activity.

[0223] The present invention further provides a method for inhibiting PRMT5, comprising administering to a subject in need thereof an effective amount of a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

[0224] The present invention further provides a method for preventing and / or treating diseases associated with PRMT5 activity, comprising administering to a subject in need thereof a preventive or therapeutically effective amount of a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

[0225] In a preferred embodiment of the present invention, the disease associated with PRMT5 activity according to the present invention may be a solid tumor, such as non-small cell lung cancer, mesothelial tumor, neurofibrosarcoma, pancreatic cancer, etc.

[0226] The compounds of the present invention can form pharmaceutically acceptable acid addition salts with acids according to conventional methods in the field of the present invention. The acids include inorganic acids and organic acids, with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, and the like being particularly preferred.

[0227] The compounds of the present invention can form pharmaceutically acceptable basic addition salts with bases according to conventional methods in the field of the present invention. The bases include inorganic bases and organic bases. Acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, and sodium hydroxide, and the like.

[0228] Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions and may contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, colorants, and preservatives to provide a pleasing and palatable pharmaceutical preparation. Tablets contain the active ingredient in admixture with nontoxic, pharmaceutically acceptable excipients suitable for tablet preparation. These excipients may include inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as microcrystalline cellulose, croscarmellose sodium, corn starch, or alginic acid; binders such as starch, gelatin, polyvinyl pyrrolidone, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. These tablets may be uncoated or may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release over a longer period of time. For example, water-soluble taste masking substances such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time-extending substances such as ethylcellulose, cellulose acetate butyrate may be used.

[0229] Oral preparations may also be provided in hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oily vehicle such as peanut oil, liquid paraffin or olive oil.

[0230] Aqueous suspensions contain the active substance and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersants or wetting agents, which may be naturally occurring phospholipids such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain fatty alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, such as polyethylene oxide sorbitan monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene oxide dehydrated sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethylparaben or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

[0231] Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. Oil suspensions can contain thickeners such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents can be added to provide a palatable preparation. These compositions can be preserved by adding antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

[0232] Dispersible powders and granules suitable for preparing aqueous suspensions can be provided with the active ingredient and a dispersant or wetting agent, a suspending agent, or one or more preservatives for mixing by the addition of water. Suitable dispersants or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavorings, and coloring agents may also be added. These compositions can be preserved by the addition of an antioxidant such as ascorbic acid.

[0233] The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of the partial esters and ethylene oxide, such as polyethylene oxide sorbitol monooleate. Emulsions can also contain sweeteners, flavorings, preservatives, and antioxidants. Syrups and elixirs prepared with sweeteners such as glycerol, propylene glycol, sorbitol, or sucrose can be used. Such preparations can also contain demulcents, preservatives, colorants, and antioxidants.

[0234] The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous solutions. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. Sterile injectable formulations may be sterile injectable oil-in-water microemulsions in which the active ingredient is dissolved in an oil phase. For example, the active ingredient may be dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerol to form a microemulsion. The injection or microemulsion may be injected into the patient's bloodstream via local, bolus injection. Alternatively, the solution or microemulsion may be administered in a manner that maintains a constant circulating concentration of the compound of the invention. To maintain this constant concentration, a continuous intravenous delivery device may be used.

[0235] The pharmaceutical compositions of the present invention may be in the form of sterile injectable aqueous or oil suspensions for intramuscular and subcutaneous administration. Such suspensions may be formulated using suitable dispersants or wetting agents and suspending agents as described above, according to known techniques. Sterile injectable formulations may also be sterile injectable solutions or suspensions prepared in a nontoxic, parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. Furthermore, sterile fixed oils may conveniently be used as solvents or suspending media. For this purpose, any blended fixed oil, including synthetic mono- or diglycerides, may be used. Furthermore, fatty acids, such as oleic acid, may also be used to prepare injectable formulations.

[0236] The compounds of this invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid in the rectum and thereby dissolves and releases the drug in the rectum. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights, and mixtures of fatty acid esters of polyethylene glycol.

[0237] It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors, including but not limited to the following: the activity of the specific compound used, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the time of administration, the route of administration, the rate of excretion, the combination of drugs, etc. In addition, the optimal treatment method, such as the mode of treatment, the daily dosage of the general formula compound or the type of pharmaceutically acceptable salt can be verified according to traditional treatment protocols.

[0238] The present invention may contain a compound represented by general formula (I), and a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and prepared into a clinically acceptable dosage form. The derivatives of the present invention can be used in combination with other active ingredients, as long as they do not produce other adverse effects, such as allergic reactions. The compounds of the present invention can be used as the sole active ingredient or in combination with other drugs for treating diseases associated with PRMT5 activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

[0239] Definition of terms

[0240] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0241] The carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopes, that is, the carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (D, also known as heavy hydrogen), tritium (T, also known as super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, nitrogen isotopes include 14 N and 15 N, fluorine isotopes include19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl, bromine include 79 Br and 81 Br.

[0242] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, an alkyl group containing 1 to 4 carbon atoms or an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment and may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylate.

[0243] The term "alkylene" refers to a divalent alkyl group, wherein alkyl is as defined above, having from 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) carbon atoms (i.e., C 1-20 The alkylene group is preferably an alkylene group having 1 to 12 carbon atoms (i.e., C 1-12alkylene), more preferably an alkylene containing 1 to 6 carbon atoms (i.e., C 1-6 Alkylene), further preferably an alkylene containing 1 to 4 carbon atoms (i.e., C 1-6 Alkylene). Non-limiting examples of alkylene include, but are not limited to, methylene (—CH—), 1,1-ethylene (—CH(CH)—), 1,2-ethylene (—CHCH)—, 1,1-propylene (—CH(CHCH)—), 1,2-propylene (—CHCH(CH)—), 1,3-propylene (—CHCHCHCH—), and 1,4-butylene (—CHCHCHCHCH—). Alkylene may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment. The substituent may be selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, alkylthio, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, and oxo.

[0244] The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, preferably an alkenyl group containing 2 to 4 carbon atoms, such as ethenyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

[0245] The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, preferably an alkynyl group containing 2 to 4 carbon atoms or preferably an alkynyl group containing 3 to 4 carbon atoms, such as ethynyl, propynyl, butynyl, etc. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

[0246] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls.

[0247] The term "spiroalkyl" refers to a polycyclic group having a carbon atom (called a spiro atom) shared between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the rings, the spiroalkyl group is divided into a single spiroalkyl group, a double spiroalkyl group or a multi-spiroalkyl group, preferably a single spiroalkyl group and a double spiroalkyl group. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiroalkyl group. Non-limiting examples of spiroalkyl groups include:

[0248] The term "fused cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl groups. Non-limiting examples of fused cycloalkyl groups include:

[0249] The term "bridged cycloalkyl" refers to a 5-20 membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6-14 members, more preferably 7-10 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged cycloalkyl groups include:

[0250] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, tetrahydrobenzofuranyl, tetrahydrobenzoxazolyl, tetrahydrobenzisoxazolyl, cyclopentathienyl, tetrahydrobenzothiazolyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0251] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 4 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 7 to 12 ring atoms, of which 1 to 4 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably 1, 2, 5-oxadiazolyl, pyranyl or morpholinyl. Polycyclic heterocyclic groups include spirocyclic, fused ring and bridged heterocyclic groups.

[0252] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. It is preferably 6 to 14 members, more preferably 7 to 12 members. According to the number of shared spiral atoms between the rings, the spiro heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral heterocyclic group. Non-limiting examples of spiro heterocyclic groups include:

[0253] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 12 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

[0254] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m(wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 12 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclic groups include:

[0255] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include: wait.

[0256] The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylate.

[0257] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. More preferably, phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring, non-limiting examples of which include:

[0258] The aryl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0259] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 10-membered, containing 1 to 3 heteroatoms; more preferably 5 or 6-membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0260] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylate.

[0261] The term "heteroalkyl" refers to a straight or branched chain alkyl group containing 1 to 20 carbon atoms and 1 to 3 heteroatoms selected from O, N, Si and S, wherein alkyl is as defined above, and wherein N and S may be optionally oxidized and N may be optionally quaternized.

[0262] The term "alkoxy" refers to -O-(alkyl), wherein the definition of alkyl is as described above. The limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents can be one or more following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0263] The term "cycloalkoxy" refers to an -O-(cycloalkyl) group, wherein cycloalkyl is as defined above.

[0264] The term "heterocycloalkoxy" refers to -O-(heterocyclyl), wherein heterocyclyl is as defined above.

[0265] The term "cycloalkylthio" refers to -S-(cycloalkyl) where cycloalkyl is as defined above.

[0266] The term "heterocycloalkylthio" refers to an -S-(heterocyclyl) group wherein heterocyclyl is as defined above.

[0267] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0268] The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0269] The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0270] The term "hydroxy" refers to an -OH group.

[0271] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0272] The term "amino" refers to -NH2.

[0273] The term "cyano" refers to -CN.

[0274] The term "nitro" refers to -NO2.

[0275] The term "oxo" refers to =0.

[0276] The term "carboxy" refers to -C(O)OH.

[0277] The term "mercapto" refers to -SH.

[0278] The term "ester group" refers to -C(O)O(alkyl) or -C(O)O(cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

[0279] The term "acyl" refers to a compound containing a -C(O)R group, where R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0280] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0281] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0282] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0283] "Pharmaceutically acceptable salts" or "pharmaceutically acceptable salts" refer to salts of the compounds of the present invention that are safe and effective when used in mammals and have the desired biological activity.

[0284] "Carrier" refers to a vehicle or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0285] Synthesis method of the compound of the present invention

[0286] In order to achieve the purpose of the present invention, the present invention adopts the following technical solutions.

[0287] In some embodiments, the compound represented by general formula (I) of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt can be prepared by Scheme 1.

[0288] Solution 1

[0289] Step 1: In the presence of an alkaline reagent, compound Ie is subjected to a condensation reaction with compound If to obtain a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the condensation reagent is preferably N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, and the alkaline reagent is preferably N-methylimidazole;

[0290] in, Ring A, R 1 、R 2 、R 3 、R 4 、m1、m2、m3、m4、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 As defined in general formula (I).

[0291] In some embodiments, the compound represented by general formula (II) of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt can be prepared by Scheme 2.

[0292] Option 2

[0293] Step 1: In the presence of an alkaline reagent, compound Ie is subjected to a condensation reaction with compound IIf to obtain a compound represented by general formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the condensation reagent is preferably N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, and the alkaline reagent is preferably N-methylimidazole;

[0294] Among them, ring A, R 1 、R 2 、R 3 、R 4 、R 5 、R 7 、m1、m2、m3、m4、X3 、X 4 、X 5 、X 6 , t are as defined in general formula (II).

[0295] In some embodiments, the compound represented by general formula (III) of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt can be prepared by Scheme 3.

[0296] Option 3

[0297] Step 1: In the presence of an alkaline reagent, compound Ie is subjected to a condensation reaction with compound IIIf to obtain a compound represented by general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the condensation reagent is preferably N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, and the alkaline reagent is preferably N-methylimidazole;

[0298] Among them, ring A, ring E, R 1 、R 2 、R 3 、R 4 、R 9 、m1、m2、m3、m4、X 3 、X 4 、X 5 、X 6 , t, and s are as defined in the general formula (III). DETAILED DESCRIPTION

[0299] The present invention is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present invention.

[0300] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker dps300 nuclear magnetic spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0301] LC-MS measurements were performed using an 1100 Series LC / MSD Trap (ESI) mass spectrometer (manufacturer: Agilent).

[0302] GC-MS was performed using a GCMS-QP2010 SE.

[0303] Preparative liquid chromatography was performed using an LC3000 high performance liquid chromatograph and an LC6000 high performance liquid chromatograph (manufacturer: Innovation Tongheng). The chromatographic column was a Daisogel C18 10 μm 60A (20 mm × 250 mm).

[0304] High performance liquid chromatography (HPLC) was performed using a Shimadzu LC-20AD high pressure liquid chromatograph (Agilent TC-C18 250×4.6 mm 5 μm column) and a Shimadzu LC-2010AHT high pressure liquid chromatograph (Phenomenex C18 250×4.6 mm 5 μm column).

[0305] The thin layer chromatography silica gel plate used was Qingdao Ocean Chemical GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15 mm to 0.2 mm, and the specification used for thin layer chromatography separation and purification products was 0.4 mm to 0.5 mm.

[0306] Column chromatography generally uses Qingdao marine silica gel 100-200 mesh and 200-300 mesh silica gel as the carrier.

[0307] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from online shopping malls, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Yinuokai, Nanjing Yaoshi, Anaiji Chemical and other companies.

[0308] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0309] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0310] A CEM Discover SP microwave reactor was used for the microwave reaction.

[0311] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0312] Unless otherwise specified in the examples, the reaction temperature is room temperature, particularly 20°C to 30°C.

[0313] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent systems used in the reactions were: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, and D: acetone. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0314] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used to purify the compound include: A: dichloromethane and methanol system, B: petroleum ether, ethyl acetate and dichloromethane system, C: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

[0315] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention.

[0316] Preparation Example

[0317] Preparation Example 1: Preparation of 4-bromo-3-(difluoromethyl)-1-(2-methyl)-1H-pyrazole (INT1)

[0318] Step 1: Preparation of 1-(4-methoxybenzyl)-1H-pyrazole-3-carbaldehyde (INT1a)

[0319] 1H-pyrazole-3-carboxaldehyde (3.00 g, 31.2 mmol), 1-(chloromethyl)-4-methoxybenzene (4.87 mg, 31.2 mmol), cesium carbonate (15.2 g, 46.8 mmol), and N,N-dimethylformamide (30 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. The reaction solution was poured into water (100 mL) and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to afford 3.50 g of the title compound as a yellow oil in a 51.9% yield.

[0320] LC-MS: m / z 216.2[M+H] + .

[0321] Step 2: Preparation of 3-(difluoromethyl)-1-(4-methoxybenzyl)-1H-pyrazole (INT1b)

[0322] Compound INT1a (3.00 g, 13.9 mmol), diethylaminosulfur trifluoride (4.47 g, 27.8 mmol), and dichloromethane (30 mL) were added to a reaction flask at 0°C and reacted at room temperature under a nitrogen atmosphere for 2 hours. The reaction solution was poured into water (100 mL) and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to obtain the title compound 2.10 as a yellow oil in a yield of 63.6%.

[0323] LC-MS: m / z 239.2[M+H] + .

[0324] Step 3: Preparation of 3-(difluoromethyl)-1H-pyrazole (INT1c)

[0325] Compound INT1b (2.10 g, 8.82 mmol) and trifluoroacetic acid (20 mL) were added to a reaction flask at room temperature and reacted at 70°C under a nitrogen atmosphere for 2 hours. The reaction mixture was poured into water (100 mL) and the pH was adjusted to 7 with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to obtain 700 mg of the title compound as a yellow oil. Yield: 67.3%.

[0326] LC-MS: m / z 119.2[M+H] + .

[0327] Step 4: Preparation of 4-bromo-3-(difluoromethyl)-1H-pyrazole (INT1d)

[0328] Compound INT1c (700 mg, 5.93 mmol), N-bromosuccinimide (1.05 g, 5.93 mmol), and acetonitrile (10 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to afford 530 mg of the title compound as a yellow oil. Yield: 45.6%.

[0329] LC-MS: m / z 196.2[M+H] + .

