PRMT5 inhibitor, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a PRMT5 inhibitor, a preparation method therefor and a use thereof. Specifically disclosed are a compound represented by formula I, a tautomer or pharmaceutically acceptable salt thereof, and a use of the compound in preparation of drugs for treating and / or preventing cancers. The cancers can be MTAP-related / mediated cancers.

Description

A PRMT5 inhibitor, its preparation method and application

[0001] This application claims priority to Chinese Patent Application No. 2023108524930 filed on July 11, 2023, Chinese Patent Application No. 2023114530286 filed on November 2, 2023, and Chinese Patent Application No. 202410865364X filed on June 28, 2024. The entire contents of the aforementioned Chinese patent applications are incorporated herein by reference. Technical Field

[0002] The present invention relates to a PRMT5 inhibitor, a preparation method and application thereof. Background Art

[0003] Protein arginine N-methyltransferases (PRMTs) are responsible for the methylation of the guanidino group of arginine. The guanidino group of arginine has two terminal nitrogen atoms that can undergo monomethylation or dimethylation. PRMTs can transfer the methyl group from S-adenosylmethionine (SAM) to the guanidino nitrogen atom of the protein arginine side chain to generate methylated arginine. PRMTs regulate arginine methylation in three different forms: monomethylarginine (MMA), asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) methylation. PRMTs mainly include 9 subtypes, and the enzymes are further divided into type I or type II based on the type of dimethylation. Type I PRMTs catalyze monomethylation or asymmetric dimethylation, while type II enzymes catalyze symmetric dimethylation.

[0004] Protein arginine N-methyltransferase 5 (PRMT5) is a type II arginine methyltransferase responsible for symmetrical dimethylation of arginine residues on histones and non-histone proteins, thereby regulating many life processes in mammalian cells. PRMT5 is overexpressed in many types of cancer, including lymphoma, lung cancer, breast cancer, colorectal cancer, leukemia, glioblastoma, prostate cancer, and ovarian cancer, and is associated with the progression and poor prognosis of most cancers. PRMT5 can also inhibit the transcription of several tumor suppressor genes, including tumorigenicity suppressor 7, non-metastatic gene 23, retinoblastoma family, and programmed cell death 4. Therefore, PRMT5 is a potential target for cancer treatment.

[0005] Methylthioadenosine phosphorylase (MTAP) is a tumor suppressor gene that is frequently deleted in many human tumors, including 53% of glioblastomas and 26% of pancreatic cancers. It is the most frequently mutated gene in cancer. Furthermore, MTAP is often co-deleted with the common tumor suppressor gene CDKN2A, with this co-deletion occurring in 9% to 15% of tumors.

[0006] Synthetic lethality refers to a phenomenon in which simultaneous mutations in two genes lead to cell death, whereas mutations in either gene alone do not cause cell death. Studies have shown that PRMT5 and MTAP form a synthetic lethal pair, and that inhibition of PRMT5 can lead to synthetic lethality in MTAP-deficient tumor cells. MTAP is involved in the metabolism of methylthioadenosine (MTA). In cancer cells with MTAP gene deletion, MTA accumulates and binds to PRMT5, forming a PRMT5-MTA complex. Therefore, PRMT5 inhibitors may be a highly selective treatment for patients with MTAP deficiency or low expression.

[0007] MRTX1719, developed by Mirati Therapeutics, is a novel PRMT5 inhibitor that selectively inhibits PRMT5 bound to MTA. Its structure is as follows:

[0008] Although the above-mentioned new PRMT5 inhibitors have entered the clinical trial stage and have made some progress in this field, they have not yet been approved for treatment. Therefore, it is still necessary to continue to develop new PRMT5 inhibitors targeting the PRMT5-MTA complex. This new PRMT5-specific small molecule inhibitor may bring new chemotherapy for cancer.

[0009] Summary of the Invention

[0010] The technical problem to be solved by the present invention is to overcome the single structure of PRMT5 inhibitors in the prior art and provide a novel PRMT5 inhibitor, its preparation method and use. The PRMT5 inhibitor provided by the present invention has good inhibitory activity or selectivity against human colon cancer cells with stable MTAP protein knockout (HCT-116-MTAP-KO-1A2), and can inhibit the proliferation of tumor cells.

[0011] The present invention provides a compound as shown in Formula I, its tautomers or pharmaceutically acceptable salts thereof,

[0012] R 1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 1a Substituted C1-C6 alkyl, one or more R 1b Substituted C2-C6 alkenyl, one or more R 1c Substituted C2-C6 alkynyl, one or more R 1d Substituted 3-7 membered cycloalkyl, one or more R 1eSubstituted 3-7 membered cycloalkenyl, one or more R 1f substituted 4-10 membered heterocyclic group, one or more R 1g Substituted C6-C 10 Aryl or one or more R 1h substituted 5-10 membered heteroaryl;

[0013] Each R 1a 、R 1b 、R 1c 、R 1d 、R 1e 、R 1f 、R 1g and R 1h are independently deuterium, halogen, -OH, =O, cyano, -NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0014] R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-OR 13 、-C(= O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 、-P(=O)(R 12 )2、-P(=O)(OR 12 )2、-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl, one or more R 2b Substituted C2-C6 alkenyl, one or more R 2c Substituted C2-C6 alkynyl, one or more R 2d Substituted 3-7 membered cycloalkyl, one or more R 2e Substituted 3-7 membered cycloalkenyl, one or more R 2f substituted 4-10 membered heterocyclic group, one or more R 2g Substituted C6-C 10 Aryl or one or more R 2h substituted 5-10 membered heteroaryl;

[0015] Each R 2a 、R 2b 、R 2c 、R 2d 、R 2e 、R 2f 、R 2g and R 2h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0016] R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-C(=O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 、-P(=O)(R 12)2、-P(=O)(OR 12 )2、-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12 S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 5a Substituted C1-C6 alkyl, one or more R 5b Substituted 3-7 membered cycloalkyl, one or more R 5c Substituted 3-7 membered cycloalkenyl, one or more R 5d substituted 4-10 membered heterocyclic group, one or more R 5e Substituted C6-C 10 Aryl or one or more R 5f substituted 5-10 membered heteroaryl;

[0017] Each R 5a 、R 5b 、R 5c 、R 5d 、R 5e and R 5f Each is independently deuterium, halogen, =O, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -OR 13 、-NR 10 R 11 、-C(=O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 、-P(=O)(R 12 )2、-P(=O)(OR12 )2、-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12 S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl, one or more R 5-b Substituted C2-C6 alkenyl, one or more R 5-c Substituted C2-C6 alkynyl, one or more R 5-d Substituted 3-7 membered cycloalkyl, one or more R 5-e Substituted 3-7 membered cycloalkenyl, one or more R 5-f substituted 4-10 membered heterocyclic group, one or more R 5-g Substituted C6-C 10 Aryl or one or more R 5-h substituted 5-10 membered heteroaryl;

[0018] Each R 5-a 、R 5-b 、R 5-c 、R 5-d 、R 5-e 、R 5-f 、R 5-g and R 5-h are independently deuterium, halogen, =O, cyano, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C1-C6 alkyl substituted by one or more halogen, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuterium, or C1-C6 alkyl substituted by one or more cyano groups;

[0019] R 7 For hydrogen, deuterium, halogen, cyano, nitro, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 7a Substituted C1-C6 alkyl, one or more R 7b Substituted C2-C6 alkenyl, one or more R 7c Substituted C2-C6 alkynyl, one or more R 7d Substituted 3-7 membered cycloalkyl, one or more R 7e Substituted 3-7 membered cycloalkenyl, one or more R 7f substituted 4-10 membered heterocyclic group, one or more R 7g Substituted C6-C 10 Aryl or one or more R 7h substituted 5-10 membered heteroaryl;

[0020] Each R 7a 、R 7b 、R 7c 、R 7d 、R 7e 、R 7f 、R 7g and R 7h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0021] Each R 8 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-OR 13 , by one or more R 8a Substituted C1-C6 alkyl, one or more R 8b Substituted C2-C6 alkenyl, one or more R 8c Substituted C2-C6 alkynyl, one or more R 8d Substituted 3-7 membered cycloalkyl, one or more R 8e Substituted 3-7 membered cycloalkenyl, one or more R 8f substituted 4-10 membered heterocyclic group, one or more R 8g Substituted C6-C 10 Aryl or one or more R 8h substituted 5-10 membered heteroaryl;

[0022] Each R 8a 、R 8b 、R 8c 、R 8d 、R 8e 、R 8f 、R 8g and R 8h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0023] R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b 、-C(=O)-OC 6-10 Aryl, -C(=O)-C 1-6 Alkyl or -(CH2) m -OP(=O)(OR 9c )2;

[0024] R 9a and R 9b are each independently hydrogen, deuterium or C 1-20 alkyl;

[0025] Each R 9c are each independently hydrogen, deuterium or C 1-20 alkyl;

[0026] Each R 10 and R 11 Each is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 10a Substituted C1-C6 alkyl, one or more R 10b Substituted C2-C6 alkenyl, one or more R 10c Substituted C2-C6 alkynyl, one or more R 10d Substituted 3-7 membered cycloalkyl, one or more R 10e Substituted 3-7 membered cycloalkenyl, one or more R 10f substituted 4-10 membered heterocyclic group, one or more R 10g Substituted C6-C 10 Aryl or one or more R 10h substituted 5-10 membered heteroaryl;

[0027] Each R10a 、R 10b 、R 10c 、R 10d 、R 10e 、R 10f 、R 10g and R 10h Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0028] Each R 12 are independently hydrogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, 4-10 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 12a Substituted C 1-6 Alkyl, one or more R 12b Substituted C 3-8 Cyclic hydrocarbon group, one or more R 12c substituted 4-10 membered heterocyclic group, one or more R 12d Substituted C 6-10 Aryl or one or more R 12e substituted 5-10 membered heteroaryl;

[0029] Each R 12a 、R 12b 、R 12c 、R 12d and R 12e Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0030] Each R 13 Each is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 13a Substituted C1-C6 alkyl, one or more R 13b Substituted C2-C6 alkenyl, one or more R 13c Substituted C2-C6 alkynyl, one or more R 13d Substituted 3-7 membered cycloalkyl, one or more R 13e Substituted 3-7 membered cycloalkenyl, one or more R 13f substituted 4-10 membered heterocyclic group, one or more R 13g Substituted C6-C 10 Aryl or one or more R13h substituted 5-10 membered heteroaryl;

[0031] Each R 13a 、R 13b 、R 13c 、R 13d 、R 13e 、R 13f 、R 13g and R 13h Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl;

[0032] n is 1 or 2;

[0033] m is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0034] The heteroatoms in the 5-10 membered heteroaryl group and the 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3.

[0035] In certain preferred embodiments of the present invention, certain groups in the compound as shown in Formula I, its tautomers or pharmaceutically acceptable salts thereof are defined as follows, and the unmentioned groups are the same as those described in any scheme of the present application (referred to as "in certain preferred embodiments of the present invention").

[0036] In certain preferred embodiments of the present invention, the compound represented by Formula I is a stereoisomer or a mixture thereof. For example, when it contains one chiral center, the compound represented by Formula I is an R configuration, an S configuration, or an R, S configuration racemate.

[0037] In certain preferred embodiments of the present invention, the compound of formula I is a compound of formula IA or formula IB:

[0038] In certain preferred embodiments of the present invention, the compound as shown in Formula I is a compound as shown in Formula I-1, Formula I-2, Formula I-3 or Formula I-4:

[0039] In certain preferred embodiments of the present invention, the compound as shown in Formula I is a compound as shown in Formula I-2-1-1, Formula I-2-1-2, Formula I-3-1-1, Formula I-3-1-2, Formula I-4-1-1 or Formula I-4-1-2:

[0040] In certain preferred embodiments of the present invention, the compound of formula I is a compound of formula I-1-1-1 or formula I-1-1-2:

[0041] In certain preferred embodiments of the present invention, R 1 wherein the C1-C6 alkyl group and the 1a The C1-C6 alkyl group in the substituted C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.

[0042] In certain preferred embodiments of the present invention, R 6 wherein the halogen is fluorine, chlorine, bromine or iodine, for example fluorine.

[0043] In certain preferred embodiments of the present invention, R 3 wherein the 4-10 membered heterocyclic group and the one or more R 2f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is a 4-10 membered heterocycloalkyl or a 4-10 membered heterocycloalkenyl group, the 4-10 membered heterocycloalkyl group may independently be a 5-6 membered heterocycloalkyl group, the 4-10 membered heterocycloalkyl group may independently be a monocyclic ring, the heteroatom of the 4-10 membered heterocycloalkyl group may independently be N, the number of heteroatoms may independently be 1, or may be For example

[0044] In certain preferred embodiments of the present invention, R 5 wherein the C1-C6 alkyl group and the 5a The C1-C6 alkyl group in the substituted C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl, ethyl or isopropyl.

[0045] In certain preferred embodiments of the present invention, R 5 wherein the 3-7 membered cycloalkyl group and the one or more R 5b The 3- to 7-membered cycloalkyl groups in the substituted 3- to 7-membered cycloalkyl groups are independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, for example, cyclopropyl.

[0046] In certain preferred embodiments of the present invention, each R 5a wherein the 4-10 membered heterocyclic group and one or more R 5-fThe 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is a 4-10 membered heterocycloalkyl or a 4-10 membered heterocycloalkenyl, and the 4-10 membered heterocycloalkyl is independently a 4-6 membered monoheterocycloalkyl or a 7-10 membered biheterocycloalkyl. The biheterocycloalkyl may be a spirocyclic, a bridged ring or a fused ring. The heteroatoms of the 4-10 membered heterocycloalkyl may independently be N and / or O, and the number of heteroatoms may independently be 1 or 2. The 4-6 membered monoheterocycloalkyl may independently be For example The 7-10 membered biheterocycloalkyl group may independently be a bridged ring, or may independently be a 7-8 membered bridged ring, for example For example

[0047] In certain preferred embodiments of the present invention, each R 5-f wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine.

[0048] In certain preferred embodiments of the present invention, each R 5-f wherein the C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.

[0049] In certain preferred embodiments of the present invention, each R 10 and R 11 wherein the C1-C6 alkyl group and the 10a The C1-C6 alkyl group in the substituted C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.

[0050] In certain preferred embodiments of the present invention, each R 13 wherein the C1-C6 alkyl group is replaced by one or more R 13a The C1-C6 alkyl group in the substituted C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.

[0051] In certain preferred embodiments of the present invention, each R 13 , the 3-7 membered cycloalkyl group and one or more R 13d The 3- to 7-membered cycloalkyl groups in the substituted 3- to 7-membered cycloalkyl groups are independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, for example, cyclopropyl.

[0052] In certain preferred embodiments of the present invention, each R 13dwherein the C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl.

[0053] In certain preferred embodiments of the present invention, each C1-C 20 Alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0054] In certain preferred embodiments of the present invention, each C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0055] In certain preferred embodiments of the present invention, each C2-C6 alkenyl group is independently ethenyl, propenyl, butenyl or pentenyl.

[0056] In certain preferred embodiments of the present invention, each C2-C6 alkynyl group is independently ethynyl, propynyl, butynyl or pentynyl.

[0057] In certain preferred embodiments of the present invention, each 3-7 membered cycloalkyl group is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0058] In certain preferred embodiments of the present invention, each 3-7 membered cycloalkenyl group is independently cyclopropenyl, cyclobutenyl, cyclopentenyl or cyclohexenyl.

[0059] In certain preferred embodiments of the present invention, each 4-10 membered heterocyclyl is independently a saturated or unsaturated heterocyclyl, such as a saturated heterocycloalkyl.

[0060] In certain preferred embodiments of the present invention, each 4-10 membered heterocyclic group is independently a monoheterocyclic group or a biheterocyclic group, and the biheterocyclic group may be spirocyclic, bridged or fused.

[0061] In certain preferred embodiments of the present invention, each 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group. For example The 7-10 membered biheterocyclic group may be a bridged ring or a 7-8 membered bridged ring, for example For example

[0062] In certain preferred embodiments of the present invention, the heteroatom in each 4-10 membered heterocyclic group may independently be N and / or O, and the number of heteroatoms may independently be 1 or 2.

[0063] In certain preferred embodiments of the present invention, each C6-C 10 Aryl is independently phenyl or naphthyl.

[0064] In certain preferred embodiments of the present invention, each 5-10 membered heteroaryl group is independently a 5-6 membered monocyclic heteroaryl group or an 8-10 membered polycyclic heteroaryl group, wherein the heteroatom species is N and the number of heteroatoms is 1, 2 or 3.

[0065] In certain preferred embodiments of the present invention, each halogen is independently fluorine, chlorine, bromine or iodine.

[0066] In certain preferred embodiments of the present invention, each C1-C6 alkyl-O- is independently methyl-O-, ethyl-O-, n-propyl-O-, isopropyl-O-, n-butyl-O-, tert-butyl-O-, isobutyl-O- or sec-butyl-O-.

