Substituted oxamide derivative as well as preparation method and application thereof

Abstract

The invention discloses a substituted oxamide derivative as shown in a formula I, a protein arginine methyltransferase 5 (PRMT5) inhibitor or a pharmaceutically acceptable salt thereof, a preparation method and application of the substituted oxamide derivative. # imgabs0 #

Description

A substituted oxamide derivative, its preparation method and use Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to a substituted oxamide derivative as a protein arginine methyltransferase 5 (PRMT5) inhibitor or a pharmaceutically acceptable salt thereof, a preparation method and use thereof. Background Art

[0002] Protein arginine methyltransferases (PRMTs) methylate a variety of proteins and play important roles in biological processes such as gene expression, splicing, and DNA repair. Currently, 11 members of the PRMT family have been discovered, with PRMT5 being a major research focus. PRMT5 participates in histone methylation, binding to SAM and transferring its methyl group to the guanidinium nitrogen atom of the arginine side chain in proteins, generating methylated arginine.

[0003] Methylthioadenosine phosphorylase (MTAP) is a key enzyme that catalyzes the synthesis of methionine in vivo. Loss of the MTAP gene hinders the downstream synthesis of methylated arginine, increasing reliance on the PRMT5 pathway for sufficient methylated arginine. Furthermore, loss of the MTAP gene leads to the accumulation of methylthioadenosine (MTA). This large amount of MTA competes with SAM, leading to reduced PRMT5 activity. Therefore, in MTAP-deficient cells, MTA-mediated small molecule inhibitors of PRMT5 can inhibit PRMT5 activity in the presence of elevated MTA concentrations.

[0004] Studies have found that MTAP gene deletion occurs in approximately 10% of all tumor types, including pancreatic cancer, lung cancer, bladder cancer, and many other tumor types. Downregulating PRMT5 expression or inhibiting its activity can significantly inhibit tumor cell growth.

[0005] Therefore, the development of an efficient and specific MTA-mediated PRMT5 small molecule inhibitor is of great significance for achieving precise treatment of tumors and reducing toxic side effects.

[0006] Summary of the Invention

[0007] The present application relates to a substituted oxamide derivative as an MTA-mediated PRMT5 small molecule inhibitor, as well as its preparation method and medical application, particularly a substituted oxamide derivative as shown in Formula I below and its use in preparing drugs for MTA-PRMT5-mediated diseases, more specifically, its use in preparing drugs suitable for tumors.

[0008] One object of the present application is to provide a compound having a structure shown in the following formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0009] in,

[0010] R 1 Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, carboxyl, hydroxyl, C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy, wherein the C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkyloxy;

[0011] R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 hydrocarbon group, C1-C6 hydrocarbon group oxy group, C3-C8 carbocyclyl group or C3-C8 carbocyclyl group oxy group, wherein C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 and R 3 Can form a 5-8 membered carbocyclic ring or a 5-8 membered heterocyclic ring;

[0012] R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclic group or 3-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino, wherein the C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic group, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino, may be further substituted with halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy or amino;

[0013] R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 aryl or 4-10 membered heterocyclic group, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 The aryl or 4-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy, C3-C 10 Cycloalkylamino, 3-10 membered heterocyclylamino,

[0014] X is selected from: a single bond, -CH2-, wherein -CH2- may be further substituted by 1 or 2 methyl groups or halogen;

[0015] n is selected from: 0 or 1.

[0016] In some embodiments, R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclic group or 3-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic group, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino,

[0017] In other embodiments, R 1 Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, carboxyl, hydroxyl, C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy, wherein the C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkyloxy;

[0018] R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 hydrocarbon group, C1-C6 hydrocarbon group oxy group, C3-C8 carbocyclyl group or C3-C8 carbocyclyl group oxy group, wherein C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 and R 3 Can form a 5-8 membered carbocyclic ring or a 5-8 membered heterocyclic ring;

[0019] R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclyl or 3-10 membered heterocyclyl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 haloalkyl, C1-C6 alkyl, C1-C6 alkyloxy or C1-C6 haloalkyloxy;

[0020] R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy or C1-C6 haloalkyloxy;

[0021] X is selected from: a single bond or -CH2-, wherein -CH2- may be further substituted by 1 or 2 methyl groups or halogen;

[0022] n is selected from: 0 or 1.

[0023] In some embodiments, R 1Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, carboxyl, hydroxyl, C1-C3 hydrocarbon, C1-C3 alkyloxy, C3-C6 cycloalkyl or C3-C6 cycloalkyloxy, wherein the C1-C3 hydrocarbon, C1-C3 alkyloxy, C3-C6 cycloalkyl or C3-C6 cycloalkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkyloxy;

[0024] Preferably, R 1 Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino;

[0025] More preferably, R 1 Selected from amino groups.

[0026] In other embodiments, R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 and R 3 Can form a 5-8 membered carbocyclic ring or a 5-8 membered heterocyclic ring;

[0027] More preferably, R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C3 alkyl or C1-C3 alkyloxy, wherein C1-C3 alkyl or C1-C3 alkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C3 alkyl or C1-C3 alkyloxy; R 2 and R 3 Can form a 5-6 membered carbocyclic ring or a 5-6 membered heterocyclic ring;

[0028] More preferably, R 2 、R 3 、R 4 Each is independently selected from: hydrogen, halogen or C1-C3 alkyl;

[0029] In some embodiments, R 5 Selected from: C3-C8 cycloalkyl, 3-10 membered heterocyclic group or C6-C 10 Aryl, wherein the C3-C8 cycloalkyl, 3-10 membered heterocyclic group or C6-C 10 The aryl group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, The C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, may be further substituted with halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy or amino;

[0030] In other embodiments, R 5 Further preferably selected from: C6-C 10 Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, The C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, It may be further substituted with 1, 2 or 3 F, Cl, Br or C1-C4 alkyl groups;

[0031] In other embodiments, R 5 Further preferably selected from: C6-C 10Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, and the C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl may be further substituted by 1, 2 or 3 F, Cl, Br, C1-C4 alkyl.

[0032] In some embodiments, R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-8 membered heterocycloalkyl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-8 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino,

[0033] In other embodiments, R 6 Preferred are: C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heterocyclic group, wherein the 5-10 membered heterocyclic group is selected from 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino,

[0034] In other embodiments, R 6 Further preferably selected from: C3-C6 cycloalkyl, C6-C 10 aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino,

[0035] In other embodiments, R 6 Further preferably selected from: C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from F, Cl, Br, cyano, C1-C3 alkyl, C1-C3 alkyloxy or C1-C3 haloalkyloxy.

[0036] In some embodiments, a compound having a structure shown in the following Formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided:

[0037] in,

[0038] R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein C1-C3 alkyl, C1-C3 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C3 alkyl or C1-C3 alkyloxy; R 2 and R 3 Can form a 5-6 membered carbocyclic ring or a 5-6 membered heterocyclic ring;

[0039] R 5 Selected from: C3-C8 cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C8 cycloalkyl, 3-10 membered heterocyclyl, C6-C 10The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, It may be further substituted with 1-3 F, Cl, Br or C1-C4 alkyl groups;

[0040] R 6 Selected from: C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heterocyclic group, wherein the 5-10 membered heterocyclic group is selected from 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino,

[0041] n is selected from: 0 or 1.

[0042] In some embodiments, R 2 、R 3 、R 4 Each is independently preferably selected from: hydrogen, halogen, cyano or C1-C3 alkyl.

[0043] Further, in some other embodiments, R 2 Selected from C1-C3 alkyl; R 3 、R 4 All are hydrogen;

[0044] More preferably, R 2 Selected from methyl or ethyl.

[0045] In some embodiments, R 5 Preferred from: C6-C10 Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, and the C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl may be further substituted by 1-3 F, Cl, Br or C1-C4 alkyl;

[0046] Furthermore, R 5 Selected from: The above substituents may be further substituted by one or more substituents independently selected from F, Cl, Br or C1-C4 alkyl.

[0047] In other embodiments, R 6 Further selected from: C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from F, Cl, Br, cyano, C1-C3 alkyl, C1-C3 alkyloxy or C1-C3 haloalkyloxy;

[0048] Furthermore, R 6 since The above substituents may be further substituted by one or more substituents independently selected from F, Cl, Br, C1-C3 alkyl, C1-C3 alkyloxy or C1-C3 haloalkyloxy.

[0049] In some specific embodiments, the compound of the structure shown in Formula I, its stereoisomers or pharmaceutically acceptable salts thereof, is selected from:

[0050] Another object of the present application is to provide a method for preparing the compound represented by the structure of the above formula I and its stereoisomers, the method comprising the following steps:

[0051] Among them, R 1 、R 2 、R 3 、R 4 、R 5 、R 6, X and n are as defined above for the compound of the structure shown in Formula I;

[0052] (1) condensing compound I-1 with compound I-2 to obtain I-3;

[0053] The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from dichloromethane, acetonitrile, N,N-dimethylformamide, and any combination thereof, preferably dichloromethane. The base can be selected from triethylamine, diisopropylethylamine, potassium carbonate, sodium carbonate, preferably triethylamine. The reaction is preferably carried out at a suitable temperature, preferably 0-50°C. The reaction is preferably carried out for a suitable time, for example, 0-4 hours.

[0054] (2) subjecting compound I-3 to a hydrolysis reaction to obtain I-4;

[0055] The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide, water, and any combination thereof, preferably aqueous tetrahydrofuran. The base can be selected from sodium hydroxide, potassium hydroxide, and lithium hydroxide, preferably lithium hydroxide. The reaction is preferably carried out at a suitable temperature, preferably 0-50°C. The reaction is preferably carried out for a suitable time, for example, 8-12 hours.

[0056] (3) condensing compound I-4 with compound I-5 to obtain a compound of formula I;

[0057] The reaction is preferably carried out in a suitable organic solvent. The organic solvent can be selected from dichloromethane, acetonitrile, tetrahydrofuran, N,N-dimethylformamide, and any combination thereof, preferably N,N-dimethylformamide. The reaction is preferably carried out in the presence of a suitable condensing agent. The condensing agent can be selected from HATU, HBTU, T3P, etc. The reaction is preferably carried out at a suitable temperature, preferably 0-70°C. The reaction is preferably carried out for a suitable time, for example, 2-8 hours.

[0058] The specific conditions of each of the above reaction steps are well known in the art and are not specifically limited in this application. Based on the teachings of this application and common knowledge in the art, those skilled in the art can select and replace the substituents in the general formula to prepare different compounds, and such selections and replacements are within the scope of protection of this application.

[0059] The present application also relates to the use of the compound of formula I in the preparation of drugs for preventing or treating diseases related to PRMT5.

[0060] In some embodiments, the PRMT5-related disease or condition is selected from a tumor or cancer, such as breast cancer, pancreatic cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, lymphoma, malignant sarcoma, cervical cancer, oral cancer, brain cancer, gastric cancer, liver cancer, skin cancer, bone cancer, kidney cancer, bladder cancer, fallopian tube tumor, mesothelioma, melanoma, glioma, glioblastoma, papillary malignant tumor, head and neck tumor, myeloma or leukemia.