[0330] Step 5: Preparation of 4-bromo-3-(difluoromethyl)-1-(2-methyl)-1H-pyrazole (INT1)

[0331] Compound INT1d (530 mg, 2.70 mmol), iodomethane (766 mg, 5.40 mmol), and N,N-dimethylformamide (10 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to afford 383 mg of the title compound as a yellow oil in a 67.3% yield.

[0332] LC-MS: m / z 212.2 [M+H] + .

[0333] Preparation Example 2: Preparation of 5-iodo-1-methyl-3-(methoxymethyl)-1H-pyrazole (INT2)

[0334] Step 1: Preparation of 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-3-acetaldehyde (INT2a)

[0335] 1H-pyrazole-3-acetaldehyde (5.29 g, 55.0 mmol) and tetrahydrofuran (80 mL) were added to a reaction flask, cooled to below 0°C, and 60% NaH (3.30 g, 82.5 mmol) was added under a nitrogen atmosphere. The mixture was stirred for 0.5 hours, followed by (2-(chloromethoxy)ethyl)trimethylsilane (9.65 g, 57.8 mmol). The mixture was allowed to return to room temperature under a nitrogen atmosphere and stirred for 2 hours. Water (200 mL) was added to the reaction mixture at 0°C, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated. The residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-15%) to afford 10.5 g of the title compound as a clear oil in an 84.4% yield.

[0336] LCMS: m / z 227.11[M+H] + .

[0337] Step 2: Preparation of (1-(2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methanol (INT2b)

[0338] Compound INT2a (10.4 g, 46.0 mmol) and ethanol (90 mL) were added to a reaction flask. Sodium borohydride (2.10 g, 55.2 mmol) was added at 0°C and stirred for 16 hours under a nitrogen atmosphere. Saturated ammonium chloride solution (10 mL) was added to the reaction mixture at 0°C to quench the mixture. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-30%) to obtain 9.23 g of the title compound as a clear oil in an 88.0% yield.

[0339] LCMS: m / z 229.13 [M+H] + .

[0340] Step 3: Preparation of 3-(methoxymethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (INT2c)

[0341] Compound INT2b (9.11 g, 40.0 mmol) and tetrahydrofuran (100 mL) were added to a reaction flask. 60% NaH (4.00 g, 100 mmol) was added at 0°C and stirred for 0.5 hours. Methyl iodide (8.52 g, 60.0 mmol) was added and the mixture was returned to room temperature under a nitrogen atmosphere and stirred for 3 hours. Water (150 mL) was added dropwise to the reaction solution at 0°C to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated to obtain 10.7 g of the title compound as a clear oil, which was used without further purification.

[0342] LCMS: m / z 243.15 [M+H] + .

[0343] Step 4: Preparation of 5-iodo-3-(methoxymethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (INT2d)

[0344] Compound INT2c (5.10 g, 17.0 mmol) and tetrahydrofuran (70 mL) were added to a reaction flask, cooled to below -70°C, and n-butyllithium (2.5 M, 7.5 mL, 18.7 mmol) was added. The mixture was stirred at this temperature for 1 hour, followed by the addition of a solution of iodine (4.75 g, 18.7 mmol) in tetrahydrofuran (20 mL), and stirred for 2 hours. The reaction mixture was quenched with water at 0°C, concentrated under reduced pressure to remove the tetrahydrofuran, and dissolved in ethyl acetate. The mixture was washed sequentially with 10% sodium bicarbonate solution and 10% sodium bisulfite solution. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated. The residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-20%) to obtain 5.60 g of the title compound as a clear oil in an 89.5% yield.

[0345] LCMS: m / z 369.04 [M+H] + .

[0346] Step 5: Preparation of 5-iodo-3-(methoxymethyl)-1H-pyrazole (INT2e)

[0347] Compound INT2d (5.41 g, 14.7 mmol) and dichloromethane (25 mL) were added to a reaction flask at room temperature, and trifluoroacetic acid (33.6 g, 294 mmol) was added dropwise. The mixture was stirred under a nitrogen atmosphere for 16 hours. The mixture was concentrated under reduced pressure, and ethanol (50 mL) and sodium acetate (6.04 g, 73.5 mmol) were added. The mixture was stirred for 4 hours, filtered, and the filtrate was collected and concentrated. The residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-40%) to obtain 4.15 g of the title compound as a clear oil, which was used without further purification.

[0348] LCMS: m / z 238.96 [M+H] + .

[0349] Step 6: Preparation of 5-iodo-1-methyl-3-(methoxymethyl)-1H-pyrazole (INT2)

[0350] Compound INT2e (595 mg, 2.50 mmol), DMF (7 mL), iodomethane (426 mg, 3.00 mmol), and cesium carbonate (1.23 g, 3.75 mmol) were added to a reaction flask at room temperature and stirred at 100°C under nitrogen for 16 hours. The mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated. The residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-20%) to obtain 462 mg of the title compound as a clear oil, in a yield of 73.6%.

[0351] LCMS: m / z 266.99 [M+H] + .

[0352] Intermediate 3: Preparation of 2-bromo-5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridine (INT3)

[0353] Step 1: Preparation of 2-bromo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine (INT3a).

[0354] 4,5,6,7-Tetrahydrothieno[3,2-c]pyridine (1 g, 5.69 mmol) and tetrahydrofuran (15 mL) were added to a reaction flask at 0°C and stirred for 5 minutes. NBS (1.1 g, 6.26 mmol) was then slowly added. After stirring at room temperature for 1 hour, the residue was separated and purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 3) to obtain 600 mg of the title compound as a yellow oil in a yield of 50.1%.

[0355] LC-MS: m / z 218[M+H]+ .

[0356] Step 2: Preparation of 2-bromo-5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridine (INT3).

[0357] Compound INT3a (600 mg, 2.76 mmol), methanol (10 mL), and paraformaldehyde (1 g, 27.6 mmol) were added to a reaction flask at room temperature. After stirring for 40 minutes, sodium cyanoborohydride (664 mg, 8.28 mmol) was slowly added and stirred for 2 hours. The reaction was quenched by adding water (100 mL) and extracted with EA. The combined organic phases were concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 2) to afford 400 mg of the title compound as a yellow oil in a 62.7% yield.

[0358] LC-MS: m / z 232[M+H] + .

[0359] Preparation Example 4: Preparation of 2-(2-methyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5-yl trifluoromethanesulfonate (INT4)

[0360] Step 1: Preparation of tert-butyl 5-(((trifluoromethyl)sulfonyl)oxy)-3,3a,4,6a-tetrahydrocyclopentyl[c]pyrrole-2(1H)-carboxylate (INT4a)

[0361] Under a nitrogen atmosphere, tert-butyl 5-oxohexahydrocyclopenta[c]pyrrole-2(1H)-carboxylate (2.3 g, 10 mmol) and 30 mL of anhydrous tetrahydrofuran were added to a reaction flask. The temperature was cooled to -75°C, and LDA (10 mL, 20.0 mmol) was added dropwise in portions. The mixture was stirred for 1 hour. 1,1,1-Trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (4.3 g, 12.0 mmol) was dissolved in 10 mL of anhydrous tetrahydrofuran and added dropwise to the reaction mixture. The mixture was allowed to warm to room temperature and allowed to react for 16 hours. The reaction mixture was poured into 300 mL of saturated NaHCO₃ solution and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was used directly in the next reaction.

[0362] Step 2: Preparation of 1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5-yl trifluoromethanesulfonate (INT4b)

[0363] Compound INT4a (714 mg, 2.0 mmol), dichloromethane (3 mL), and a 4 M dioxane hydrochloride solution (3 mL) were added to a reaction flask at room temperature and stirred under a nitrogen atmosphere for 1 h. The reaction solution was concentrated, and the resulting crude product was used directly in the next reaction.

[0364] LCMS: m / z 258.1[M+H] + .

[0365] Step 3: Preparation of 2-(2-methyl)-1,2,3,3a,4,6a-hexahydrocyclopenta[c]pyrrol-5-yl trifluoromethanesulfonate (INT4)

[0366] Compound INT4b (257 mg, 1.0 mmol), iodomethane (213 mg, 1.5 mmol), anhydrous ethanol (3 mL), and potassium carbonate (414 mg, 3.0 mmol) were added to a reaction flask and reacted at 50°C overnight under a nitrogen atmosphere. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 5%-20%) to obtain 210 mg of the title compound as a brown liquid.

[0367] Preparation Example 5: Preparation of 3-bromo-5,6,7,8-tetrahydro-1,6-naphthyridine (INT5)

[0368] Step 1: Preparation of tert-butyl 3-nitro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (INT5a)

[0369] At room temperature, 1-methyl-3,5-dinitropyridin-2(1H)-one (8.01 g, 40.2 mmol), a 7.0 M methanolic ammonia solution (81 mL, 563 mmol), and tert-butyl 4-oxopiperidine-1-carboxylate (8.80 g, 44.3 mmol) were added to a reaction flask and heated to 120°C with stirring for 1 hour. The mixture was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (eluent: ethyl acetate / dichloromethane = 0-20%) to afford 10.1 g of the title compound as a light yellow oil in an 89.5% yield.

[0370] LCMS: m / z 280.12 [M+H] + .

[0371] Step 2: Preparation of tert-butyl 3-amino-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (INT5b)

[0372] At room temperature, compound INT5a (8.92 g, 32.0 mmol), DMF (100 mL), and 4,4'-bipyridine (375 mg, 2.40 mmol) were added to a reaction flask. The temperature was cooled to below 0°C, and tetrahydroxydiboron (8.62 g, 96.0 mmol) was added portionwise. The mixture was stirred for 5 minutes. The mixture was returned to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated to obtain 11.0 g of the title compound as a yellow oil, which was used without further purification.

[0373] LCMS: m / z 250.15 [M+H] + .

[0374] Step 3: Preparation of tert-butyl 3-bromo-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (INT5c)

[0375] At room temperature, compound INT5b (11.0 g, 28.2 mmol), acetonitrile (140 mL), and copper bromide (9.48 g, 42.3 mmol) were added to a reaction flask. The temperature was lowered to below 0°C, and tert-butyl nitrite (3.52 g, 33.9 mmol) was added dropwise. The mixture was stirred for 1 hour, then returned to room temperature and stirred for 16 hours. The mixture was concentrated under reduced pressure, diluted with water, filtered, and the filtrate was extracted with ethyl acetate. The organic phases were combined and concentrated, and the resulting residue was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-30%) to obtain 8.30 g of the title compound as a yellow oil, which was used without further purification.

[0376] LCMS: m / z 313.05 [M+H] + .

[0377] Step 4: Preparation of 3-bromo-5,6,7,8-tetrahydro-1,6-naphthyridine (INT5)

[0378] Compound INT5c (2.50 g, 8.00 mmol), ethyl acetate (10 mL), and a solution of hydrogen chloride in 1,4-dioxane (4.0 M, 10 mL, 40.0 mmol) were added to a reaction flask at room temperature and stirred under a nitrogen atmosphere for 16 hours. The mixture was concentrated under reduced pressure, dissolved in dichloromethane, and the pH was adjusted to 9-10 with aqueous potassium carbonate. The aqueous phase was extracted three times with dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and concentrated to afford 1.65 g of the title compound as a yellow solid in a 97.2% yield.

[0379] LCMS: m / z 212.99 [M+H] + .

[0380] Example

[0381] Example 1: Preparation of (4-amino-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)(7-methoxy-3,4,4a,9b-tetrahydrobenzofurano[3,2-b]pyridin-1(2H)-yl)methanone (1)

[0382] Step 1: Preparation of 5-bromo-1-methyl-1H-pyrazole-4-carbonitrile (1b)

[0383] At room temperature, tert-butyl nitrite (6.33 g, 61.41 mmol), acetonitrile (50 mL), and copper bromide (10.97 g, 49.13 mmol) were added to a reaction flask and reacted at 50°C for 1 hour. 5-Amino-1-methyl-1H-pyrazole-4-carbonitrile (1a) was dissolved in acetonitrile (50 mL) and added dropwise to the reaction system. The reaction was allowed to proceed at 45°C for 2 hours. The mixture was filtered, added with water (100 mL), extracted with ethyl acetate (100 mL), washed with saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and concentrated. The concentrate was dissolved in ethyl acetate (5 mL), and petroleum ether (25 mL-30 mL) was added dropwise. The mixture was filtered to obtain 5 g of compound 1b as a gray solid in a 66% yield.

[0384] Step 2: Preparation of methyl 4-amino-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1d)

[0385] Methyl 4-amino-3-bromobenzoate (1c) (5 g, 21.74 mmol), 1.4-dioxane (8 mL), water (2 mL), potassium acetate (6.38 g, 65.20 mmol), and pinacol diboron (8.28 g, 32.61 mmol) were added to a reaction flask at room temperature. The atmosphere was purged with nitrogen 2-3 times. DPPF palladium dichloride (0.79 g, 1.06 mmol) was added, and the atmosphere was purged with nitrogen 2-3 times. The reaction was allowed to proceed at 80°C for 2 hours. The mixture was filtered and concentrated, and the residue was washed twice with dichloromethane (50 mL) and filtered to obtain 3.1 g of compound 1d as an off-white solid in a 50% yield.

[0386] Step 3: Preparation of methyl 4-amino-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylate (1e): Compound 1d (3.0 g, 10.82 mmol), 1.4-dioxane (32 mL), water (8 mL), potassium phosphate (4.58 g, 21.64 mmol), and compound 1b (3.01 g, 16.23 mmol) were added to a reaction flask at room temperature. The atmosphere was purged with nitrogen 2-3 times. Dichlorodi-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II) (77 mg, 0.11 mmol) was added, and the atmosphere was purged with nitrogen 2-3 times. The reaction was allowed to proceed at 90°C for 2 hours. The mixture was filtered, extracted with water (50 mL), and ethyl acetate (50 mL). The mixture was washed with saturated aqueous sodium chloride (50 mL), dried over anhydrous sodium sulfate, and concentrated to afford compound 1e, 2.0 g, in a yield of 72%.

[0387] Step 4: Preparation of 4-amino-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (1f)

[0388] Compound 1e (4.05 g, 15.81 mmol) was dissolved in THF:MeOH:H2O = 1:1:1 (40 mL) at room temperature. Solid lithium hydroxide (1.14 g, 47.43 mmol) was added and reacted at 80°C for 2 hours. Water (120 mL) was added to adjust the pH to 3-4, resulting in the precipitation of a solid. The resulting solid was rinsed twice with water (20 mL) and dried to afford 1f as a gray powder (3 g, yield: 78%).

[0389] Step 5: Preparation of 6-(2-bromo-4-methoxyphenyl)-5-nitropiperidin-2-one (1h)

[0390] Under a nitrogen atmosphere, 2-bromo-4-methoxybenzaldehyde (1 g) (10.0 g, 46.7 mmol), methyl 4-nitrobutyrate (6.86 g, 46.7 mmol), ammonium acetate (7.21 g, 93.4 mmol), and ethanol (150 mL) were added to a reaction flask at room temperature. The temperature was raised to 90°C and the reaction was allowed to proceed for 2 hours. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 3:1 to 1:1) to obtain 11.4 g of the title compound as a yellow liquid in a yield of 74.5%.

[0391] LC-MS: m / z 329 [M+H] + .

[0392] Step 6: Preparation of 6-(2-bromo-4-methoxyphenyl)piperidine-2,5-dione (1i)

[0393] Under a nitrogen atmosphere, ammonium acetate (7.21 g, 93.4 mmol) and titanium trichloride solution (150 mL) were added to a reaction flask at room temperature and stirred for 30 minutes. Compound 1h (3.28 g, 9.34 mmol) was then dissolved in methanol (20 mL) and potassium tert-butoxide (1.57 g, 14.1 mmol) was added portionwise. The methanol solution was added dropwise to the reaction flask at 0°C and allowed to warm to room temperature for 2 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1-30:1) to obtain 1.45 g of the title compound as a yellow liquid in a yield of 52.3%.