[0067] In certain preferred embodiments of the present invention, R 1 is hydrogen, C1-C6 alkyl or is replaced by one or more R 1a Substituted C1-C6 alkyl; preferably, R 1 It is a C1-C6 alkyl group.

[0068] In certain preferred embodiments of the present invention, each R 1a are independently deuterium, halogen, -OH, cyano or -NR 10 R 11 .

[0069] In certain preferred embodiments of the present invention, R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl or one or more R 2f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 2f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is 1, 2 or 3; preferably, R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 , the heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1.

[0070] In certain preferred embodiments of the present invention, R 3 is a 4-10 membered heterocyclic group or -OR 13 ; The heteroatom in the 4-10 membered heterocyclic group is N, O or S, and the number of heteroatoms is 1, 2 or 3.

[0071] In certain preferred embodiments of the present invention, R 6 It is a halogen.

[0072] In certain preferred embodiments of the present invention, each R 2a and R 2f are independently deuterium, halogen, =O, cyano, -NR 10 R 11 or C1-C6 alkyl.

[0073] In certain preferred embodiments of the present invention, R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 5a Substituted C1-C6 alkyl, one or more R 5b Substituted 3-7 membered cycloalkyl, one or more R 5d substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted one or more R 5d The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is independently 1 or 2; preferably, R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or one or more R 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl.

[0074] In certain preferred embodiments of the present invention, R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl.

[0075] In certain preferred embodiments of the present invention, each R 5a 、R 5b and R 5d Each is independently deuterium, halogen, cyano, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl, one or more R5-f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; preferably, each R 5a and R 5b Each independently represents a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2.

[0076] In certain preferred embodiments of the present invention, each R 5a Each independently represents a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5- f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2. Preferably, each R 5a Each independently represents a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 The heteroatoms in the 4-6 membered heterocyclic group are N and O, the number of heteroatoms is 2, and more preferably each R 5a Each is independently a 4-6 membered monoheterocyclic group; the heteroatoms in the 4-6 membered monoheterocyclic group are N and O, and the number of heteroatoms is 2.

[0077] In certain preferred embodiments of the present invention, each R 5b Each is independently -NH2.

[0078] In certain preferred embodiments of the present invention, each R 5-f and R 5-a are independently deuterium, halogen, =O, cyano, -OR13 、-NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuteriums or C1-C6 alkyl substituted by one or more cyano groups; preferably, each R 5-f and R 5-a are each independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl, more preferably, each R 5-f are independently halogen or C1-C6 alkyl.

[0079] In certain preferred embodiments of the present invention, R 7 is hydrogen, deuterium, halogen, cyano, nitro, -OH, -NH2, C1-C6 alkyl or is replaced by one or more R 7a Substituted C1-C6 alkyl; preferably, R 7 For hydrogen.

[0080] In certain preferred embodiments of the present invention, each R 7a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl.

[0081] In certain preferred embodiments of the present invention, each R 8 are independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, -NR 10 R 11 、-OR 13 or by one or more R 8a Substituted C1-C6 alkyl; preferably, each R 8 are each independently hydrogen or -NR 10 R 11 ; For example, R 8 -NR 10 R 11 .

[0082] In certain preferred embodiments of the present invention, each R 8a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl.

[0083] In certain preferred embodiments of the present invention, R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b or -(CH2) m -OP(=O)(OR9c )2; preferably, R 9 are independently hydrogen.

[0084] In certain preferred embodiments of the present invention, R 9a and R 9b are each independently hydrogen or C 1-20 alkyl.

[0085] In certain preferred embodiments of the present invention, each R 9c are each independently hydrogen or C 1-20 alkyl.

[0086] In certain preferred embodiments of the present invention, R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 10a Substituted C1-C6 alkyl, one or more R 10d substituted 3-7 membered cycloalkyl or one or more R 10f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 10f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; preferably, R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl.

[0087] In certain preferred embodiments of the present invention, each R 10a 、R 10d and R 10f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; preferably, each R 10a each independently deuterium.

[0088] In certain preferred embodiments of the present invention, each R 13 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 13a Substituted C1-C6 alkyl, one or more R 13d substituted 3-7 membered cycloalkyl or one or more R 13f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 13f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; preferably, each R13 Each independently represents hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl. More preferably, each R 13 Each is independently hydrogen or a 3-7 membered cycloalkyl group.

[0089] In certain preferred embodiments of the present invention, each R 13a 、R 13d and R 13f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl.

[0090] In certain preferred embodiments of the present invention, n is 1 or 2;

[0091] R 1 is hydrogen, C1-C6 alkyl or is replaced by one or more R 1a Substituted C1-C6 alkyl;

[0092] Each R 1a are independently deuterium, halogen, -OH, cyano or -NR 10 R 11 ;

[0093] R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl or one or more R 2f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 2f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is independently 1, 2 or 3;

[0094] Each R 2a and R 2f are independently deuterium, halogen, =O, cyano, -NR 10 R 11 or C1-C6 alkyl;

[0095] R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 5a Substituted C1-C6 alkyl, one or more R 5b substituted 3-7 membered cycloalkyl or one or more R 5dsubstituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and said substituted by one or more R 5d The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0096] Each R 5a 、R 5b and R 5d Each is independently deuterium, halogen, cyano, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl or one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0097] Each R 5-f and R 5-a are independently deuterium, halogen, =O, cyano, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl substituted by one or more halogen, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuterium, or C1-C6 alkyl substituted by one or more cyano groups;

[0098] R 7 For hydrogen, deuterium, halogen, cyano, nitro, -OR 13 、-NR 10 R 11 , C1-C6 alkyl or one or more R 7a Substituted C1-C6 alkyl;

[0099] Each R 7a are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 or C1-C6 alkyl;

[0100] Each R 8 are independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, -NR 10 R 11 、-OR 13 or by one or more R 8a Substituted C1-C6 alkyl;

[0101] Each R 8a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl;

[0102] R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b or -(CH2) m -OP(=O)(OR 9c )2;

[0103] R 9a and R 9b are each independently hydrogen or C 1-20 alkyl;

[0104] Each R 9c are each independently hydrogen or C 1-20 alkyl;

[0105] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 10a Substituted C1-C6 alkyl, one or more R 10d substituted 3-7 membered cycloalkyl or one or more R 10f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 10f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3;

[0106] Each R 10a 、R 10d and R 10f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl;

[0107] Each R 13 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, one or more R 13a Substituted C1-C6 alkyl, one or more R 13d substituted 3-7 membered cycloalkyl or one or more R 13f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 13f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3;

[0108] Each R 13a 、R 13d and R 13f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl.

[0109] In certain preferred embodiments of the present invention, n is 1 or 2;

[0110] R 1 is a C1-C6 alkyl group;

[0111] R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0112] R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0113] Each R 5a and R 5b Each independently represents a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0114] Each R 5-f are each independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl;

[0115] R 7 is hydrogen;

[0116] Each R 8 are each independently hydrogen or -NR 10 R 11 ;

[0117] R 9 are independently hydrogen;

[0118] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0119] Each R 10a Each independently is deuterium;

[0120] Each R 13 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, or is replaced by one or more R 13d substituted 3-7 membered cycloalkyl;

[0121] Each R 13d Each is independently a C1-C6 alkyl group.

[0122] In certain preferred embodiments of the present invention, n is 1 or 2;

[0123] R 1 is a C1-C6 alkyl group;

[0124] R 2 、R 3 、R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0125] R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or one or more R 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0126] Each R 5a and R 5b Each independently represents a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted one or more R 5-fThe heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0127] Each R 5-f are each independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl;

[0128] R 7 is hydrogen;

[0129] Each R 8 are each independently hydrogen or -NR 10 R 11 ;

[0130] R 9 are independently hydrogen;

[0131] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0132] Each R 10a Each independently is deuterium;

[0133] Each R 13 Each is independently hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl.

[0134] In certain preferred embodiments of the present invention, n is 1 or 2;

[0135] R 1 is a C1-C6 alkyl group;

[0136] R 2 is cyano;

[0137] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0138] R 4 is hydrogen;

[0139] R 6 is a halogen;

[0140] R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0141] Each R 5a and R 5b Each independently represents a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0142] Each R 5-f are each independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl;

[0143] R 7 is hydrogen;

[0144] Each R 8 are each independently hydrogen or -NR 10 R 11 ;

[0145] R 9 are independently hydrogen;

[0146] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0147] Each R 10a Each independently is deuterium;

[0148] Each R 13 Each independently represents hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl or substituted by one or more R 13d Substituted 3-7 membered cycloalkyl; each R 13d Each is independently a C1-C6 alkyl group.

[0149] In certain preferred embodiments of the present invention, n is 1 or 2;

[0150] R 1 is a C1-C6 alkyl group;

[0151] R 2 is cyano;

[0152] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0153] R 4 is hydrogen;

[0154] R 6 is a halogen;

[0155] R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or one or more R 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0156] Each R 5a and R 5b Each independently represents a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0157] Each R 5-f are each independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl;

[0158] R 7 is hydrogen;

[0159] Each R 8 are each independently hydrogen or -NR 10 R 11 ;

[0160] R 9 are independently hydrogen;

[0161] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0162] Each R10a Each independently is deuterium;

[0163] Each R 13 Each is independently hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl.

[0164] In certain preferred embodiments of the present invention, n is 1;

[0165] R 1 is a C1-C6 alkyl group;

[0166] R 2 is cyano;

[0167] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0168] R 4 is hydrogen;

[0169] R 6 is a halogen;

[0170] R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0171] Each R 5a Each independently represents a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0172] Each R 5b each independently -NH2;

[0173] Each R 5-f are independently halogen or C1-C6 alkyl;

[0174] R 7 is hydrogen;

[0175] R 8 -NR10 R 11 ;

[0176] R 9 are independently hydrogen;

[0177] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0178] Each R 10a independently for deuterium;

[0179] Each R 13 Each independently represents a 3-7 membered cycloalkyl group, or is replaced by one or more R 13d substituted 3-7 membered cycloalkyl;

[0180] Each R 13d Each is independently a C1-C6 alkyl group.

[0181] In certain preferred embodiments of the present invention, n is 1;

[0182] R 1 is a C1-C6 alkyl group;

[0183] R 2 is cyano;

[0184] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0185] R 4 is hydrogen;

[0186] R 6 is a halogen;

[0187] R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or one or more R 5a Substituted C1-C6 alkyl or one or more R 5b substituted 3-7 membered cycloalkyl;

[0188] Each R 5a Each independently represents a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or by one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-fThe 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, wherein the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2;

[0189] Each R 5b each independently -NH2;

[0190] Each R 5-f are independently halogen or C1-C6 alkyl;

[0191] R 7 is hydrogen;

[0192] R 8 -NR 10 R 11 ;

[0193] R 9 are independently hydrogen;

[0194] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0195] Each R 10a independently for deuterium;

[0196] Each R 13 Each is independently a 3-7 membered cycloalkyl group.

[0197] In certain preferred embodiments of the present invention, n is 1;

[0198] R 1 is a C1-C6 alkyl group;

[0199] R 2 is cyano;

[0200] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0201] R 4 is hydrogen;

[0202] R 6 is a halogen;

[0203] R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl or is replaced by one or more R 5aSubstituted C1-C6 alkyl;

[0204] Each R 5a Each independently represents a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 The heteroatoms in the 4-6 membered monoheterocyclic group are N and O, and the number of heteroatoms is 2;

[0205] R 7 is hydrogen;

[0206] R 8 -NR 10 R 11 ;

[0207] R 9 are independently hydrogen;

[0208] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0209] Each R 10a Each independently is deuterium;

[0210] Each R 13 Each independently represents a 3-7 membered cycloalkyl group, or is replaced by one or more R 13d substituted 3-7 membered cycloalkyl;

[0211] Each R 13d Each is independently a C1-C6 alkyl group.

[0212] In certain preferred embodiments of the present invention, n is 1;

[0213] R 1 is a C1-C6 alkyl group;

[0214] R 2 is cyano;

[0215] R 3 is a 5-6 membered monoheterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1;

[0216] R 4 is hydrogen;

[0217] R 6 is a halogen;

[0218] R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl or is replaced by one or more R 5a Substituted C1-C6 alkyl;

[0219] Each R 5a Each independently represents a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 The heteroatoms in the 4-6 membered monoheterocyclic group are N and O, and the number of heteroatoms is 2;

[0220] R 7 is hydrogen;

[0221] R 8 -NR 10 R 11 ;

[0222] R 9 are independently hydrogen;

[0223] R 10 and R 11 Each independently represents hydrogen, C1-C6 alkyl or is replaced by one or more R 10a Substituted C1-C6 alkyl;

[0224] Each R 10a Each independently is deuterium;

[0225] Each R 13 Each is independently a 3-7 membered cycloalkyl group.

[0226] In certain preferred embodiments of the present invention, R 1 It is a methyl group.

[0227] In certain preferred embodiments of the present invention, R 2 、R 3 、R 4 and R 6 are independently hydrogen, cyano, F,

[0228] In certain preferred embodiments of the present invention, R 2 、R 3 、R 4 and R 6 are independently hydrogen, cyano, F,

[0229] In certain preferred embodiments of the present invention, R 2 、R 3 、R 4 and R 6 Each independently

[0230] In certain preferred embodiments of the present invention, R 2 It is a cyano group.

[0231] In certain preferred embodiments of the present invention, R 3 for

[0232] In certain preferred embodiments of the present invention, R 3 for

[0233] In certain preferred embodiments of the present invention, R 3 for

[0234] In certain preferred embodiments of the present invention, R 4 For hydrogen.

[0235] In certain preferred embodiments of the present invention, R 6 For F.

[0236] In certain preferred embodiments of the present invention, R 5 For hydrogen, methyl, Preferably, hydrogen, methyl,

[0237] In certain preferred embodiments of the present invention, R 5 for methyl, Preferably methyl,

[0238] In certain preferred embodiments of the present invention, R 5 For hydrogen.

[0239] In certain preferred embodiments of the present invention, R 7 For hydrogen.

[0240] In certain preferred embodiments of the present invention, R 8 Independently

[0241] In certain preferred embodiments of the present invention, R 9 For hydrogen.

[0242] In certain preferred embodiments of the present invention, the compound represented by formula I is any one of the following compounds:

[0243] The present invention also provides a compound as shown below,

[0244] The present invention also provides a pharmaceutical composition comprising the compound as shown in Formula I, its tautomers or pharmaceutically acceptable salts thereof, and at least one pharmaceutical excipient.

[0245] The selection of the pharmaceutical excipients varies depending on the route of administration and the characteristics of the action, and can generally be fillers, diluents, binders, wetting agents, disintegrants, lubricants, emulsifiers, suspending agents, etc. conventional in the art.

[0246] The present invention also provides the use of the compound represented by Formula I, its tautomer, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above, in the preparation of a PRMT5 inhibitor, such as a PRMT5·MTA inhibitor. In such applications, the PRMT5 inhibitor can be used in mammalian organisms; it can also be used in vitro, primarily for experimental purposes, for example, as a standard or control sample for comparison, or prepared according to conventional methods in the art to provide a kit for rapid detection of the effect of inhibiting PRMT5.

[0247] In the present invention, PRMT5·MTA refers to a complex formed by the binding of MTA (methylthioadenosine) and PRMT5 (protein arginine N-methyltransferase 5) in cancer cells lacking the MTAP (methylthioadenosine phosphorylase) gene.

[0248] The present invention also provides the use of the compound as shown in Formula I, its tautomer or pharmaceutically acceptable salt, or the pharmaceutical composition as described above in the preparation of a drug, for example, in the preparation of a drug for treating and / or preventing cancer. The cancer may be an MTAP-related / mediated cancer, preferably, the MTAP-related / mediated cancer is head and neck cancer (such as thyroid cancer, meningeal cancer, intracranial metastasis or glioblastoma), respiratory system cancer (such as lung cancer or nasopharyngeal cancer), digestive system cancer (esophageal cancer, gastric cancer, liver cancer, bile duct cancer, pancreatic cancer, colorectal cancer, rectal cancer or colon cancer), urinary system cancer (such as kidney cancer, bladder cancer, prostate cancer or testicular cancer), bone cancer, gynecological cancer (such as breast cancer, endometrial cancer, cervical cancer or ovarian cancer), blood system cancer (such as leukemia, lymphoma or myeloma) or other types of cancer (such as melanoma or skin cancer), more preferably lymphoma, lung cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, leukemia, glioblastoma, prostate cancer or ovarian cancer.