[0061] This application reports a novel class of PRMT5 inhibitors having a structure as shown in Formula I, which have good activity. DETAILED DESCRIPTION

[0062] In order to make the purpose and technical scheme of this application clearer, the present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, the specific experimental methods not mentioned in the following examples are all carried out according to conventional experimental methods.

[0063] In this application, when a chemical name and a structural formula are inconsistent, the structural formula shall prevail unless the context indicates that the chemical name rather than the structural formula is correct.

[0064] Definition and Description

[0065] Unless otherwise specified, the terms used in this application have the following meanings: A specific term should not be considered as uncertain or unclear if it is not specifically defined, but should be understood according to its ordinary meaning in the art.

[0066] As used herein, unless otherwise indicated, "alkoxy" refers to an -O-alkyl group.

[0067] As used herein, unless otherwise indicated, "halogen" refers to fluorine, chlorine, bromine or iodine.

[0068] As used herein, unless otherwise indicated, a "cycloalkyloxy" group refers to an -O-cycloalkyl group.

[0069] Herein, unless otherwise specified, "substituted or unsubstituted amino group" encompasses unsubstituted amino group or amino group substituted with a group selected from C1-C6 alkyl group or C3-C8 cycloalkyl group.

[0070] As used herein, unless otherwise specified, "carbocyclyl," also referred to as "carbocycle," refers to a cyclic group having multiple carbon atoms and no ring heteroatoms, including single or multiple rings (fused, bridged, or spiro); including saturated cycloalkyl, unsaturated cycloalkyl, and aryl. An unsaturated cycloalkyl refers to a cyclic group with an unsaturated double bond; an aryl is typically a monocyclic, bicyclic, or tricyclic aromatic hydrocarbon group having multiple carbon atoms.

[0071] As used herein, unless otherwise indicated, "carbocyclyloxy" refers to -O-carbocyclyl.

[0072] As used herein, unless otherwise specified, "heterocyclyl", also referred to as "heterocycle", refers to a cyclic group containing at least one heteroatom, which may be a single ring or multiple rings (fused, bridged, spiro), and which may be a saturated heterocycloalkyl, cycloalkyl, unsaturated heterocycloalkyl, aryl or heteroaryl, wherein a heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom, and when multiple rings (fused, bridged, spiro) form a cyclic group, the heteroatom may be on any ring, or multiple rings may contain heteroatoms at the same time; the term "heterocycloalkyl" also includes a partially saturated cyclic group formed by the fusion of an aromatic ring containing at least one heteroatom and a non-aromatic ring, and the attachment site may be located at a non-aromatic carbon atom, an aromatic carbon atom or a heteroatom. Heteroaryl groups include, for example, pyridyl, indolyl, quinoxalinyl, quinolyl, isoquinolyl, benzothiophenyl, benzofuranyl, benzothiophenyl, benzopyranyl, benzothiapyranyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, oxadiazolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, and the like.

[0073] In this document, unless otherwise specified, the term “C m -C n " means that the part modified by the term has mn carbon atoms (n is greater than m, and both are integers). For example, C1-C6 means that the part modified by the term has 1-6 carbon atoms, such as 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms.

[0074] The abbreviations used in this document have the following meanings:

[0075] The structures of the compounds were determined by mass spectrometry (MS) or nuclear magnetic resonance (NMR). 1 H NMR).

[0076] Nuclear magnetic resonance (NMR) 1 H NMR) shifts (δ) are given in parts per million (ppm); nuclear magnetic resonance ( 1 H NMR) was measured using a Bruker AVANCE-400 nuclear magnetic spectrometer. The solvent was deuterated dimethyl sulfoxide (DMSO-d6), the internal standard was tetramethylsilane (TMS), and the chemical shift was 10 -6 The units are given in ppm.

[0077] Mass spectrometry (MS) was performed using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Therm, model: Finnigan LCQ advantage MAX).

[0078] Thin layer silica gel uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate.

[0079] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0080] Unless otherwise specified in this application, all reactions mentioned in the present invention were carried out under a nitrogen atmosphere.

[0081] The term "nitrogen atmosphere" in this application refers to, for example, connecting a reaction bottle to a 1 L nitrogen balloon.

[0082] The term "hydrogen atmosphere" in the present application refers to, for example, connecting the reaction bottle to a 1 L hydrogen balloon.

[0083] Unless otherwise specified in the present application, the solution mentioned in the reaction of the present invention is an aqueous solution.

[0084] The term "room temperature" in this application refers to a temperature between 10°C and 25°C.

[0085] Example 1 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((3-chloropyridin-2-yl)methyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (1)

[0086] Step 1: Preparation of compound 1b

[0087] Compound 1a (5 g, 32.7 mmol) was dissolved in dichloromethane (100 mL). 4-Dimethylaminopyridine (0.8 g, 6.5 mmol) and di-tert-butyl dicarbonate (15.7 g, 71.9 mmol) were added sequentially. The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 10 g of the title compound in an 86.7% yield.

[0088] LC-MS (ESI) m / z (M+H) + :354.2

[0089] Step 2: Preparation of compound 1c

[0090] Compound 1b (10 g, 28.3 mmol) was dissolved in methanol (300 mL), followed by the addition of 10% palladium on carbon (0.5 g). The reaction system was stirred at room temperature under a hydrogen atmosphere for 12 hours. The palladium on carbon was filtered off, and the filtrate was concentrated under reduced pressure to afford 8.8 g of the title compound in a 96.5% yield.

[0091] LC-MS (ESI) m / z (M+H) + :324.2

[0092] Step 3: Preparation of compound 1d

[0093] Compound 1c (8.8 g, 27.3 mmol) was dissolved in dichloromethane (150 mL). Ethyl oxalyl chloride (5.6 g, 41.0 mmol) and triethylamine (2.8 g, 27.3 mmol) were added sequentially. The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 10.5 g of the title compound in a yield of 90.8%.

[0094] LC-MS (ESI) m / z (M+H) + :424.2

[0095] Step 4: Preparation of compound 1e

[0096] Compound 1d (10.5 g, 24.8 mmol) was dissolved in 300 mL of a tetrahydrofuran-water (1:10) mixture. Lithium hydroxide (0.7 g, 27.3 mmol) was added, and the reaction system was stirred at room temperature for 12 hours. The reaction solution was adjusted to pH 6-7 with 2% dilute hydrochloric acid and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 9.2 g of the title compound in a yield of 94.3%.

[0097] LC-MS (ESI) m / z (M+H) + :396.2

[0098] Step 5: Preparation of compound 1h

[0099] Compound 1f (500 mg, 3.5 mmol) and compound 1g (625 mg, 3.5 mmol) were dissolved in methanol (30 mL). Glacial acetic acid (0.01 mL) was added dropwise, and the reaction system was stirred at room temperature for 0.5 hours. Sodium triacetoxyborohydride (2.3 g, 10.6 mmol) was then added, and the reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 420 mg of the title compound in a yield of 39.4%.

[0100] LC-MS (ESI) m / z (M+H) + :302.1

[0101] Step 6: Preparation of compound 1i

[0102] Compound 1h (100 mg, 0.3 mmol) and compound 1e (132 mg, 0.3 mmol) were dissolved in N,N-dimethylformamide (5 mL). 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (165 mg, 0.4 mmol) and triethylamine (68 mg, 0.7 mmol) were then added. The reaction system was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 32 mg of the title compound in a yield of 14.2%.

[0103] LC-MS (ESI) m / z (M+H) + :679.2

[0104] Step 7: N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((3-chloropyridin-2-yl)methyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (1)

[0105] Compound 1i (32 mg, 0.05 mmol) was dissolved in a 1:2 dichloromethane-trifluoroacetic acid mixture (6 mL). The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 21 mg of the title compound in a 93.5% yield.

[0106] LC-MS (ESI) m / z (M+H) + :479.1

[0107] 1 H NMR (400MHz, DMSO-d6) δ10.43(s,1H),8.90(s,1H),8.49(d,J=4.8,Hz,1H),8.19(d,J=8.0Hz,1H),7.97(s,1H),7.90(d,J=8.0 Hz,1H),7.66(d,J=8.3Hz,1H),7.48–7.40(m,1H),7.36(d,J=8.1Hz,1H),5.63(s,2H),5.16(s,2H),4.86(s,2H),1.99(s,3H).

[0108] Example 2 N 1 -(6-amino-5-methylpyridin-3-yl)-N2 -(3,3-difluorocyclobutyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (2)

[0109] Using compound 2a and compound 2b as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0110] LC-MS (ESI) m / z (M+H) + :444.2

[0111] 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.88(s,1H),8.18(d,J=9.2Hz,1H),8.01(s,1H),7.56(d,J=9. 2Hz,1H),7.45(s,1H),5.63(s,2H),4.90(s,2H),4.66–4.30(m,1H),2.02(s,3H),1.25–1.17(m,4H).

[0112] Example 3 N 1 -(6-amino-5-trifluoromethylpyridin-3-yl)-N 2 -((3-chloropyridin-2-yl)methyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (3)

[0113] Using compound 3a as the starting material, the preparation process was the same as in Example 1 to obtain the title compound.

[0114] LC-MS (ESI) m / z (M+H) + :533.1

[0115] 1 H NMR (400MHz, DMSO-d6) δ10.75(s,1H),8.90(s,1H),8.47(d,J=4.8Hz,1H),8.37(d,J=4.8Hz,1H),8.18(s,1H) ,7.96(s,1H),7.89(d,J=6.6Hz,1H),7.64(s,1H),7.39–7.33(m,1H),6.39(s,2H),5.19(s,2H),4.87(s,2H).

[0116] Example 4 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(cyclopropylmethyl)-N2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (4)

[0117] Using compound 4a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0118] LC-MS (ESI) m / z (M+H) + :408.2

[0119] 1 H NMR (400MHz, DMSO-d6) δ10.60(s,1H),8.94(s,1H),8.22(d,J=8.4Hz,1H),8.04(s,1H),7.62(d,J=8.4Hz,1H),7.53(s,1H ),5.73(s,2H),4.85(s,2H),3.30(d,J=7.0Hz,2H),2.04(s,3H),1.06–1.02(m,1H),0.45–0.32(m,2H),0.18–0.14(m,2H).

[0120] Example 5 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxycyclohexyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (5)

[0121] Compound 5a and compound 2a were used as starting materials and the preparation process was the same as in Example 1 to obtain the title compound, which was then purified by column chromatography to obtain compound 5-2. Compound 5-2 was then purified by SFC to obtain compounds 5-2-P1 (Rt = 3.789 min) and 5-2-P2 (Rt = 4.987 min).

[0122] 5-2-P2:

[0123] LC-MS (ESI) m / z (M+H) + :466.2

[0124] 1H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.91(s,1H),8.20(d,J=11.2Hz,1H),8. 06(s,1H),7.57–7.51(m,2H),5.65(s,2H),4.78(d,J=16.8Hz,1H),4.66(d,J=1 6.8Hz,1H),3.96–3.88(m,1H),3.24–3.19(m,1H),3.08(s,3H),2.04(s,3H),1 .65-1.59(m,2H),1.56-1.41(m,2H),1.36-1.25(m,2H),1.14(d,J=8.2Hz,2H).