[0394] LC-MS: m / z 298 [M+H] + .

[0395] Step 7: Preparation of 6-(2-bromo-4-methoxyphenyl)-5-hydroxypiperidin-2-one (1j)

[0396] Under a nitrogen atmosphere, compound 1i (1.45 g, 4.87 mmol) and tetrahydrofuran (20.0 mL) were added to a reaction flask, cooled to -40°C, and lithium isopropylborohydride (9.73 mL, 9.73 mmol) was added dropwise with stirring. The reaction mixture was allowed to react for 1 hour. Saturated aqueous ammonium chloride (10.0 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (20.0 mL x 3). The organic phase was washed with saturated brine (20.0 mL x 1), dried over anhydrous sodium sulfate, and filtered. The residue was isolated and purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1-30:1) to obtain 1.21 g of the title compound as a yellow liquid in a yield of 82.3%.

[0397] LC-MS: m / z 301[M+H] + .

[0398] Step 8: Preparation of 2-(2-bromo-4-methoxyphenyl)piperidin-3-ol (1k)

[0399] Under a nitrogen atmosphere, compound 1j (1.21 g, 4.02 mmol) and tetrahydrofuran (10.0 mL) were added to a reaction flask at room temperature. Borane dimethyl sulfide solution (20.0 mL) was added dropwise, and the temperature was raised to 65°C with stirring for 3 hours. The reaction mixture was added with 5.00 mL of water, extracted with ethyl acetate (10.0 mL x 3), washed with saturated brine (30.0 mL x 1), dried over anhydrous sodium sulfate, and filtered. The residue was isolated and purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1 to 30:1) to obtain 500 mg of the title compound as a yellow liquid in a yield of 43.6%.

[0400] LC-MS: m / z 286 [M+H] + .

[0401] Step 9: Preparation of tert-butyl 2-(2-bromo-4-methoxyphenyl)-3-hydroxypiperidine-1-carboxylate (11)

[0402] Under a nitrogen atmosphere, compound 1k (500 mg, 1.75 mmol), di-tert-butyl dicarbonate (571 mg, 2.62 mmol), triethylamine (265 mg, 2.62 mmol), 4-methylaminopyridine (20 mg), and dichloromethane (10.0 mL) were added to a reaction flask at room temperature and stirred for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1-30:1) to obtain 270 mg of the title compound as a yellow liquid in a 40.0% yield.

[0403] LC-MS: m / z 386[M+H] + .

[0404] Step 10: Preparation of tert-butyl 7-methoxy-3,4,4a,9b-tetrahydrobenzofuro[3,2-b]pyridine-1(2H)-carboxylate (1m)

[0405] Under a nitrogen atmosphere, sodium tert-butoxide (389 mg, 4.05 mmol) and 1,4-dioxane (2.00 mL) were added to a reaction flask at room temperature and stirred for 30 minutes. Compound 11 (270 mg, 0.699 mmol) was then added and stirred for 10 minutes. Cuprous iodide (10.0 mg) was then added, and the temperature was raised to 120°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1-30:1) to afford 110 mg of the title compound as a yellow liquid in a 51.5% yield.

[0406] LC-MS: m / z 306 [M+H] + .

[0407] Step 11: Preparation of 7-methoxy-1,2,3,4,4a,9b-hexahydrobenzofuro[3,2-b]pyridine (1n): Under a nitrogen atmosphere, compound 1m (110 mg, 0.361 mmol), dichloromethane (2.00 mL), and trifluoroacetic acid (1.00 mL) were added to a reaction flask at room temperature and allowed to react for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 50:1-30:1) to obtain 60.0 mg of the title compound as a yellow liquid in a yield of 40.4%.

[0408] LC-MS: m / z 206 [M+H] + .

[0409] Step 12: Preparation of (4-amino-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)(7-methoxy-3,4,4a,9b-tetrahydrobenzofuro[3,2-b]pyridin-1(2H)-yl)methanone (1)

[0410] Under a nitrogen atmosphere, compound 1n (60 mg, 0.292 mmol), DMF (3.00 mL), compound 1f (153 mg, 0.490 mmol), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (137 mg, 0.621 mmol), and N-methylimidazole (40.0 mg, 0.488 mmol) were added to a reaction flask and stirred at 30°C for 3 hours. The reaction mixture was added with 5.00 mL of water, extracted with ethyl acetate (5.00 mL x 3), washed with saturated brine (5.00 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was isolated and purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 5.00 mg of the title compound as a white solid, in a yield of 3.98%.

[0411] LC-MS: m / z 430 [M+H] + .

[0412] 1 H NMR (400MHz, DMSO-d6) δ13.82(s,1H),9.61(s,1H),9.06(s,1H),8.62(s,1H),8.40(s,1H),7.84(d,J=8.1Hz,1H),7.30(d,J= 8.2Hz,1H),6.51(dd,J=20.1,11.5Hz,2H),5.07(s,1H),4.50(s,3H),3.73(s,3H),2.17–1.45(m,5H),1.24(d,J=3.0Hz,2H).

[0413] Example 2: Preparation of (4-amino-(3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)(7-methoxy-3,4,4a,9b-tetrahydrobenzofuro[3,2-b]pyridin-1(2H)-yl)methanone (2)

[0414] Step 1: Preparation of 2-methyl-3-cyano-4-oxotetrahydrofuran (2b)

[0415] At room temperature, sodium hydride (5.55 g, 60%, 138.77 mmol), ultra-dry tetrahydrofuran (250 mL), and methyl 2-hydroxyacetate 2a (25 g, 277.54 mmol) were added to a reaction flask. The temperature was raised to 65°C, and trans-crotononitrile (22.34 g, 333.04 mmol) was slowly added. After 2 hours, the temperature was lowered to 25°C and quenched with 2N aqueous sodium hydroxide solution (250 mL). The mixture was extracted with diethyl ether (500 mL). The aqueous phase was retained, the pH adjusted to 2 with 2M hydrochloric acid, and extracted with dichloromethane (250 mL). The product was washed with saturated aqueous sodium chloride solution (250 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (mobile phase: PE:EA = 90:10) to obtain the title compound as a brown solid (14 g, yield: 40.31%).

[0416] Step 2: Preparation of 4-cyano-5-methyl-2,5-dihydrofuran-3-yl trifluoromethanesulfonate (2c)

[0417] Compound 2b (1.1 g, 8.79 mmol) was dissolved in ultra-dry dichloromethane (20 ml) at room temperature, cooled to -78°C, and N,N-diisopropylethylamine (2.27 g, 17.58 mmol) and trifluoromethanesulfonic anhydride (3.47 g, 12.31 mmol) were added. After stirring for 30 minutes, the mixture was returned to room temperature and water (250 ml) was added. The mixture was extracted with dichloromethane (25 mL), washed with saturated aqueous sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Diethyl ether was added to the residue, stirred for 10 minutes, and filtered. The mother liquor was concentrated to give the brown title compound, 1.8 g, yield: 79%.

[0418] Step 3: Preparation of methyl 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (2d)

[0419] Compound 2c (1.0 g, 3.89 mmol), 1.4-dioxane (10 mL), water (2 mL), potassium phosphate (2.47 g, 11.67 mmol), and compound 1d (1.08 g, 3.89 mmol) were added to a reaction flask at room temperature. The atmosphere was purged with nitrogen 2-3 times. DPPF palladium dichloride (500.94 mg, 0.39 mmol) was added and the atmosphere was purged with nitrogen 2-3 times. After stirring at 80°C for 16 hours, the mixture was filtered, added with water (10 mL), extracted with ethyl acetate (10 mL), washed with saturated aqueous sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: PE:EA = 1:1) to give a brown solid compound, 0.8 g, yield: 80%.

[0420] Step 4: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid (2e)

[0421] Compound 2d (1.0 g, 3.87 mmol) was dissolved in THF:MeOH:H2O = 1:1:1 (10 mL) at room temperature. Solid lithium hydroxide (278.64 mg, 11.61 mmol) was added and stirred at 50°C for 2 hours. Water (30 mL) was added and the pH was adjusted to 3-4 with 2M hydrochloric acid to precipitate a solid. The resulting solid was rinsed twice with water (20 mL) and dried to afford 0.5 g of the compound as a gray powdery solid (yield: 53%).

[0422] The remaining steps were the same as those in Example 1, except that compound 2e was used instead of compound 1f to prepare the title compound 2.

[0423] LC-MS: m / z 432[M+H] + .

[0424] 1 H NMR(400MHz,DMSO-d6)δ7.61(d,J=14.2Hz,3H),7.26(dd,J=8.3,4.2Hz,1H),6.7 1(d,J=12.9Hz,2H),6.52(dd,J=11.3,9.0Hz,1H),6.47(s,1H),6.01–5.95(m,1H ),5.47–5.23(m,4H),5.03(s,1H),3.75(d,J=15.8Hz,3H),2.77–2.71(m,1H),1. 94(t,J=7.3Hz,2H),1.77(s,1H),1.59(d,J=12.6Hz,1H),1.41(d,J=5.9Hz,3H).

[0425] Example 3: Preparation of 4-amino-1-methylpyrazolo[4,3-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazin-1-yl)methanone (3)

[0426] Step 1: Preparation of tert-butyl (((3,6-dibromo-2,3-dihydrobenzofuran-2-yl)oxy)methyl)carbamate (3b)

[0427] At -30°C under a nitrogen atmosphere, N-bromosuccinimide (2.92 g, 11.2 mmol) and tert-butyl (2-hydroxyethyl)carbamate (1.75 g, 10.9 mmol) were dissolved in dichloromethane (50.0 mL). After stirring for 10 minutes, 6-bromobenzofuran 3a (2.14 g, 10.9 mmol) was added dropwise. The mixture was allowed to react at room temperature for 24 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 20:1-10:1) to obtain 2.35 g of the title compound as a yellow solid in a yield of 55.6%.

[0428] LC-MS: m / z 424 [M+H] + .

[0429] Step 2: Preparation of tert-butyl 7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine-1-carboxylate (3c)

[0430] Under a nitrogen atmosphere, compound 3b (2.35 g, 5.56 mmol) and silver oxide (6.45 g, 27.8 mmol) were dissolved in ethyl acetate (50.0 mL) and reacted at 50°C for 3 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 3:1-1:1) to obtain 1.42 g of the title compound as a yellow liquid in a yield of 71.1%.

[0431] LC-MS: m / z 356 [M+H] + .

[0432] Step 3: Preparation of tert-butyl 7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine-1-carboxylate (3d)

[0433] At 0°C under a nitrogen atmosphere, compound 3c (1.42 g, 3.99 mmol) and compound 3f (1.24 g, 5.98 mmol), Pd(dppf)Cl2 (0.251 g, 0.342 mmol), and potassium carbonate (1.65 g, 12.0 mmol) were dissolved in dioxane (50.0 mL) and reacted at 100°C for 12 hours. Water (50.0 mL) was added, and the mixture was extracted with EA (50.0 mL x 3). The mixture was washed with saturated brine (50.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 3:1 to 1:1) to obtain 730 mg of the title compound as a yellow liquid in a yield of 51.2%.

[0434] LC-MS: m / z 358 [M+H]+ .

[0435] Step 4: Preparation of 7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine (3e)

[0436] At 0°C under a nitrogen atmosphere, compound 3d (730 mg, 2.04 mmol) was dissolved in dichloromethane (5.00 mL). 0.100 mL of concentrated hydrochloric acid was added dropwise with stirring. The mixture was reacted at 60°C for 3 hours. 10.0 mL of saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted with EA (20.0 mL x 3). The mixture was washed with saturated brine (20.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 340 mg of the title compound as a yellow liquid in a yield of 64.8%.

[0437] LC-MS: m / z 258 [M+H] + .

[0438] Step 5: Preparation of (4-amino-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)(7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazin-1-yl)methanone (3)

[0439] Under a nitrogen atmosphere, compound 3e (150 mg, 0.584 mmol) was dissolved in DMF (5.00 mL). Compound 1f (155 mg, 0.642 mmol), TCFH (304 mg, 1.08 mmol), and NMI (88.4 mg, 1.08 mmol) were added with stirring, and the mixture was stirred at room temperature for 3 hours. 5.00 mL of water was added, and the mixture was extracted with EA (5.00 mL x 3). The mixture was washed with saturated brine (5.00 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was isolated and purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 67.0 mg of the title compound as a white solid, in a yield of 23.8%.

[0440] LC-MS: m / z 482[M+H] + .

[0441] 1H NMR (400MHz, DMSO-d6) δ8.37(s,1H),8.27(s,1H),8.16(s,2H),7.88(s,1H),7.70(d,J=8.9Hz,1H),7.65(d,J=8.5Hz,1H),7.43( d,J=7.7Hz,1H),7.28–7.19(m,4H),7.15(s,1H),6.05(d,J=6.1Hz,1H),4.42(s,3H),3.86(s,3H),3.66(dd,J=11.6,5.5Hz,1H).

[0442] Example 4: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (4)

[0443] The title compound 4 was prepared in the same manner as in Example 3, except that compound 2e was used instead of compound 1f.

[0444] LC-MS: m / z 484 [M+H] + .

[0445] 1 H NMR (400MHz, DMSO-d6) δ8.15 (d, J=6.8Hz, 2H), 7.88 (s, 1H), 7.72 (s, 2H), 7.68–7. 58(m,2H),7.37(d,J=8.3Hz,1H),7.22(dt,J=7.9,1.9Hz,1H),7.15(s,1H),6.69( s,2H),6.05(s,1H),5.77(s,1H),5.43(tt,J=8.8,5.1Hz,1H),5.34(s,1H),4.16( s,1H),3.86(s,3H),3.84–3.77(m,1H),3.64(s,1H),1.41(dd,J=6.2,2.3Hz,3H).

[0446] Examples 5-1 and 5-2: Preparation of ((S)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone and ((R)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (5-1 and 5-2)

[0447] Step 1: Preparation of (R)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate and (S)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (2d-1 and 2d-2)

[0448] 5 g of compound 2d was added to 167 mL of a 19 / 1 (v / v) THF / H₂O solution and stirred thoroughly until clear. 5.098 g of chiral acid R-binaphthol phosphate was then added to 167 mL of a 19 / 1 (v / v) THF / H₂O solution and stirred thoroughly until clear. The two solutions were mixed and stirred at 30°C for 18 hours. The solution gradually changed from a clear brownish-yellow solution to a turbid beige solution. The solution was filtered to obtain a solid, which was dried at 50°C for 4 hours to yield 3.501 g of a white solid 2d-1 with an ee value of 96.54%.

[0449] The mother liquor was completely dried by spin drying, and 50 mL of sodium bicarbonate solution (pH = 8) was added and stirred thoroughly. 40 mL of DCM solution was then added, stirred thoroughly, and transferred to a 30°C oven and stirred overnight. The solution was filtered to remove R-binaphthol phosphate. The filtrate was passed through a separatory funnel, and the DCM phase was concentrated to dryness to obtain 2.09 g of solid. 70 mL of THF / H2O = 19 / 1 (v / v) was added and stirred thoroughly until clear. 2.1 g of chiral acid S-binaphthol phosphate was then added to 70 mL of THF / H2O = 19 / 1 (v / v) and stirred thoroughly until clear. The two solutions were mixed and stirred in a 30°C forced air drying oven until the solution became brownish-yellow and clear. After stirring at 30°C overnight, the solution became beige and turbid. The solution was filtered and the solid was dried at 60°C for 2 h to obtain 2.865 g of white solid 2d-2 with an ee value of 94.86%.