[0249] The present invention also provides the use of a compound as shown in Formula I as described above, its tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above in the preparation of a medicament, for example, in the preparation of a medicament for treating and / or preventing a disease associated with MTAP / mediated. The disease may be cancer. The cancer may be head and neck cancer (e.g., thyroid cancer, meningeal cancer, intracranial metastasis or glioblastoma), respiratory cancer (e.g., lung cancer or nasopharyngeal cancer), digestive cancer (esophageal cancer, gastric cancer, liver cancer, bile duct cancer, pancreatic cancer, colorectal cancer, rectal cancer or colon cancer), urinary cancer (e.g., kidney cancer, bladder cancer, prostate cancer or testicular cancer), bone cancer, gynecological cancer (e.g., breast cancer, endometrial cancer, cervical cancer or ovarian cancer), hematological cancer (e.g., leukemia, lymphoma or myeloma) or other types of cancer (e.g., melanoma or skin cancer), preferably lymphoma, lung cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, leukemia, glioblastoma, prostate cancer or ovarian cancer.

[0250] The present invention also provides a method for treating and / or preventing a disease associated with or mediated by MTAP, comprising administering to a patient an effective amount of a compound as described in Formula I, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above. The MTAP-associated / mediated cancer is selected from the group consisting of head and neck cancer (e.g., thyroid cancer, meningeal cancer, intracranial metastasis, or glioblastoma), respiratory cancer (e.g., lung cancer or nasopharyngeal cancer), digestive cancer (e.g., esophageal cancer, gastric cancer, liver cancer, bile duct cancer, pancreatic cancer, colorectal cancer, rectal cancer, or colon cancer), urinary cancer (e.g., kidney cancer, bladder cancer, prostate cancer, or testicular cancer), bone cancer, gynecological cancer (e.g., breast cancer, endometrial cancer, cervical cancer, or ovarian cancer), hematologic cancer (e.g., leukemia, lymphoma, or myeloma), or other types of cancer (e.g., melanoma or skin cancer), more preferably lymphoma, lung cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, leukemia, glioblastoma, prostate cancer, or ovarian cancer.

[0251] The present invention also provides a method for treating and / or preventing cancer, comprising administering to a patient an effective amount of a compound as shown in Formula I, a tautomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above. The cancer may be head and neck cancer (e.g., thyroid cancer, meningeal cancer, intracranial metastasis, or glioblastoma), respiratory cancer (e.g., lung cancer or nasopharyngeal cancer), digestive cancer (e.g., esophageal cancer, gastric cancer, liver cancer, bile duct cancer, pancreatic cancer, colorectal cancer, rectal cancer, or colon cancer), urinary cancer (e.g., kidney cancer, bladder cancer, prostate cancer, or testicular cancer), bone cancer, gynecological cancer (e.g., breast cancer, endometrial cancer, cervical cancer, or ovarian cancer), hematologic cancer (e.g., leukemia, lymphoma, or myeloma), or other types of cancer (e.g., melanoma or skin cancer), preferably lymphoma, lung cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, leukemia, glioblastoma, prostate cancer, or ovarian cancer.

[0252] The compound of formula I, its tautomer or pharmaceutically acceptable salt, or the pharmaceutical composition described above can be in a therapeutically effective amount.

[0253] The compound of formula I, its tautomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above can be administered to a subject by any suitable route, preferably orally or by injection (intravenous, intramuscular, subcutaneous and coronary).

[0254] The present invention also provides a method for preparing the compounds of Formulae I, IA, and IB as described above, comprising the following steps: starting from intermediates Ia and Ib, undergoing a Suzuki coupling reaction to obtain compound Ic; undergoing a Sonogashira coupling reaction of compound Ic with a corresponding alkynyl compound to obtain compound Id; further removing the protecting group of compound Id to obtain a compound of Formula I; and subjecting the obtained compound of Formula I to chiral preparative resolution to obtain a pair of atropisomers, compound IA and compound IB.

[0255] Among them, n, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 、R 8 and R 9 As defined above.

[0256] In some embodiments of the present invention, the preparation method of the intermediate Ia comprises the following steps: starting from compound Ia-1, performing carbonyl insertion reaction and halogen migration to obtain compound Ia-2, esterifying compound Ia-2, and performing Still coupling reaction to obtain compound Ia-4; compound Ia-4 is reacted with the corresponding R-containing 8 The compound of the group undergoes a substitution reaction to obtain compound Ia-5; compound Ia-5 is protected to obtain compound Ia-6, and compound Ia-6 is further brominated, substituted and hydrolyzed to obtain compound Ia-8; compound Ia-8 is then hydrazinolyzed and cyclized to obtain compound Ia-10; compound Ia-10 is halogenated, deprotected, substituted with the amino group, and then protected to obtain intermediate Ia;

[0257] Among them, R 8 NHR 11 , hydroxyl or C 2-6 Alkynyl, R 8-1 -NR 11 -, -O- or C 2-6 Alkynylidene, P1 is amino, hydroxyl or C 2-6 Alkynyl protecting group; R 11 and R 9 As defined above.

[0258] In some embodiments of the present invention, the preparation method of the intermediate Ia comprises the following steps: starting from compound Ia-1, performing carbonyl insertion reaction and halogen migration to obtain compound Ia-2, esterifying compound Ia-2, and performing Still coupling reaction to obtain compound Ia-4; compound Ia-4 is reacted with the corresponding R-containing 8 The compound of the group undergoes a substitution reaction to obtain compound Ia-5'; compound Ia-5' is further brominated, substituted and hydrolyzed to obtain compound Ia-8'; compound Ia-8' is further hydrazinolyzed and cyclized to obtain compound Ia-9'; compound Ia-9' is halogenated, substituted with an amino group and then protected to obtain intermediate Ia;

[0259] Among them, R 8 and R 9 As defined above.

[0260] In some embodiments of the present invention, the method for preparing intermediate Ib comprises the following steps: compound Ib-1 is substituted or coupled to obtain compound Ib-2; compound Ib-2 is iodinated to obtain compound Ib-3; compound Ib-3 is further subjected to a Suzuki coupling reaction to obtain compound Ib-4; compound Ib-4 is iodinated to generate the corresponding borate ester to obtain intermediate Ib;

[0261] Among them, R 1 、R 2 、R 3 、R 4 、R 6 and R 7 As defined above.

[0262] The present invention also provides a PRMT5-MEP50-MTA-compound 2-P2 complex crystal, wherein the compound 2-P2 is

[0263] The X-ray data of the composite crystals are shown in the following table:

[0264] Definition of terms

[0265] In addition to the foregoing, when used in the specification and claims of this application, the following terms have the meanings indicated below unless otherwise specifically stated.

[0266] As used herein, the compounds of Formula I of the present invention may contain one or more chiral centers and exist in different optically active forms. When a compound contains one chiral center, the compound comprises enantiomers. The present invention includes both isomers and mixtures of isomers, such as racemic mixtures. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When a compound of Formula I contains more than one chiral center, diastereomers may exist. The present invention includes resolved optically pure specific isomers and mixtures of diastereomers. Diastereomers can be resolved by methods known in the art, such as crystallization and chiral chromatography.

[0267] The term "stereoisomer" includes conformational isomers and configurational isomers, wherein configurational isomers mainly include cis-trans isomers and optical isomers. The compounds described in the present invention may exist in the form of stereoisomers, and therefore encompass all possible stereoisomeric forms, including but not limited to cis-trans isomers, enantiomers, diastereomers, atropisomers, etc. The compounds described in the present invention may also exist in the form of any combination or any mixture of the aforementioned stereoisomers, such as meso-, racemic-, and equal mixtures of atropisomers, or single enantiomers, single diastereomers, or mixtures thereof, or single atropisomers or mixtures thereof.

[0268] The term "tautomer" refers to functional isomers that result from the rapid shift of an atom between two positions in a molecule.

[0269] When the compounds described herein contain olefinic double bonds, unless otherwise specified, they include cis-isomers and trans-isomers, and any combination thereof.

[0270] The compound as shown in Formula I, its tautomer or its pharmaceutically acceptable salt is intended to cover any isotopically labeled (or "radiolabeled") variant of the compound as shown in Formula I, its tautomer or its pharmaceutically acceptable salt. Such a variant can be a compound as shown in Formula I, its tautomer or its pharmaceutically acceptable salt in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. The radionuclide used will depend on the specific application of the radiolabeled variant. For example, for in vitro receptor labeling and competition assays, 3 H or 14 C is often useful. For radiographic applications, 11 C or 18 F is often useful.

[0271] Certain isotopic variants of the compounds of the invention, particularly those into which one or more radioactive isotopes have been incorporated, can be useful, for example, in studying the mechanism of action or the distribution of the active ingredient in the body; because of the relative ease of preparation and detectability, isotopic variants labeled with 3 H or 14 Compounds containing a C isotope are particularly suitable for this purpose. Furthermore, the incorporation of an isotope, such as deuterium, can provide particular therapeutic benefits due to improved metabolic stability of the compound, for example, by extending the half-life in vivo or reducing the required effective dose; thus, such modifications of the compounds of the present invention may also constitute preferred embodiments of the present invention in some cases. Isotopic variants of the compounds of the present invention can be prepared by methods known to those skilled in the art, such as those described below and in the working examples, by using corresponding isotopically modified specific reagents and / or starting compounds.

[0272] As used herein, a "pharmaceutical composition" refers to a formulation comprising a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism, thereby facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0273] In this application, "pharmaceutically acceptable" refers to substances (such as pharmaceutical excipients) that do not affect the biological activity or properties of the compounds of the present invention and are relatively non-toxic, that is, the substance can be administered to a subject without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.

[0274] The term "pharmaceutical excipient" or "pharmaceutically acceptable carrier" refers to the excipients and additives used in the production of drugs and the preparation of prescriptions. It is all substances contained in pharmaceutical preparations in addition to the active ingredients. Please refer to the Pharmacopoeia of the People's Republic of China (2015 Edition) Part IV, or the Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition). Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition. They can also provide methods to dissolve the active ingredient at a desired rate after the subject receives the drug, or to promote the effective absorption of the active ingredient after the subject receives the composition. The pharmaceutical excipients can be inert fillers, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0275] The pharmaceutical compositions of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0276] When used as a drug, the compound as shown in Formula I, its stereoisomers, its tautomers or pharmaceutically acceptable salts thereof can be administered in any form of a pharmaceutical composition. These compositions can be administered in various ways, depending on the need for local or systemic treatment and the area to be treated. Administration can be topical (including epidermal and transdermal, ocular and mucosal, including intranasal, vaginal and rectal delivery), pulmonary, oral or parenteral. Oral administration can include dosage forms formulated for once-daily or twice-daily (BID) administration. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or injection or infusion; or intracranial such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dose, or can be by continuous infusion pump. Conventional pharmaceutical carriers, water, powders or oily bases, thickeners, etc. may be necessary or required. The pharmaceutical composition of the present invention can also be a controlled release or delayed release dosage form (e.g., liposomes or microspheres).

[0277] The term "treat" refers to therapeutic treatment or palliative measures. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition, or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition. "Treatment" may also mean prolonging survival as compared to expected survival if not receiving treatment.

[0278] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0279] The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat a disease or condition described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art.

[0280] The term "patient" refers to any animal, preferably a mammal, and most preferably a human, that is about to receive or has received a compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans, with humans being the most preferred.

[0281] Unless otherwise indicated, the following definitions as used herein shall apply. For purposes of the present invention, the chemical elements are referred to in accordance with the Periodic Table of the Elements, CAS version, and Handbook of Chemistry and Physics, 75th edition, 1994. In addition, general principles of organic chemistry may be found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, the entire contents of which are incorporated herein by reference.

[0282] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for use by patients) acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, bismuth salts, ammonium salts, and the like. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. The pharmaceutically acceptable acid includes inorganic acids and organic acids. For details, see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977), or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0283] In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural moieties and compounds. When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents obtained when the structural formula is written from right to left.

[0284] Certain chemical groups defined herein are preceded by a simplified symbol to indicate the total number of carbon atoms present in the group. For example, a C1-C4 alkyl or a C 1-4 Alkyl refers to an alkyl group as defined below having a total of 1, 2, 3 or 4 carbon atoms. The total number of carbon atoms in the shorthand notation does not include carbons that may be present in substituents of the group being described.

[0285] As used herein, numerical ranges defined in substituents, such as 0 to 10, 1-6, 1-3, etc., indicate integers within the range, such as 1-6 represents 1, 2, 3, 4, 5, 6.

[0286] The term "comprising" is an open expression, that is, including the contents specified in the present invention, but not excluding other contents.

[0287] The term "substituted with" or "substituted with" means that any one or more hydrogen atoms on a particular atom are replaced with a substituent, as long as the valence of the particular atom is normal and the compound after the substitution is stable.

[0288] In general, the term "substituted" or "substituted" means that one or more hydrogen atoms in a given structure are replaced by a specified substituent. Furthermore, when a group is substituted with one or more of the substituents, the substituents are independent of each other, that is, the one or more substituents may be different or the same. Unless otherwise indicated, a substituent group may be substituted at every substitutable position of the substituted group. When more than one position in a given structure is substituted by one or more substituents selected from the specified group, the substituents may be the same or different at each position.

[0289] In various parts of this specification, substituents of compounds disclosed herein are disclosed in terms of group types or ranges. It is specifically noted that the present invention includes each independent subcombination of the individual members of these group types and ranges. The term "C x -C y Alkyl" or "C x-y "Alkyl" refers to a straight or branched chain saturated hydrocarbon containing x to y carbon atoms. For example, the term "C1-C6 alkyl" or "C 1-6 "Alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl disclosed independently; "C 1-4 The term "alkyl" specifically refers to the independently disclosed methyl, ethyl, C3 alkyl (i.e., propyl, including n-propyl and isopropyl), and C4 alkyl (i.e., butyl, including n-butyl, isobutyl, sec-butyl, and tert-butyl).

[0290] As used herein, the terms "moiety," "moiety," "chemical moiety," "group," and "chemical group" refer to specific segments or functional groups in a molecule. A chemical moiety is generally considered to be a chemical entity embedded in or attached to a molecule.

[0291] When a substituent is listed without indicating the atom via which it is bonded in a chemical formula (including but not specifically mentioned compounds), such substituent may be bonded via any atom thereof. Combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0292] When any variable (such as R 1-a ) appears multiple times in the definition of a compound, the definition of each position of the variable is independent of the definition of the other positions, and their meanings are independent of each other and do not affect each other. Therefore, if a group is replaced by 1, 2 or 3 R 1-a group substituted, that is, the group may be replaced by up to 3 R 1-a Substitution, where a position R 1-a Definition and other positions R 1-aIn addition, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0293] When a group is listed without specifying that it has a substituent, such group is only meant to be unsubstituted. For example, when "C 1-6 When "alkyl" is not limited to "substituted or unsubstituted", it only refers to "C 1-6 Alkyl" itself or "unsubstituted C 1-6 alkyl".

[0294] In various parts of the present invention, linking substituents are described. When the structure clearly requires a linking group, the Markush variable listed for that group should be understood to be a linking group. For example, if the structure requires a linking group and the Markush group definition for that variable lists "alkyl", it should be understood that the "alkyl" represents a linking alkylene group.

[0295] In some specific structures, when an alkyl group is explicitly indicated as a linking group, the alkyl group represents a linking alkylene group, for example, the group "halo-C 1-6 C in "alkyl-" 1-6 Alkyl should be understood as C 1-6 Alkylene.

[0296] The term "halogen" refers to fluorine, chlorine, bromine or iodine, in particular to F, Cl or Br.

[0297] In this application, the term "alkyl" as a group or part of another group (for example, in a group such as a haloalkyl group, a deuterated alkyl group, etc.) refers to a saturated aliphatic hydrocarbon group including branched and straight chains having the specified number of carbon atoms, consisting only of carbon atoms and hydrogen atoms, having, for example, 1 to 20 (preferably 1 to 8, more preferably 1 to 6, and most preferably 1 to 4) carbon atoms, and connected to the rest of the molecule by a single bond, wherein propyl is a C3 alkyl group (including isomers such as n-propyl or isopropyl); butyl is a C4 alkyl group (including isomers such as n-butyl, sec-butyl, isobutyl or tert-butyl); butyl); pentyl is a C5 alkyl group (including isomers such as n-pentyl, 1-methyl-butyl, 1-ethyl-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, isopentyl, tert-pentyl or neopentyl); hexyl is a C6 alkyl group (including isomers such as 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). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, n-octyl, nonyl, decyl, and the like.

[0298] In the present application, as a group or a part of other groups, the alkyl group in the term "alkyl-O-" is as defined above.

[0299] In the present application, the term "alkenyl" as a group or part of another group refers to a straight-chain or branched hydrocarbon group having at least one double bond, consisting only of carbon atoms and hydrogen atoms, having, for example, 2 to 12 (preferably 2 to 8, more preferably 2 to 6, most preferably 2 to 4) carbon atoms, and connected to the rest of the molecule by a single bond, for example, including but not limited to vinyl, 1-propenyl, n-allyl, but-1-enyl, but-2-enyl, pent-1-enyl or pent-1,4-dienyl, etc.