[0125] Example 6 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(1-(pyrimidin-2-yl)ethyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (6)

[0126] Using compound 6a and compound 1g as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0127] LC-MS (ESI) m / z (M+H) + :460.2

[0128] 1 H NMR (400MHz, DMSO-d6) δ10.43(s,1H),8.78(d,J=12.2Hz,1H),8.74(s,1H),8.72(s,1H),8.10(d,J=8.4Hz,1H),7.94(s,1H),7.66(d,J= 8.4Hz,1H),7.50(s,1H),7.39–7.32(m,1H),5.69(t,J=7.0Hz,1H),5.61(s,2H),5.17–4.50(m,2H),1.97(s,3H),1.57(d,J=6.9Hz,3H).

[0129] Example 7 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(1-methyl-1H-pyrazol-4-yl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (7)

[0130] Using compound 7a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0131] LC-MS (ESI) m / z (M+H) + :434.2

[0132] 1 H NMR (400MHz, DMSO-d6) δ10.47(s,1H),9.01(s,1H),8.24(d,J=8.4Hz,1H),8.04(s,1H),7.78(s,1H), 7.64(d,J=8.4Hz,1H),7.56(s,1H),7.40(s,1H),5.65(s,2H),5.08(s,2H),3.71(s,3H),2.00(s,3H).

[0133] Example 8 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxycyclohexyl)-N 2 Preparation of -((5-bromopyridin-2-yl)methyl)oxalamide (8)

[0134] Compound 5a and compound 8a were used as starting materials and the preparation process was the same as in Example 1 to obtain the title compound, which was then purified by column chromatography to obtain compound 8-2. Compound 8-2 was then purified by SFC to obtain compounds 8-2-P1 (RT = 3.525 min) and 8-2-P2 (RT = 4.732 min).

[0135] 8-2-P2:

[0136] LC-MS (ESI) m / z (M+H) + :476.38

[0137] 1 H NMR (400MHz, DMSO-d6) δ10.28(s,1H),8.64(s,1H),8.03(d,J=12.2Hz,2H),7.52(s,1H),7.32(s,1H),5.60(d ,J=17.2Hz,2H),4.69(d,J=16.4Hz,1H),4.51(d,J=16.4Hz,1H),3.95–3.78(m,1H),3.18(td,J=10.2,4.4Hz, 1H),3.08(s,3H),2.04(s,3H),1.63–1.59(m,2H),1.52–1.35(m,2H),1.23–1.07(m,4H).

[0138] Example 9 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -cyclohexyl-N 2 Preparation of -((5-bromopyridin-2-yl)methyl)oxalamide (9)

[0139] Using compound 9a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0140] LC-MS (ESI) m / z (M+H) + :436.3

[0141] 1 H NMR (400MHz, DMSO-d6) δ10.50(s,1H),8.92(s,1H),8.20(dd,J=8.4,2.2Hz,1H),8.03(s,1H),7.53(d,J=8.4Hz,1H),7.50(d,J= 3.0Hz,1H),5.67(s,2H),4.71(s,2H),3.85–3.71(m,1H),2.06(s,3H),1.78–1.73(m,2H),1.60–1.49(m,4H),1.24–1.13(m,4H).

[0142] Example 10 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-fluoro-2,3-dihydrobenzofuran-4-yl)methyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (10)

[0143] Using compound 10a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0144] LC-MS (ESI) m / z (M+H) + :504.1

[0145] 1H NMR (400MHz, DMSO-d6) δ10.42(s,1H),8.88(s,1H),8.15(d,J=5.6Hz,1H),7.95(d,J=2.5Hz,1H),7.57–7.39(m,2H),6.88–6.81(m,1H),6 .68–6.60(m,1H),5.63(d,J=15.2Hz,2H),4.91(d,J=2.9Hz,2H),4.65(s,2H),4.53(t,J=8.8Hz,2H),3.18(t,J=8.8Hz,2H),1.99(s,3H).

[0146] Example 11 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(cyclopentylmethyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (11)

[0147] Using compound 11a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0148] LC-MS (ESI) m / z (M+H) + :436.3

[0149] 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H),8.92(s,1H),8.22(t,J=9.6Hz,1H),7.98(s,1H),7.58(d,J=8.4Hz,1H),7.51–7.38(m,1H),5.63 (s,2H),4.86(s,2H),3.47(d,J=7.6Hz,2H),2.28–2.16(m,1H),2.02(s,3H),1.62–1.58(m,2H),1.53–1.43(m,3H),1.24–1.18(m,3H).

[0150] Example 12 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -cyclopentyl-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (12)

[0151] Using compound 12a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0152] LC-MS (ESI) m / z (M+H) + :422.2

[0153] 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H),8.90(s,1H),8.23–8.17(m,1H),7.96(s,1H),7.54(s,1H),7.42(s, 1H),5.66(s,2H),4.77(s,2H),3.55–3.51(m,1H),2.01(s,3H),1.91–1.81(m,2H),1.65–1.47(m,6H).

[0154] Example 13 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxycyclohexyl)-N 2 Preparation of -((5-cyclopropylpyridin-2-yl)methyl)oxalamide (13)

[0155] Compound 8-2 (47.5 mg, 0.1 mmol) and compound 13a (25.8 mg, 0.3 mmol) were dissolved in 1,4-dioxane (20 mL). 1,1-Bis(diphenylphosphino)diphenylferric palladium chloride (7.3 mg, 0.01 mmol) and potassium carbonate (41.5 mg, 0.3 mmol) were added sequentially. Under nitrogen, the reaction system was stirred at 100°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 21 mg of the title compound in a yield of 47.9%.

[0156] LC-MS (ESI) m / z (M+H) + :438.2

[0157] 1 H NMR(400MHz,DMSO-d6)δ10.25(s,1H),8.31(s,1H),8.01(s,1H),7.49(s,1H), 7.41–7.35(m,1H),7.19(s,1H),5.59(s,2H),4.71(d,J=16.0Hz,1H),4.45(d, J=16.0Hz,1H),3.93–3.81(m,1H),3.22–3.15(m,1H),3.10(s,3H),2.01(s,3H) ),1.97–1.88(m,1H),1.61–1.42(m,4H),1.16–0.89(m,6H),0.77–0.64(m,2H).

[0158] Example 14 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(5,6,7,8-tetrahydroquinolin-8-yl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (12)

[0159] Using compound 14a and compound 2a as starting materials, the preparation process was the same as in Example 1 to obtain the title compound.

[0160] LC-MS (ESI) m / z (M+H) + :485.2

[0161] 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.81(s,1H),8.40(s,1H),8.16(s,1H),8.00(s,1H),7.67–7.48(m,2H),7.35–7.20(m,2 H),5.64(s,2H),5.50–5.35(m,1H),5.12–4.68(m,2H),2.84–2.69(m,2H),2.00(s,3H),1.98–1.91(m,2H),1.81–1.70(m,2H).

[0162] Example 15 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (15)

[0163] Step 1: Preparation of compound 15b

[0164] Compound 15a (500 mg, 4.1 mmol) and compound 2a (711 mg, 4.1 mmol) were dissolved in methanol (30 mL). Glacial acetic acid (0.01 mL) was added dropwise, and the reaction system was stirred at room temperature for 0.5 hours. Sodium triacetoxyborohydride (2.6 g, 12.3 mmol) was then added, and the reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 740 mg of the title compound in a yield of 64.0%.

[0165] LC-MS (ESI) m / z (M+H) + :283.2

[0166] Step 2: Preparation of compound 15c

[0167] Compound 15b (740 mg, 2.6 mmol) was dissolved in dichloromethane (50 mL). Ethyl oxalyl chloride (425 mg, 3.1 mmol) and triethylamine (316 mg, 2.6 mmol) were added sequentially. The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 934 mg of the title compound in a 93.2% yield.

[0168] LC-MS (ESI) m / z (M+H) + :383.2

[0169] Step 3: Preparation of compound 15d

[0170] Compound 15c (934 mg, 2.4 mmol) was dissolved in a tetrahydrofuran-water (1:10) mixture (50 mL). Lithium hydroxide (65 mg, 2.7 mmol) was added, and the reaction system was stirred at room temperature for 12 hours. The reaction solution was adjusted to pH 6-7 with 2% dilute hydrochloric acid and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 848 mg of the title compound in a yield of 91.3%.

[0171] LC-MS (ESI) m / z (M+H) + :355.1

[0172] Step 4: Preparation of compound 15e

[0173] Compound 15d (100 mg, 0.3 mmol) and compound 1c (92 mg, 0.3 mmol) were dissolved in N,N-dimethylformamide (5 mL). 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (152 mg, 0.4 mmol) and triethylamine (70 mg, 0.7 mmol) were then added. The reaction system was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 69 mg of the title compound in a 34.7% yield.

[0174] LC-MS (ESI) m / z (M+H) + :660.3

[0175] Step 5: N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (15)

[0176] Compound 15e (69 mg, 0.1 mmol) was dissolved in a 1:2 mixture of dichloromethane and trifluoroacetic acid (6 mL). The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 46 mg of the title compound in a 90.7% yield.

[0177] LC-MS (ESI) m / z (M+H) + :460.2

[0178] 1 H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.88(s,1H),8.22(dd,J=8.4,2.4Hz,1H),7.83(d,J=2.5Hz,1H),7.71(d,J=8.4Hz,1H),7.34–7.2 1(m,3H),7.06–7.01(m,1H),6.91–6.82(m,1H),5.57(s,2H),5.26(d,J=16.4Hz,1H),4.80(d,J=16.4Hz,1H),3.74(s,3H),1.96(s,3H).

[0179] Example 16 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((1R,2R)-2-(trifluoromethoxy)cyclohexyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (16)

[0180] Step 1: Preparation of compound 16b

[0181] Compound 16a (1 g, 8.7 mmol) was dissolved in dichloromethane (100 mL). 4-Dimethylaminopyridine (212 mg, 1.7 mmol) and di-tert-butyl dicarbonate (5.7 g, 26.0 mmol) were added sequentially. The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 2.3 g of the title compound in a yield of 83.2%.

[0182] LC-MS (ESI) m / z (M+H) + :316.2

[0183] Step 2: Preparation of compound 16c

[0184] To compound 16b (2.3 g, 7.3 mmol) in ethyl acetate (100 mL) were added trifluoromethyltrimethylsilane (3.1 g, 21.9 mmol), silver trifluoromethanesulfonate (3.7 g, 14.6 mmol), 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane bis(tetrafluoroborate) salt (3.9 g, 10.9 mmol), potassium fluoride (1.3 g, 21.9 mmol), and 2-fluoropyridine (2.1 g, 21.9 mmol). Under nitrogen protection, the reaction system was stirred at room temperature for 24 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography to obtain 1.9 g of the title compound in a yield of 67.8%.