[0450] Step 2: Preparation of (R)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid and (S)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid (2e-1 and 2e-2)

[0451] The title compound was prepared in the same manner as compound 2e, except that compound 2d-1 and 2d-2 were used instead of compound 2d, respectively.

[0452] 2e-1:

[0453] LC-MS: m / z 244.9 [M+H] + .

[0454] 2e-2:

[0455] LC-MS: m / z 245.0 [M+H] + .

[0456] Step 3: Preparation of ((S)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone and ((R)-4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (5-1 and 5-2)

[0457] The title compounds 5-1 and 5-2 were prepared in the same manner as in Example 4, except that compounds 2e-1 and 2e-2 were used instead of compound 2e, respectively.

[0458] 5-1:

[0459] LC-MS: m / z 484 [M+H] + .

[0460] 1 H NMR (400MHz, DMSO-d6) δ8.15 (d, J=6.8Hz, 2H), 7.88 (s, 1H), 7.72 (s, 2H), 7.68–7. 58(m,2H),7.37(d,J=8.3Hz,1H),7.22(dt,J=7.9,1.9Hz,1H),7.15(s,1H),6.69( s,2H),6.05(s,1H),5.77(s,1H),5.43(tt,J=8.8,5.1Hz,1H),5.34(s,1H),4.16( s,1H),3.86(s,3H),3.84–3.77(m,1H),3.64(s,1H),1.41(dd,J=6.2,2.3Hz,3H).

[0461] 5-2:

[0462] LC-MS: m / z 484 [M+H] + .

[0463] 1 H NMR(400MHz,DMSO-d6)δ8.75(s,2H),8.17(s,1H),8.07–7.80(m,4H),7.38(s,1H),7.28–7.12(m,2H),6 .13(s,1H),5.63–5.34(m,4H),3.86(s,3H),3.21(d,J=33.0Hz,1H),1.44(d,J=5.9Hz,3H),1.24(s,1H).

[0464] 2.73g of compound 5-2 was subjected to chiral separation (chromatographic column model: CHIRALPAK IF 5cm*25cm, 10μm, mobile phase: MtBE (0.5% 2mM NH3-MeOH): (MeOH:DCM=1:1), 95mL / min) to obtain 0.81g of white solid compound 5-2-A (first peak) and 1.36g of white solid compound 5-2-B (second peak). Its structure is selected from:

[0465] 5-2-A:

[0466] LC-MS: m / z 484.2 [M+H] + ;

[0467] 5-2-B:

[0468] LC-MS: m / z 484.2 [M+H] + .

[0469] Example 6: Preparation of 4-amino-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (6)

[0470] Step 1: Preparation of 4-amino-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid (6e)

[0471] The title compound was prepared in the same manner as intermediate 2e, except that acrylonitrile was used instead of trans-crotononitrile.

[0472] LCMS: m / z 230.96 [M+H] + .

[0473] The title compound 6 was prepared in the same manner as in Example 3, except that compound 6e was used instead of compound 1f.

[0474] LC-MS: m / z 470 [M+H] + .

[0475] 1 H NMR(400MHz,DMSO-d6)δ8.94(s,2H),8.17(s,1H),8.02(s,1H),7.94(s,1H),7.91–7.81(m,2H),7. 25–7.11(m,2H),6.13(s,1H),5.50(s,2H),5.11(t,J=3.8Hz,2H),3.86(s,4H),3.10–2.99(m,1H).

[0476] Example 7: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (7)

[0477] The title compound 7 was prepared in the same manner as in Example 4, except that 5-bromobenzofuran was used instead of compound 3a.

[0478] LC-MS: m / z 484 [M+H] + .

[0479] 1 H NMR(400MHz,DMSO-d6)δ14.27(s,1H),8.88(s,3H),8.26–8.07(m,2H),7.98(s,2H),7.89–7.81(m,2H),7.55–7.44(m,2H),7.10–6.79(m,1H) ,6.10(dd,J=55.9,49.3Hz,2H),5.52(s,4H),5.33(s,1H),3.87(s,4H),3.76(s,1H),3.61(s,2H),3.37–2.98(m,2H),1.45(d,J=6.1Hz,3H).

[0480] Example 8: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9a-tetrahydro-4H-pyrido[2,3':4,5]furo[2,3-b][1,4]oxazin-4-yl)methanone (8)

[0481] Step 1: Preparation of 5-chloro-2-iodopyridin-3-ol (8b)

[0482] 5-Chloro-3-hydroxypyridine 8a (15 g, 116.3 mmol), iodine (30 g, 116.3 mmol) and sodium carbonate (24.65 g, 232.6 mmol) were dissolved in water (150.0 mL) at room temperature and reacted for 24 h. The reaction solution was adjusted to pH 3-4 with 10% dilute hydrochloric acid and extracted with EA. The organic phase was concentrated under reduced pressure to give 28.12 g of the title compound as a yellow solid in a yield of 94.4%.

[0483] LC-MS: m / z 256 [M+H] + .

[0484] Step 2: Preparation of 6-chloro-2-(trimethylsilyl)furo[3,2-B]pyridine (8c)

[0485] Under a nitrogen atmosphere, compound 8b (28.12 g, 110.3 mmol) and trimethylethynylsilane (21.61 g, 220.5 mmol) were dissolved in dioxane (100.0 mL) and reacted at 100°C for 12 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 10:1-5:1) to obtain 12.09 g of the title compound as a yellow liquid in a yield of 48.7%.

[0486] LC-MS: m / z 226 [M+H] + .

[0487] Step 3: Preparation of 6-chlorofuro[3,2-B]pyridine (8d)

[0488] Compound 8c (12.09 g, 53.7 mmol) and tetrabutylammonium fluoride solution in tetrahydrofuran (214.8 ml, 107.4 mmol) were dissolved in dichloromethane (100.0 mL) at room temperature and allowed to react for 2 hours. Water (50.0 mL) was added, and the mixture was extracted with EA (100.0 mL x 3). The mixture was washed with saturated brine (50.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 4.96 g of the title compound as a yellow liquid in a 60.4% yield.

[0489] LC-MS: m / z 154 [M+H] + .

[0490] Step 4: Preparation of tert-butyl (2-((3-bromo-6-chloro-2,3-dihydrofuro[3,2-b]pyridin-2-yl)oxy)ethyl)carbamate (8e)

[0491] Compound 8d (4.96 g, 32.4 mmol), N-Boc-ethanolamine, and NBS were dissolved in acetonitrile (50.0 mL) at room temperature and reacted for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 5:1-1:1) to obtain 2.89 g of the title compound as a yellow solid, yield: 22.8%.

[0492] LC-MS: m / z 393 [M+H] + .

[0493] Step 5: Preparation of tert-butyl 7-chloro-2,3,4a,9a-tetrahydro-4H-pyrido[2',3':4,5]furo[2,3-b][1,4]oxazine-4-carboxylate (8f)

[0494] At room temperature, compound 8e (2.89 g, 7.37 mmol) was dissolved in EA (20.0 mL) and silver oxide (8.55 g, 36.85 mmol) was added. The mixture was stirred at 80°C for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 3:1 to 1:1) to obtain 828.0 mg of the title compound as a yellow solid, in a yield of 36.0%.

[0495] LC-MS: m / z 313 [M+H] + .

[0496] Step 6: Preparation of tert-butyl 7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9a-tetrahydro-4H-pyrido[2',3':4,5]furo[2,3-b][1,4]oxazine-4-carboxylate (8 g)

[0497] Under a nitrogen atmosphere, compound 8f (0.828 g, 2.65 mmol), compound 3f (0.552 g, 2.65 mmol), Pd(dtbpf)Cl2 (0.173 g, 0.265 mmol), and sodium carbonate (0.562 g, 5.3 mmol) were dissolved in dioxane (10.0 mL) at 0°C and reacted at 100°C for 12 hours. 20.0 mL of water was added, and the mixture was extracted with EA (50.0 mL x 3). The mixture was washed with saturated brine (50.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (mobile phase: PE / EA = 3:1 to 1:1) to obtain 450 mg of the title compound as a yellow liquid in a yield of 47.4%.

[0498] LC-MS: m / z 359[M+H] + .

[0499] Step 7: Preparation of 7-(1-methyl-1H-pyrazol-4-yl)-3,4a,9a-tetrahydro-2H-pyrido[2',3':4,5]furo[2,3-b][1,4]oxazine (8h)

[0500] At room temperature, 8 g (450 mg, 1.26 mmol) of compound 5 was dissolved in 5 mL of 2 M hydrochloric acid in dioxane. The mixture was reacted at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain 220 mg of the title compound as a brown solid. Yield: 67.7%.

[0501] LC-MS: m / z 259 [M+H] + .

[0502] Step 8: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9a-tetrahydro-4H-pyrido[2,3':4,5]furo[2,3-b][1,4]oxazin-4-yl)methanone (8)

[0503] Under a nitrogen atmosphere, compound 8h (220 mg, 0.86 mmol) was dissolved in DMF (3.00 mL), and compound 2e (166 mg, 0.682 mmol), TCFH (479 mg, 1.71 mmol), and NMI (140 mg, 1.71 mmol) were added with stirring. The filtrate was concentrated under reduced pressure, and the residue was purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 15.0 mg of the title compound as a white solid, in a yield of 3.6%.

[0504] LC-MS: m / z 485 [M+H] + .

[0505] 1H NMR (400MHz, DMSO-d6) δ8.81(t,J=5.5Hz,1H),8.59(d,J=1.8Hz,1H),8.19(s,1H),8.07(d,J=2.1Hz,1H),8.00–7.93(m,3H),7.56(d,J=8.8Hz ,1H),6.69(s,2H),6.09(d,J=0.9Hz,1H),5.47–5.25(m,4H),4.45(t,J=5.4Hz,2H),3.87(s,3H),3.76(d,J=5.5Hz,1H),1.41(d,J=6.1Hz,3H).

[0506] Example 9: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-pyrido[3,4':4,5]furo[2,3-b][1,4]oxazine-1-methanone (9)

[0507] Step 1: Preparation of 2-chloro-5-iodo-4-methoxypyridine (9b)

[0508] At room temperature, 2-chloro-4-methoxypyridine 9a (20 g, 139.50 mmol) and p-toluenesulfonic acid (200 mL) were added to a reaction flask. The temperature was lowered to -20°C, and NIS (34.52 g, 153.45 mmol) was added. The mixture was stirred at room temperature for 16 hours. The mixture was quenched with ice water, the pH was adjusted to 10, and the mixture was extracted with ethyl acetate. The organic phase was washed with aqueous sodium thiosulfate and aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated. 22 g of the title compound was obtained as a red oil, with a yield of 23.6%.

[0509] Step 2: Preparation of 2-chloro-5-iodopyridin-4-ol (9c)

[0510] Compound 9b (22 g, 81.81 mmol), glacial acetic acid (110 mL), and hydrobromic acid (110 mL) were added to a reaction flask at room temperature. The mixture was stirred at 110°C for 4 hours, cooled to 25°C, and filtered to obtain a filter cake. THF (100 mL) was added to dissolve the solid, which was then washed with 1N sodium hydroxide solution, extracted with tetrahydrofuran, and the organic phase was washed with sodium chloride solution. The mixture was dried over anhydrous sodium sulfate and concentrated. This afforded 15 g of the title compound as a red solid in a 72.1% yield.

[0511] Step 3: Preparation of 6-chloro-2-(trimethylsilyl)furo[3,2-c]pyridine (9d)

[0512] At room temperature, compound 9c (20 g, 78.43 mmol), iodide (1.19 g, 6.27 mmol), trimethylsilyl chloride (11.55 g, 117.64 mmol), triethylamine (11.88 g, 117.64 mmol), and DMF (200 mL) were added to a reaction flask. After nitrogen was purged 2-3 times, DPPF palladium dichloride (1 g) was added and the nitrogen was purged 2-3 times. The temperature was raised to 80°C and stirred for 16 hours. Water (100 mL) and EA (100 mL) were added, the mixture was filtered, and then extracted with EA. The organic phase was washed with aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated. 8 g of the title compound was obtained as a yellow solid in a 45.4% yield.

[0513] Step 4: Preparation of 6-chlorofuro[3,2-c]pyridine (9e)

[0514] Compound 9d (3.3 g, 12.94 mmol) and dichloromethane (40 mL) were added to a reaction flask at room temperature. The temperature was cooled to 0°C, TBFA (6.76 g, 25.88 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (50 mL), extracted with dichloromethane (50 mL), and the organic phase was washed with sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated. Purification by silica gel column chromatography (mobile phase: PE:EA = 95:5) afforded 2.2 g of the title compound as a white solid in a 98.2% yield.

[0515] Step 5: Preparation of N-(2-((3-bromo-6-chloro-2,3-dihydrofuro[3,2-c]pyridin-2-yl)oxy)ethyl)-4-nitrobenzenesulfonamide (9f)

[0516] Compound 9e (5.72 g, 37.14 mmol), acetonitrile (60 mL), and NBS (10.96 g, 44.567 mmol) were added to a reaction flask at room temperature. After stirring at room temperature for 30 minutes, N-(2-hydroxyethyl)-4-nitrobenzenesulfonamide (5.72 g, 37.14 mmol) was added and stirred at room temperature for 2 hours. Water (50 mL) was added, and the mixture was extracted with dichloromethane (50 mL). The organic phase was washed with aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated. Purification by silica gel column chromatography (mobile phase: PE:EA = 9:1) afforded 2.2 g of the title compound as a red oil in a yield of 12.3%.

[0517] Step 6: Preparation of 7-chloro-1-((4-nitrophenyl)sulfonyl)-2,3,4a,9b-tetrahydro-1H-pyrido[3',4':4,5]furo[2,3-b][1,4]oxazine (9 g)

[0518] Compound 9f (2.2 g, 4.61 mmol), acetonitrile (20 mL), and anhydrous potassium carbonate (0.95 g, 6.92 mmol) were added to a reaction flask at room temperature and stirred at 50°C for 16 hours. Water (50 mL) was added, followed by extraction with EA (50 mL). The organic phase was washed with aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to yield 0.56 g of the crude title compound as a yellow oil.

[0519] Step 7: Preparation of 7-(1-methyl-1H-pyrazol-4-yl)-1-((4-nitrophenyl)sulfonyl)-2,3,4a,9b-tetrahydro-1H-pyrido[3',4':4,5]furo[2,3-b][1,4]oxazine (9h)

[0520] Compound 9g (0.46 g, 1.16 mmol), compound 3f (362.04 mg, 1.74 mmol), and anhydrous sodium carbonate (245.92 mg, 2.32 mmol) were added to 1.4-dioxane (1 mL) and water (0.2 mL) at room temperature. The atmosphere was purged with nitrogen 2-3 times, and then DPPF palladium dichloride (92 mg) was added. The atmosphere was purged with nitrogen 2-3 times, and the temperature was raised to 100°C with stirring for 2 hours. The reaction system was filtered, and water (10 mL) was added. Extraction was performed with ethyl acetate (10 mL), and the mixture was washed with saturated aqueous sodium chloride solution (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: DCM:MeOH = 93:7) to obtain 250 mg of the title compound as a yellow solid in a yield of 49.0%.