[0300] In this application, the term "alkynyl" as a group or part of another group refers to a straight or branched hydrocarbon group having at least one triple bond, consisting only of carbon atoms and hydrogen atoms, having, for example, 2 to 12 (preferably 2 to 8, more preferably 2 to 6, most preferably 2 to 4) carbon atoms, and connected to the rest of the molecule by a single bond, for example, including but not limited to ethynyl, 1-propynyl, n-propargyl, but-1-ynyl, but-2-ynyl, pent-1-ynyl or pent-1,4-diynyl, etc.

[0301] In this application, the term "cycloalkyl" as a group or part of another group means a saturated monocyclic or polycyclic (e.g., bicyclic, tricyclic or more cyclic bridged ring, fused ring or spirocyclic ring system) carbocyclic substituent, which can be connected to the rest of the molecule by a single bond through any suitable carbon atom; such as a 3-15 membered cycloalkyl having 3 to 15 carbon atoms, preferably a 3-12 membered cycloalkyl having 3 to 12 carbon atoms, more preferably a 3-7 membered cycloalkyl having 3 to 7 carbon atoms, and most preferably a 3-6 membered cycloalkyl having 3 to 6 carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, and the like.

[0302] The term "cycloalkyl" means a saturated or unsaturated monocyclic or polycyclic ring, wherein the saturated monocyclic or polycyclic ring is the same as defined in "cycloalkyl", wherein the unsaturated monocyclic or polycyclic ring is a monocyclic or polycyclic ring containing one or more double bonds, and the definition of the monocyclic or polycyclic ring is the same as that of the saturated monocyclic or polycyclic ring.

[0303] In the present application, the term "heterocyclyl" as a group or part of another group refers to a stable saturated or partially unsaturated monocyclic or polycyclic (e.g., bicyclic, tricyclic or more cyclic bridged, fused or spirocyclic) non-aromatic cyclic group consisting of carbon atoms and 1, 2, 3, 4, 5 or 6 heteroatoms selected from N, O and S; preferably a 3-12 membered heterocyclyl containing 1, 2, 3 or 4 heteroatoms selected from N, O and S, more preferably a 4-10 membered heterocyclyl containing 1, 2, 3 or 4 heteroatoms selected from N, O and S, and most preferably a 4-6 membered monoheterocyclyl or 7-10 membered biheterocyclyl containing 1, 2, 3 or 4 heteroatoms selected from N, O and S. When it is a bicyclic, tricyclic or higher ring cycloheterocyclic group, it may also include the formation of a cycloheterocyclic group (condensed ring) with a cycloalkyl, aryl or heteroaryl group as defined herein, provided that the heterocyclyl group is connected to the rest of the molecule by a single bond via any suitable atom in the saturated or partially unsaturated heterocycle. When it is a bicyclic, tricyclic or higher ring spirocyclic heterocyclyl group, it may also include the formation of a spirocyclic group with a cycloalkyl group as defined herein, provided that the heterocyclyl group is connected to the rest of the molecule by a single bond via any suitable atom in the saturated or partially unsaturated heterocycle. In a certain embodiment of the present invention, heterocyclic groups include "heterocycloalkyl" and "heterocycloalkenyl". Heterocycloalkyl refers to a cyclic group consisting of a stable saturated monocyclic or polycyclic ring (e.g., a bicyclic, tricyclic or more cyclic bridged ring, a cyclic (condensed ring) or spiro ring system) of carbon atoms and 1, 2, 3, 4, 5 or 6 heteroatoms selected from N, O and S, and a non-aromatic cyclic group consisting of a stable partially unsaturated monocyclic or polycyclic ring (e.g., a bicyclic, tricyclic or more cyclic bridged ring, a cyclic (condensed ring) or spiro ring system) of carbon atoms and 1, 2, 3, 4, 5 or 6 heteroatoms selected from N, O and S. For example, the 4-10 membered heterocyclic group includes a 4-10 membered heterocycloalkyl or a 4-10 membered heterocycloalkenyl. In some embodiments, "heterocycloalkyl" is a 4 to 6 membered monocyclic heterocycloalkyl, a 4-8 membered heterocycloalkyl, a 7 to 10 membered heterocycloalkyl or a 5-10 membered spiro ring. Exemplary 4-membered heterocycloalkyl groups include, but are not limited to, azetidinyl (e.g. ) or isomers and stereoisomers thereof; exemplary 5-membered heterocycloalkyl groups include, but are not limited to, pyrrolidinyl (For example ), or isomers and stereoisomers thereof. Exemplary 6-membered heterocycloalkyl groups include, but are not limited to, piperidinyl (e.g. ), tetrahydropyranyl, morpholinyl (e.g. ), piperazinyl (e.g. ), or isomers and stereoisomers thereof. Exemplary 7-membered heterocycloalkyl groups include, but are not limited to, (For example )or (For example For example ), (For example ), or isomers and stereoisomers thereof. Exemplary 8-membered heterocycloalkyl groups include, but are not limited to, (For example ), or isomers and stereoisomers thereof.

[0304] In this application, the term "aryl" as a group or part of another group refers to an aromatic group consisting of a conjugated hydrocarbon ring system of carbon atoms that satisfies the 4n+2 rule, with each ring having aromaticity. In one embodiment, "aryl" refers to an aromatic group having 6 to 18 (preferably 6 to 10) carbon atoms. Examples of aryl groups include, but are not limited to, phenyl or naphthyl.

[0305] In the present application, the term "heteroaryl" as a group or part of another group means a conjugated ring system radical having carbon atoms and 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur in the ring. Unless otherwise specified in the specification, the heteroaryl group can be a monocyclic, bicyclic, tricyclic or higher ring system. When it is a bicyclic, tricyclic or higher ring fused ring, it can also include a cycloalkyl or heterocyclyl group as defined herein, provided that the heteroaryl group is connected to the rest of the molecule by a single bond through an atom on the aromatic ring.

[0306] The "-" at the end or both ends of a group means that the group is connected to other fragments in the molecule through this site.

[0307] It is to be understood that as used herein, the singular forms, such as "a," "an," and the like, include plural referents unless otherwise specified.

[0308] The term "one or more" or "one or more than two" means 1, 2, 3, 4, 5, 6, 7, 8, 9 or more.

[0309] Unless otherwise stated, the present invention adopts conventional methods of mass spectrometry and elemental analysis, and each step and condition can refer to conventional operating steps and conditions in the art.

[0310] Unless otherwise indicated, the present invention employs standard nomenclature and standard laboratory procedures and techniques of analytical chemistry, synthetic organic chemistry, and optics. In some instances, standard techniques were used for chemical syntheses and chemical analyses.

[0311] Additionally, it should be noted that, unless explicitly stated otherwise, the term "independently" used in the present invention should be broadly interpreted to mean that the individual entities described are independent of each other and can independently represent the same or different specific groups. More specifically, the term "independently" can mean that the specific options represented by the same symbol in different groups do not affect each other, or that the specific options represented by the same symbol in the same group do not affect each other.

[0312] Unless otherwise specified, all technical and scientific terms used herein have the standard meanings in the art to which the claimed subject matter belongs. If there are multiple definitions for a term, the definition herein shall prevail.

[0313] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0314] The reagents and raw materials used in the present invention are commercially available.

[0315] The positive progress of the present invention is that: the present invention provides a class of PRMT5 inhibitors, which have good inhibitory activity or selectivity against human colon cancer stably knocked-out MTAP protein cells (HCT-116-MTAP-KO-1A2) and can inhibit the proliferation of tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0316] Figure 1 is the co-crystal structure of the PRMT5-MEP50-MTA-Compound 2-P2 complex. DETAILED DESCRIPTION

[0317] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0318] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). Nuclear magnetic resonance chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3) as the solvent and tetramethylsilane (TMS) as the internal standard. Mass spectrometry measurements were performed using an Agilent 1260-6125B single quadrupole liquid chromatography-mass spectrometer employing an electrospray ionization (ESI) source.

[0319] For silica gel column chromatography, a Biotage Selekt flash preparative chromatograph and a Biotage BK-SIL silica gel prepacked column or an Agela Claricep Flash silica gel prepacked column of appropriate specifications were used.

[0320] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the specification of which is 0.15mm-0.20mm, and the specification of which is 0.4mm-0.5mm for preparative thin layer chromatography.

[0321] Preparative high-performance liquid chromatography (HPLC) was performed using a Waters AutoPurification LC system equipped with an ACQUITY QDa mass spectrometer. A SunFire C18 5μm 19x250mm OBD preparative column was used as the preparative column. Compounds were eluted using a gradient of water (containing 0.1% formic acid)-acetonitrile as the mobile phase.

[0322] The Chinese and English abbreviations are as follows:

[0323] X-PHOS: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl; Pd2(dba)3: tris(dibenzylideneacetone)dipalladium;

[0324] Boc: tert-butyloxycarbonyl; CDCl3: deuterated chloroform; DMSO: dimethyl sulfoxide; DCM: dichloromethane.

[0325] Synthesis part

[0326] Intermediate a1

[0327] 2-Methylpropan-2-yl [({7-bromo-4-oxylidene-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl}methyl)amino]methane

[0328] Step 1: 6-Bromo-2-fluoro-3-iodopyridine

[0329] Under argon at -70°C, lithium diisopropylamide (41.82 mL, 83.65 mmol) was added to a solution of 2-bromo-6-fluoropyridine (10 g, 76.04 mmol) in tetrahydrofuran (100 mL). The reaction mixture was stirred for 2 hours, followed by the addition of a solution of iodine (21.23 g, 83.65 mmol) in tetrahydrofuran (100 mL). The reaction mixture was stirred at -70°C for 1 hour. After completion, saturated sodium sulfite (100 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (200 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether = 100%, volume ratio) to afford the product as a white solid (16 g, yield 81.76%). ESI-MS m / z: 257.8 [M+1] + . 1 H NMR (400MHz, CDCl3) δ8.09 (t, J = 8.0 Hz, 1H), 7.03 (dd, J = 7.9, 1.2 Hz, 1H).

[0330] Step 2: 6-Bromo-2-fluoro-4-iodonicotinic acid

[0331] Under argon protection and at -70°C to -60°C, lithium diisopropylamide (29.96 mL, 59.94 mmol) was added to a solution of 6-bromo-2-fluoro-3-iodopyridine (14 g, 54.74 mmol) in tetrahydrofuran (20 mL). The reaction mixture was stirred for 2 hours. The reaction system was purged with a carbon dioxide balloon and stirred for another 5 hours. After completion of the reaction, water (100 mL) was added to the reaction mixture, and the reaction system was extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH 6-7 with dilute hydrochloric acid (2 M) and then extracted with ethyl acetate (100 mL x 3). The organic phase was concentrated to yield the product (16 g, crude) as a red solid. ESI-MS m / z: 301.9 [M+1] + .

[0332] Step 3: Methyl 6-bromo-2-fluoro-4-iodonicotinate

[0333] To a solution of 6-bromo-2-fluoro-4-iodonicotinic acid (1.4 g, 4.64 mmol) and potassium carbonate (0.963 g, 6.97 mmol) in N,N-dimethylformamide (15 mL) was added iodomethane (433 μL, 6.97 mmol) at 0°C. The reaction mixture was stirred for 3 hours. After completion of the reaction, water (100 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (100 mL x 3). The organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-2%, volume ratio) to afford the product as a yellow solid (1.12 g, yield 76.45%). ESI-MS m / z: 315.9 [M+1] + . 1 H NMR (400MHz, CDCl3) δ7.77(s,1H),3.99(s,3H).

[0334] Step 4: Methyl 6-bromo-4-(1-ethoxyvinyl)-2-fluoronicotinate

[0335] Under argon, a reaction mixture of methyl 6-bromo-2-fluoro-4-iodonicotinate (5 g, 15.873 mmol), tetrakistriphenylphosphine palladium (917 mg, 0.7936 mmol), bis(triphenylphosphine)palladium dichloride (557.02 mg, 0.7936 mmol), and tributyl(1-ethoxyethylene)tin (5.732 g, 15.873 mmol) was stirred at 110°C for 24 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-3%, volume ratio) to afford the product as a colorless oil (3.4 g, 82.62% yield). ESI-MS m / z: 260.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ7.35 (s, 1H), 4.68 (d, J = 3.5Hz, 1H), 4.47 (d, J = 3.5Hz, 1H), 3.91 (s, 3H), 3.86 (q, J = 7.0Hz, 2H), 1.35 (t, J = 7.0Hz, 3H).

[0336] Step 5: Methyl 6-bromo-4-(1-ethoxyvinyl)-2-((methyl-d3)amino)nicotinate

[0337] To a solution of methyl 6-bromo-4-(1-ethoxyvinyl)-2-fluoronicotinate (5.0 g, 16.5 mmol) in tetrahydrofuran (50 mL) were added triethylamine (10 mL) and deuterated methylamine hydrochloride (5.82 g, 82.5 mmol). The mixture was stirred at 80°C in a sealed tube for 1 hour and 15 minutes. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20:1, volume ratio) to afford the product as a pale yellow oil (5.2 g, 99.0% yield). ESI-MS m / z: 317.9; 319.9 [M+1] + .

[0338] Step 6: 6-bromo-2-(tert-butoxycarbonyl)(methyl-d3)amino)-4-(1-ethoxyvinyl)nicotinate

[0339] Methyl 6-bromo-4-(1-ethoxyvinyl)-2-((methyl-d3)amino)nicotinate (5.2 g, 16.3 mmol) was dissolved in tetrahydrofuran (50 mL). Triethylamine (10 mL), 4-dimethylaminopyridine (4.0 g, 32.6 mmol), and di-tert-butyl dicarbonate (42.8 g, 196.1 mmol) were added. The mixture was refluxed under nitrogen for 1.5 hours, cooled to room temperature, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, volume ratio) to obtain the product as a yellow oil (5.74 g, yield 83.9%). ESI-MS m / z: 317.9; 319.9 [M-Boc+1] + . 1 H NMR (400MHz, CDCl3) δ7.43 (s, 1H), 4.50 (d, J = 3.2Hz, 1H), 4.37 (d, J = 3.2Hz, 1H), 3.81 (m, 5H), 1.41 (s, 9H), 1.32 (t, J = 7.0Hz, 3H).

[0340] Step 7: 6-bromo-4-(2-bromoacetyl)-2-(tert-butyloxycarbonyl)(methyl-d3)amino)nicotinate

[0341] Methyl 6-bromo-2-(tert-butoxycarbonyl)(methyl-d3)amino)-4-(1-ethoxyvinyl)nicotinate (5.74 g, 13.7 mmol) was dissolved in tetrahydrofuran (75 mL). Water (25 mL) and N-bromosuccinimide (2.44 g, 13.7 mmol) were added. After stirring at 25°C for 1.5 hours, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was washed three times with water and then with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1, volume ratio) to obtain the product as a yellow oil (6.2 g, yield 96.3%). ESI-MS m / z: 367.7; 369.7; 371.7 [M-Boc+1] + . 1 H NMR (400MHz, CDCl3) δ7.27(s,1H),4.33(s,2H),3.82(s,3H),1.44(s,9H).

[0342] Step 8: 4-(2-acetoxyacetyl)-6-bromo-2-((tert-butyloxycarbonyl)(methyl-d3)amino)nicotinate

[0343] To a solution of potassium acetate (3.9 g, 39.6 mmol) and acetic acid (1.19 mg, 19.8 mmol) in acetonitrile (50 mL) was added dropwise a solution of methyl 6-bromo-4-(2-bromoacetyl)-2-(tert-butoxycarbonyl)(methyl-d3)amino)nicotinate (6.2 g, 13.2 mmol) dissolved in acetonitrile (50 mL). After stirring at 25°C for 1.5 hours, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was washed three times with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the product as a yellow oil (5.92 g, 100.0% yield). ESI-MS m / z: 347.8; 349.8 [M-Boc+1] + . 1 H NMR (400MHz, CDCl3) δ7.33(s,1H),5.02(s,2H),3.81(s,3H),2.16(s,3H),1.45(s,9H).

[0344] Step 9: tert-Butyl (6-bromo-3-(hydrazinecarbonyl)-4-(2-hydroxyacetyl)pyridin-2-yl)(methyl-d3)carbamate

[0345] Methyl 4-(2-acetoxyacetyl)-6-bromo-2-((tert-butoxycarbonyl)(methyl-d3)amino)nicotinate (5.92 g, 13.2 mmol) was dissolved in ethanol (80 mL), and hydrazine hydrate (1.55 g, 26.4 mmol, 85%) was added dropwise. The mixture was stirred at 25°C for 6 hours and concentrated under reduced pressure to give the product as a yellow solid (5.37 g, 100.0% yield). ESI-MS m / z: 305.8; 307.8 [M-Boc+1] + . 1 H NMR (400MHz, DMSO-d6) δ8.93 (s, 1H), 7.75 (s, 1H), 4.81 (t, J = 6.3Hz, 1H), 4.40 (s, 2H), 1.34 (s, 9H).