[0185] LC-MS (ESI) m / z (M+H) + :384.2

[0186] Step 3: Preparation of compound 16d

[0187] Compound 16c (1.9 g, 4.9 mmol) was dissolved in a 1:2 mixture of dichloromethane and trifluoroacetic acid (6 mL). The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 849 mg of the title compound in a 93.7% yield.

[0188] LC-MS (ESI) m / z (M+H) + :184.1

[0189] Step 4: Preparation of compound 16e

[0190] Compound 16d (849 mg, 4.6 mmol) and compound 2a (811 mg, 4.6 mmol) were dissolved in methanol (30 mL). Glacial acetic acid (0.01 mL) was added dropwise, and the reaction system was stirred at room temperature for 0.5 hours. Sodium triacetoxyborohydride (2.9 g, 13.9 mmol) was then added, and the reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 600 mg of the title compound in a yield of 38.1%.

[0191] LC-MS (ESI) m / z (M+H) + :343.1

[0192] Step 5: Preparation of compound 16f

[0193] Compound 16e (600 mg, 1.8 mmol) was dissolved in dichloromethane (50 mL). Ethyl oxalyl chloride (286 mg, 2.1 mmol) and triethylamine (213 mg, 2.1 mmol) were added sequentially. The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 732 mg of the title compound in a 94.2% yield.

[0194] LC-MS (ESI) m / z (M+H) + :443.1

[0195] Step 6: Preparation of compound 16g

[0196] Compound 16f (732 mg, 1.7 mmol) was dissolved in a tetrahydrofuran-water (1:10) mixture (50 mL). Lithium hydroxide (44 mg, 1.8 mmol) was added, and the reaction system was stirred at room temperature for 12 hours. The reaction solution was adjusted to pH 6-7 with 2% dilute hydrochloric acid and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 630 mg of the title compound in a yield of 92.1%.

[0197] LC-MS (ESI) m / z (M+H) + :415.1

[0198] Step 7: Preparation of compound 16h

[0199] Compound 15g (125 mg, 0.3 mmol) and compound 1c (92 mg, 0.3 mmol) were dissolved in N,N-dimethylformamide (5 mL). 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (152 mg, 0.4 mmol) and triethylamine (70 mg, 0.7 mmol) were then added sequentially. The reaction system was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 73 mg of the title compound in a yield of 33.8%.

[0200] LC-MS (ESI) m / z (M+H) + :720.3

[0201] Step 8: N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((1R,2R)-2-(trifluoromethoxy)cyclohexyl)-N 2 Preparation of -((5-(trifluoromethyl)pyridin-2-yl)methyl)oxalamide (16)

[0202] Compound 16h (73 mg, 0.1 mmol) was dissolved in a 1:2 mixture of dichloromethane and trifluoroacetic acid (6 mL). The reaction system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain 49 mg of the title compound in a 93.2% yield.

[0203] LC-MS (ESI) m / z (M+H) + :520.2

[0204] 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),8.93(s,1H),8.19(dd,J=8.4,2.4Hz,1H),8.07(d,J=2.4Hz,1H),7.56–7.52(m,1H),7.48(d,J=8.4Hz,1H), 5.66(s,2H),4.80(t,J=17.6Hz,2H),4.73–4.65(m,1H),4.56–4.46(m,1H ),2.04(s,3H),1.67–1.58(m,4H),1.55–1.44(m,2H),1.35–1.27(m,2H).

[0205] Example 17 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-bromo-pyridin-2-yl)methyl)oxalamide (17)

[0206] Using compound 15a and compound 17a as starting materials, the preparation process was the same as in Example 15 to obtain the title compound.

[0207] LC-MS (ESI) m / z (M+H) + :470.1

[0208] 1H NMR(400MHz,DMSO-d6)δ10.32(s,1H),8.60(d,J=2.4Hz,1H),8.07–8.03(m,1 H),7.82(d,J=2.4Hz,1H),7.44(d,J=8.4Hz,1H),7.28(d,J=2.4Hz,1H),7.25 –7.18(m,2H),7.02(d,J=8.4Hz,1H),6.84(td,J=7.6,1.2Hz,1H),5.57(s,2H ),5.19(d,J=15.6Hz,1H),4.64(d,J=15.6Hz,1H),3.74(s,3H),1.96(s,3H).

[0209] Example 18 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-cyclopropyl-pyridin-2-yl)methyl)oxalamide (18)

[0210] Using compound 17 and compound 13a as starting materials, the preparation process was the same as Example 13 to obtain the title compound.

[0211] LC-MS (ESI) m / z (M+H) + :432.2

[0212] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.27(s,1H),7.86(s,1H),7.38(dd,J=8.4,2.4Hz,1H),7.3 1(d,J=8.4Hz,1H),7.28(d,J=2.4Hz,1H),7.21(t,J=8.0Hz,1H),7.15(dd,J=7.6,1.6Hz,1H),7.01 (d,J=8.0Hz,1H),6.81(t,J=7.6Hz,1H),5.56(s,2H),5.21(d,J=15.6Hz,1H),4.56(d,J=15.6Hz, 1H),3.74(s,3H),1.96(s,3H),1.91(td,J=8.4,4.4Hz,1H),0.99–0.92(m,2H),0.72–0.68(m,2H).

[0213] Example 19 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-isopropoxyphenyl)-N2 Preparation of -((5-trifluoromethyl-pyridin-2-yl)methyl)oxalamide (19)

[0214] Using compound 19a and compound 2a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0215] LC-MS (ESI) m / z (M+H) + :488.2

[0216] 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.87(s,1H),8.23(d,J=9.0Hz,1H),7.89(d,J=2.5Hz ,1H),7.73(d,J=8.2Hz,1H),7.36(d,J=2.5Hz,1H),7.31(dd,J=7.8,1.7Hz,1H),7.19(t,J=7 .8Hz,1H),6.99(d,J=8.3Hz,1H),6.84(t,J=7.6Hz,1H),5.56(s,2H),5.11(d,J=16.1Hz,1H ), 4.96 (d, J = 16.1Hz, 1H), 4.59 (p, J = 5.9Hz, 1H), 1.97 (s, 3H), 1.15 (dd, J = 17.1, 6.0Hz, 6H).

[0217] Example 20 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-cyclopentyl-pyridin-2-yl)methyl)oxalamide (20)

[0218] Step 1: Using compound 17 and compound 20a as starting materials, the preparation process was the same as Example 13 to obtain the title compound 20b.

[0219] LC-MS (ESI) m / z (M+H) + :458.2

[0220] 1H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.35(d,J=2.4Hz,1H),7.82(d,J=2.4Hz,1H),7.64(dd,J=8.0, 2.4Hz,1H),7.37(d,J=8.0Hz,1H),7.28(d,J=2.4Hz,1H),7.21(t,J=8.4Hz,2H),7.05–6.97(m,1H),6 .92–6.77(m,1H),5.56(s,2H),5.21(d,J=15.6Hz,1H),4.58(d,J=15.6Hz,1H),3.74(s,3H),3.01–2. 92(m,1H),2.02–1.99(m,2H),1.96(s,3H),1.80–1.72(m,2H),1.69–1.59(m,2H),1.56–1.46(m,2H).

[0221] Step 2: Compound 20b was used as the starting material and the title compound 20 was obtained by palladium-carbon hydrogenation.

[0222] LC-MS (ESI) m / z (M+H) + :460.2

[0223] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.35(d,J=2.4Hz,1H),7.82(d,J=2.4Hz,1H),7.64(dd,J=8.0,2.4H z,1H),7.37(d,J=8.0Hz,1H),7.28(d,J=2.4Hz,1H),7.21(t,J=8.4Hz,2H),7.05–6.97(m,1H),6.92–6.77 (m,1H),5.56(s,2H),5.21(d,J=15.6Hz,1H),4.58(d,J=15.6Hz,1H),3.74(s,3H),3.05–2.86(m,1H),2.0 2(d,J=6.8Hz,2H),1.96(s,3H),1.76(d,J=7.6Hz,2H),1.65(dd,J=7.6,4.7Hz,2H),1.52(d,J=9.8Hz,2H).

[0224] Example 21 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((6-(3,3-difluoropyrrolidin-1-yl)-pyridin-3-yl)methyl)oxalamide (21)

[0225] Using compound 21a and compound 15a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0226] LC-MS (ESI) m / z (M+H) + :497.2

[0227] 1 H NMR (400MHz, DMSO-d6) δ10.24(s,1H),7.91(d,J=2.4Hz,1H),7.79(d,J=2.4Hz,1H),7.40(dd,J=8.6 ,2.4Hz,1H),7.24(dd,J=5.0,2.0Hz,1H),7.23–7.18(m,1H),7.00(td,J=8.0,1.6Hz,2H),6.81(td, J=7.6,1.6Hz,1H),6.47(d,J=8.6Hz,1H),5.54(s,2H),5.03(d,J=14.4Hz,1H),4.44(d,J=14.4Hz,1 H),3.78(t,J=13.4Hz,2H),3.72(s,3H),3.56(t,J=7.2Hz,2H),1.95(s,3H),1.25(d,J=9.4Hz,2H).

[0228] Example 22 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxy-5-methyl-phenyl)-N 2 Preparation of -((5-trifluoromethyl-pyridin-2-yl)methyl)oxalamide (22)

[0229] Using compound 22a and compound 2a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0230] LC-MS (ESI) m / z (M+H) + :474.2

[0231] 1H NMR(400MHz,DMSO-d6)δ10.31(s,1H),8.89(s,1H),8.23(dd,J=8.4,2.4Hz,1H ),7.83(d,J=2.4Hz,1H),7.71(d,J=8.4Hz,1H),7.29(d,J=1.8Hz,1H),7.13(d, J=2.2Hz,1H),7.06–7.01(m,1H),6.91(d,J=8.4Hz,1H),5.58(s,2H),5.19(d, J=15.6Hz,1H),4.84(d,J=15.6Hz,1H),3.68(s,3H),2.14(s,3H),1.97(s,3H).

[0232] Example 23 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-Methoxy-5-chloro-phenyl)-N 2 Preparation of -((5-trifluoromethyl-pyridin-2-yl)methyl)oxalamide (23)

[0233] Using compound 23a and compound 2a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0234] LC-MS (ESI) m / z (M+H) + :494.1

[0235] 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.90(s,1H),8.23(d,J=8.4Hz,1H),7.85(d,J=2.0Hz,1H),7.70(d,J=8.4Hz,1H),7.45(d,J=2.8H z,1H),7.32–7.28(m,2H),7.06(d,J=8.6Hz,1H),5.60(s,2H),5.20(d,J=15.0Hz,1H),4.88(d,J=15.0Hz,1H),3.71(s,3H),1.98(s,3H).

[0236] Example 24 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-Methoxy-4-chloro-phenyl)-N 2 Preparation of -((5-trifluoromethyl-pyridin-2-yl)methyl)oxalamide (24)

[0237] Using compound 24a and compound 2a as starting materials, the preparation process was the same as in Example 15 to obtain the title compound.

