CN118702697A - Benzoyl pyrrolopyrimidine derivatives and their use and preparation method - Google Patents

Abstract

The present invention relates to the field of medicine and is a class of benzoyl pyrrolopyrimidine derivatives as shown in formula I and applications thereof. It is a highly active and specific BTK inhibitor that can enhance the activity and selectivity of the drug, improve toxic side effects and physicochemical properties, and cope with drug-resistant mutations caused by treatment. The compound, its stereoisomer or pharmaceutically acceptable salt and its pharmaceutical composition are used in the preparation of drugs for the prevention and treatment of BTK-related diseases. A series of tumor cell test results show that the compound of the present invention has good anti-BTK kinase activity.

Description

Benzoyl pyrrolopyrimidine derivatives and their use and preparation method

Technical Field

The present invention relates to the field of medicine, and includes a class of benzoylpyrrolopyrimidine derivatives and a preparation method thereof; the present invention also relates to the application of the compound in the preparation of anticancer drugs.

Background Art

BTK, Bruton's tyrosine kinase, is a key kinase in the B cell receptor (BCR) signal transduction pathway. It is widely expressed in different types of malignant blood diseases and is involved in the proliferation, differentiation and apoptosis of B cells. Due to the high specificity of BTK small molecule inhibitors, it has shown great advantages in the treatment of B cell malignancies and some B cell immune diseases. Therefore, BTK inhibitors have become the most promising drugs in the blood tumor market.

Ibrutinib is a first-generation covalent BTK inhibitor with an IC50 of 0.5nM. In 2013, the FDA approved it for the treatment of mantle cell lymphoma, and later approved it for chronic lymphocytic leukemia, Waldenstrom's macroglobulinemia, small lymphocytic lymphoma, marginal zone lymphoma, graft-host disease, etc. Ibrutinib still has many shortcomings, such as the selectivity of the target, which may lead to related bleeding, diarrhea, rash, atrial fibrillation and other problems; another example is the drug resistance caused by the mutation of cysteine at position 481 of the BTK protein to serine.

Acalabrutinib is a second-generation covalent BTK inhibitor with an IC50 of 3nM, which greatly improves the selectivity problem of the first-generation BTK inhibitor. It was approved by the FDA in 2017 for the treatment of mantle cell lymphoma and later approved for chronic lymphocytic leukemia. However, it has not yet solved the drug resistance problem caused by Cys481 mutation.

Pirtobrutinib is a third-generation BTK inhibitor and the first non-covalent BTK inhibitor. It was approved by the FDA in 2023 for adult relapsed and refractory mantle cell lymphoma who have received third-line or higher systemic treatment (including treatment with other BTK inhibitors). Unlike the currently marketed covalent BTK inhibitors, Pirtobrutinib does not covalently bind to C481, so it will not be inhibited by the activity of drug-resistant C481 mutations, and is expected to overcome the resistance of covalent BTK inhibitors.

HCK is a hematopoietic stem cell kinase of the SRC family. Its abnormal activation drives the formation of blood malignancies. Studies have shown that increased HCK levels shorten the survival of colorectal cancer patients. Inhibiting HK activity can inhibit the selective activation of tumor-associated macrophages and the growth of colon cancer xenografts. Therefore, HCK can be used as a therapeutic target for malignant solid tumors.

Summary of the invention

The present invention provides a compound represented by formula I or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

in,

X is NH, C or O;

X1, X2, W, W2, W3 are independently selected from C, N, O;

a, b, and m represent the number of W, W2, and W3, respectively, and are independently selected from 0, 1, 2, or 3;

n is 0 or 1;

e is a chemical bond, representing any reasonable conformation;

R1 is selected from H, halogen, C1-3 alkyl; the number of R1 is 1, 2 or 3, and R1 is located at any reasonable position of the benzene ring;