[0346] Step 10: tert-Butyl (7-bromo-1-(hydroxymethyl)-4-oxo-3,4-dihydropyridin[3,4-d]pyridazin-5-yl)(methyl-d3)carbamate

[0347] Tert-butyl (6-bromo-3-(hydrazinocarbonyl)-4-(2-hydroxyacetyl)pyridin-2-yl)(methyl-d3)carbamate (5.37 g, 13.2 mmol) was dissolved in toluene (125 mL) and stirred under reflux for 12 hours. The mixture was then cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 100:3, volume ratio) to afford the product as a yellow solid (4.3 g, yield 83.8%). ESI-MS m / z: 287.9; 289.8 [M-Boc+1] + . 1 H NMR (400MHz, DMSO-d6) δ12.85(s,1H),8.00(s,1H),5.61(t,J=5.8Hz,1H),4.63(d,J=5.6Hz,2H),1.27(s,9H).

[0348] Step 11: 2-Methylpropan-2-yl {[7-bromo-1-(chloromethyl)-4-oxylidene-3H-pyrido[4,3-d][1,2]diazin-5-yl](trideuteriomethyl)amino}methanoate

[0349] 2-Methylpropan-2-yl {[7-bromo-1-(hydroxymethyl)-4-oxo-3H-pyridinium[4,3-d][1,2]diazin-5-yl](trideuteriomethyl)amino}carboxylate (2 g, 5.15 mmol) and thionyl chloride (3.06 g, 25.76 mmol) were dissolved in DCM (60 mL). N,N-dimethylformamide (0.083 mL, 1.03 mmol) was added dropwise and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the solvent was concentrated under reduced pressure to obtain a yellow solid (2 g, 4.92 mmol, 95.47%), which was used directly in the next reaction. MS m / z: 406 [M+1] + .

[0350] Step 12: 7-Bromo-1-(chloromethyl)-5-[(trideuteriomethyl)amino]-3,4-dihydropyrido[4,3-d][1,2]diazin-4-one

[0351] Dissolve 2-methylpropan-2-yl {[7-bromo-1-(chloromethyl)-4-oxyylidene-3H-pyrido[4,3-d][1,2]diazin-5-yl](trideuteriomethyl)amino}methane (2 g, 4.92 mmol) in DCM (50 mL). Add trifluoroacetic acid (11.298 mL, 147.54 mmol) at room temperature. Stir the mixture at room temperature for 30 minutes. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10:1, volume ratio) to afford the product as a yellow solid (1.5 g, 99.49%). ESI-MS m / z: 306 [M+1] + .

[0352] Step 13: 1-(Aminomethyl)-7-bromo-5-[(trideuteriomethyl)amino]-3,4-dihydropyrido[4,3-d][1,2]diazin-4-one

[0353] Dissolve 7-bromo-1-(chloromethyl)-5-[(trideuteriomethyl)amino]-3,4-dihydropyrido[4,3-d][1,2]diazin-4-one (1.5 g, 4.89 mmol) in dichloromethane (50 mL) and add dropwise to ammonia-methanol solution (20.970 mL, 7 M) at room temperature. Stir and react at room temperature for half an hour. After completion of the reaction, concentrate under reduced pressure to obtain a yellow solid (1.4 g, 4.88 mmol, 99.65%). ESI-MS m / z: 287 [M+1] + .

[0354] Step 14: 2-Methylpropan-2-yl [({7-bromo-4-oxylidene-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl}methyl)amino]methanoate

[0355] 1-(Aminomethyl)-7-bromo-5-[(trideuteriomethyl)amino]-3,4-dihydropyrido[4,3-d][1,2]diazin-4-one (1.5 g, 5.22 mmol), di-tert-butyl dicarbonate (1.37 g, 6.27 mmol), and triethylamine (1.089 mL, 7.84 mmol) were dissolved in tetrahydrofuran (40 mL) and stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / ethyl acetate = 5:1, volume ratio) to obtain the product as a yellow solid (1.5 g, 75% yield). ESI-MS m / z: 387 [M+1] + .

[0356] Intermediate b1

[0357] 4-Chloro-6-cyclopropyloxy-3-fluoro-2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-5-yl)benzonitrile

[0358] Step 1: 4-Chloro-6-(cyclopropyloxy)-3-fluoro-2-(4-iodo-2-methylpyrazol-3-yl)benzene-1-carbonitrile

[0359] To a solution of 4-chloro-6-(cyclopropyloxy)-3-fluoro-2-(2-methylpyrazol-3-yl)benzene-1-carbonitrile (235 mg, 0.806 mmol, the synthesis of this intermediate is referred to in the Journal of Medicinal Chemistry, 2022, vol. 65, (3) p. 1749–1766) in acetic acid (10 mL) was added N-iodosuccinimide (271.9 mg, 1.208 mmol), and the reaction mixture was stirred at 80° C. for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-25%, volume ratio) to obtain the product as a red oil (400 mg, crude product). ESI-MS m / z: 417.7 [M+1] + .

[0360] Step 2: 4-Chloro-6-cyclopropyloxy-3-fluoro-2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-5-yl)benzonitrile

[0361] 4-Chloro-6-(cyclopropyloxy)-3-fluoro-2-(4-iodo-2-methylpyrazol-3-yl)benzene-1-carbonitrile (45 mg, 0.108 mmol), pinacol borane (138.13 mg, 1.08 mmol), and triethylamine (54.50 mg, 0.540 mmol) were dissolved in dioxane (10 mL). Tris(dibenzylideneacetone)dipalladium (19.75 mg, 0.022 mmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (20.58 mg, 0.043 mmol) were added under argon atmosphere. The mixture was stirred at 100°C for 2 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 0-2%, volume ratio) to obtain the product as a yellow solid (100 mg, crude). ESI-MS m / z: 417.9[M+1] + .

[0362] Intermediate b2

[0363] 4-Chloro-3-fluoro-2-[2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-3-yl]-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0364] Step 1: 4-Chloro-5-fluoro-2-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0365] To a solution of 4-chloro-2,5-difluorobenzene-1-carbonitrile (2 g, 11.52 mmol) and potassium carbonate (3.98 g, 28.81 mmol) in N,N-dimethylformamide (20 mL) was added tetrahydropyrrole (1.446 mL, 17.29 mmol). The reaction mixture was stirred at 75°C for 2 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined and washed with saturated brine (3 × 50 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (20 g Flash Silica, ethyl acetate-petroleum ether, gradient elution 0-10%) to afford the product as a light green solid (2.5 g, yield 96.56%). MS m / z: 225.0 [M+1] + . 1 H NMR (400MHz, CDCl3) δ7.23(d,J=8.6Hz,1H),6.69(d,J=6.1Hz,1H),3.62–3.54(m,4H),2.09–2.00(m,4H).

[0366] Step 2: 4-Chloro-3-fluoro-2-iodo-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0367] Under argon protection and -78°C to -60°C, to a solution of 4-chloro-5-fluoro-2-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile (1 g, 4.45 mmol) in tetrahydrofuran (10 mL) was added a solution of lithium diisopropylamide in tetrahydrofuran (2.671 mL, 5.34 mmol). The reaction solution was stirred at this temperature for 0.5 hour, and then a solution of iodine (1.36 g, 5.34 mmol) in tetrahydrofuran (5 mL) was added. The reaction solution was warmed to room temperature and stirred for 0.5 hour. After the reaction was complete, saturated sodium chloride solution (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and washed with saturated sodium chloride solution (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (20 g Flash Silica, ethyl acetate-petroleum ether, 0-10% gradient elution) to obtain the product as a light green solid (1.5 g, yield 96.13%). ESI-MS m / z: 350.9 [M+1] + . 1 H NMR (400MHz, CDCl3) δ6.75 (d, J = 6.1Hz, 1H), 3.65–3.51 (m, 4H), 2.08–1.96 (m, 4H).

[0368] Step 3: 4-Chloro-3-fluoro-2-(2-methylpyrazol-3-yl)-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0369] Under nitrogen, 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) dichloride complex (0.35 g, 0.43 mmol) was added to a solution of 4-chloro-3-fluoro-2-iodo-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile (1.5 g, 4.28 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.78 g, 8.56 mmol), and potassium carbonate (1.77 g, 12.84 mmol) in dioxane (20 mL) and water (2 mL). The reaction mixture was stirred at 100°C for 12 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-20%, volume ratio) to obtain the product as a yellow solid (1.7 g, crude product). ESI-MS m / z: 305.0[M+1] + .

[0370] 1 H NMR (400MHz, CDCl3) δ7.62(d,J=2.0Hz,1H),6.83(d,J=6.1Hz,1H),6.47(d,J=2.0Hz,1H),3.83(d,J=1.0Hz,3H),3.64–3.60(m,4H),2.08–2.03(m,4H).

[0371] Step 4: 4-Chloro-3-fluoro-2-(4-iodo-2-methylpyrazol-3-yl)-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0372] Iodine (587.16 mg, 2.31 mmol) was added to a solution of 4-chloro-3-fluoro-2-(2-methylpyrazol-3-yl)-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile (470 mg, 1.54 mmol) and silver nitrate (130.99 mg, 0.77 mmol) in ethanol (15 mL). The reaction mixture was stirred at 70°C for 2 hours. After completion of the reaction, the reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-14%, volume ratio) to obtain the product as a yellow solid (200 mg, yield 30.11%). ESI-MS m / z: 430.9 [M+1] + . 1 H NMR (400MHz, CDCl3) δ7.68 (s, 1H), 6.88 (d, J = 6.1Hz, 1H), 3.86 (s, 3H), 3.70–3.55 (m, 4H), 2.12–2.02 (m, 4H).

[0373] Step 5: 4-Chloro-3-fluoro-2-[2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-3-yl]-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0374] The same method as in step 2 of step b1 was used to obtain the above-mentioned boronate compound. ESI-MS m / z: 431.1 [M+1] + .

[0375] Referring to the synthetic route of b2, the following intermediates were synthesized

[0376] Intermediate b3-1

[0377] 4-Chloro-3-fluoro-6-[(methylcyclopropyl)oxy]-2-[2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-3-yl]benzene-1-carbonitrile

[0378] Step 1: 4-Chloro-5-fluoro-2-[(methylcyclopropyl)oxy]benzene-1-carbonitrile

[0379] 1-Methylcyclopropane-1-ol (1.66 g, 23.02 mmol) was dissolved in dimethylformamide (40 mL), nitrogen was protected, the temperature was lowered to 0 ° C, sodium hydride (1.38 g, 34.53 mmol) was added, and the mixture was stirred for 1 hour. Then, 4-chloro-2,5-difluorobenzene-1-carbonitrile (4 g, 23.02 mmol) was added and the mixture was stirred for 3 hours. The reaction solution was quenched with aqueous ammonium chloride (15 mL), the reaction solution was diluted with ethyl acetate, the organic phase was washed three times with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1, volume ratio) to obtain the product as a white solid (1.5 g, yield: 28.9%). ESI-MS m / z: 226.0 [M+1] + . 1 H NMR (400MHz, CDCl3) δ7.34 (dd, J = 7.0, 6.0Hz, 2H), 1.60 (s, 3H), 1.12–1.07 (m, 2H), 0.85–0.80 (m, 2H).

[0380] Step 2: 4-Chloro-3-fluoro-2-iodo-6-[(methylcyclopropyl)oxy]benzene-1-carbonitrile

[0381] 4-Chloro-5-fluoro-2-[(methylcyclopropyl)oxy]benzene-1-carbonitrile (2 g, 8.86 mmol) was dissolved in tetrahydrofuran (100 mL) under nitrogen protection and cooled to -78 ° C. Lithium diisopropylamide (2 mol / L, 4.88 mL) was added and stirred for 1 hour. Iodine (3.37 g, 23.02 mmol) was added and reacted at room temperature for 3 hours. The reaction solution was quenched with saturated sodium sulfite aqueous solution (10 mL) and diluted with ethyl acetate. The organic phase was washed three times with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. It was further purified by silica gel column chromatography (petroleum ether / dichloromethane = 9:1, volume ratio) to obtain the compound (2.2 g, yield: 72%). ESI-MS m / z: 351.9 [M+1] + . 1H NMR (400MHz, CDCl3) δ7.34 (d, J = 5.6Hz, 1H), 1.60 (s, 3H), 1.12–1.07 (m, 2H), 0.85–0.81 (m, 2H).

[0382] Step 3: 4-Chloro-3-fluoro-6-[(methylcyclopropyl)oxy]-2-(2-methylpyrazol-3-yl)benzene-1-carbonitrile

[0383] Under argon, 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (93.15 mg, 0.11 mmol) was added to a solution of 4-chloro-3-fluoro-2-iodo-6-[(methylcyclopropyl)oxy]benzene-1-carbonitrile (400 mg, 1.14 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (474.40 mg, 2.28 mmol), and potassium carbonate (471.75 mg, 3.41 mmol) in dioxane (10 mL) and water (1 mL). The reaction mixture was stirred at 100°C for 12 hours. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (ethyl acetate-petroleum ether, gradient elution 0-15%) to obtain the product as a colorless oil (245 mg, 70.29%). ESI-MS m / z: 306.0[M+1] + . 1 H NMR (400MHz, CDCl3) δ7.64(d,J=2.0Hz,1H),7.44(d,J=5.9Hz,1H),6.52(d,J =2.0Hz,1H),3.84(s,3H),1.65(s,3H),1.17–1.11(m,2H),0.89–0.85(m,2H).

[0384] Step 4: 4-Chloro-3-fluoro-2-(4-iodo-2-methylpyrazol-3-yl)-6-[(methylcyclopropyl)oxy]benzene-1-carbonitrile

[0385] To a solution of 4-chloro-3-fluoro-6-[(methylcyclopropyl)oxy]-2-(2-methylpyrazol-3-yl)benzene-1-carbonitrile (245 mg, 0.8 mmol) in acetic acid (10 mL) was added N-iodosuccinimide (270.44 mg, 1.2 mmol). The reaction mixture was stirred at 80°C for 1.5 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was diluted with saturated sodium chloride (50 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic phases were washed with saturated sodium sulfite solution (1 × 50 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate-petroleum ether, gradient elution, 0-17%) to afford the product as a yellow solid (290 mg, 83.84%). ESI-MS m / z: 431.9 [M+1] + . 1 H NMR (400MHz, CDCl3) δ7.67(s,1H),7.50(d,J=5.9Hz,1H),3.85(s,3H),1.67(s,3H),1.20–1.14(m,2H),0.90–0.83(m,2H).

[0386] Step 5: 4-Chloro-3-fluoro-6-[(methylcyclopropyl)oxy]-2-[2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-3-yl]benzene-1-carbonitrile

[0387] Under argon, to a solution of 4-chloro-3-fluoro-2-(4-iodo-2-methylpyrazol-3-yl)-6-[(methylcyclopropyl)oxy]benzene-1-carbonitrile (290 mg, 0.67 mmol) and triethylamine (0.280 mL, 2.02 mmol) in dioxane (10 mL) were added trisdibenzylideneacetone dipalladium (61.53 mg, 0.07 mmol), 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (64.06 mg, 0.13 mmol), and pinacol borane (859.86 mg, 6.72 mmol). The reaction mixture was stirred at 100°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate-petroleum ether, gradient elution, 0-15%) to afford the product as a yellow oil (180 mg, 62.06%). ESI-MS m / z: 432.1[M+1] + .

[0388] Example 1

[0389] 2-{4-[1-(aminomethyl)-4-carbonyl-5-[(trideuterated methyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-6-(cyclopropyloxy)-3-fluoro-4-[3-(1,4-oxazin-4-yl)prop-1-ynyl]benzene-1-carbonitrile (Compound 1)

[0390] Step 1: tert-Butyl (7-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-1-(hydroxymethyl)-4-oxo-3,4-dihydropyridin[3,4-d]pyridazin-5-yl)(methyl-d3)carbamate

[0391] 4-Chloro-6-cyclopropyloxy-3-fluoro-2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzofuran-2-yl)-1H-pyrazol-5-yl)benzonitrile (intermediate b1) (537 mg, 1.29 mmol), tert-butyl (7-bromo-1-(hydroxymethyl)-4-oxo-3,4-dihydropyridin[3,4-d]pyridazin-5-yl)(methyl-d3)carbamate (intermediate a1) (498 mg, 1.29 mmol) and potassium carbonate (355.5 mg, 2.58 mmol) were dissolved in dioxane (30 mL) and water (3 mL). Under argon atmosphere, 1,1'-di-tert-butylphosphinoferrocenepalladium dichloride (83.9 mg, 0.129 mmol) was added and the reaction was stirred at 90°C for 12 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 100:3, volume ratio) to obtain the product as a yellow solid (370 mg, yield 48.0%). ESI-MS m / z: 598.2 [M+1] + . 1 HNMR (400MHz, CDCl3) δ10.25(s,1H),8.66(s,1H),8.19(s,1H),7.50(d,J=5.8Hz,1H),7.07(s,1 H),4.54–4.47(m,2H),3.87(dt,J=6.0,3.1Hz,1H),3.79(s,3H),1.49(s,9H),1.00–0.81(m,4H).