[0521] Step 8: Preparation of 7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-pyrido[3',4':4,5]furo[2,3-b][1,4]oxazine (9i)

[0522] Compound 9h (250 mg, 0.56 mmol), THF (4 mL), water (4 mL), dodecyl mercaptan (228.72 mg, 1.13 mmol), and anhydrous potassium carbonate (156.17 mg, 1.13 mmol) were added to a reaction flask at room temperature and stirred at 50°C for 16 hours. Water (50 mL) was then added and extracted with EA (50 mL). The organic phase was washed with aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to obtain 0.2 g of the crude title compound as a yellow oil.

[0523] Step 9: Preparation of (4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)(7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-pyrido[3',4':4,5]furo[2,3-b][1,4]oxazin-1-yl)methanone (9)

[0524] Compound 9i (70 mg, 0.27 mmol) was dissolved in N,N-dimethylformamide (2 mL) at room temperature. Compound 2e (65.9 mg, 0.27 mmol) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (227.27 mg, 0.81 mmol) were added. After stirring for 10 minutes, a methylimidazole solution (110.84 mg, 1.35 mmol) was added dropwise. The mixture was reacted at room temperature for 16 hours. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL). The organic phase was concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (Daisogei 30 mm*250 mm, C18, 10 μm, 100 Å, mobile phase: acetonitrile / water, gradient: 30%-80%) to give 7.0 mg of the compound as a white solid powder.

[0525] LC-MS: m / z 485.2 [M+H] + .

[0526] 1 H NMR(400MHz,DMSO-d6)δ8.46(s,1H),8.29(s,1H),8.01(s,1H),7.74(s,1H),7.67(s,1H),7.30(s,1H),6.67(s,2H),6.20(s,1H),5.43(d d,J=6.7,3.6Hz,1H),5.34(t,J=17.5Hz,2H),3.88(s,3H),3.62(d,J=6.2Hz,1H),3.32(s,2H),1.41(dd,J=6.0,1.7Hz,3H),1.24(s,1H).

[0527] Example 10: Preparation of (4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,10b-tetrahydrochromeno[3,4-b][1,4]oxazin-1(5H)-yl)methanone (10)

[0528] Step 1: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinoline-8-carbonyl chloride (10k)

[0529] Compound 2e (2.43 g, 10.0 mmol), dichloromethane (40 mL), and thionyl chloride (20.00 mL) were added to a reaction flask at room temperature and reacted at 50°C for two hours. The reaction solution was concentrated under reduced pressure to obtain the title compound as a light yellow solid, 2.17 g, in a yield of 83.1%.

[0530] Step 2: Preparation of 2-(allyloxy)-4-bromobenzaldehyde (10b)

[0531] 4-Bromo-2-hydroxybenzaldehyde 10a (10.0 g, 50.0 mmol), 3-bromopropene (6.00 g, 50.0 mmol), potassium carbonate (8.34 g, 50.0 mmol), and acetonitrile (100 mL) were added to a reaction flask at room temperature. The reaction was allowed to proceed at 60°C under a nitrogen atmosphere for 2 hours. After completion of the reaction, the reaction mixture was filtered, the filtrate collected, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 5:1) to afford the title compound as a pale yellow solid, 1.5 g, in a 95.9% yield.

[0532] LC-MS: m / z 239.1[M+H] + .

[0533] Step 3: Preparation of 2-(allyloxy)-4-bromo-1-vinylbenzene (10c)

[0534] Compound 10b (1.5 g, 47.9 mmol), triphenylphosphine hydrobromide (24.6 g, 71.8 mmol), sodium hydride (2.87 g, 71.8 mmol), and tetrahydrofuran (200 mL) were added to a reaction flask at 0°C. The reaction was allowed to proceed at room temperature under a nitrogen atmosphere for 2 hours. After completion of the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 10:1) to obtain the title compound as a pale yellow oil, 8.50 g, in a yield of 74.6%.

[0535] LC-MS: m / z 239.3 [M+H] + .

[0536] Step 4: Preparation of 7-bromo-2H-chromene (10d)

[0537] Compound 10c (8.50 g, 35.7 mmol), phenylmethylenebis(tricyclohexylphosphine)ruthenium dichloride (2.94 g, 3.10 mmol), and dichloromethane (60.0 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. After the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 10:1) to obtain the title compound as a pale yellow oil, 6.70 g, in a yield of 90.6%.

[0538] LC-MS: m / z 210.1 [M+H] + .

[0539] Step 5: Preparation of 4-(2H-chromen-7-yl)-1-methyl-1H-pyrazole (10e)

[0540] Compound 10d (6.70 g, 31.9 mmol), compound 3f (6.63 g, 31.9 mmol), 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (2.33 g, 3.19 mmol), sodium carbonate (5.07 g, 47.9 mmol), dioxane (100 mL), and water (5.00 mL) were added to a reaction flask at room temperature and reacted at 80°C under a nitrogen atmosphere for 2 hours. After completion of the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 5:1) to obtain the title compound as a pale yellow oil, 4.70 g, in a yield of 69.5%.

[0541] LC-MS: m / z 212.1 [M+H] + .

[0542] Step 6: Preparation of 3-bromo-7-(1-methyl-1H-pyrazol-4-yl)chroman-4-ol (10f)

[0543] Compound 10e (4.70 g, 22.2 mmol), N-bromosuccinimide (7.90 g, 44.4 mmol), dimethyl sulfoxide (50.0 mL), and water (10.0 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 12 hours. After completion of the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 3:1) to obtain the title compound as a pale yellow oil, 2.70 g, in a yield of 39.4%.

[0544] LC-MS: m / z 309.1[M+H] + .

[0545] Step 7: Preparation of 4-amino-7-(1-methyl-1H-pyrazol-4-yl)chroman-3-ol (10 g)

[0546] Compound 10f (2.70 g, 8.77 mmol), aqueous ammonia (5.00 mL), and isopropanol (10.0 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to afford the title compound as a pale yellow solid (1.70 g, 79.4% yield).

[0547] LC-MS: m / z 245.3 [M+H] + .

[0548] Step 8: Preparation of 2-chloro-N-(3-hydroxy-7-(1-methyl-1H-pyrazol-4-yl)chroman-4-yl)acetamide (10h)

[0549] Compound 10g (1.70g, 6.94mmol), chloroacetyl chloride (0.932g, 8.33mmol), triethylamine (0.841g, 8.33mmol), and dichloromethane (10.0mL) were added to a reaction flask at room temperature under a nitrogen atmosphere for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to afford the title compound as a pale yellow oil (1.35g, 60.5% yield).

[0550] LC-MS: m / z 322.4 [M+H] + .

[0551] Step 9: Preparation of 8-(1-methyl-1H-pyrazol-4-yl)-1.4a.5.10b-tetrahydrochromeno[3.4-b][1.4]oxazin-2(3H)-one (10i)

[0552] Compound 10h (1.35 g, 4.19 mmol), sodium hydride (0.23 g, 6.29 mmol), and dichloromethane (10.0 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. After completion of the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (mobile phase: dichloromethane:methanol = 10:1) to obtain the title compound as a light yellow oil, 700 mg, in a yield of 3.9%.

[0553] LC-MS: m / z 286.1 [M+H] + .

[0554] Step 10: Preparation of 8-(1-methyl-1H-pyrazol-4-yl)-1.2.3.4.5.10b-hexahydrochromeno[3.4-b][1.4]oxazine (10j)

[0555] Compound 10i (700 mg, 2.45 mmol), lithium aluminum hydride (0.12 g, 2.94 mmol), and tetrahydrofuran (10.0 mL) were added to a reaction flask at room temperature and refluxed under a nitrogen atmosphere for 2 hours. After completion of the reaction, water (50.0 mL) was added and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (mobile phase: dichloromethane:methanol = 10:1) to obtain the title compound as a pale yellow oil, 400 mg, in a yield of 61.1%.

[0556] LC-MS: m / z 272.2 [M+H] + .

[0557] Step 11: Preparation of (4-amino-3-methyl-1,3-dihydrofuro[3.4-c]quinolin-8-yl)(8-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,10b-tetrahydrochromeno[3.4-b][1.4]oxazin-1(5H)-yl)methanone (10)

[0558] Compound 10k (387 mg, 1.42 mmol), compound 10j (400 mg, 1.42 mmol), sodium hydride (85.2 mg, 2.13 mmol), and tetrahydrofuran (6.00 mL) were added to a reaction flask at room temperature under a nitrogen atmosphere and allowed to react for 2 hours. After completion of the reaction, the mixture was filtered, and the mother liquor residue was separated by preparative liquid chromatography (column model: Daisogei 30 mm x 250 mm, C18, 10 μm, 100 Å, mobile phase: acetonitrile / water, gradient: 10%-100%) to obtain 15.0 mg of the title compound as a brown solid. Yield: 2.05%.

[0559] LC-MS: m / z 497.2 [M+H] + .

[0560] 1 H NMR (400MHz, DMSO-d6) δ8.86 (d, J = 2.3Hz, 1H), 8.37 (s, 1H), 8.13–8.04 (m, 2H), 7. 94–7.81(m,2H),7.75(d,J=11.4Hz,1H),7.64(d,J=8.3Hz,1H),4.94(dd,J=8.4,4. 5Hz,1H),4.24(t,J=6.7Hz,1H),2.37(s,3H),2.19–2.08(m,1H),1.91(d,J=37.4Hz ,2H),1.75(t,J=14.3,9.2Hz,1H),1.61–1.46(m,2H),1.23(dd,J=9.6,5.2Hz,3H).

[0561] Example 11: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (11)

[0562] Step 1: Preparation of 7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine (11a)

[0563] Under nitrogen atmosphere, compound 3c (200 mg, 0.563 mmol) was dissolved in 2.0 mL of hydrochloric acid-dioxane at 0°C. After addition, the mixture was reacted at 40°C for 3 hours. The filtrate was concentrated under reduced pressure to obtain 140 mg of the title compound as a yellow liquid. Yield: 97.5%.

[0564] LC-MS: m / z 255 [M+H] + .

[0565] Step 2: Preparation of (4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)(7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (11)

[0566] Under a nitrogen atmosphere, compound 11a (140 mg, 0.549 mmol) was dissolved in DMF (5.00 mL). Compound 2e (152 mg, 0.624 mmol), TCFH (304 mg, 1.08 mmol), and NMI (88.4 mg, 1.08 mmol) were added with stirring, and the mixture was stirred at room temperature for 3 hours. 5.00 mL of water was added, and the mixture was extracted with EA (5.00 mL x 3). The mixture was washed with saturated brine (5.00 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was isolated and purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 8.0 mg of the title compound as a white solid, in a yield of 3.0%.

[0567] LC-MS: m / z 482[M+H] + .

[0568] 1 H NMR (400MHz, DMSO-d6) δ8.74 (s, 2H), 8.07–7.74 (m, 3H), 7.31 (d, J = 63.6Hz, 3H), 6.13 (s, 1H) ),5.71–5.34(m,4H),3.84(d,J=25.8Hz,2H),1.44(d,J=6.0Hz,3H),1.24(d,J=3.4Hz,2H).

[0569] Example 12: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(2-methylpyrimidin-5-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (12)

[0570] The title compound 12 was prepared by the same method as in Example 4, except that (2-methylpyrimidin-5-yl)boronic acid was used instead of compound 3f.

[0571] LC-MS: m / z 496 [M+H] + .

[0572] 1H NMR(400MHz,DMSO-d6)δ14.35(s,1H),9.03(s,2H),8.75(s,2H),8.09–7.88(m,3H),7.68–7.33(m,4H ), 6.20 (d, J = 26.1Hz, 1H), 5.64–5.41 (m, 3H), 3.98–3.53 (m, 2H), 2.67 (s, 3H), 1.45 (d, J = 6.0Hz, 3H).

[0573] Example 13: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-methoxy-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (13)

[0574] The title compound 13 was prepared in the same manner as in Example 4, except that methanol was used instead of compound 3f.

[0575] LC-MS: m / z 434 [M+H] + .

[0576] 1 H NMR (400MHz, DMSO-d6) δ8.72(s,2H),7.92(s,1H),7.82(d,J=9.0Hz,1H),7.30(d,J=8.8Hz,1H),6.57(d,J= 7.3Hz,2H),6.08(s,1H),5.58–5.46(m,1H),4.16(s,1H),3.75(s,3H),3.62(s,3H),1.44(d,J=6.0Hz,3H).

[0577] Example 14: Preparation of 2-amino-3-methylquinolin-6-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazin-1-yl)methanone (14)

[0578] The title compound 14 was prepared in the same manner as in Example 3, except that 2-amino-3-methyl-6-carboxyquinoline was used instead of compound 1f.

[0579] LC-MS: m / z 442 [M+H] + .

[0580] 1H NMR (400MHz, DMSO-d6) δ8.84(s,2H),8.32(s,1H),8.16(s,1H),8.10(s,1H),7.88(s,1H),7.77(d,J=8.6Hz,1H),7.36 (s,1H),7.22(s,1H),7.17(s,1H),6.12(s,1H),3.86(s,3H),3.64(s,1H),3.13(d,J=85.7Hz,1H),2.35–2.31(m,3H).

[0581] Example 15: Preparation of 2-amino-3-methylquinolin-6-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (15)

[0582] The title compound 15 was prepared in the same manner as in Example 13, except that 5-bromobenzofuran was used instead of compound 3a.

[0583] LC-MS: m / z 434 [M+H] + .

[0584] 1 H NMR(400MHz,DMSO-d6)δ7.86–7.49(m,3H),6.90(d,J=44.0Hz,3H),6.65(s,2H),5 .97(s,1H),5.55–5.24(m,2H),3.75(d,J=1.6Hz,3H),1.41(dd,J=6.2,2.1Hz,3H).

[0585] Example 16: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-cyclopropyl-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (16)

[0586] The title compound 16 was prepared in the same manner as in Example 7, except that cyclopropylboronic acid was used instead of compound 3f.

[0587] LC-MS: m / z 443 [M+H] + .

[0588] 1H NMR(400MHz, DMSO-d6)δ8.67(s,2H),8.05–7.80(m,3H),7.13(s,1H),7.01–6.68(m,2H),6.04(t,J=36.5Hz,2H),5.61– 5.40(m,4H),1.93(td,J=8.1,4.3Hz,1H),1.44(dd,J=6.3,2.1Hz,3H),0.92(dd,J=8.4,2.8Hz,2H),0.76–0.53(m,2H).

[0589] Example 17: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(pyrrolidin-1-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (17)

[0590] The title compound 17 was prepared in the same manner as in Example 4, except that pyrrolidine was used instead of compound 3f.

[0591] LC-MS: m / z 473 [M+H] + .

[0592] 1 H NMR(400MHz, DMSO-d6)δ9.36–8.71(m,3H),8.30–7.61(m,4H),7.35–6.97(m,1H),6.05(br,2H),5.65–5.40( m,3H),4.38–4.04(m,4H),3.28–3.04(m,4H),1.93(dq,J=13.4,6.9,5.9Hz,3H),1.46(p,J=5.2,4.1Hz,4H).

[0593] Example 18: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)(7-(2,5-dihydrofuran-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (18)

[0594] The title compound 18 was prepared by the same method as in Example 4, except that 2-(2,5-dihydrofuran-3-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane was used instead of compound 3f.

[0595] LC-MS: m / z 472[M+H] + .

[0596] 1 H NMR(400MHz,DMSO-d6)δ7.71(s,1H),7.67–7.57(m,2H),7.40(d,J=7.6Hz,1H) ,7.09–7.01(m,2H),6.65(s,2H),6.55(s,1H),6.05(s,1H),5.66(s,1H),5.45 –5.36(m,2H),5.30(d,J=20.3Hz,1H),4.88(s,2H),4.73(d,J=7.1Hz,2H),3.8 0(s,1H),3.61(s,2H),3.30(s,1H),1.41(dd,J=6.2,2.2Hz,3H),1.24(s,2H).