[0238] LC-MS (ESI) m / z (M+H) + :494.1

[0239] 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),8.89(s,1H),8.23(dd,J=8.4,2.4Hz,1 H),7.86(d,J=2.4Hz,1H),7.69(d,J=8.4Hz,1H),7.32(d,J=2.4Hz,1H),7.29( d,J=8.4Hz,1H),7.14(d,J=2.4Hz,1H),6.96(dd,J=8.4,2.3Hz,1H),5.60(s,2 H),5.24(d,J=16.0Hz,1H),4.79(d,J=16.0Hz,1H),3.76(s,3H),1.98(s,3H).

[0240] Example 25 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-Chloro-phenyl)-N 2 Preparation of -((5-trifluoromethyl-pyridin-2-yl)methyl)oxalamide (25)

[0241] Using compound 25a and compound 2a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0242] LC-MS (ESI) m / z (M+H) + :464.2

[0243] 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),8.91(d,J=1.1Hz,1H),8.23(dd,J=8.4,2.2Hz,1H),7.84(d,J=2.5Hz ,1H),7.73(d,J=8.4Hz,1H),7.53(dd,J=7.6,1.8Hz,1H),7.44(dd,J=7.6,2.0Hz,1H),7.34(dd,J=7.5,2.0 Hz,1H),7.30(dd,J=7.2,2.0Hz,2H),5.60(s,2H),5.42(d,J=15.6Hz,1H),4.70(d,J=15.6Hz,1H),1.96(s,3H).

[0244] Example 26 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -(pyridin-2-ylmethyl)oxamide (26)

[0245] Using compound 26a and compound 15a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0246] LC-MS (ESI) m / z (M+H) + :392.2

[0247] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.46(d,J=4.8Hz,1H),7.83(d,J=2.5Hz,1 H),7.77(td,J=7.8,1.8Hz,1H),7.46(d,J=7.8Hz,1H),7.29–7.27(m,1H),7.27– 7.23(m,1H),7.22–7.17(m,2H),7.02(d,J=8.0Hz,1H),6.85–6.79(m,1H),5.56( s,2H),5.26(d,J=15.6Hz,1H),4.62(d,J=15.6Hz,1H),3.75(s,3H),1.96(s,3H).

[0248] Example 27 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-(cyclopent-1-en-1-yl)pyridin-2-yl)methyl)oxalamide (27)

[0249] Using compound 17 and compound 20a as starting materials, the preparation process was the same as Example 20 to obtain the title compound.

[0250] LC-MS (ESI) m / z (M+H) + :458.2

[0251] 1H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.55(d,J=1.8Hz,1H),7.85–7.79(m,2H),7.41(d,J= 8.4Hz,1H),7.28(d,J=3.0Hz,1H),7.24–7.19(m,1H),7.17(dd,J=7.8,1.8Hz,1H),7.02(dd, J=8.4,1.3Hz,1H),6.82(td,J=7.8,1.2Hz,1H),6.44–6.36(m,1H),5.56(s,2H),5.25(d,J= 15.4Hz,1H),4.60(d,J=15.4Hz,1H),3.75(s,3H),2.66(t,J=6.2Hz,2H),2.01–1.92(m,7H).

[0252] Example 28 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((quinolin-2-yl)methyl)oxalamide (28)

[0253] Using compound 28a and compound 15a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0254] LC-MS (ESI) m / z (M+H) + :442.2

[0255] 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),8.37(d,J=8.4Hz,1H),7.95(d,J=8.6Hz,1H),7.92(d,J=8.2Hz,1H),7.84 (d,J=2.4Hz,1H),7.73(ddd,J=8.4,6.9,1.5Hz,1H),7.64(d,J=8.6Hz,1H),7.58(ddd,J=8.2,6.8,1.2Hz,1H),7 .29(d,J=1.7Hz,1H),7.27(dd,J=7.6,1.7Hz,1H),7.20(td,J=8.0,1.7Hz,1H),7.02(dd,J=8.4,1.3Hz,1H),6.8 1(td,J=7.6,1.3Hz,1H),5.57(s,2H),5.44(d,J=15.6Hz,1H),4.84(d,J=15.6Hz,1H),3.76(s,3H),1.97(s,3H).

[0256] Example 29 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((Benzothiazol-5-yl)methyl)oxalamide (29)

[0257] Using compound 29a and compound 15a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0258] LC-MS (ESI) m / z (M+H) + :448.1

[0259] 1 H NMR(400MHz,DMSO-d6)δ10.90(s,1H),9.38(s,1H),8.09(d,J=8.4Hz,1H),8.0 2(s,1H),7.97(s,1H),7.73(s,1H),7.37(d,J=8.4Hz,1H),7.21(t,J=7.8Hz,1H ),7.11(d,J=7.8Hz,1H),7.00(d,J=8.2Hz,1H),6.82(t,J=7.6Hz,1H),5.61(s, 2H), 5.29 (d, J = 14.8Hz, 1H), 4.86 (d, J = 14.8Hz, 1H), 3.74 (s, 3H), 2.12 (s, 3H).

[0260] Example 30 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((6-fluoropyridin-3-yl)methyl)oxalamide (30)

[0261] Using compound 21a and compound 15a as starting materials, the preparation process was the same as Example 15 to obtain the title compound.

[0262] LC-MS (ESI) m / z (M+H) + :410.2

[0263] 1H NMR (400MHz, DMSO-d6) δ10.28(s,1H),8.08(d,J=2.4Hz,1H),7.83(td,J=8.4,2.4Hz,1H),7.79(d,J=2.4Hz,1H),7.27–7.21(m,2H),7.16–7.09(m,2 H),7.01(dd,J=8.4,1.2Hz,1H),6.86(td,J=7.6,1.2Hz,1H),5.56(s,2H), 5.04(d,J=15.0Hz,1H),4.74(d,J=15.0Hz,1H),3.69(s,3H),1.96(s,3H).

[0264] Example 31 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-cyclopropylpyridin-2-yl)methyl)-N 2 Preparation of -(5-fluoro-2-methoxyphenyl)oxamide (31)

[0265] Using compound 31a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0266] LC-MS (ESI) m / z (M+H) + :450.2

[0267] 1 H NMR (400MHz, DMSO-d6) δ10.34 (s, 1H), 8.30 (d, J = 2.4Hz, 1H), 7.86 (s, 1H) ,7.43–7.36(m,1H),7.35–7.27(m,2H),7.18–7.13(m,1H),7.10–6.97(m,2 H),5.58(s,2H),5.14(d,J=15.4Hz,1H),4.67(d,J=15.4Hz,1H),3.69(s,3 H),1.97(s,3H),1.94–1.88(m,1H),1.00–0.95(m,2H),0.72–0.68(m,2H).

[0268] Example 32 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(Isoquinolin-3-ylmethyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (32)

[0269] Using compound 32a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0270] LC-MS (ESI) m / z (M+H) + :442.2

[0271] 1 H NMR (400MHz, DMSO-d6) δ10.35(s,1H),9.24(d,J=1.1Hz,1H),8.10(d,J=8.4Hz,1H),7.92(d ,J=8.4Hz,1H),7.87–7.82(m,2H),7.81–7.76(m,1H),7.65(ddd,J=8.2,6.8,1.2Hz,1H),7.3 0(d,J=2.4Hz,1H),7.26–7.17(m,2H),7.02(dd,J=8.2,1.2Hz,1H),6.80(td,J=7.6,1.2Hz, 1H),5.56(s,2H),5.42(d,J=15.6Hz,1H),4.80(d,J=15.6Hz,1H),3.78(s,3H),1.97(s,3H).

[0272] Example 33 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((2',6'-dimethyl-[3,4'-bipyridyl]-6-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (33)

[0273] Using compound 17 as the starting material, the preparation process was the same as in Example 20 to obtain the title compound.

[0274] LC-MS (ESI) m / z (M+H) + :497.2

[0275] 1H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.86(d,J=2.4Hz,1H),8.16(dd,J=8.2,2.4 Hz,1H),7.84(d,J=2.4Hz,1H),7.58(d,J=8.2Hz,1H),7.43(s,2H),7.29(d,J=2.4H z,1H),7.26–7.21(m,2H),7.05–6.98(m,1H),6.88–6.81(m,1H),5.56(s,2H),5.28 (d,J=15.6Hz,1H),4.73(d,J=15.6Hz,1H),3.76(s,3H),2.48(s,6H),1.97(s,3H).

[0276] Example 34 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-(3,6-dihydro-2H-pyran-4-yl)pyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (34)

[0277] Using compound 17 as the starting material, the preparation process was the same as in Example 20 to obtain the title compound.

[0278] LC-MS (ESI) m / z (M+H) + :474.2

[0279] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.56(d,J=2.4Hz,1H),7.87–7.80(m,2H),7.43(d ,J=8.4Hz,1H),7.28(d,J=2.4Hz,1H),7.23–7.18(m,2H),7.02(d,J=8.4Hz,1H),6.83(t, J=7.6Hz,1H),6.36(s,1H),5.56(s,2H),5.25(d,J=15.6Hz,1H),4.62(d,J=15.6Hz,1H) ,4.22(d,J=2.9Hz,2H),3.85–3.81(m,2H),3.75(s,3H),2.46–2.43(m,2H),1.96(s,3H).

[0280] Example 35 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2Preparation of -((5-(1-methyl-1H-pyrazol-5-yl)pyridin-2-yl)methyl)oxalamide (35)

[0281] Using compound 17 as the starting material, the preparation process was the same as in Example 20 to obtain the title compound.

[0282] LC-MS (ESI) m / z (M+H) + :472.2

[0283] 1 H NMR (400MHz, DMSO-d6) δ10.25(s,1H),7.97(d,J=2.4Hz,1H),7.79(d,J=2.4Hz,1 H),7.55(dd,J=8.4,2.4Hz,1H),7.27–7.16(m,3H),7.07–7.00(m,2H),6.87(d,J =17.4Hz,1H),6.82(d,J=7.6Hz,1H),6.75(d,J=8.4Hz,1H),5.55(s,2H),5.03(d ,J=14.8Hz,1H),4.57(d,J=14.8Hz,1H),3.80(s,3H),3.70(s,3H),1.95(s,3H).

[0284] Example 36 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-methylpyrazin-2-yl)methyl)-N 2 Preparation of 2-naphthalene-2-yl)oxamide (36)

[0285] Using compound 36a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0286] LC-MS (ESI) m / z (M+H) + :427.2

[0287] 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),8.60(s,1H),8.46(s,1H),7.92–7.85(m,3H),7.82–7.78(m,1H),7.71(d ,J=2.4Hz,1H),7.55–7.49(m,3H),7.15(d,J=2.4Hz,1H),5.56(s,2H),5.20(s,2H),2.45(s,3H),1.90(s,3H).

[0288] Example 37 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-(4-fluorophenyl)pyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (37)

[0289] Compound 17 was used as the starting material to obtain the title compound through Suziki coupling reaction.