[0392] Step 2: 2-Methylpropan-2-yl {[(7-{5-[6-cyano-5-(cyclopropyloxy)-2-fluoro-3-[3-(1,4-oxazin-4-yl)prop-1-ynyl]phenyl]-1-methylpyrazol-4-yl}-4-carbonyl-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl)methyl]amino}methanoate

[0393] Tert-butyl (7-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-1-yl)methyl)carbamate (100 mg, 0.17 mmol) was dissolved in dioxane (10 mL) and 4-(prop-1-yn-3 -yl)-1,4-oxazepine (251.16 mg, 2.01 mmol), potassium carbonate (69.33 mg, 0.50 mmol), X-PHOS (7.97 mg, 0.02 mmol), Pd2(dba)3 (7.66 mg, 0.01 mmol), and N,N-dimethylformamide (5 mL) were stirred at 100°C under argon for 0.5 h. The reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined and washed with saturated brine (1 × 50 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanolic ammonia solution in dichloromethane, gradient elution from 0-3%) to obtain the product as a red oil (80 mg, 69.67%). ESI-MS m / z: 687.3 [M+1] +

[0394] Step 3: 2-{4-[1-(aminomethyl)-4-carbonyl-5-[(trideuterated methyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-6-(cyclopropyloxy)-3-fluoro-4-[3-(1,4-oxazin-4-yl)prop-1-ynyl]benzene-1-carbonitrile

[0395] 2-Methylpropan-2-yl ({[(7-{5-[6-cyano-5-(cyclopropyloxy)-2-fluoro-3-[3-(1,4-oxazin-4-yl)prop-1-ynyl]phenyl]-1-methylpyrazol-4-yl}-4-carbonyl-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl)methyl]amino}methane (55 mg, 0.08 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (0.5 mL) was added dropwise. The mixture was stirred at 25°C for 3 hours, concentrated under reduced pressure, and analyzed by preparative high performance liquid chromatography to give the product as a light yellow solid (3 mg, yield 6.4%). ESI-MS m / z: 587.2 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δ12.98(s,1H),8.79(s,1H),8.52(s,1H),8.28(s,2H),7.73(d,J=5.6Hz,1H),7.16(s,1H),4.34(d,J=2.4Hz,2H) ,4.18(tt,J=6.1,2.9Hz,1H),3.72(s,3H),3.66(s,2H),3.60(t,J=4.7Hz,4H),2.54–2.52(m,4H),0.95–0.79(m,3H),0.69~0.65(m,1H).

[0396] Example 2

[0397] 2-(4-(1-(aminomethyl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-cyclopropyloxy-3-fluoro-4-(prop-1-yn-1-yl)benzene-1-carbonitrile (Compound 2)

[0398] (M / P)-2-(4-(1-(aminomethyl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-cyclopropyloxy-3-fluoro-4-(prop-1-yn-1-yl)benzene-1-carbonitrile (Compound 2-P1 and Compound 2-P2)

[0399] Step 1: Tert-butyl (7-(5-(2-cyano-3-cyclopropyloxy-6-fluoro-5-(prop-1-yn-1-yl)phenyl)-1-methyl-1H-pyrazol-4-yl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-1-yl)methyl)carbamate

[0400] Tert-butyl (7-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-1-yl)methyl)carbamate (from Example 1, Step 1) (100 mg, 0.17 mmol) was dissolved in dioxane (10 mL), 1-propynyl-tri-n-butyltin (220.13 mg, 0.67 mmol) was added, and tetrakis(triphenylphosphine)palladium (38.65 mg, 0.03 mmol) was added under argon atmosphere. The reaction was stirred at reflux for 18 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100:40, volume ratio) to give the product as a yellow solid (60 mg, 59.6% yield). ESI-MS m / z: 602.2[M+1] + . 1 H NMR (400MHz, CDCl3) δ10.33 (s, 1H), 8.61 (s, 1H), 8.17 (s, 1H), 7.41 (d, J = 5.3Hz, 1H), 7.04 (s, 1H) ),4.50(d,J=5.0Hz,2H),3.85(m,1H),3.77(s,3H),2.15(s,3H),1.49(s,9H),0.98–0.79(m,4H).

[0401] Step 2: 2-(4-(1-(aminomethyl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-cyclopropyloxy-3-fluoro-4-(prop-1-yn-1-yl)benzene-1-carbonitrile (Compound 2)

[0402] Tert-butyl (7-(5-(2-cyano-3-cyclopropyloxy-6-fluoro-5-(prop-1-yn-1-yl)phenyl)-1-methyl-1H-pyrazol-4-yl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-1-yl)methyl)carbamate (54 mg, 0.09 mmol) was dissolved in dioxane (2 mL), and a solution of hydrogen chloride in 1,4-dioxane (3 mL, 12.00 mmol, 4 M) was added dropwise. The mixture was stirred at 25°C for 1.5 hours, concentrated under reduced pressure, and analyzed by preparative high-performance liquid chromatography to obtain the product as a light yellow solid (20 mg, 44.4% yield). ESI-MS m / z: 502.1 [M+1] + . 1H NMR (400MHz, DMSO-d6) δ12.6(s,1H), δ8.79(s,1H),8.50(s,1H),7.68(d,J=5.6Hz,1H),7.17(s,1H),4.13(dt,J=6.0,3.1Hz,1H),4.08 (s,2H),3.69(s,3H),2.16(s,3H),0.89(dd,J=10.9,5.1Hz,2H),0.82(dp,J=9.4,4.2,2.9Hz,1H),0.66(ddd,J=10.2,5.4,2.8Hz,1H). 13 C NMR(101MHz,DMSO-d6)δ160.90,158.16,157.19,155.05,154.29,152.63,139.16,136.89,131.22,124.05,123.8 4,122.35,118.59,118.25,118.07,113.77,103.63,102.68,99.67,98.04,72.48,53.20,37.08,6.33,6.29,4.52.

[0403] Step 3: (M / P)-2-(4-(1-(aminomethyl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-cyclopropyloxy-3-fluoro-4-(prop-1-yn-1-yl)benzene-1-carbonitrile

[0404] 2-(4-(1-(aminomethyl)-5-((methyl-d3)amino)-4-oxo-3,4-dihydropyrido[3,4-d]pyridazin-7-yl)-1-methyl-1H-pyrazol-5-yl)-6-cyclopropyloxy-3-fluoro-4-(prop-1-yn-1-yl)benzene-1-carbonitrile was further subjected to chiral preparative separation (ChiralPak IE, 250×40mm ID, 10μm column, mobile phase A: n-hexane; mobile phase B: ethanol containing 0.1% ammonia; gradient: B volume proportion in the mobile phase is 60%, elution time: 43.5 minutes) to obtain two products, compound 2-P1 and compound 2-P2.

[0405] Compound 2-P1: Chiral analysis method: chromatographic column ChiralPak IE, HS, 150×4.6 mm ID, 5 μm; mobile phase A: n-hexane; mobile phase B: ethanol containing 0.1% ammonia; gradient: B accounts for 60% by volume in the mobile phase; retention time: 10.917 minutes.

[0406] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),8.80(d,J=4.8Hz,1H),8.43(d,J=46.4Hz,1H),7.69(dd,J=5.7,2.4Hz,1H),7.15(d,J=12.9Hz, 1H),4.65(s,1H),4.14(tt,J=6.3,2.9Hz,1H),3.97(s,1H),3.70(d,J=2.1Hz,3H),2.17(s,3H),0.93–0.80(m,3H),0.70–0.64(m,1H).

[0407] Compound 2-P2: Chiral analysis method: chromatographic column ChiralPak IE, HS, 150×4.6 mm ID, 5 μm; mobile phase: A: n-hexane; mobile phase B: ethanol containing 0.1% ammonia; gradient: B accounts for 60% by volume in the mobile phase; retention time: 15.487 minutes.

[0408] 1 H NMR (400MHz, DMSO-d6) δ12.63(s,1H),8.80(d,J=3.0Hz,1H),8.43(d,J=48.5Hz,1H),7.69(dd,J=5.7,1.9Hz,1H),7.15(d,J=12.5Hz, 1H),4.65(s,1H),4.14(tt,J=6.1,2.9Hz,1H),4.02(s,1H),3.70(d,J=1.5Hz,3H),2.17(s,3H),0.92–0.79(m,3H),0.70–0.63(m,1H).

[0409] Intermediate a1 and intermediate b1 or intermediate a1 and intermediate b3-1 were reacted with corresponding alkynyl compounds according to the method of Example 1 and Example 2 to obtain the following target compounds.

[0410] The characterization data of Examples 3-18, 21 and 22 are as follows:

[0411] Example 19

[0412] 2-{4-[1-(Aminomethyl)-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-3-fluoro-4-(prop-1-ynyl)-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile (Compound 19)

[0413] Step 1: 2-Methylpropan-2-yl({[7-(5-{5-chloro-2-cyano-3-[(methyl)aminoethyl]-6-fluorophenyl}-1-methylpyrazol-4-yl)-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl]methyl}amino)formate

[0414] The coupling product was obtained by using intermediate b2 instead of intermediate b1 in the same manner as in step 1 of Example 1 above.

[0415] ESI-MS m / z: 611.2[M+1] + .

[0416] Step 2: 2-Methylpropan-2-yl {[(7-{5-[2-cyano-6-fluoro-5-(prop-1-ynyl)-3-(tetrahydro-1H-pyrrol-1-yl)phenyl]-1-methylpyrazol-4-yl}-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl)methyl]amino}methanoate

[0417] 2-Methylpropan-2-yl ({[(7-{5-[5-chloro-2-cyano-6-fluoro-3-(tetrahydro-1H-pyrrol-1-yl)phenyl]-1-methylpyrazol-4-yl}-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl)methyl]amino}methanoate (104 mg, 0.17 mmol) was dissolved in dioxane (10 mL). 1-Propyne-tri-n-butyltin (220.13 mg, 0.67 mmol) was added, followed by tetrakis(triphenylphosphine)palladium (38.65 mg, 0.03 mmol) under argon atmosphere. The mixture was stirred under reflux for 18 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / dichloromethane = 0-20%, volume ratio) to obtain the product as a yellow solid (70 mg, yield 67.3%). ESI-MS m / z: 615.7 [M+1] + .

[0418] Step 3: 2-{4-[1-(aminomethyl)-4-carbonyl-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-3-fluoro-4-(prop-1-ynyl)-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile

[0419] The same method as in step 3 of Example 1 was used to remove the protecting group to obtain the product.

[0420] ESI-MS m / z: 515.3[M+1] + ; 1 H NMR(400MHz,DMSO-d6)δ12.94(s,1H),8.74(s,1H),8.46(s,1H),8.32(s,2H),7.11(s,1H),6.9 5(d,J=5.9Hz,1H),4.32(s,2H),3.70(s,3H),3.49(s,4H),2.11(s,3H),1.93(d,J=6.7Hz,4H).

[0421] Example 20

[0422] 2-{4-[1-(Aminomethyl)-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-3-fluoro-4-[3-(1,4-oxazepan-4-yl)prop-1-ynyl]-6-(tetrahydro-1H-pyrrol-1-yl)benzene-1-carbonitrile (Compound 20)

[0423] Step 1: 2-Methylpropan-2-yl({[7-(5-{6-cyano-2-fluoro-3-[3-(1,4-oxazinan-4-yl)prop-1-ynyl]-5-(tetrahydro-1H-pyrrol-1-yl)phenyl}-1-methylpyrazol-4-yl)-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl]methyl}amino)formate

[0424] Under argon protection, the reaction mixture was stirred at room temperature in the presence of 2-methylpropan-2-yl{[(7-{5-[3-chloro-6-cyano-2-fluoro-5-(tetrahydro-1H-pyrrol-1-yl)phenyl]-1-methylpyrazol-4-yl}-4-carbonyl-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl)methyl]amino}formate (70 mg, 0.11 mmol), 4-propynyl-1-morpholine To a solution of 1,4-dimethylformamide (143.38 mg, 1.15 mmol) and potassium carbonate (47.49 mg, 0.34 mmol) in N,N-dimethylformamide (5 mL) were added tris(dibenzylideneacetone)dipalladium (10.49 mg, 0.01 mmol) and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (10.49 mg, 0.02 mmol). The reaction mixture was stirred in a microwave at 110°C for 0.5 hour. After completion of the reaction, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ammonia methanol / dichloromethane = 0-5%) to afford the product as a red oil (24 mg, 29.94% yield). ESI-MS m / z: 700.3 [M+1] +

[0425] Step 2: 2-{4-[1-(aminomethyl)-4-carbonyl-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-7-yl]-2-methylpyrazol-3-yl}-6-(cyclopropenyloxy)-3-fluoro-4-[3-(piperazin-1-yl)prop-1-ynyl]benzene-1-carbonitrile

[0426] 2-Methylpropan-2-yl ({[7-(5-{6-cyano-2-fluoro-3-[3-(1,4-oxazin-4-yl)prop-1-ynyl]-5-(tetrahydro-1H-pyrrol-1-yl)phenyl}-1-methylpyrazol-4-yl)-4-oxo-5-[(trideuteriomethyl)amino]-3H-pyrido[4,3-d][1,2]diazin-1-yl]methyl}amino)formate (24 mg, 0.03 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (0.5 mL) was added. The reaction solution was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was concentrated under reduced pressure and analyzed by preparative high-performance liquid chromatography to obtain the product as a light yellow solid (11 mg, yield 53.48%). ESI-MS m / z: 600.2 [M+1] + . 1H NMR (400MHz, DMSO-d6) δ12.96(s,1H),8.77(s,1H),8.48(s,1H),8.35(s,2H),7.13(s,1H),7.05(d,J=5.8Hz,1H ),4.38–4.31(m,2H),3.93(s,2H),3.74–3.63(m,7H),3.57–3.42(m,4H),3.04–2.72(m,4H),1.99–1.90(m,4H).

[0427] Example 23: Formation of PRMT5-MEP50-MTA-Compound 2-P2 Complex Crystals and Analysis of the Crystal Structure

[0428] Protein expression and purification

[0429] The PRMT5-MEP50 protein was expressed using the baculovirus expression system. The PRMT5 (Uniprot O14744 1-637) gene with an N-terminal flag tag and the MEP50 (Uniprot Q9BQA1 1-342) gene with an N-terminal His tag were cloned into the baculovirus transfer vector pFastBac to construct recombinant plasmids. The recombinant plasmids were transformed into DH10Bac competent cells containing the shuttle vector Bacmid. After transposition, recombinant Bacmid containing the target gene was obtained. The recombinant Bacmid DNA was transfected into Sf9 insect cells to obtain recombinant baculovirus. The PRMT5 and MEP50 recombinant baculoviruses were co-infected into Sf9 cells. After 48-72 hours of infection, the cells were harvested by centrifugation and lysed in buffer A (25 mM Tris-HCl, 300 mmol / L NaCl, 5% glycerol, 14 mM BME). Cell debris was removed by centrifugation at 14,000 rpm for 1 hour. The supernatant was purified by AntiFlag affinity chromatography. 0.2 mg / mL of α-aminobutyric acid was added to buffer A. The flag polypeptide was eluted, and the eluate was further purified by Ni-column affinity chromatography using 250 mmol / L imidazole in buffer A. This fraction was concentrated and further purified by gel filtration chromatography (25 mmol / L Hepes pH 7.5, 150 mmol / L NaCl, 5% glycerol, 1 mmol / L TCEP). The purified protein was finally analyzed by SDS-PAGE. The PRMT5-MEP50 complex proteins were combined and concentrated to 10 mg / mL.

[0430] Protein crystallization

[0431] After incubation of 8.8 mg / mL PRMT5-MEP50 with 1 mmol / L MTA and 0.5 mmol / L Compound 2-P2, crystals were grown by vapor diffusion. The first PRMT5-MEP50-MTA-Compound 2-P2 protein crystal was grown by sitting drop method at 8°C in 15% PEG 4000, 100 mmol / L sodium citrate pH 5.0, and 150 mmol / L sodium acetate. The final crystal used for data collection was grown by adding 200 nL of serially diluted seed crystals to the protein crystal culture medium.

[0432] Structural analysis

[0433] Structural data of PRMT5-MEP50-MTA-Compound 2-P2 complex crystals ( The data were collected at the Shanghai Synchrotron Radiation Facility (SSRF) at 1.5-1.5 resolution. Data were processed using the HKL2000 software package, resulting in a space group of P21212. Phase solution was performed using the molecular replacement phaser in CCP4, and the structure was further refined using CCP4.refine and Coot. The main parameters of the protein cocrystal are shown in Table 1, and the structure of the protein cocrystal is shown in Figure 1.