[0597] Example 19: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-morpholinyl-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (19)

[0598] The title compound 19 was prepared in the same manner as in Example 4, except that morpholine was used instead of compound 3f.

[0599] LC-MS: m / z 489[M+H] + .

[0600] 1 H NMR(400MHz,DMSO-d6)δ9.36–8.71(m,3H),8.30–7.61(m,4H),7.35–6.97(m,1H),6.05(br,2H),5.65–5 .40(m,3H),4.38–4.04(m,4H),3.82–3.64(m,4H),3.24-3.03(m,4H),1.93(dq,J=13.4,6.9,5.9Hz,3H).

[0601] Example 20: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(3,6-dihydropyran-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (20)

[0602] The title compound 20 was prepared in the same manner as in Example 4, except that 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester was used instead of compound 3f.

[0603] LCMS: m / z 486.20 [M+H] + .

[0604] 1 H NMR (400MHz, DMSO-d6) δ8.85–8.56(m,2H),7.94(s,2H),7.82(d,J=9.0Hz,1H),7.39(s,1H),7.17–6.89(m,2H),6.28(s,1H),6.12(d,J=24.9Hz,1H ),5.61–5.21(m,3H),4.28–4.17(m,2H),3.81(t,J=5.5Hz,3H),3.65(d,J =25.4Hz, 2H), 2.43 (s, 2H), 1.44 (d, J = 6.0Hz, 3H), 1.24 (d, J = 3.6Hz, 1H).

[0605] Example 21: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(4-methylpiperazin-1-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (21)

[0606] The title compound 21 was prepared in the same manner as in Example 4, except that N-methylpiperazine was used instead of compound 3f.

[0607] LCMS: m / z 502.20 [M+H] + .

[0608] 1 H NMR (400MHz, DMSO-d6) δ7.74–7.54(m,3H),7.20(d,J=8.5Hz,1H),6.71–6.46(m,4H),5.99(s,1H),5.38(d,J=38.3Hz,4H),3.79 (s,1H),3.62(s,1H),3.14(d,J=5.3Hz,4H),2.42(t,J=5.0Hz,3H),2.21(s,3H),1.41(dt,J=6.3,3.1Hz,3H),1.30–1.10(m,3H).

[0609] Example 22: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(3,6-dihydropyran-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (22)

[0610] The title compound 22 was prepared in the same manner as in Example 20, except that 5-bromobenzofuran was used instead of compound 3a.

[0611] LC-MS: m / z 486[M+H] + .

[0612] 1 H NMR (400MHz, DMSO-d6)8.71(s,2H),7.96(s,2H),7.83(d,J=8.8Hz,1H),7.46(s,1H),7.36(d,J=8.5Hz,1H),6.92(s,1H),6.21(s,1H),6.0 5(d,J=31.6Hz,1H),5.62–5.26(m,3H),4.23(q,J=2.9Hz,2H),3.83(t,J=5.5Hz,3H),2.40(d,J=17.3Hz,1H),1.45(dd,J=6.3,2.2Hz,3H).

[0613] Example 23: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(2,5-dihydrofuran-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (23)

[0614] The title compound 23 was prepared in the same manner as in Example 18, except that 5-bromobenzofuran was used instead of compound 3a.

[0615] LC-MS: m / z 472.1[M+H] + .

[0616] 1H NMR (400MHz, DMSO-d6)8.82–8.62(m,1H),8.15–7.89(m,2H),7.83(d,J=8.7Hz,1H),7. 49(d,J=26.8Hz,1H),7.29(d,J=15.6Hz,1H),6.95(s,1H),6.47(d,J=40.8Hz,1H),6.2 3–5.97(m,1H),5.64–5.47(m,2H),4.93(dt,J=4.9,2.6Hz,1H),4.87(q,J=6.8,6.1Hz, 1H), 4.74 (d, J = 5.1Hz, 2H), 3.82 (d, J = 39.3Hz, 3H), 3.03 (s, 4H), 1.45 (d, J = 6.2Hz, 3H).

[0617] Example 24: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-morpholinyl-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (24)

[0618] The title compound 24 was prepared in the same manner as in Example 17, except that 5-bromobenzofuran was used instead of compound 3a.

[0619] LCMS: m / z 489.1[M+H] + .

[0620] 1 H NMR(400MHz, DMSO-d6)δ8.91(s,2H),8.01(br,1H),7.95(s,1H),7.84(d,J=8.7Hz,1H),7.05(s,1H),6.93-6.80(m,2H),6.15–5.9 1(m,1H),5.54–5.35(m,4H),3.76(t,J=4.8Hz,4H),3.62(ddt,J=12.8,6.2,3.2Hz,2H),3.16–3.01(m,6H),1.44(d,J=5.6Hz,3H).

[0621] Example 25: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(pyrrolidin-1-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (25)

[0622] The title compound 25 was prepared in the same manner as in Example 19, except that 5-bromobenzofuran was used instead of compound 3a.

[0623] LC-MS: m / z 473[M+H] + .

[0624] 1H NMR (400MHz, DMSO-d6) δ8.88(s,2H),7.98(d,J=42.1Hz,2H),7.85(d,J=8.8Hz,1H),6.78(d,J=23.1Hz,1H),6.63-6.45(m,2H),6 .18-5.84(m,2H),5.57-5.40(m,3H),3.45(s,1H),3.32-3.15(m,6H),3.02(s,1H),1.97(d,J=5.8Hz,4H),1.45(d,J=5.9Hz,3H).

[0625] Example 26: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-(4-methylpiperazin-1-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (26)

[0626] The title compound 26 was prepared in the same manner as in Example 21, except that 5-bromobenzofuran was used instead of compound 3a.

[0627] LC-MS: m / z 502[M+H] + .

[0628] 1 H NMR (400MHz, DMSO-d6) δ9.87(s,1H),8.73(s,2H),7.96(d,J=38.4Hz,2H),7.80(d,J=8.8Hz,1H),7.10(d,J=28.4Hz,1H),6 .89(s,2H),6.20-5.87(m,2H),5.49(d,J=27.3Hz,3H),3.82-3.65(m,4H),3.18(s,4H),2.89(s,6H),1.45(d,J=5.6Hz,3H).

[0629] Example 27: Preparation of 8-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (27)

[0630] The title compound 27 was prepared in the same manner as in Example 11, except that 5-bromobenzofuran was used instead of compound 3a.

[0631] LC-MS: m / z 482.1[M+H] + .

[0632] 1 H NMR(400MHz,Chloroform-d)δ7.92(d,J=8.5Hz,1H),7.81–7.71(m,2H),7.50(s,1H),7.41(dd,J=8.6,2.1Hz,1H),6. 84(d,J=8.5Hz,1H),5.64–5.50(m,2H),5.43(dt,J=13.5,2.6Hz,2H),3.74(d,J=64.2Hz,4H),1.61(d,J=6.1Hz,3H).

[0633] Example 28: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)8-piperazin-1-yl-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (28)

[0634] The title compound 28 was prepared in the same manner as in Example 7, except that the piperazine compound 3f was used.

[0635] LC-MS: m / z 488.2[M+H] + .

[0636] 1 H NMR (400MHz, DMSO-d6) δ7.73(s,1H),7.67(d,J=8.9Hz,1H),7.60(d,J=8.6Hz,1H),7.00–6.92(m,1H),6.80(q,J=8.8Hz,2H),6.65(s ,2H),5.94(s,1H),5.52–5.28(m,3H),3.70(d,J=57.0Hz,4H),3.04–2.92(m,4H),2.84(t,J=5.1Hz,4H),1.41(dd,J=6.1,3.5Hz,3H).

[0637] Example 29: Preparation of 4-amino-7-fluoro-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(1-methylpyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (29)

[0638] Step 1: Preparation of methyl 4-amino-5-bromo-2-fluorobenzoate (29b)

[0639] Under ice bath, methyl 4-amino-2-fluorobenzoate 29a (3.00 g, 17.1 mmol) and 1-bromopyrrolidine-2,5-dione (3.21 g, 17.5 mmol) were dissolved in chloroform (30.0 mL) and reacted at room temperature for 5 hours. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 20:1-10:1) to obtain 3.48 g of the title compound as a yellow solid in a yield of 82.1%.

[0640] LC-MS: m / z 248 [M+H] + .

[0641] Step 2: Preparation of methyl 4-amino-2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (29c)

[0642] Under a nitrogen atmosphere, compound 29b (3.48 g, 14.0 mmol) and 4,4,4',4',5,5',5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (3.21 g, 17.5 mmol), potassium acetate (2.74 g, 28.0 mmol), and Pd(dppf)Cl2 (219 mg, 0.280 mmol) were dissolved in dioxane (30.0 mL). The mixture was added and reacted at 100°C for 12 hours. The filtrate was concentrated under reduced pressure and the residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 10:1-1:1) to give 2.87 g of the title compound as a yellow solid in a yield of 69.5%.

[0643] LC-MS: m / z 296 [M+H] + .

[0644] Step 3: Preparation of methyl 4-amino-7-fluoro-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylate (29d)

[0645] Under a nitrogen atmosphere, compound 29c (2.87 g, 9.63 mmol), compound 6b (5.78 g, 23.7 mmol), potassium carbonate (2.66 g, 19.3 mmol), and Pd(PPh3)4 (226 mg, 0.192 mmol) were dissolved in dioxane-water (33.0 mL). The mixture was added and reacted at 100°C for 2 hours. The filtrate was concentrated under reduced pressure and the residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 10:1-1:1) to give 2.23 g of the title compound as a yellow solid in a yield of 88.1%.

[0646] LC-MS: m / z 263 [M+H] + .

[0647] Step 4: Preparation of 4-amino-7-fluoro-1,3-dihydrofuro[3,4-c]quinoline-8-carboxylic acid (29e)

[0648] At room temperature, compound 29d (2.23 g, 8.47 mmol) and lithium hydroxide (1.02 g, 42.4 mmol) were dissolved in methanol-water-tetrahydrofuran (12.0 mL). The mixture was reacted at 50°C for 3 hours. The filtrate was concentrated under reduced pressure and the acidity was adjusted with hydrochloric acid. The solid was washed with water and dried to obtain 1.45 g of the title compound as a yellow solid, with a yield of 69.1%.

[0649] LC-MS: m / z 249 [M+H] + .

[0650] Step 5: Preparation of (4-amino-7-fluoro-1,3-dihydrofuro[3,4-c]quinolin-8-yl)(7-(1-methyl-1H-pyrazol-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (29)

[0651] Under a nitrogen atmosphere, compound 3e (200 mg, 0.778 mmol) was dissolved in DMF (3.00 mL). Compound 29e (193 mg, 0.778 mmol), TCFH (304 mg, 1.08 mmol), and NMI (88.4 mg, 1.08 mmol) were added with stirring, and the mixture was stirred at room temperature for 3 hours. 5.00 mL of water was added, and the mixture was extracted with EA (5.00 mL x 3). The mixture was washed with saturated brine (5.00 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was isolated and purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 31.0 mg of the title compound as a white solid in a yield of 8.17%.

[0652] LC-MS: m / z 488[M+H] + .

[0653] 1 H NMR(400MHz, DMSO-d6)δ8.82(s,3H),8.24–7.80(m,3H),7.63(d,J=10.4Hz,1H),7.33–7.04(m,3H),6 .23–5.96(m,2H),5.61–5.36(m,3H),5.09(t,J=3.7Hz,3H),3.85(d,J=9.3Hz,4H),3.49–3.02(m,2H).

[0654] Example 30: Preparation of 4-amino-3-methyl-1,3-dihydrofuro[3,4-c]quinolin-8-yl)7-(piperazin-1-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (30)

[0655] The title compound 30 was prepared in the same manner as in Example 4, except that piperazine was used instead of compound 3f.

[0656] LC-MS: m / z 488[M+H] + .

[0657] According to the preparation method of Example 3 or 4, the following compound was prepared from the corresponding raw materials:

[0658] According to the preparation method of Example 7, the following compounds were prepared from the corresponding raw materials:

[0659] Example 66: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (66)

[0660] Step 1: Preparation of (4aS,9bR)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine or (4aR,9bS)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazine (11a-1 or 11a-2)

[0661] 38g of compound 11a was subjected to chiral separation (chromatographic column model: CHIRALPAK IG 5cm*25cm, 5μm, mobile phase: CO2:MeOH (5% 2mM NH3-MeOH), 200mL / min) to obtain 14.36g of light yellow solid compound 11a-1 (first peak) and 15.39g of white solid compound 11a-2 (second peak).

[0662] 11a-1:

[0663] LC-MS: m / z 255.95 [M+H] + ;

[0664] 11a-2:

[0665] LC-MS: m / z 255.95 [M+H] + .

[0666] Step 2: Preparation of 4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinoline-8-carboxylic acid (66a)

[0667] The title compound 66a was prepared in the same manner as compound 1f, except that compound 29c was used instead of compound 1k.

[0668] Step 3: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-bromo-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (66)

[0669] Under a nitrogen atmosphere, compound 11a-1 or 11a-2 (3 mg, 0.20 mmol), compound 66a (57 mg, 0.22 mmol), 1 mL of DMF, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (84 mg, 0.30 mmol), and 1-methylimidazole (49 mg, 0.60 mmol) were added to a reaction flask and reacted at room temperature for 2 h. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was separated by preparative liquid chromatography (Daisogei 30 mm x 250 mm, C18, 10 μm, 100 Å column, mobile phase: acetonitrile / water, gradient: 30%-80%) to afford 37 mg of the title compound as a white solid.

[0670] LCMS: m / z 497.9 [M+H] + .

[0671] 1 H NMR (400MHz, DMSO-d6) δ8.20(d,J=45.2Hz,2H),7.40–7.27(m,4H),7.23(d,J=9.1Hz,1H),7.18(d,J=8.2Hz,1 H), 6.16 (q, J = 7.1Hz, 1H), 6.05–5.51 (m, 1H), 4.40 (s, 3H), 3.78–3.57 (m, 2H), 3.10 (dd, J = 56.1, 10.2Hz, 2H).

[0672] Example 67: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-(3,6-dihydro-2H-pyran-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-(3,6-dihydro-2H-pyran-4-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (67)

[0673] Under nitrogen atmosphere, compound 66 (120 mg, 0.241 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (60.7 mg, 0.289 mmol), Pd(dppf)Cl2-DCM (18.9 mg, 0.0211 mmol), cesium carbonate (73.3 mg, 0.201 mmol), 1,4-dioxane (2.50 mL), and water (0.50 mL) were added to a reaction flask. The reaction was carried out at 100°C for 12 hours. The filtrate was concentrated under reduced pressure and dried over anhydrous sodium sulfate. The residue was separated and purified by C18 silica gel column chromatography (mobile phase: ACN / water = 1:2) to obtain 40.0 mg of the title compound as a white solid, yield: 33.1%.

[0674] LC-MS: m / z 502 [M+H] + .

[0675] 1H NMR (400MHz, DMSO-d6) δ8.58(d,J=3.7Hz,1H),7.67(d,J=10.2Hz,1H),7.18–6.95(m,2H),6.33–5.90(m,3H ),4.49(d,J=3.0Hz,3H),4.30–4.16(m,2H),3.93–3.58(m,4H),3.20–3.00(m,1H),2.43(d,J=21.8Hz,3H).

[0676] According to the preparation method of Example 67, the following compound was prepared from the corresponding raw materials:

[0677] Example 118: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (118)

[0678] Step 1: Preparation of (4aS,9bR)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazine or (4aR,9bS)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazine (118a).