[0290] LC-MS (ESI) m / z (M+H) + :486.3

[0291] 1 H NMR(400MHz, DMSO-d6)δ10.35(s,1H),8.77(s,1H),8.07(d,J=8.2Hz,1H), 7.84(s,1H),7.80–7.74(m,2H),7.55(d,J=8.2Hz,1H),7.36–7.28(m,3H),7 .27–7.19(m,2H),7.04(d,J=8.2Hz,1H),6.85(t,J=7.6Hz,1H),5.57(s,2H) ,5.28(d,J=15.4Hz,1H),4.69(d,J=15.4Hz,1H),3.76(s,3H),1.97(s,3H).

[0292] Example 38 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(4-bromo-2-fluorobenzyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (38)

[0293] Using compound 38a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0294] LC-MS (ESI) m / z (M+H) + :487.1

[0295] 1H NMR (400MHz, DMSO-d6) δ10.28(s,1H),7.79(d,J=2.5Hz,1H),7.46(dd,J=9.6,1.7Hz,1H),7.40–7.33(m,2H),7.26–7.20(m,2H),7.09(dd,J=7.8,1.7Hz ,1H),7.01(dd,J=8.3,1.3Hz,1H),6.83(td,J=7.6,1.3Hz,1H),5.56(s,2H) ,5.07(d,J=14.9Hz,1H),4.67(d,J=15.0Hz,1H),3.72(s,3H),1.95(s,3H).

[0296] Example 39 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((4-bromothien-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (39)

[0297] Using compound 39a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0298] LC-MS (ESI) m / z (M+H) + :475.0

[0299] 1 H NMR (400MHz, DMSO-d6) δ10.29(s,1H),7.81(d,J=2.6Hz,1H),7.55(d,J=1.5Hz,1H),7.29–7.23(m,2H),7.11(dd,J=7.7,1.7Hz,1H),7.07–7.02(m ,1H),6.93(d,J=1.5Hz,1H),6.88(td,J=7.6,1.3Hz,1H),5.56(s,2H),5 .19(d,J=15.3Hz,1H),4.76(d,J=15.3Hz,1H),3.72(s,3H),1.95(s,3H).

[0300] Example 40 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methyl-2H-indazol-4-yl)-N 2 Preparation of -((4-methylthiazol-5-yl)methyl)oxamide (40)

[0301] LC-MS (ESI) m / z (M+H)+ :436.2

[0302] 1 H NMR(400MHz,Chloroform-d)δ8.66(s,1H),7.88(d,J=2.6Hz,1H),7.71(d,J=8.8Hz,1H),7.63(s,1H),7.59(d,J=2.0Hz,1H),7.20–7. 16(m,1H),6.59(d,J=7.0Hz,1H),5.26(d,J=14.7Hz,1H),4.99(d,J=14.3Hz,1H),4.34(s,2H),4.18(s,3H),2.02(s,3H),1.88(s,3H).

[0303] Example 41 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(1-(5-cyclopropylpyridin-2-yl)ethyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (41)

[0304] Using compound 41a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0305] LC-MS (ESI) m / z (M+H) + :446.2

[0306] 1 H NMR(400MHz,DMSO-d6)δ10.09(s,1H),8.37(s,1H),7.77(s,1H),7.44–7.41(m ,1H),7.37–7.35(m,1H),7.33–7.30(m,1H),7.23–7.21(m,2H),7.02–7.00(m, 1H),6.79(t,J=7.8Hz,1H),5.75(q,J=7.2Hz,1H),5.53(s,2H),3.74(s,3H),1 .96–1.88(m,4H),1.27(d,J=7.2Hz,3H),1.02–0.97(m,2H),0.78–0.72(m,2H).

[0307] Example 42 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((6-cyclopropylpyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (42)

[0308] Using compound 42a as the starting material, the preparation process was the same as in Example 15 to obtain intermediate 42c, which was then subjected to Suziki coupling and further followed by reference to Example 15 to obtain the title compound.

[0309] LC-MS (ESI) m / z (M+H) + :432.2

[0310] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),7.83(d,J=2.6Hz,1H),7.60(t,J=7.8Hz,1H) ,7.29(s,1H),7.21(td,J=7.2,5.6Hz,2H),7.13(d,J=7.8Hz,1H),7.03–6.99(m,1H ),6.90(s,1H),6.86–6.82(m,1H),5.55(s,2H),5.14(d,J=15.6Hz,1H),4.52(d,J= 15.6Hz,2H),3.74(s,3H),2.03–1.96(m,4H),1.04–0.99(m,2H),0.80–0.74(m,2H).

[0311] Example 43 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((4-cyclopropylpyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (43)

[0312] Using compound 43a as the starting material, the preparation process was the same as in Example 15 to obtain intermediate 43c, which was then subjected to Suziki coupling and further followed by reference to Example 15 to obtain the title compound.

[0313] LC-MS (ESI) m / z (M+H) + :432.2

[0314] 1H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.25(d,J=5.1Hz,1H),7.85(s,1H),7.31(s, 1H),7.24–7.18(m,2H),7.11(s,1H),7.01(d,J=8.0Hz,1H),6.93(d,J=5.2Hz,1H),6 .85(t,J=7.6Hz,1H),,5.56(s,2H),5.14(d,J=15.6Hz,1H), 4.68(d,J=15.6Hz,1H), 3.74(s,3H),1.97(s,3H),1.93–1.88(m,1H),1.08–1.04(m,2H),0.77–0.69(m,2H).

[0315] Example 44 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-cyclopropyl-3-methylpyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (44)

[0316] Using compound a as the starting material, the preparation process was the same as in Example 15 to obtain the title compound.

[0317] LC-MS (ESI) m / z (M+H) + :446.21

[0318] 1 H NMR(400MHz,DMSO-d6)δ10.17(s,1H),8.09(s,1H),7.78(s,1H),7.24(s,1H),7.18(t ,J=7.2Hz,1H),7.14(s,1H),7.06(d,J=7.6Hz,1H),6.97(d,J=7.6Hz,1H),6.75(t,J= 7.2Hz,1H),5.53(s,2H),5.32(d,J=14.8Hz,1H),4.48(d,J=14.8Hz,1H),3.73(s,3H) ,2.23(s,3H),1.95(s,3H),1.89–1.82(m,1H),0.97–0.93(m,2H),0.88–0.83(m,2H).

[0319] Example 45 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(4-cyclopropyl-2-fluorobenzyl)-N 2Preparation of -(2-methoxyphenyl)oxalamide (45)

[0320] Compound 38e was used as the starting material to obtain the title compound through Suziki coupling reaction and BOC removal.

[0321] LC-MS (ESI) m / z (M+H) + :449.2

[0322] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),7.79(s,1H),7.27–7.18(m,3H),7.04–6.98(m,2H),6.87(d,J=8.0Hz,1H),6.83–6.75(m,2H),5.55( s,2H),5.10(d,J=14.8Hz,1H),4.59(d,J=14.8Hz,1H),3.72(s,3H),1.95(s,3H),1.91–1.85(m,1H),0.97–0.90(m,2H),0.68–0.63(m,2H).

[0323] Example 46 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(4-cyclopentyl-2-fluorobenzyl)-N 2 Preparation of -(2-methoxyphenyl)oxalamide (46)

[0324] Compound 38e was used as the starting material to obtain the title compound through Suziki coupling reaction, hydrogen reduction and BOC removal.

[0325] LC-MS (ESI) m / z (M+H) + :477.2

[0326] 1H NMR (400MHz, DMSO-d6) δ10.26 (s, 1H), 7.80 (s, 1H), 7.30 (t, J = 8.0Hz, 1H), 7.27 –7.19(m,2H),7.07(d,J=7.8Hz,1H),7.01(td,J=8.4,1.5Hz,2H),6.97(d,J=11 .6Hz,1H),6.81(t,J=7.6Hz,1H),5.55(s,2H),5.09(d,J=14.9Hz,1H),4.62(d, J=14.9Hz,1H),3.72(s,3H),3.00–2.87(m,1H),1.95(s,3H),1.76–1.14(m,8H).

[0327] Example 47 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(4-cyclohexyl-2-fluorobenzyl)-N 2 Preparation of -(2-methoxyphenyl)oxalamide (47)

[0328] Compound 38e was used as the starting material to obtain the title compound through Suziki coupling reaction, hydrogen reduction and BOC removal.

[0329] LC-MS (ESI) m / z (M+H) + :491.2

[0330] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),7.79(d,J=2.5Hz,1H),7.30(t,J=8.0Hz,1H),7.26–7.19(m,2H),7.08( dd,J=7.8,1.7Hz,1H),7.00(d,J=8.1Hz,2H),6.95(dd,J=11.5,1.7Hz,1H),6.81(td,J=7.6,1.3Hz,1H),5.55 (s,2H),5.07(d,J=14.9Hz,1H),4.63(d,J=14.9Hz,1H),2.69–2.63(m,1H),1.95(s,3H),1.81–1.63(m,6H),1.38–1.30(m,4H).

[0331] Example 48 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-fluoro-4-(1-methylpiperidin-4-yl)benzyl)-N 2Preparation of -(2-methoxyphenyl)oxamide (48)

[0332] Compound 38e was used as the starting material to obtain the title compound through Suziki coupling reaction, hydrogen reduction and BOC removal.

[0333] LC-MS (ESI) m / z (M+H) + :506.3

[0334] 1 H NMR (400MHz, DMSO-d6) δ10.26(s,1H),7.79(s,1H),7.33(t,J=8.0Hz,1H),7.26–7.19(m,2H),7.09–6.96(m,4H),6.82(t,J=7.6Hz,1H),5.55(s,2H ),5.08(d,J=14.8Hz,1H),4.63(d,J=14.8Hz,1H),3.71(s,3H),3.00–2.8 8(m,2H),2.48–2.44(m,1H),2.27(s,3H),1.95(s,3H),1.77–1.53(m,6H).

[0335] Example 49 N 1 -(6-amino-5-methylpyridin-3-yl)-N 1 -(2-methoxyphenyl)-N 2 Preparation of -((5-(1-methylpiperidin-4-yl)pyridin-2-yl)methyl)oxalamide (49)

[0336] Compound 17 was used as the starting material to obtain the title compound through Suziki coupling reaction and hydrogen reduction.

[0337] LC-MS (ESI) m / z (M+H) + :489.3

[0338] 1H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.35(s,1H),7.82(s,1H),7.65(d,J=8.2Hz,1H),7.38 (d,J=8.2Hz,1H),7.28(s,1H),7.20(d,J=7.6Hz,2H),7.02(d,J=8.2Hz,1H),6.83(t,J=7.6H z,1H),5.55(s,2H),5.20(d,J=15.6Hz,1H),4.59(d,J=15.6Hz,1H),3.73(s,3H),2.87–2.84 (m,2H),2.48–2.43(m,1H),2.18(s,3H),1.96(s,3H),1.95–1.90(m,2H),1.71–1.62(m,4H).

[0339] Example 50 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-cyclohexylpyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (50)

[0340] Compound 17 was used as the starting material to obtain the title compound through Suziki coupling reaction and hydrogen reduction.