[0434] Table 1 X-ray data collection and optimization statistics of PRMT5-MEP50-MTA-Compound 2-P2 complex crystals

[0435] Conclusion: From Table 1 and Figure 1, we can see that the structural formula of compound 2-P2 is as follows:

[0436] Biological Test Examples

[0437] Test Example 1: Proliferation inhibitory activity on human colon cancer cells HCT-116 and human colon cancer cells with stable MTAP protein knockout HCT-116-MTAP-KO-1A2

[0438] The PrestoBlue assay was used to determine the inhibitory activity of the compounds on the proliferation of human colon cancer cells HCT-116 and HCT-116-MTAP-KO-1A2, in which MTAP protein was stably knocked out in HCT-116, in vitro.

[0439] Cell sources: HCT-116 cells were purchased from the Institute of Cell Biology, Chinese Academy of Sciences; HCT-116-MTAP-KO-1A2 cells were purchased from Kangyuan Broad Biotechnology (Beijing) Co., Ltd.

[0440] HCT-116 and HCT-116-MTAP-KO-1A2 cells were cultured in RPMI1640 complete medium supplemented with 10% fetal bovine serum. HCT-116 and HCT-116-MTAP-KO-1A2 cells in the logarithmic growth phase were seeded in 96-well plates at a density of 500 cells / 135 μl complete medium / well and incubated at 37°C in a 5% CO2 incubator for 24 hours. Each compound was dissolved in dimethyl sulfoxide (DMSO) to a 10 mM stock solution and then diluted in DMSO and complete medium, sequentially. Remove the 96-well plate seeded with cells and use one plate as a no-growth control (a medium control with no cell growth at 0 hours). Add 15 μL of each compound to each well of the remaining 96-well plates, resulting in final concentrations of 10,000, 2,500, 625, 156.3, 39.1, 9.8, 2.4, 0.6, 0.15, and 0.04 nM. Each compound concentration was replicated in triplicate. A negative control (medium control containing cells but no compound) was also established. The DMSO concentration in each well was 0.5%. The remaining no-growth control was immediately incubated with PrestoBlue HS Cell Viability Assay Reagent and subsequently assayed. The remaining 96-well plates were incubated in a 37°C, 5% CO2 incubator for 6 days before incubation with PrestoBlue HS Cell Viability Assay Reagent and subsequently assayed.

[0441] The following steps were used to add PrestoBlue HS Cell Viability Assay Reagent and perform the following assays: Remove the 96-well cell culture plate from the CO2 incubator and add 15 μl of PrestoBlue HS Cell Viability Assay Reagent (Cat. No. P50201) to each well. Incubate the plate in a 37°C incubator with 5% CO2 for 3 hours. Remove the 96-well cell culture plate and measure the relative fluorescence intensity (RFU) using a microplate reader at an excitation wavelength of 560 nm and an emission wavelength of 590 nm. Calculate the cell inhibition rate at each concentration using the following formula.

[0442] Data were analyzed using GraphPad Prism 8.3 software. Nonlinear S-curve regression was used to fit the data to obtain the dose-effect curve, from which the IC was calculated. 50 The results are shown in Table 1.

[0443] Table 1

[0444] MRTX1719 is a positive control compound (prepared by the method described in J. Med. Chem. 2022, 65, 1749–1766). A represents <25 nM, B represents 25–200 nM, C represents 201 nM–1 μM, and D represents >1 μM. The selectivity value is the IC value for HCT-116 cells. 50 The IC values ​​of HCT-116-MTAP-KO-1A2 cells were compared with those of 50 The ratio of values.

[0445] The results showed that the compound of the present invention exhibited good proliferation inhibition activity and selectivity against human colon cancer stably knocked-out MTAP protein cell line HCT-116-MTAP-KO-1A2.

[0446] Test Example 2: Liver microsome metabolic stability experiment

[0447] 1. Purpose of the experiment

[0448] The compounds of the invention were tested for their metabolic stability in liver microsomes.

[0449] 2. Experimental Materials

[0450] Liver microsomes were purchased from Xenotech.

[0451] 3. Experimental content

[0452] 3.1 Preparation of stock solution

[0453] The compounds of the present invention were dissolved in DMSO to obtain 10 mM stock solutions.

[0454] 3.2 Preparation of working solution

[0455] (1) Add appropriate amount of 10 mM stock solution to PBS buffer to obtain 10 μM working solution of the test compound.

[0456] (2) Weigh an appropriate amount of NADPH and dilute it with an appropriate amount of PBS buffer to obtain a 20 mM NADPH working solution.

[0457] (3) Take an appropriate amount of liver microsomes and dilute them with an appropriate amount of PBS buffer to a final concentration of 0.625 mg / mL of liver microsome working solution.

[0458] 3.3 Experimental steps

[0459] 400 μL of liver microsome working solution and 50 μL of test compound working solution were added to a 96-well plate in sequence and pre-incubated at 37°C for 10 minutes. Subsequently, 50 μL of NADPH working solution was added to initiate the reaction. 50 μL of incubation solution was removed at time points of 0, 5, 10, 20, 30, and 60 minutes, and 500 μL of acetonitrile solution containing an internal standard (containing 50 ng / mL tolbutamide) was added to terminate the reaction. The plate was centrifuged at 10°C and 8000 rpm for 10 minutes. Subsequently, 50 μL of the supernatant was transferred to a 96-well plate, diluted with 500 μL of ultrapure water, and vortexed to mix. The plate was centrifuged again for 5 minutes and injected into LC-MS / MS. The test results are shown in Table 2.

[0460] 4. Experimental Results

[0461] Table 2: In vitro liver microsome metabolic stability test results

[0462] 5. Experimental Conclusion

[0463] The compounds of the present invention exhibit moderate metabolism in liver microsomes in both rat and human species.

[0464] Test Example 3: Rat Pharmacokinetic Experiment

[0465] 1. Purpose of the experiment

[0466] The pharmacokinetic properties of the compounds were tested in SD rats.

[0467] 2. Experimental Materials

[0468] SD rats (female, 180-220 g, 6-8 weeks old, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.).

[0469] 3. Experimental Procedure

[0470] The pharmacokinetic properties of the compounds were tested after single tail vein injection and oral administration after an overnight fast. The compounds were formulated as clear solutions or homogenous suspensions. The intravenous dose was 1 mg / kg, and the oral dose was 12.5 mg / kg. The solvent was 10% dimethyl sulfoxide / 10% polyethylene glycol (15)-hydroxystearate / 80% saline. Whole blood samples were collected from the submandibular vein at 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h, and plasma samples were obtained by centrifugation at 4°C and 8000 rpm for 5 minutes.

[0471] 40 μL of the plasma sample was transferred to a 96-well plate, and 300 μL of an acetonitrile solution containing the internal standard (50 ng / mL tolbutamide) was added. The sample was thoroughly vortexed for 10 minutes and centrifuged at 4°C, 6000 rpm, for 20 minutes. 100 μL of the supernatant was diluted with 300 μL of ultrapure water and injected into the plate for quantitative analysis of plasma drug concentrations by LC-MS / MS. Key pharmacokinetic parameters were calculated. The results are shown in Table 3.

[0472] 4. Experimental Results

[0473] Table 3: Pharmacokinetic test results in SD rats

[0474] 5. Experimental Conclusion

[0475] Compared with the reference product, the compound of the present invention has better oral bioavailability and plasma exposure, and shows good pharmacokinetic properties.

[0476] Test Example 4: Beagle dog pharmacokinetic experiment

[0477] 1. Purpose of the experiment

[0478] The pharmacokinetic properties of the test compounds were determined in beagle dogs.

[0479] 2. Experimental Materials

[0480] Beagle dogs (male, ≥6 months old, purchased from Beijing Masi Biotechnology Co., Ltd.)

[0481] 3. Experimental Procedure

[0482] The pharmacokinetic properties of the compounds were tested following single tail vein injection and oral administration after an overnight fast. Compounds were formulated as clear solutions or homogeneous suspensions. The intravenous dose was 2 mg / kg in 20% sulfobutyl-β-cyclodextrin / 50 mM citric acid (pH 5); the oral dose was 10 mg / kg in 0.5% methylcellulose (4000 cps) / 0.2% Tween 80. Whole blood samples were collected from the submandibular vein at 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. Plasma samples were obtained by centrifugation at 8000 rpm for 5 minutes at 4°C.

[0483] Forty microliters of plasma sample were transferred to a 96-well plate, and 300 microliters of acetonitrile solution containing the internal standard was added. The sample was thoroughly vortexed for 10 minutes and centrifuged at 6,000 rpm for 20 minutes at 4°C. A hundred microliters of supernatant was diluted with 300 microliters of ultrapure water and injected into the plate for quantitative analysis of plasma drug concentrations by LC-MS / MS. Key pharmacokinetic parameters were calculated. The results are shown in Table 4.

[0484] 4. Experimental Results

[0485] Table 4: Pharmacokinetic test results in beagle dogs

[0486] 5. Experimental Conclusion

[0487] The compound of the present invention has good oral bioavailability and plasma exposure in beagle dogs and good pharmacokinetic properties.

Claims

1. A compound as shown in formula I, its tautomer or a pharmaceutically acceptable salt thereof, R 1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 1a Substituted C1-C6 alkyl, one or more R 1b Substituted C2-C6 alkenyl, substituted by one or more R 1c Substituted C2-C6 alkynyl, substituted by one or more R 1d Substituted 3-7 membered cycloalkyl, 1e Substituted 3-7 membered cycloalkenyl, one or more R 1f substituted 4-10 membered heterocyclic group, substituted by one or more R 1g Substituted C6-C 10 Aryl or one or more R 1h substituted 5-10 membered heteroaryl; Each R 1a , R 1b , R 1c , R 1d , R 1e , R 1f , R 1g and R 1h are independently deuterium, halogen, -OH, =O, cyano, -NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-OR 13 、-C(=O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 ,-P(=O)(R 12 )2.-P(=O)(OR 12 )2.-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12 S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl, one or more R 2b Substituted C2-C6 alkenyl, substituted by one or more R 2c Substituted C2-C6 alkynyl, substituted by one or more R 2d Substituted 3-7 membered cycloalkyl, 2e Substituted 3-7 membered cycloalkenyl, one or more R 2f substituted 4-10 membered heterocyclic group, substituted by one or more R 2g Substituted C6-C 10 Aryl or one or more R 2h substituted 5-10 membered heteroaryl; Each R 2a , R 2b , R 2c , R 2d , R 2e , R 2f , R 2g and R 2h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-C(=O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 ,-P(=O)(R 12 )2.-P(=O)(OR 12 )2.-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12 S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 5a Substituted C1-C6 alkyl, one or more R 5b Substituted 3-7 membered cycloalkyl, 5c Substituted 3-7 membered cycloalkenyl, or multiple R 5d substituted 4-10 membered heterocyclic group, substituted by one or more R 5e Substituted C6-C 10 Aryl or one or more R 5f substituted 5-10 membered heteroaryl; Each R 5a , R 5b , R 5c , R 5d , R 5e and R 5f Each is independently deuterium, halogen, =O, cyano, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -OR 13 、-NR 10 R 11 、-C(=O)R 12 、-C(=O)OR 12 、-OC(=O)R 12 、-C(=O)NR 10 R 11 、-NR 12 C(=O)R 12 、-NR 12 C(=O)NR 10 R 11 、-S(=O)R 12 、-S(=O)2R 12 ,-P(=O)(R 12 )2.-P(=O)(OR 12 )2.-NR 12 S(=O)R 12 、-NR 12 S(=O)2R 12 、-NR 12 S(=O)NR 10 R 11 、-NR 12 S(=O)2NR 10 R 11 、-S(O)NR 10 R 11 、-S(O)2NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl, one or more R 5-b Substituted C2-C6 alkenyl, substituted by one or more R 5-c Substituted C2-C6 alkynyl, substituted by one or more R 5-d Substituted 3-7 membered cycloalkyl, 5-e Substituted 3-7 membered cycloalkenyl, one or more R 5-f substituted 4-10 membered heterocyclic group, substituted by one or more R 5-g Substituted C6-C 10 Aryl or one or more R 5-h substituted 5-10 membered heteroaryl; Each R 5-a , R 5-b , R 5-c , R 5-d , R 5-e , R 5-f , R 5-g and R 5-h are independently deuterium, halogen, =O, cyano, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuteriums, or C1-C6 alkyl substituted by one or more cyano groups; R 7 is hydrogen, deuterium, halogen, cyano, nitro, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 7a Substituted C1-C6 alkyl, one or more R 7b Substituted C2-C6 alkenyl, substituted by one or more R 7c Substituted C2-C6 alkynyl, substituted by one or more R 7d Substituted 3-7 membered cycloalkyl, 7e Substituted 3-7 membered cycloalkenyl, one or more R 7f substituted 4-10 membered heterocyclic group, substituted by one or more R 7g Substituted C6-C 10 Aryl or one or more R 7h substituted 5-10 membered heteroaryl; Each R 7a , R 7b , R 7c , R 7d , R 7e , R 7f , R 7g and R 7h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; Each R 8 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, -NR 10 R 11 、-OR 13 , by one or more R 8a Substituted C1-C6 alkyl, one or more R 8b Substituted C2-C6 alkenyl, substituted by one or more R 8c Substituted C2-C6 alkynyl, substituted by one or more R 8d Substituted 3-7 membered cycloalkyl, 8e Substituted 3-7 membered cycloalkenyl, one or more R 8f substituted 4-10 membered heterocyclic group, substituted by one or more R 8g Substituted C6-C 10 Aryl or one or more R 8h substituted 5-10 membered heteroaryl; Each R 8a , R 8b , R 8c , R 8d , R 8e , R 8f , R 8g and R 8h are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 , C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b 、-C(=O)-OC 6-10 Aryl, -C(=O)-C 1-6 Alkyl or -(CH2) m -OP(=O)(OR 9c )2; R 9a and R 9b are each independently hydrogen, deuterium or C 1-20 alkyl; Each R 9c are each independently hydrogen, deuterium or C 1-20 alkyl; Each R 10 and R 11 Each is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 10a Substituted C1-C6 alkyl, R 10b Substituted C2-C6 alkenyl, substituted by one or more R 10c Substituted C2-C6 alkynyl, substituted by one or more R 10d Substituted 3-7 membered cycloalkyl, 10e Substituted 3-7 membered cycloalkenyl, one or more R 10f substituted 4-10 membered heterocyclic group, substituted by one or more R 10g Substituted C6-C 10 Aryl or one or more R 10h substituted 5-10 membered heteroaryl; Each R 10a , R 10b , R 10c , R 10d , R 10e , R 10f , R 10g and R 10h Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; Each R 12 are independently hydrogen, C 1-6 Alkyl, C 3-8 Cycloalkyl, 4-10 membered heterocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 12a Substituted C 1-6 Alkyl, with one or more R 12b Substituted C 3-8 Cyclic hydrocarbon group, one or more R 12c substituted 4-10 membered heterocyclic group, substituted by one or more R 12d Substituted C 6-10 Aryl or one or more R 12e substituted 5-10 membered heteroaryl; Each R 12a , R 12b , R 12c , R 12d and R 12e Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; Each R 13 Each is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, 3-7 membered cycloalkyl, 3-7 membered cycloalkenyl, 4-10 membered heterocyclyl, C6-C 10 Aryl, 5-10 membered heteroaryl, substituted by one or more R 13a Substituted C1-C6 alkyl, one or more R 13b Substituted C2-C6 alkenyl, substituted by one or more R 13c Substituted C2-C6 alkynyl, substituted by one or more R 13d Substituted 3-7 membered cycloalkyl, 13e Substituted 3-7 membered cycloalkenyl, one or more R 13f substituted 4-10 membered heterocyclic group, substituted by one or more R 13g Substituted C6-C 10 Aryl or one or more R 13h substituted 5-10 membered heteroaryl; Each R 13a , R 13b , R 13c , R 13d , R 13e , R 13f , R 13g and R 13h Each is independently deuterium, halogen, =O, cyano, -OH, -NH2, C1-C6 alkyl, C1-C6 alkyl-O- or 3-7 membered cycloalkyl; n is 1 or 2; m is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; The heteroatoms in the 5-10 membered heteroaryl group and the 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3.