[0679] Compound 11a-2 (306 mg, 1.20 mmol), boron trifluoride-ether complex (6 mL), and N-chlorosuccinimide (193 mg, 1.44 mmol) were added to a reaction flask at room temperature and stirred under a nitrogen atmosphere for 16 hours. Saturated ammonium chloride solution was added to the reaction mixture at 0°C, and the mixture was extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: methanol / dichloromethane = 0-10%) to afford 279 mg of the title compound as a yellow oil in an 80.4% yield.

[0680] LCMS: m / z 289.95 [M+H] + .

[0681] Step 2: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-bromo-8-chloro-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (118).

[0682] Compound 66a (70.0 mg, 0.267 mmol), N,N-dimethylacetamide (2 mL), HATU (153 mg, 0.401 mmol), and N,N-diisopropylethylamine (104 mg, 0.801 mmol) were added to a reaction flask at room temperature and stirred for 5 minutes. Compound 118a (92.5 mg, 0.320 mmol) was then added, and the mixture was stirred under a nitrogen atmosphere for 16 hours. The reaction solution was concentrated under reduced pressure and separated by preparative liquid chromatography (Daisogei column, 30 mm × 250 mm, C18, 10 μm, 100 Å, mobile phase: acetonitrile / water, gradient: 30%-80%) to afford 63.0 mg of the title compound as a white solid, in a yield of 44.4%.

[0683] LCMS: m / z 532.01[M+H] + .

[0684] 1H NMR (400MHz, DMSO) δ8.60(d,J=3.1Hz,1H),8.51(d,J=6.7Hz,1H),7.69(d,J=10.4Hz,1H),7.50(s,1H),7.41 (s,1H),6.22(s,1H),6.04(d,J=7.1Hz,1H),5.51(s,2H),4.49(s,3H),3.78–3.53(m,2H),3.32–3.04(m,2H).

[0685] Example 119: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-(1-methyl-1H-pyrazol-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofuran[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-(1-methyl-1H-pyrazol-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofuran[2,3-b][1,4]oxazin-1-yl)methanone (119)

[0686] Step 1: ((4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone or (( Preparation of 4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3,4a,9b-tetrahydro-1H-benzofuro[2,3-b][1,4]oxazin-1-yl)methanone (119a)

[0687] Under a nitrogen atmosphere, compound 66 (409 mg, 0.85 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (123 mg, 0.483 mmol), dioxane (3 ml), KOAc (80.0 mg, 0.800 mmol), and Pd(dppf)Cl2 (29.5 mg, 0.040 mmol) were added to a reaction flask and reacted at 80°C for 1 hour. The mixture was returned to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH = 10:1) to afford 398 mg of the title compound as a white solid in an 88.6% yield.

[0688] LCMS: m / z 546.22 [M+H] + .

[0689] Step 2: Preparation of (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aS,9bR)-7-(1-methyl-1H-pyrazol-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazin-1-yl)methanone or (4-amino-7-fluoro-1-methyl-1H-pyrazolo[4,3-c]quinolin-8-yl)((4aR,9bS)-7-(1-methyl-1H-pyrazol-3-yl)-2,3,4a,9b-tetrahydro-1H-benzofurano[2,3-b][1,4]oxazin-1-yl)methanone (119)

[0690] Under nitrogen atmosphere, compound 119a (98 mg, 0.180 mmol), 3-bromo-1-methyl-1H-pyrazole (87.0 mg, 0.540 mmol), 1,4-dioxane (4 mL) and water (400 μL) were added to a reaction flask, followed by potassium carbonate (125 mg, 0.900 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (44.2 mg, 0.054 mmol), and the reaction was carried out at 80°C for 1.5 h. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography (column model: Daisogei 30 mm × 250 mm, C18, 10 μm, 100 Å, mobile phase: acetonitrile / water, gradient: 30%-80%) to give 76 mg of the title compound as a white solid, in a yield of 84.4%.

[0691] LCMS: m / z 500.2 [M+H] + .

[0692] 1H NMR (400MHz, DMSO) δ8.27(s,1H),7.73(d,J=8.1Hz,1H),7.38(s,1H),7.34(d ,J=7.9Hz,3H),6.71(d,J=13.8Hz,1H),6.23(d,J=6.0Hz,1H),6.13(d,J=6.3 Hz,1H),6.02(d,J=5.5Hz,1H),5.58(s,2H),4.42(s,3H),3.88(d,J=9.4Hz,3 H), 3.65 (s, 1H), 3.48 (s, 1H), 3.20 (d, J = 7.0Hz, 1H), 3.06 (d, J = 13.6Hz, 1H).

[0693] According to the preparation method of Example 119, the following compound was prepared from the corresponding raw materials:

[0694] Biological evaluation

[0695] Test Example 1: Inhibitory effect of the compounds of the present invention on PRMT5-MTA binding Reagent preparation:

[0696] The biochemical activity of the compounds was evaluated by measuring their inhibitory effects on PRMT5-MTA binding. A PRMT5 (BPS, Cat#51045)-2.6 μM MTA (Sigma, Cat#D5011) solution was prepared in assay buffer. The reference compound MRTX9768 (MCE, Cat#HY-138684) was diluted in DMSO at concentrations of 1000 μM, 8 μM, 64 nM, and 0.51 nM. The test compounds were diluted in DMSO starting at 9.901 μM and then diluted 5-fold over a 10-fold dilution series. Using an ECHO pipette (Labcyte), 0.1 μL of the compound was transferred to a 384-well plate (Corning, 3657). 5 μL of the PRMT5-MTA mixture was added and the plate was centrifuged at 1000 rpm for 30 seconds. The plate was incubated at 25°C for 30 minutes. SAM solution was prepared in assay buffer, 5 μL of SAM solution was added to the assay plate, and the plate was centrifuged at 1000 rpm for 30 seconds. The plate was then incubated at 25°C for 90 minutes. ULight-streptavidin detection solution was prepared in 1x concentration of LANCE buffer (PE, Cat#CR97-100). After 90 minutes, 10 μL of detection solution (ULight-streptavidin / peptide-biotin 1:6) was added to the assay plate. After 60 minutes, fluorescence data was read at a wavelength of 665 nm (excitation light was 320 or 340 nm) using an Envision Multilabel Analyzer (2104).

[0697] The raw data were converted into inhibition rate using the equation: (1-(sample-SignalAve_PC) / (SignalAve_VC-SignalAve_PC))×100, where SignalAve_PC is the mean fluorescence intensity of the positive control and SignalAve_VC is the mean fluorescence intensity of the negative control. 50 The values ​​of were obtained by four-parameter curve fitting ("log (inhibitor) vs. response - variable slope" mode in GraphPad Prism 8).

[0698] Table 1 provides the in vitro enzymatic inhibitory activity (IC 50 In the table, A refers to the IC value of the compound's in vitro enzymatic inhibition activity against PRMT5+2.6μM MTA. 50 <100nM; B refers to 100nM≤IC 50 <1 μM; C refers to IC 50 ≥1μM.

[0699] Table 1. Enzymatic activity of the compounds of the present invention on PRMT5 in the presence of MTA

[0700] Conclusion: The above data results show that the compounds of the present invention have a significant inhibitory effect on the PRMT5 / MTA complex.

[0701] Experimental Example 2: Inhibitory effect of the compounds of the present invention on proliferation of wild-type HCT-116 and MTAP knockout HCT-116 cells

[0702] The cell activity and selectivity of the compounds were evaluated by detecting the inhibitory effects of the compounds on the proliferation of wild-type and MTAP gene knockout HCT-116 cells.

[0703] On day 0, 100 μl of cell suspension containing 500 HCT-116 parental cells (Horizon, HD PAR-034) or HCT-116 MTAP knockout cells (Horizon, HD R02-033) was added to each well of a white 96-well plate and cultured overnight in a 37°C, 5% CO2 incubator.

[0704] On day 1, dilute the test compound starting at 10,000 μM. For paclitaxel (Taxol Injection, Shuanglu Pharmaceutical) starting at 1,000 μM, perform a three-fold serial dilution in 100% DMSO, with 9+0 concentrations, in duplicate. Add culture medium to the middle plate, then transfer the serially diluted compound to each well according to the corresponding position. After mixing, transfer 50 μL of the compound to each well of the cell plate. Incubate the cell plate in a 37°C, 5% CO2 incubator for 5 days.

[0705] On day 6, cells were digested by adding 75 μL of Trypsin-EDTA solution (Invitrogen, 25200056) to each well. The 96-well plate was placed in a 37°C, 5% CO2 incubator for 3 minutes. Digestion was terminated by adding 125 μL of culture medium to each well. 135 μL of fresh culture medium was added to a new 96-well plate, and 10 μL of the cell suspension was transferred to a new 96-well plate (1:20 dilution). After adding the same concentration of compound as on day 1, the 96-well plate was placed in a 37°C, 5% CO2 incubator for 5 days.

[0706] On day 11, Promega CellTiter-Glo assay was performed. The 96-well plate was removed and equilibrated at room temperature for 20 minutes. 40 μL of CTG (Promega, G7572) was added to each well, the plate was shaken to mix, and the plate was incubated at room temperature for 30 minutes. The fluorescence signal was read using an Envision multi-label analyzer.

[0707] The raw data were converted to inhibition rate using the equation: 100-(sample-SignalAve_PC) / (SignalAve_VC-SignalAve_PC)×100, where SignalAve_PC is the mean fluorescence intensity of the positive control (blank), and SignalAve_VC is the mean fluorescence intensity of the negative control (0.1% DMSO). 50 The values ​​of were obtained by four-parameter curve fitting ("log (inhibitor) vs. response - variable slope" mode in GraphPad Prism 8).

[0708] Table 2 provides the inhibitory activity of the compounds of the present application on the proliferation of HCT-116 parent cells and HCT-116MTAP knockout cells. In the table, a and aa refer to the inhibitory activity IC of the compounds on the proliferation of HCT-116 parent cells and HCT-116MTAP knockout cells, respectively. 50 <100nM; b and bb refer to 100nM ≤ IC 50 <1000nM; c and cc refer to IC 50 ≥1000nM.

[0709] Table 2. Inhibitory activity of the compounds of the present invention on the proliferation of HCT-116 parental cells and HCT-116 MTAP knockout cells

[0710] Conclusion: The above data show that the compounds of the present invention have significant proliferation inhibitory activity on HCT-116MTAP knockout cells, and the inhibitory activity on HCT-116MTAP knockout cells is higher than that on HCT-116 parental cells.

[0711] Experimental Example 3: Pharmacokinetic evaluation of the compound of the present invention in ICR mice

[0712] Male 7-8-week-old ICR mice (Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) were orally administered with the compounds of the present invention in a solvent containing 0.5% methylcellulose and 0.2% Tween 80 at a dosing volume of 10 mL / kg. Blood was collected from the canthal venous plexus of the mice before and at 0.25, 0.50, 1.00, 2.00, 4.00, 6.00, 8.00, and 24.00 hours after dosing. Blood was anticoagulated with sodium heparin and centrifuged at 3500 rpm for 10 minutes at 4°C. Plasma was obtained and stored at -20°C until testing. 10 μL of the plasma sample to be tested was placed in a 96-well plate, and 100 μL of acetonitrile working solution containing 5 ng / mL verapamil hydrochloride (internal standard) (100223-200102, China National Institute for the Control of Pharmaceutical and Biological Products) was added. The mixture was thoroughly mixed by vortexing for 5 minutes and then centrifuged at 4000 rpm for 10 minutes. 50 μL of the supernatant was removed, 150 μL of acetonitrile was added and mixed, and the mixture was centrifuged at 4000 rpm for 10 min. The supernatant was taken and placed in a 96-well sample plate. The blood drug concentration was analyzed by LC / MS (Waters UPLC I Class / LC 30AD, Waters). The pharmacokinetic parameters were analyzed using MassLynx V4.2 SCN977 data processing software. The main pharmacokinetic parameters of the compounds of the present invention are shown in Table 3 below.

[0713] Table 3. Pharmacokinetic parameters of the compounds of the present invention after single oral administration to male ICR mice

[0714] Test Example 4: Inhibitory activity of the compounds of the present invention on hERG

[0715] HEK-293 cells stably expressing hERG potassium channel (Creacell, A-0320) were cultured in DMEM medium (Gibco, 11995-065) containing 10% fetal bovine serum (AvantoR, 76294-180) and 0.8 mg / mL G418 (GPC, AK108) at 37°C and 5% CO2. Before patch clamping, cells were stained with TrypLE TM Express (Gibco, 12604-013) separation, 4 × 10 3 The cells were plated on coverslips and cultured in 24-well plates (final volume: 500 μL). After 18 hours, the assay was performed.

[0716] The membrane was depolarized to +30 mV for 4.8 seconds, then the voltage was restored to -50 mV for 5.2 seconds to remove inactivation and measure the inactivation tail current. The sampling interval was 15 seconds. The maximum tail current magnitude was used to determine the hERG current amplitude. A blank vector was applied to the cells to establish a baseline. After the hERG current was stabilized for at least 5 minutes, the specimen was perfused. The experimental data were collected by an IPA amplifier (Sutter Instrument) and stored in SutterPatch (with Igor Pro) software.

[0717] Drug administration was initiated after the whole-cell hERG currents were stable, and the drug concentration of 10 μM was allowed to act for 5 min (or until the currents stabilized). The assay was repeated three times using at least three cells at each concentration. All electrophysiological experiments were performed at room temperature.

[0718] The current after each drug concentration was normalized to the current of the blank control (0.3% DMSO). Then calculate the inhibition rate corresponding to each drug concentration The mean (Mean), standard deviation (SD) and standard error (SE) of the inhibition rate were calculated for each concentration.

[0719] Table 4 provides the inhibition rate (inhibition %) of the compounds of the present invention on hERG at a concentration of 10 μM. In the table, AA means the inhibition rate of the compound on hERG is <50%; BB means 50%≤inhibition%<100%.

[0720] Table 4. Inhibition rate of hERG by the compounds of the present invention at a concentration of 10 μM

[0721] Test Example 5: Liver microsomal metabolic stability of the compounds of the present invention

[0722] Experimental Materials:

[0723] Liver microsome reaction system:

[0724] The above-mentioned liver microsome reaction system was pre-incubated in a 37°C water bath for 10 minutes. 40 μL of 10mM NADPH solution was added to the reaction system to start the reaction. The final concentration of NADPH was 1mM. 40 μL of ultrapure water was used instead of NADPH as a negative control. One replicate was set up for each time point. At 0, 15, 30, 45, and 60 minutes, 50 μL of reaction sample was taken out and quenched by adding 200 μL of acetonitrile with a final concentration of 200nM. After the sample was mixed, it was centrifuged at 4000 rpm for 30 minutes. After centrifugation, 100 μL of supernatant was taken and added with 100 μL of pure water, mixed, vortexed, and centrifuged for 5 minutes for HPLC-MS / MS analysis to determine the content of the compound.

[0725] HPLC-MS / MS analysis method:

[0726] Mobile phase A: acetonitrile; mobile phase B: 1 mM ammonium acetate aqueous solution (containing 0.1% formic acid);

[0727] Chromatographic column: Waters UPLC C18 1.7 μm, 2.1 × 50 mm, L1-185.

[0728] Table 5 provides the residual rate of the prototype of the compounds of the present invention after incubation in liver microsomes for 60 min.