R2 is selected from substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C3-8 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group; the substituent is selected from C1-6 alkyl, C1-6 alkoxy, 5-8 membered heterocyclic group, wherein the 5-8 membered heterocyclic group may be further substituted by C1-6 alkyl;

Said Ra and Rb are independently selected from C1-6 alkyl;

R3 is selected from H, substituted or unsubstituted C1-6 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the substituent of the C1-6 alkyl is selected from OH, NH2, -OC(=O)Rc; the Rc is selected from C1-6 alkyl, C2-6 alkenyl, C3-8 cycloalkyl; R4 is selected from H, halogen, C1-6 alkyl or C1-6 alkoxy;

The heterocyclic group contains 1-3 heteroatoms, and the heteroatoms are selected from N, O, and S.

The compound described in Formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, W is C, a is 1, as shown in Formula II, Wherein, X is NH or O, b is 1, m is 0 or 1, W2 and W3 are independently selected from C, N, O; R1 is selected from H or halogen, the number of R1 is 1 or 2, and R1 is located at any reasonable position of the benzene ring; X1 and X2 are independently selected from C or N;

n is 0 or 1;

R2 is selected from substituted or unsubstituted C1-6 alkyl, C3-6 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclyl;

The substituent of the C1-6 alkyl group is selected from C1-4 alkoxy, 5-8 membered heterocyclic group, The substituent of the 5-8 membered heterocyclic group is a C1-6 alkyl group;

Said Ra and Rb are independently selected from C1-4 alkyl;

R3 is selected from H, substituted or unsubstituted C1-4 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the Rc is selected from C1-4 alkyl, C2-4 alkenyl, C3-6 cycloalkyl, and the substituent of the C1-6 alkyl is selected from OH, NH2;

R4 is selected from H, halogen or C1-3 alkoxy.

The compound described in Formula I, its stereoisomer or pharmaceutically acceptable salt thereof, W is C, a is 1, X is NH, as shown in Formula III, wherein b is 1, m is 0 or 1, W2 and W3 are independently selected from C, N, and O;

R1 is selected from H or halogen, the number of R1 is 1 or 2, and R1 is located at any reasonable position of the benzene ring;

X1 and X2 are independently selected from C or N;

n is 0 or 1;

R2 is selected from substituted or unsubstituted C1-6 alkyl, C3-6 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclyl;

The substituent of the C1-6 alkyl group is selected from C1-4 alkoxy, 5-8 membered heterocyclic group, The substituent of the 5-8 membered heterocyclic group is a C1-6 alkyl group;

Said Ra and Rb are independently selected from C1-4 alkyl;

R3 is selected from H, substituted or unsubstituted C1-4 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the Rc is selected from C1-4 alkyl, C2-4 alkenyl, C3-6 cycloalkyl, and the substituent of the C1-6 alkyl is selected from OH, NH2;

R4 is selected from H, halogen or C1-3 alkoxy.

The compound described in Formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, R4 is H, n is 0, W is C, a is 1, X is N, as shown in Formula IV, in,

R1 is selected from H, halogen, C1-3 alkyl;

R2 is selected from substituted or unsubstituted C1-4 alkyl, substituted or unsubstituted C1-4 alkoxy; the substituent is selected from C1-3 alkyl, C1-3 alkoxy, 5-8 membered heterocyclic group; the heterocyclic group contains 1-2 heteroatoms selected from N, O, and S; the Ra and Rb are independently selected from C1-3 alkyl;

X1, X2, W2, W3 are independently selected from C, N, O;

b and m are independently 1 or 2;

R3 is selected from substituted or unsubstituted C1-3 alkyl, and the substituent is OH, NH2.

Further, R1 is selected from H or halogen.

The compound, stereoisomer or pharmaceutically acceptable salt thereof as shown in Formula IV, R1 is selected from H, halogen, R1 is located in the para position of the benzene ring; R2 is selected from substituted or unsubstituted C1-4 alkyl, substituted or unsubstituted C1-4 alkoxy; the substituent is selected from C1-3 alkyl, 5-6 membered heterocyclic group; the heterocyclic group contains 1-2 heteroatoms selected from N, O, and S; the Ra and Rb are independently selected from C1-3 alkyl; X1 and X2 are N; W2 and W3 are C; b and m are independently 1 or 2.