2. The compound of formula I as claimed in claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 1 wherein the C1-C6 alkyl group and the 1a The C1-C6 alkyl in the substituted C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl; (2)R 6 wherein the halogen is fluorine, chlorine, bromine or iodine, for example fluorine; (3)R 3 wherein the 4-10 membered heterocyclic group and the one or more R 2f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is a 4-10 membered heterocycloalkyl or a 4-10 membered heterocycloalkenyl, the 4-10 membered heterocycloalkyl may independently be a 5-6 membered heterocycloalkyl, the 4-10 membered heterocycloalkyl may independently be a monocyclic ring, the heteroatom of the 4-10 membered heterocycloalkyl may independently be N, the number of heteroatoms may independently be 1, or may be For example (4)R 5 wherein the C1-C6 alkyl group and the 5a The C1-C6 alkyl in the substituted C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl, ethyl or isopropyl; (6)R 5 wherein the 3-7 membered cycloalkyl group and the one or more R 5b The 3-7 membered cycloalkyl in the substituted 3-7 membered cycloalkyl is is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, for example cyclopropyl; (7) Each R 5a wherein the 4-10 membered heterocyclic group and one or more R 5-f The 4-10-membered heterocyclic group in the substituted 4-10-membered heterocyclic group is a 4-10-membered heterocycloalkyl or a 4-10-membered heterocycloalkenyl, and the 4-10-membered heterocycloalkyl is independently a 4-6-membered monoheterocycloalkyl or a 7-10-membered biheterocycloalkyl; the biheterocycloalkyl may be a spiro ring, a bridged ring or a fused ring, and the heteroatoms of the 4-10-membered heterocycloalkyl may independently be N and / or O, and the number of heteroatoms may independently be 1 or 2; (8) Each R 5-f wherein the halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine; (9) Each R 5-f wherein the C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl; (10) Each R 10 and R 11 wherein the C1-C6 alkyl group and the 10a The C1-C6 alkyl in the substituted C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl; (11) Each R 13 wherein the C1-C6 alkyl group and one or more R 13a The C1-C6 alkyl in the substituted C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl; and (12) each R 13 , the 3-7 membered cycloalkyl group and one or more R 13d The 3-7-membered cycloalkyl in the substituted 3-7-membered cycloalkyl is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, for example cyclopropyl; Preferably, the 4-6 membered monoheterocycloalkyl is For example And / or, the 7-10 membered biheterocycloalkyl is a bridged ring, which may be a 7-8 membered bridged ring, for example For example 3. The compound as shown in formula I according to claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1) Each C1-C 20 Alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (2) each C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (3) each C2-C6 alkenyl group is independently ethenyl, propenyl, butenyl or pentenyl; (4) each C2-C6 alkynyl group is independently ethynyl, propynyl, butynyl or pentynyl; (5) each 3-7 membered cycloalkyl group is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; (6) each 3-7 membered cycloalkenyl group is independently cyclopropenyl, cyclobutenyl, cyclopentenyl or cyclohexenyl; (7) Each 4-10 membered heterocyclic group is independently a saturated or unsaturated heterocyclic group, such as a saturated heterocycloalkyl group; (8) Each 4-10 membered heterocyclic group is independently a monocyclic heterocyclic group or a bicyclic heterocyclic group, and the bicyclic heterocyclic group may be a spirocyclic ring, a bridged ring or a fused ring; (9) Each 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group; (10) The heteroatoms in each 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms can be independently 1 or 2; (11) Each C6-C 10 Aryl is independently phenyl or naphthyl; (12) Each 5-10 membered heteroaryl group is independently a 5-6 membered monocyclic heteroaryl group or an 8-10 membered polycyclic heteroaryl group, wherein the heteroatom species is N and the number of heteroatoms is 1, 2 or 3; (13) Each halogen is independently fluorine, chlorine, bromine or iodine; (14) each C1-C6 alkyl-O- is independently methyl-O-, ethyl-O-, n-propyl-O-, isopropyl-O-, n-butyl-O-, tert-butyl-O-, isobutyl-O- or sec-butyl-O-; and (15) the compound represented by formula I is any stereoisomer or a mixture thereof, for example, when containing one chiral center, the compound represented by formula I is an R configuration, an S configuration, or an R, S configuration racemate; Preferably, the 4-6 membered monoheterocyclic group is For example And / or, the 7-10 membered biheterocyclic group is a bridged ring, which may be a 7-8 membered bridged ring, for example For example 4. The compound of formula I as described in any one of claims 1 to 3, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 1 is hydrogen, C1-C6 alkyl or is replaced by one or more R 1a Substituted C1-C6 alkyl; preferably, R 1 is a C1-C6 alkyl group; (2) Each R 1a are independently deuterium, halogen, -OH, cyano or -NR 10 R 11 ; (3)R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl or one or more R 2f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 2f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is 1, 2 or 3; preferably, R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 , the heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is independently 1; (4)R 6 is a halogen; (5) Each R 2a and R 2f are independently deuterium, halogen, =O, cyano, -NR 10 R 11 or C1-C6 alkyl; (6)R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, or one or more R 5a Substituted C1-C6 alkyl, one or more R 5b Substituted 3-7 membered cycloalkyl, 5d substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and the 5d The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is independently 1 or 2; preferably, R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or is replaced by one or more R 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; (7) Each R 5a , R 5b and R 5d Each is independently deuterium, halogen, cyano, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl, one or more R 5-f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; preferably, each R 5a and R 5b Each is independently a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; (8) Each R 5-f and R 5-a are independently deuterium, halogen, =O, cyano, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuteriums, or C1-C6 alkyl substituted by one or more cyano groups; preferably, each R 5-f and R 5-a are independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl, more preferably, each R 5-f are independently halogen or C1-C6 alkyl; (9)R 7 is hydrogen, deuterium, halogen, cyano, nitro, -OH, -NH2, C1-C6 alkyl, or is replaced by one or more R 7a Substituted C1-C6 alkyl; preferably, R 7 is hydrogen; (10) Each R 7a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; (11) Each R 8 are independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, -NR 10 R 11 、-OR 13 or one or more R 8a Substituted C1-C6 alkyl; preferably, each R 8 are each independently hydrogen or -NR 10 R 11 ; For example, R 8 For-NR 10 R 11 ; (12) Each R 8a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; (13)R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b or -(CH2) m -OP(=O)(OR 9c )2; preferably, R 9 are independently hydrogen; (14)R 9a and R 9b are independently hydrogen or C 1-20 alkyl; (15) Each R 9c are independently hydrogen or C 1-20 alkyl; (16)R 10 and R 11 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, 10a Substituted C1-C6 alkyl, one or more R 10d substituted 3-7 membered cycloalkyl or one or more R 10f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 10f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; preferably, R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; (17) Each R 10a , R 10d and R 10f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; preferably Select a location, each R 10a Each independently is deuterium; (18) Each R 13 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, 13a Substituted C1-C6 alkyl, one or more R 13d substituted 3-7 membered cycloalkyl or one or more R 13f substituted 4-10 membered heterocyclic group, wherein the 4-10 membered heterocyclic group and one or more R 13f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; preferably, each R 13 Each R is independently hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl. More preferably, each R 13 Each is independently hydrogen or a 3-7 membered cycloalkyl; and (19) each R 13a , R 13d and R 13f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; Preferably, R 3 is a 4-10 membered heterocyclic group or -OR 13 ; The heteroatom in the 4-10 membered heterocyclic group is N, O or S, and the number of heteroatoms is 1, 2 or 3; and / or, each R 5a Each is independently a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2. Preferably, each R 5a Each is independently a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 The heteroatoms in the 4-6 membered monoheterocyclic group are N and O, the number of heteroatoms is 2, and more preferably each R 5a Each is independently a 4-6 membered monoheterocyclic group; the heteroatoms in the 4-6 membered monoheterocyclic group are N and O, and the number of heteroatoms is 2; and / or, each R 5b Each is independently -NH2.

5. The compound as shown in formula I according to claim 4, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: Each R 13d wherein the C1-C6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl; and / or, R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl.

6. The compound of formula I as claimed in claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I is a compound as shown in formula IA or formula IB:

7. The compound of formula I as claimed in claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I is a compound as shown in formula I-1, formula I-2, formula I-3 or formula I-4: Preferably, the compound as shown in formula I is a compound as shown in formula I-2-1-1, formula I-2-1-2, formula I-3-1-1, formula I-3-1-2, formula I-4-1-1 or formula I-4-1-2: Preferably, the compound as shown in formula I is a compound as shown in formula I-1-1-1 or I-1-1-2:

8. The compound of formula I as claimed in claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in Formula I is Scheme 1, Scheme 2, Scheme 3, Scheme 4, Scheme 5, Scheme 6, Scheme 7, Scheme 8 or Scheme 9: Solution 1: n is 1 or 2; R 1 is hydrogen, C1-C6 alkyl or is replaced by one or more R 1a Substituted C1-C6 alkyl; Each R 1a are independently deuterium, halogen, -OH, cyano or -NR 10 R 11 ; R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 2a Substituted C1-C6 alkyl or one or more R 2f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 2f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N, O or S, and the number of heteroatoms is independently 1, 2 or 3; Each R 2a and R 2f are independently deuterium, halogen, =O, cyano, -NR 10 R 11 or C1-C6 alkyl; R 5 is hydrogen, deuterium, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, or one or more R 5a Substituted C1-C6 alkyl, one or more R 5b substituted 3-7 membered cycloalkyl or one or more R 5d substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and the one or more R 5d The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5a , R 5b and R 5d Each is independently deuterium, halogen, cyano, C1-C6 alkyl, 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 , by one or more R 5-a Substituted C1-C6 alkyl or one or more R 5-f substituted 4-10 membered heterocyclic group; said 4-10 membered heterocyclic group and one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5-f and R 5-a are independently deuterium, halogen, =O, cyano, -OR 13 、-NR 10 R 11 , C1-C6 alkyl, C1-C6 alkyl substituted by one or more halogen, C1-C6 alkyl substituted by one or more -OH, C1-C6 alkyl substituted by one or more deuterium, or C1-C6 alkyl substituted by one or more cyano; R 7 is hydrogen, deuterium, halogen, cyano, nitro, -OR 13 、-NR 10 R 11 , C1-C6 alkyl or one or more R 7a Substituted C1-C6 alkyl; Each R 7a are independently deuterium, halogen, =O, cyano, -NR 10 R 11 、-OR 13 or C1-C6 alkyl; Each R 8 are independently hydrogen, deuterium, halogen, cyano, nitro, C1-C6 alkyl, -NR 10 R 11 、-OR 13 or one or more R 8a Substituted C1-C6 alkyl; Each R 8a Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; R 9 are independently hydrogen, deuterium, -C(=O)-O-CH(R 9a )-OC(O)-R 9b or -(CH2) m -OP(=O)(OR 9c )2; R 9a and R 9b are independently hydrogen or C 1-20 alkyl; Each R 9c are independently hydrogen or C 1-20 alkyl; R 10 and R 11 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, 10a Substituted C1-C6 alkyl, one or more R 10d substituted 3-7 membered cycloalkyl or one or more R 10f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 10f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; Each R 10a , R 10d and R 10f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; Each R 13 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 4-10 membered heterocyclic group, 13a Substituted C1-C6 alkyl, one or more R 13d substituted 3-7 membered cycloalkyl or one or more R 13f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 13f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; Each R 13a , R 13d and R 13f Each is independently deuterium, halogen, =O, cyano, -OH, -NH2 or C1-C6 alkyl; Solution 2 n is 1 or 2; R 1 is a C1-C6 alkyl group; R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or is replaced by one or more R 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a and R 5b Each is independently a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The substituted 4-10 membered heterocyclic group The heteroatoms are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5-f are independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl; R 7 is hydrogen; Each R 8 are each independently hydrogen or -NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each is independently hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl; Solution 3 n is 1 or 2; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or is replaced by one or more R 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a and R 5b Each is independently a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5-f are independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl; R 7 is hydrogen; Each R 8 are each independently hydrogen or -NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each is independently hydrogen, C1-C6 alkyl or 3-7 membered cycloalkyl; Solution 4: n is 1; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl, or is replaced by one or more R 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a Each is independently a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5b Each is independently -NH2; Each R 5-f are independently halogen or C1-C6 alkyl; R 7 is hydrogen; R 8 For-NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a independently for deuterium; Each R 13 Each is independently a 3-7 membered cycloalkyl group; Solution 5: n is 1; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is C1-C6 alkyl, 3-7 membered cycloalkyl or is replaced by one or more R 5a Substituted C1-C6 alkyl; Each R 5a Each is independently a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 ; The heteroatoms in the 4-6 membered monoheterocyclic group are N and O, and the number of heteroatoms is 2; R 7 is hydrogen; R 8 For-NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each is independently a 3-7 membered cycloalkyl group; Solution 6: n is 1 or 2; R 1 is a C1-C6 alkyl group; R 2 , R 3 , R 4 and R 6 Each is independently hydrogen, halogen, cyano, 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a and R 5b Each is independently a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5-f are independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl; R 7 is hydrogen; Each R 8 are each independently hydrogen or -NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, or substituted by one or more R 13d Substituted 3-7 membered cycloalkyl; Each R 13d Each is independently a C1-C6 alkyl group; Solution 7: n is 1 or 2; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a and R 5b Each is independently a 4-10 membered heterocyclic group, -OR 13 、-NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5-f are independently halogen, -OR 13 、-NR 10 R 11 or C1-C6 alkyl; R 7 is hydrogen; Each R 8 are each independently hydrogen or -NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each is independently hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl or substituted by one or more R 13d Substituted 3-7 membered cycloalkyl; each R 13d Each is independently a C1-C6 alkyl group; Solution 8: n is 1; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl, 5a Substituted C1-C6 alkyl or one or more R 5b Substituted 3-7 membered cycloalkyl; Each R 5a Each is independently a 4-10 membered heterocyclic group, -OH, -NR 10 R 11 or one or more R 5-f substituted 4-10 membered heterocyclic group; the 4-10 membered heterocyclic group and one or more R 5-f The 4-10 membered heterocyclic group in the substituted 4-10 membered heterocyclic group is independently a 4-6 membered monoheterocyclic group or a 7-10 membered biheterocyclic group, and the 4-10 membered heterocyclic group and the substituted 4-10 membered heterocyclic group are independently substituted by one or more R 5-f The heteroatoms in the substituted 4-10 membered heterocyclic group are independently N and / or O, and the number of heteroatoms is independently 1 or 2; Each R 5b Each is independently -NH2; Each R 5-f are independently halogen or C1-C6 alkyl; R 7 is hydrogen; R 8 For-NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a independently for deuterium; Each R 13 Each independently represents a 3-7 membered cycloalkyl group, or is replaced by one or more R 13d Substituted 3-7 membered cycloalkyl; Each R 13d Each is independently a C1-C6 alkyl group; Solution 9: n is 1; R 1 is a C1-C6 alkyl group; R 2 is cyano; R 3 is a 5-6 membered heterocyclic group or -OR 13 ; The heteroatom in the 5-6 membered monoheterocyclic group is N, and the number of heteroatoms is 1; R 4 is hydrogen; R 6 is a halogen; R 5 is hydrogen, C1-C6 alkyl, 3-7 membered cycloalkyl or is replaced by one or more R 5a Substituted C1-C6 alkyl; Each R 5a Each is independently a 4-6 membered monoheterocyclic group, -OH or -NR 10 R 11 The heteroatom in the 4-6 membered heterocyclic group is N and O, the number of heteroatoms is 2; R 7 is hydrogen; R 8 For-NR 10 R 11 ; R 9 are independently hydrogen; R 10 and R 11 Each independently is hydrogen, C1-C6 alkyl or substituted by one or more R 10a Substituted C1-C6 alkyl; Each R 10a Each independently is deuterium; Each R 13 Each independently represents a 3-7 membered cycloalkyl group, or is replaced by one or more R 13d Substituted 3-7 membered cycloalkyl; Each R 13d Each is independently a C1-C6 alkyl group.

9. The compound of formula I as claimed in claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 1 is methyl; (2)R 2 , R 3 , R 4 and R 6 are independently hydrogen, cyano, F, (3)R 2 is cyano; (4)R 3 for (5)R 4 is hydrogen; (6)R 6 is F; (7)R 5 for methyl, (8)R 7 is hydrogen; (9)R 8 Independently (10)R 9 For hydrogen.

10. The compound of formula I according to claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 2 , R 3 , R 4 and R 6 are independently hydrogen, cyano, F, (2)R 3 for (3)R 5 For hydrogen, methyl, 11. The compound of formula I according to claim 1, its tautomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I is any of the following compounds:

12. A pharmaceutical composition comprising the compound of formula I as described in any one of claims 1 to 11, its tautomer or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

13. Use of the compound of formula I as claimed in any one of claims 1 to 11, its tautomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as claimed in claim 12 in the preparation of a PRMT5 inhibitor, for example, in the preparation of a PRMT5·MTA inhibitor.

14. Use of the compound of formula I as described in any one of claims 1 to 11, its tautomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 12 in the preparation of a drug, for example, in the preparation of a drug for treating and / or preventing cancer.

15. The use according to claim 14, characterized in that The application meets one or more of the following conditions: (1) The cancer may be a MTAP-related / mediated cancer, preferably, the MTAP-related / mediated cancer is head and neck cancer, respiratory cancer, digestive cancer, urinary cancer, bone cancer, gynecological cancer, blood cancer, melanoma or skin cancer, more preferably lymphoma, lung cancer, breast cancer, colorectal cancer, colon cancer, rectal cancer, leukemia, glioblastoma, prostate cancer or ovarian cancer; (2) The compound of formula I, its tautomer or its pharmaceutically acceptable salt, or the pharmaceutical composition is in a therapeutically effective amount.

16. A compound as shown below, 17. A PRMT5-MEP50-MTA-Compound 2-P2 composite crystal, characterized in that: The compound 2-P2 is The X-ray data of the composite crystals are shown in the following table:

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