[0729] Table 5

[0730] Test Example 6: Pharmacodynamics study of the compounds of the present invention

[0731] (1)HCT116 MTAP - / - In vivo drug efficacy experiments on subcutaneous xenograft tumors

[0732] Female Nu / Nu nude mice (6-8 weeks old, Beijing Weitonglihua Experimental Animal Technology Co., Ltd.) were housed in an SPF animal room at a temperature of 20-25°C, a relative humidity of 40-70%, and a light and dark illumination cycle of 12 hours each. The animals had free access to water and food. The animals were adaptively raised for 5 days before the start of the experiment.

[0733] HCT116 MTAP - / - Cells (from Kyinno Biotechnology) were cultured and expanded in vitro. Cells in the logarithmic growth phase were collected and resuspended in serum-free McCoy's 5A medium. The cell density was adjusted to 5×10 7cells / mL, 100 μL of cell suspension was injected into the right anterior axilla of each mouse, and the animal status was observed regularly to monitor the tumor growth. When the tumor volume reached 150 mm 3 Around 40 days, animals with excessively large, small, or irregularly shaped tumors were removed. Tumor-bearing mice with uniform weight, good condition, and similar tumor volumes were randomly divided into groups of six animals each. The model group received a vehicle (3% DMSO + 0.5% methylcellulose + 0.2% Tween 80). The compound was suspended in the vehicle and then administered once daily via gavage at a volume of 10 mL / kg. Tumor diameters were measured twice weekly with a vernier caliper, and tumor volumes were calculated. Animal body weights were also recorded.

[0734] The formula for calculating tumor volume (TV) is: TV = 1 / 2 × a × b 2 , where a represents the long diameter of the tumor and b represents the short diameter of the tumor. The formula for calculating tumor growth inhibition (TGI (100%)) is: TGI = [1-(TV t(T) -TV 初始(T) ) / TV t(C) -TV 初始(C) ]×100%, of which TV t(T) Indicates the tumor volume of the drug-treated group measured each time, TV 初始(T) Indicates the tumor volume of the drug-treated group at the time of drug administration, TV t(C) represents the tumor volume of the vehicle group at each measurement, TV 初始(C) The tumor volume when the vehicle group was administered is shown.

[0735] Table 6 provides the effects of the compounds of the present invention on HCT116 MTAP. - / - Growth inhibition rate of subcutaneous xenograft tumors.

[0736] Table 6

[0737] (2) In vivo efficacy study of NCI-H2228 subcutaneous xenograft tumors

[0738] Female NOG mice (6 weeks old, Beijing Weitonglihua Experimental Animal Technology Co., Ltd.) were housed in an SPF animal room at a temperature of 20-25°C, a relative humidity of 40-70%, and 12 hours of light and dark lighting. The animals had free access to water and food. The animals were adaptively fed for 5 days before the start of the experiment.

[0739] NCI-H2228 cells (from ATCC) were cultured and expanded in vitro. Cells in the logarithmic growth phase were collected and resuspended in serum-free RPMI-1640 medium. The cell density was adjusted to 5 × 107 cells / mL, 100 μL of cell suspension was injected into the right anterior axilla of each mouse, and the animal status was regularly observed to monitor the tumor growth. When the tumor volume reached 150-200 mm 3 Around 40 days, animals with excessively large, small, or irregularly shaped tumors were removed. Tumor-bearing mice with uniform weight, good condition, and similar tumor volumes were randomly divided into groups of six animals each. The model group received a vehicle (3% DMSO + 0.5% methylcellulose + 0.2% Tween 80). The compound was suspended in the vehicle and then administered once daily via gavage at a volume of 10 mL / kg. Tumor diameters were measured twice weekly with a vernier caliper, and tumor volumes were calculated. Animal body weights were also recorded.

[0740] The formula for calculating tumor volume (TV) is: TV = 1 / 2 × a × b 2 The calculation formula of tumor growth inhibition (TGI (100%)) is: TGI = [1-(TV t(T) -TV 初始 (T) ) / TV t(C) -TV 初始(C) ]×100%, of which TV t(T) Indicates the tumor volume of the drug-treated group measured each time, TV 初始(T) Indicates the tumor volume of the drug-treated group at the time of drug administration, TV t(C) represents the tumor volume of the vehicle group at each measurement, TV initial(C) The tumor volume when the vehicle group was administered is shown.

[0741] Table 7 provides the growth inhibition rate of NCI-H2228 subcutaneous xenograft tumors by the compounds of the present invention.

[0742] Table 7

Claims

1. A compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in: is a single bond or a double bond; X 1 CR 5 , CR 5 R 6 、N、NR 5 or C=O; X 2 CR 7 , CR 7 R 8 、N、NR 7 or C=O; X 3 、X 4 are each independently selected from CH2, O, S or NH; X 5 、X 6 are each independently selected from a bond, CH2, O, S or NH; Ring A is selected from saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclyl, phenyl or heteroaryl; Each R 1 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkenyl, alkynyl, heteroalkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a , -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a wherein the alkyl, alkenyl, alkynyl, heteroalkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted by one or more R 1a replace; Each R 2 are each independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, heteroalkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein said alkyl, heteroalkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally replaced by one or more R 2a replace; Each R 3 each independently selected from hydrogen, halogen, amino, nitro, cyano, oxo, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl; Each R 4 Each is independently selected from hydrogen, halogen, hydroxy, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; Each R 1a are independently selected from hydrogen, deuterated, halogen, hydroxy, amino, cyano, alkyl, heteroalkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a , -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-O-(CH2) p -NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-C(=O)NR b -(CH2) p -NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a The alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups; Each R 2a independently selected from hydrogen, halogen, hydroxy, amino, cyano, alkyl, alkoxy, heteroalkyl, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -L-haloalkyl, -(CH2) p -OR a , -L-cycloalkyl, -L-heterocyclyl, -L-aryl, -L-heteroaryl, -SF 6 、HC(=O)-、-NR a R b 、-SO2R a 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-NHC(=O)OR a 、-OC(=O)NR a R b 、-OC(=O)R a 、-NHC(=O)NR a R b 、-NHSO2R a 、-(CH2) p -NR a R b 、-(CH2) p -C(=O)R a 、-(CH2) p -C(=O)NR a R b 、-(CH2) p -NHC(=O)R a 、-(CH2) p -NHC(=O)OR a 、-(CH2) p -OC(=O)NR a R b 、-(CH2) p -OC(=O)R a -(CH2) p -NHC(=O)NR a R b 、-(CH2) p -NHSO2R a ; Each L is independently -O-, -NH-, alkylene, wherein the alkylene is optionally substituted with one or more groups selected from hydroxy, hydroxyalkyl and heteroaryl; R 5 、R 6 Each is independently selected from hydrogen, deuterium, halogen, hydroxy, alkyl, hydroxyalkyl, aminoalkyl, haloalkyl, cyano, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NR a R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R b The alkyl, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; R 7 、R 8 Each is independently selected from hydrogen, deuterium, halogen, hydroxy, alkyl, hydroxyalkyl, aminoalkyl, haloalkyl, cyano, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)OR a 、-NR a C(=O)R b 、-NR a C(=O)OR b 、-C(=O)NR a R b 、-OC(=O)NR a R b 、-S(=O)R a 、-S(=O)2R a 、-SR a 、-S(=O)(=NR a )R b 、-NR a S(=O)2R b and -S(=O)2NR a R b The alkyl, -CH2-aryl, -CH2-heteroaryl, -CH2-cycloalkyl, -CH2-heterocyclyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, sulfhydryl, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; Or, R 5 or R 6 With R 7 or R 8 and the atoms to which they are attached together form a saturated or partially saturated cycloalkyl, a saturated or partially saturated heterocyclyl, an aryl or a heteroaryl, wherein the cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more groups selected from deuterated, halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, deuterated alkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; R a and R b Each is independently selected from hydrogen, halogen, hydroxy, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, thiol, carboxyl, ester, oxo, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl; or, R a and R b together with the nitrogen atom to which it is attached, form a nitrogen-containing heterocyclic group, which is optionally substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl; m1 is 0, 1, 2, 3, or 4; m2 is 0, 1, 2, or 3; m3 is 0, 1, 2 or 3; m4 is 0, 1, or 2; t is 0, 1, or 2; p is an integer from 1 to 6.

2. The compound of general formula (I) according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound of general formula (III) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in: Ring E is selected from saturated or partially saturated 5-6 membered cycloalkyl, saturated or partially saturated 5-6 membered heterocyclyl, phenyl or 5-6 membered heteroaryl; Each R 9 are each independently selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl; s is 0, 1, 2, or 3; Ring A, X 3 、X 4 、X 5 、X 6 、R 1 、R 2 、R 3 、R 4 , m1, m2, m3, m4, t as defined in claim 1.

3. The compound of general formula (I) according to claim 1 or 2, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from phenyl or 5- to 6-membered heteroaryl, preferably phenyl or pyridyl.

4. The compound of general formula (I) according to any one of claims 1 to 3, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Selected from in: X 3 Selected from CH2, O or NH, preferably CH2 and O; X 4 selected from CH2; X 5 Selected from CH2, O, S or NH, preferably O; X 6 Selected from CH2 or a bond, preferably a bond; t is 0, 1, or 2, preferably 1; Among them, R 1 、R 2 、R 3 , m1, m2, m3 as defined in claim 1.

5. The compound of general formula (I) according to any one of claims 1 to 4, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Selected from Among them, R 1 、R 2 、R 3 , m1, m2, m3 as defined in claim 1.

6. The compound of general formula (I) according to any one of claims 1 to 5, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring E is selected from R 10a and R 10b Each independently selected from hydrogen, halogen, amino, hydroxyl, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl; R 10c Selected from halogen, amino, hydroxy, thiol, carboxyl, oxo, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Alkoxy, C 1-6 Cyanoalkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-6 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl.

7. A compound of the general formula (I) according to any one of claims 1 to 6, or a tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, which is a compound of the general formula (IVA) or general formula (IVB), or a tautomer, mesoform, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, in: X 3 Selected from O or CH2, preferably O; R 10a Selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl; preferably C 1-6 Alkyl, C 1-6 deuterated alkyl; R 10b Selected from hydrogen, C 1-6 Alkyl, C 1-6 Deuterated alkyl; preferably C 1-6 alkyl; R 10c Selected from hydrogen, C 1-6 Alkyl; preferably hydrogen; R 1 、R 3 、R 4 , m3, m4 as defined in claim 1.

8. The compound of general formula (I) according to any one of claims 1 to 7, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Each R 1 are each independently selected from halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -NHC(=O)R a , where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocyclyl, phenyl, 5-6 membered heteroaryl are optionally substituted with one or more R 1a replace; Each R 1a independently selected from halogen, cyano, C 1-6 alkyl, 4 to 8 membered heteroalkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a 、-(CH2) p -NR a R b ; R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl; R b Selected from hydrogen, C 1-6 alkyl; m1 is 1; p is an integer from 1 to 4.

9. The compound of general formula (I) according to any one of claims 1 to 8, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Each R 1 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-10 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl, wherein C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 5-10 membered heterocyclyl, 5-10 membered aryl, 5-10 membered heteroaryl are optionally substituted by one or more R 1a replace; Each R 1a Independently selected from deuterated, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-O-(CH2) p -NR a R b 、-C(=O)NR a R b 、-C(=O)NR b -(CH2) p -NR a R b 、-(CH2) p -NR a R b ; The 4-6 membered heterocyclic group is optionally selected from C 1-6 substituted with one or more groups of an alkyl group; R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, C 3-6 Heterocyclic group; said C 3-6 The heterocyclic group is selected from halogen, C 1-6 One or more groups are substituted on the alkyl group; R b Selected from hydrogen, C 1-6 Alkyl; or R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group; m1 is 1; p is an integer from 1 to 4.

10. The compound of general formula (I) according to any one of claims 1 to 8, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is a 5-6 membered heteroaryl group, preferably a pyrazolyl group, a thiazolyl group, an oxazolyl group, a thienyl group, which is optionally further selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 3-6 Heteroalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -C(=O)R a 、-C(=O)NR a R b 、-NHC(=O)R a is substituted by one or more groups; R a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl; R b Selected from hydrogen, C 1-6 alkyl.

11. The compound of general formula (I) according to any one of claims 1 to 8, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is a 5-6 membered heteroaryl or phenyl group, preferably pyrazolyl, thiazolyl, oxazolyl, thienyl, pyridyl, phenyl, which is optionally substituted by one or more R 1a replace; Each R 1a Independently selected from deuterated, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 1-6 aminoalkyl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic group, -OR a 、-(CH2) p -OR a 、-NR a R b 、-O-(CH2) p -NR a R b 、-C(=O)NR b -(CH2) p -NR a R b 、-(CH2) p -NR a R b The 4-6 membered heterocyclic group is optionally C 1-6 Alkyl substituted; R a Selected from C 1-6 Alkyl, C 3-6 Heterocyclic group; said C 3-6 The heterocyclic group is selected from halogen, C 1-6 One or more groups are substituted on the alkyl group; R b Selected from hydrogen, C 1-6 Alkyl; or R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group; m1 is 1; p is an integer of 1 to 4, preferably 1 or 2.

12. A compound of the general formula (I) according to any one of claims 1 to 8, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is a 9-10 membered heterocyclic group or a 9-10 membered heteroaryl group, preferably It is optionally C 1-6 Alkyl substitution; m1 is 1.

13. A compound of formula (I) according to any one of claims 1 to 8, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is a 5-6 membered heterocyclic group, preferably a dihydrofuranyl group, a tetrahydrofuranyl group, a piperidinyl group, which is optionally substituted by C 1-6 Alkyl substitution; m1 is 1.

14. The compound of general formula (I) according to any one of claims 1 to 8, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 C 2-6 Alkynyl, preferably propynyl, which is optionally replaced by R 1a replace; Each R 1a Independently selected from C 3-6 Cycloalkyl, -OR a 、-NR a R b ; R a Selected from C 1-6 Alkyl, C 3-6 Heterocyclic group; said C 3-6 Heterocyclic group is C 1-6 Alkyl substitution; R b Selected from hydrogen, C 1-6 Alkyl; or R a and R b Together with the nitrogen atom to which it is attached, it forms a 4-6 membered nitrogen-containing heterocyclic group; the 4-6 membered nitrogen-containing heterocyclic group is optionally selected from C 1-6 One or more groups are substituted on the alkyl group; m1 is 1.

15. The compound of general formula (I) according to any one of claims 1 to 14, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 2 Selected from hydrogen.

16. The compound of general formula (I) according to any one of claims 1 to 15, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 3 Selected from hydrogen.

17. A compound of formula (I) according to any one of claims 1 to 16, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 4 Selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, m4 is 0 or 1.

18. A compound according to any one of claims 1 to 17, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, selected from:

19. A method for preparing a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, comprising the steps of: In the presence of a condensation agent and an alkaline agent, compound Ie is subjected to a condensation reaction with compound If to obtain a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, Among them, the condensation reagent is preferably N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, and the alkaline reagent is preferably N-methylimidazole; in, Ring A, R 1 、R 2 、R 3 、R 4 、m1、m2、m3、m4、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 As defined in claim 1.

20. A pharmaceutical composition comprising a compound of general formula (I) according to any one of claims 1 to 18, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

21. Use of a compound of formula (I) according to any one of claims 1 to 18 or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 17 in the preparation of a PRMT5 inhibitor.

22. Use of a compound of formula (I) according to any one of claims 1 to 18 or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 20 in the preparation of a medicament for preventing and / or treating diseases associated with PRMT5 activity.

23. The use according to claim 22, wherein the disease associated with PRMT5 activity is a solid tumor, such as non-small cell lung cancer, mesothelial tumor, neurofibrosarcoma, and pancreatic cancer.

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