[0341] LC-MS (ESI) m / z (M+H) + :474.2

[0342] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),8.33(s,1H),7.83(s,1H),7.62(d,J=8.1Hz,1 H),7.37(d,J=8.1Hz,1H),7.29(s,1H),7.20(d,J=7.4Hz,2H),7.01(d,J=8.2Hz,1H) ,6.83(t,J=7.6Hz,1H),5.55(s,2H),5.20(d,J=15.6Hz,1H),4.59(d,J=15.6Hz,1H) ,3.73(s,3H),2.04–1.99(m,1H),1.96(s,3H),1.84–1.66(m,6H),1.41–1.33(m,4H).

[0343] Example 51 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2-(2-Fluoro-4-(tetrahydro-2H-pyran-4-yl)benzyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (51)

[0344] Compound 38e was used as the starting material to obtain the title compound through Suziki coupling reaction and hydrogen reduction.

[0345] LC-MS (ESI) m / z (M+H) + :493.2

[0346] 1 H NMR (400MHz, DMSO-d6) δ10.27(s,1H),7.79(s,1H),7.34(t,J=8.0Hz,1H),7.25(s,1H), 7.22(t,J=7.8Hz,1H),7.08(d,J=7.8Hz,1H),7.05(d,J=8.0Hz,1H),7.02(s,1H),6.99( s,1H),6.82(t,J=7.6Hz,1H),5.55(s,2H),5.09(d,J=15.0Hz,1H),4.64(d,J=14.9Hz,1 H),4.02–3.84(m,4H),3.71(s,3H),2.79–2.71(m,1H),1.95(s,3H),1.69–1.60(m,4H).

[0347] Example 52 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -(2-methoxyphenyl)-N 2 Preparation of -((5-(tetrahydro-2H-pyran-4-yl)pyridin-2-yl)methyl)oxalamide (52)

[0348] Compound 17 was used as the starting material to obtain the title compound through Suziki coupling reaction and hydrogen reduction.

[0349] LC-MS (ESI) m / z (M+H) + :476.2

[0350] 1H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.37(s,1H),7.83(s,1H),7.67(d,J=8.0Hz, 1H),7.40(d,J=8.0Hz,1H),7.30(s,1H),7.28–7.24(m,2H),7.02(d,J=8.2Hz,1H),6 .84(t,J=7.8Hz,1H),5.60(s,2H),5.21(d,J=15.6Hz,1H),4.60(d,J=15.6Hz,1H), 4.05–3.84(m,4H),3.74(s,3H),2.85–2.74(m,1H),1.97(s,3H),1.71–1.62(m,4H).

[0351] Example 53 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-(4,4-difluorocyclohex-1-en-1-yl)pyridin-2-yl)methyl)-N 2 Preparation of -(2-methoxyphenyl)oxamide (53)

[0352] Compound 17 was used as the starting material to obtain the title compound through Suziki coupling reaction.

[0353] LC-MS (ESI) m / z (M+H) + :508.2

[0354] 1 H NMR(400MHz,DMSO-d6)δ10.32(s,1H),8.56(s,1H),7.87–7.80(m,2H),7.43(d, J=8.2Hz,1H),7.28(s,1H),7.24–7.14(m,2H),7.02(d,J=7.6Hz,1H),6.82(t,J =7.6Hz,1H),6.11(s,1H),5.56(s,2H),5.25(d,J=15.6Hz,1H),4.62(d,J=15.6 Hz,1H),3.75(s,3H),2.76–2.64(m,2H),2.22–2.13(m,2H),2.05–1.93(m,5H).

[0355] Example 54 N 1 -(6-amino-5-methylpyridin-3-yl)-N 2 -((5-(4,4-difluorocyclohexyl)pyridin-2-yl)methyl)-N 2Preparation of -(2-methoxyphenyl)oxalamide (54)

[0356] Compound 53 was used as the starting material and reduced with hydrogen to obtain the title compound.

[0357] LC-MS (ESI) m / z (M+H) + :510.2;

[0358] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),8.37(s,1H),7.82(s,1H),7.67(d,J=8.2Hz,1H),7 .39(d,J=8.2Hz,1H),7.29(s,1H),7.21(d,J=7.6Hz,2H),7.01(d,J=7.6Hz,1H),6.83(t, J=7.6Hz,1H),5.56(s,2H),5.20(d,J=15.4Hz,1H),4.61(d,J=15.4Hz,1H),3.73(s,3H), 2.16–2.05(m,2H),2.05–1.99(m,2H),1.96(s,3H),1.89–1.83(m,2H),1.73–1.60(m,2H).

[0359] Biological evaluation

[0360] Test Example 1: HCT116 cell proliferation inhibition test

[0361] 1. Purpose of the test

[0362] This experiment tested the inhibitory effect of the compounds on the proliferation of HCT116 mutant (MTAP-null) and wild-type (WT) cells.

[0363] 2. Test materials

[0364] 2.1 Compounds

[0365] The test compounds were prepared in sequence according to the corresponding examples.

[0366] 2.2 Reagents and Instruments

[0367] Cell-Titer Glo reagent, Promega;

[0368] RPMI1640 medium, Hyclone;

[0369] DMSO, Sigma;

[0370] Fetal bovine serum, Gibco;

[0371] Microplate reader (BMG), POLARstar;

[0372] Pipettes, Eppendorf;

[0373] Cell counter, Countstar;

[0374] CO2 constant temperature incubator, Thermo;

[0375] Biological safety cabinet, Thermo;

[0376] Inverted microscope, OLYMPUS;

[0377] 3. Test methods

[0378] 3.1 Compound Preparation

[0379] Preparation of 10 mM compound stock solutions: Dissolve compound powder in 100% DMSO to prepare 10 mM compound stock solutions.

[0380] 3.2 Test methods

[0381] (1) For cells in the logarithmic growth phase, remove the culture medium and wash twice with PBS;

[0382] (2) Digest the cells with 0.25% trypsin for 2 min until the cells become round and fall off, then add complete medium to terminate the digestion;

[0383] (3) Transfer the digested cells to a centrifuge tube and centrifuge at 1000 rpm for 3 min;

[0384] (4) Remove the supernatant. Add 10 ml of complete culture medium to resuspend the cells. Take 20 μL of the cell suspension and add 20 μL of trypan blue, mix well, and count using a cell counter. Select three fields of view to count and record the viable cell density, cell viability, and cell generation number.

[0385] (5) Diluted cell density is 5.56*10 3 cells / mL, 90 μL was inoculated into a 96-well plate, 500 cells per well;

[0386] (6) The 96-well plate was placed at 37°C and 5% CO2 and incubated overnight;

[0387] (7) On the second day, 10 mM stock solution of the compound was diluted with complete medium containing 1% DMSO to make the final concentrations 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3 nM; 30000, 10000, 3000, 1000, 300, 100, 30, 10, 3 nM;

[0388] (8) After the compound is prepared, remove the cell culture plate and add 10 μL of the diluted compound to the cells;

[0389] (9) After gentle mixing, culture in a 37°C 5% CO2 incubator;

[0390] (10) After 5 days of culture, 50 μL of CTG detection solution was added to each well, and the cells were lysed in a horizontal shaker in the dark for 10 min. 20 μL was pipetted into a 384-well white plate;

[0391] (11) The luminescence module of a BMG microplate reader was used to read the results and calculate the inhibition rate.

[0392] 3.3 Data Analysis

[0393] Cell viability = (As-Ab) / (Ac-Ab)*100%

[0394] As: absorbance of experimental wells (containing cells, culture medium, CTG solution and drug solution)

[0395] Ac: Absorbance of control well (containing cells, culture medium, CTG solution, but no drug)

[0396] Ab: absorbance of blank well (containing culture medium and CTG solution, but not cells or drugs)

[0397] The logarithm of the concentration was used as the X-axis and the percentage survival rate was used as the Y-axis. The log(inhibitor) vs. response-Variable slop(Four parameters) formula of the analysis software Graphpad Prism 9 was used to fit the dose-effect curve to obtain the IC of each compound. 50 value.

[0398] 4. Test results

[0399] Table 1 The inhibitory activity of the patented compounds on HCT116 mutant (MTAP-null) cell proliferation and HCT116 wild type / mutant (WT / MTAP-null) selectivity data

[0400] From the experimental data of the inhibitory activity of the exemplary compounds on HCT116 mutant (MTAP-null) cells and HCT116 wild-type / mutant (WT / MTAP-null) selectivity in Table 1, it can be seen that the compounds of the present application have obvious inhibitory activity on HCT116 mutant (MTAP-null) cells, and have weak inhibitory activity on HCT116 wild-type (WT) cells, and have high selectivity for MTAP gene-deficient cells.

[0401] Test Example 2: PRMT5-MTA enzymatic inhibition test

[0402] 1. Purpose of the test

[0403] The MTase-Glo Assay is a bioluminescent assay that monitors the formation of the reaction product, s-adenosylhomocysteine ​​(SAH), and can detect changes in the activity of multiple methyltransferases, including DNA, protein, RNA, and small molecule methyltransferases. This study tested the effects of test compounds on PRMT5-MTA enzyme activity and used the inhibition rate to calculate the IC value of the test compounds on PRMT5-MTA enzyme. 50 value.

[0404] 2. Test materials

[0405] 2.1 Compounds

[0406] The test compounds were prepared in sequence according to the corresponding examples.

[0407] 2.2 Reagents and Instruments

[0408] PRMT5 / MEP50, active motif;

[0409] 5'-Deoxy-5'-(methylthio)adenosine(MTA), Sigma-Aidrich;

[0410] Bio-H4(1-21), GenScript;

[0411] MTase_GloTM Methyltransferase Assay, Promega;

[0412] 96-well plate, Nunc;

[0413] 384-well plates, Greiner;

[0414] Centrifuge, Xiangyi;

[0415] Microplate reader, BMG.

[0416] 3. Test methods

[0417] 3.1 Compound Preparation

[0418] Preparation of 10 mM compound stock solutions: Dissolve compound powder in 100% DMSO to prepare 10 mM compound stock solutions.

[0419] 3.2 Test methods

[0420] (1) Compound concentration configuration: Compound IC 50 The final test concentration was 1000 nM, 3-fold dilution, 10 concentrations, and duplicate well detection was set for each concentration.

[0421] (2) Pipette 20 nL of compound into a 384-well plate, and centrifuge the 384-well plate at 1000 rpm.

[0422] (3) Add 2 μL of PRMT5-MEP50 / MTA working solution to each well of a 384-well plate and incubate at 25°C for 15 min.

[0423] (4) Add 2 μL of Bio-H4(1-21)&SAM working solution to each well and incubate at 25°C for 180 min.

[0424] (5) Add 1 μL of 5X MTase-Glo reagent to each well, centrifuge the 384-well plate at 1000 rpm, and incubate at 25°C for 30 min.

[0425] (6) Add 5 μL of MTase-Glo detection solution to each well, centrifuge the 384-well plate at 1000 rpm, and incubate at 25°C for 30 min.

[0426] (7) Detect the bioluminescence value using an enzyme-labeled instrument.

[0427] 3.3 Data Processing

[0428] Calculation formula

[0429] Inhibition percentage (%) = (ave high control – compounds) / (ave high control – ave low control) * 100%

[0430] The logarithm of the concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slop (Four parameters) formula of the analysis software Graphpad Prism 5 was used to fit the dose-effect curve to obtain the IC of each compound on the enzyme activity. 50 value.

[0431] 4. Test results

[0432] The inhibitory activities of the compounds of the present invention on PRMT5-MTA enzyme are shown in Table 2.

[0433] Table 2 Inhibitory activity of the compounds of the present invention on PRMT5-MTA enzyme

[0434] From the test data of the PRMT5-MTA enzyme inhibitory activity of the compounds in Table 2, it can be seen that the exemplary compounds of the present application have strong PRMT5-MTA enzyme inhibitory activity.

[0435] Test Example 3: Oral pharmacokinetic study in rats

[0436] 1. Test principle

[0437] SD rats were used as test animals, and the LC-MS / MS method was used to determine the plasma concentration of the compound of the present invention at different time points after oral administration to obtain the pharmacokinetic parameters of the compound of the present invention in rats and study its pharmacokinetic characteristics.

[0438] 2. Test materials

[0439] 2.1 Compounds

[0440] The test compounds were prepared in sequence according to the corresponding examples.

[0441] 2.2 Test instruments:

[0442] Shimadzu LC-30A AB API4500 tandem mass spectrometer, vacuum blood collection tubes, blood collection needles, filter paper, syringes, etc.

[0443] 2.3 Experimental Animals

[0444] Female SD rats weighing 180-220 g were used in each group. The animals were housed in an animal room after purchase and allowed to adapt for at least 3 days before being used in the experiment after passing quarantine.

[0445] 3. Test methods

[0446] 3.1. Grouping: The rats were randomly divided into groups according to Table 6. After grouping, there was no statistical difference in body weight between the groups.

[0447] 3.2. Solvent PO: 0.5% MC+0.2% Tween 80+5% DMSO

[0448] Table 3 Experimental groups and dosing regimens

[0449] 3.3. Blood sample collection and measurement:

[0450] According to Table 3, each group was gavaged with the corresponding test drug. Before administration and 0.25 h, 0.5 h, 0.75 h, 1 h, 1.5 h, 2 h, 4 h, 6 h, and 24 h after administration, a fixed volume of blood was collected from the jugular vein, placed in an EDTA-K2 anticoagulant tube, and centrifuged at 4500 rpm for 10 min. The plasma was separated and placed in a centrifuge tube and frozen in a -80°C refrigerator.

[0451] 3.4 Analysis Methods

[0452] Plasma at each time point stored at -80°C was taken out, and a fixed volume of methanol was added. After vortexing at 1500 rpm for 2 min, the cells were centrifuged for 15 min (3500 r / min), and a fixed volume of the supernatant was taken for LC-MS / MS analysis.

[0453] 4. Calculation of pharmacokinetic parameters:

[0454] The pharmacokinetic behavior of the test compounds was fitted with a non-compartmental model, and the main pharmacokinetic parameters (T 1 / 2 、T max 、C max , AUC last wait).

[0455] 5. Test results:

[0456] Table 4 Pharmacokinetic parameters of the compounds in the examples

[0457] It can be seen from the test results in Table 4 that the exemplary compounds of the present application have good pharmacokinetic properties.

Claims

1. A compound having the structure of formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in, R 1 Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, carboxyl, hydroxyl, C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy, wherein the C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 hydrocarbon group, C1-C6 hydrocarbon group oxy group, C3-C8 carbocyclyl group or C3-C8 carbocyclyl group oxy group, wherein C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 and R 3 Can form a 5-8 membered carbocyclic ring or a 5-8 membered heterocyclic ring; R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclic group or 3-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic group, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino, wherein the C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic group, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino, may be further substituted with halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy or amino; R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 aryl or 4-10 membered heterocyclic group, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 The aryl or 4-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C8 cycloalkyl, C3-C8 cycloalkyloxy, C3-C 10 Cycloalkylamino, 3-10 membered heterocyclylamino, X is selected from: a single bond or -CH2-, wherein -CH2- may be further substituted by 1 or 2 methyl groups or halogen; n is selected from: 0 or 1.

2. The compound of formula I according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclic group or 3-10 membered heterocyclic group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy, C1-C6 alkylamino, C1-C6 haloalkyloxy, C3-C 10 Carbocyclic group, C3-C 10 Cycloalkyloxy, C3-C 10 cycloalkylamino, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, 3-10 membered heterocyclylamino, 3. The compound of formula I according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: R 1 Selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, carboxyl, hydroxyl, C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy, wherein the C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C6 hydrocarbon group, C1-C6 hydrocarbon group oxy group, C3-C8 carbocyclyl group or C3-C8 carbocyclyl group oxy group, wherein C1-C6 alkyl, C1-C6 alkyloxy, C3-C8 carbocyclyl or C3-C8 carbocyclyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl or C1-C6 alkyloxy; R 2 and R 3 Can form a 5-8 membered carbocyclic ring or a 5-8 membered heterocyclic ring; R 5 Selected from: C3-C 10 Carbocyclic or 3-10 membered heterocyclic, wherein the C3-C 10 The carbocyclyl or 3-10 membered heterocyclyl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 haloalkyl, C1-C6 alkyl, C1-C6 alkyloxy or C1-C6 haloalkyloxy; R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkyloxy or C1-C6 haloalkyloxy; X is selected from: a single bond or -CH2-, wherein -CH2- may be further substituted by 1 or 2 methyl groups or halogen; n is selected from: 0 or 1.

4. The compound of formula I according to any one of claims 1 to 3, its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that: R 5 Selected from: C3-C8 cycloalkyl, 3-10 membered heterocyclic group or C6-C 10 Aryl, wherein the C3-C8 cycloalkyl, 3-10 membered heterocyclic group or C6-C 10 The aryl group is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, The C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, may be further substituted with halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy or amino; Preferably, R 5 Selected from: C6-C 10 Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, The C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, It may be further substituted with 1, 2 or 3 F, Cl, Br or C1-C4 alkyl groups; More preferably, R 5 Selected from: C6-C 10 Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or It is substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, and the C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl may be further substituted by 1, 2 or 3 F, Cl, Br, C1-C4 alkyl.

5. The compound of formula I according to any one of claims 1 to 4, its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that: R 6 Selected from: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-8 membered heterocycloalkyl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C8 cycloalkyl, C6-C 10 Aryl, 4-8 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino, R 6 Preferred are: C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heterocyclic group, wherein the 5-10 membered heterocyclic group is selected from 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino, R 6 Further preferably selected from: C3-C6 cycloalkyl, C6-C 10 aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10 Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino, R 6 Further preferably selected from: C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from F, Cl, Br, cyano, C1-C3 alkyl, C1-C3 alkyloxy or C1-C3 haloalkyloxy.

6. The compound of formula I according to any one of claims 1 to 4, its stereoisomers or pharmaceutically acceptable salts thereof, characterized in that: R 1 is selected from hydrogen, halogen, cyano, and substituted or unsubstituted amino.

7. The compound of formula I according to any one of claims 1 to 5, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, substituted or unsubstituted amino, C1-C3 alkyl or C1-C3 alkyloxy, wherein C1-C3 alkyl or C1-C3 alkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C3 alkyl or C1-C3 alkyloxy; R 2 and R 3 It can form a 5-6 membered carbocyclic ring or a 5-6 membered heterocyclic ring.

8. A compound having the structure of formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in, R 2 、R 3 、R 4 are independently selected from: hydrogen, halogen, cyano, C1-C3 alkyl, C1-C3 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy, wherein C1-C3 alkyl, C1-C3 alkyloxy, C3-C8 cycloalkyl or C3-C8 cycloalkyloxy is unsubstituted or substituted by one or more substituents selected from halogen, cyano, hydroxy, amino, C1-C3 alkyl or C1-C3 alkyloxy; R 2 and R 3 Can form a 5-6 membered carbocyclic ring or a 5-6 membered heterocyclic ring; R 5 Selected from: C3-C8 cycloalkyl, 3-10 membered heterocyclic group, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C8 cycloalkyl, 3-10 membered heterocyclyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkyloxy, C1-C3 alkylamino, C1-C3 haloalkyloxy, C3-C6 carbocyclyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclyl, 3-6 membered heterocyclyloxy, 3-6 membered heterocyclylamino, It may be further substituted with 1-3 F, Cl, Br or C1-C4 alkyl groups; R 6 Selected from: C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heterocyclic group, wherein the 5-10 membered heterocyclic group is selected from 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl, wherein the C1-C3 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, 4-6 membered heterocycloalkyl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkyloxy, C1-C4 alkylamino, C1-C4 haloalkyloxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, C3-C6 cycloalkylamino, 3-6 membered heterocyclylamino, n is selected from: 0 or 1; Preferably, R 2 、R 3 、R 4 Each is independently selected from: hydrogen, halogen, cyano or C1-C3 alkyl; More preferably, R 2 Selected from C1-C3 alkyl; R 3 、R 4 All are hydrogen; Preferably, R 5 Selected from: C6-C 10 Aryl or 5-8 membered heteroaryl, wherein the C6-C 10 The aryl or 5-8 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from halogen, cyano, C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, and the C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 3-6 membered heterocyclyl may be further substituted by 1-3 F, Cl, Br or C1-C4 alkyl; Preferably, R 6 Further selected from: C3-C6 cycloalkyl, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C3-C6 cycloalkyl, C6-C 10 The aryl or 5-10 membered heteroaryl is unsubstituted or substituted by one or more substituents selected from F, Cl, Br, cyano, C1-C3 alkyl, C1-C3 alkyloxy or C1-C3 haloalkyloxy.

9. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is selected from:

10. A method for preparing the compound according to any one of claims 1 to 9 or a stereoisomer thereof, comprising the following steps: (1) Compound I-1 and compound I-2 are condensed to obtain I-3; (2) Compound I-3 is hydrolyzed to obtain I-4; (3) Compound I-4 and Compound I-5 are condensed to obtain a compound having a structure shown in Formula I; Among them, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 , X and n are as defined for each substituent in claims 1-9.

11. A pharmaceutical composition, characterized in that The invention comprises the compound according to any one of claims 1 to 9, its stereoisomer or pharmaceutically acceptable salt, and pharmaceutically acceptable excipients.

12. Use of a compound according to any one of claims 1 to 9, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating a disease mediated by protein arginine methyltransferase 5 (PRMT5); preferably, the disease mediated by protein arginine methyltransferase 5 (PRMT5) is selected from a tumor or a cancer; more preferably, the disease of PRMT5 is selected from a tumor or a cancer; more preferably, the disease is selected from breast cancer, pancreatic cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, lymphoma, malignant sarcoma, cervical cancer, oral cancer, brain cancer, gastric cancer, liver cancer, skin cancer, bone cancer, kidney cancer, bladder cancer, fallopian tube tumor, mesothelioma, melanoma, glioma, glioblastoma, papillary malignant tumor, head and neck tumor, myeloma, or leukemia.

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