Furthermore, the R1 is selected from H, F, Br, and I.

In some embodiments, R1 is H or F.

Further, the R2 is selected from substituted or unsubstituted C1-3 alkyl, substituted or unsubstituted C1-3 alkoxy, substituted or unsubstituted C3-6 cycloalkyl, substituted or unsubstituted 5-6 membered heterocyclic group; the substituent is selected from C1-3 alkyl, C1-36 alkoxy, 5-6 membered heterocyclic group, wherein the 5-6 membered heterocyclic group may be further substituted by C1-3 alkyl; said Ra and Rb are independently selected from C1-3 alkyl.

Further, the R2 is selected from substituted or unsubstituted C1-3 alkyl, C3-6 cycloalkyl, substituted or unsubstituted 5-6 membered heterocyclic group; the substituent of the C1-3 alkyl is selected from C1-3 alkoxy, 5-6 membered heterocyclic group, The substituent of the 5-6 membered heterocyclic group is C1-alkyl; and Ra and Rb are independently selected from C1-3 alkyl.

Further, the R2 is selected from substituted methyl, ethyl, propyl, methoxy, ethoxy, propoxy, piperidinyl, piperazinyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydropyranyl, oxetane; the substituent is selected from methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, -N(CH3)2, -N(CH2CH3)2.

In some embodiments, R2 is selected from methyl, methylpiperazine, cyclopropyl, cyclobutyl, ethylpiperazinyl, -CH2CH2N(CH3)2, Methoxy, -N(CH3)2, -CH2CH2N(CH2CH3)2, -CH2CH2-O-CH3.

In certain embodiments, the compound of Formula I, its stereoisomers or pharmaceutically acceptable salts thereof are selected from the following compounds: (4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-fluorophenyl)(4-((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; )-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(3-morpholinopropyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone [2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (2-((1-(2-(diethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-(dimethylamino)piperidin-1-yl)phenyl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (2-((4-(4-(dimethylamino)piperidin-1-yl) -2-methoxyphenyl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; N-((1s,4s)-4-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; N-((1s,4s)-4-((2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-(-4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (3,4-difluorophenyl)(4-(( 1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((1-cyclobutyl-1H-pyrazol-4-yl)amino)-4-((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl ) ketone; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H- (R)-1-(3-((2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(2,4-difluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidine -1-yl)prop-2-en-1-one; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)butyramide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)cyclopropanecarboxamide; (2-fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro- (2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(2-(dimethylamino)ethoxy)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone pyrimidine-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)acetamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)acetamide; 4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propane-1-sulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propane-1-sulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propane )cyclopropanesulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((5-methyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((5-isopropyl-1H-pyrazol-3-yl)amino)-7H -pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((5-cyclopropyl-1H-pyrazol-3-yl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; N-((1r,4r)-4-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; N-((1r,4r)-4-((2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)cyclohexyl)isobutyramide (((1r,4r)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; N-((1r,4r)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; ((((1r,4r)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; N-((1r,4r)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; ((((1r,4r)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (R)-(4-fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (S)-(4-fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydrofuran-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone.

In certain embodiments, the compound of Formula I, its stereoisomers or pharmaceutically acceptable salts thereof are selected from the following compounds: (4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-fluorophenyl)(4-((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; ((((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; ((((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; ((((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; )-2-((1-(3-morpholinopropyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (2-((1-(2-(diethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone (((4-(dimethylamino)piperidin-1-yl)phenyl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-(dimethylamino)piperidin-1-yl)phenyl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (2-(( 4-(4-(dimethylamino)piperidin-1-yl)-2-methoxyphenyl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; N-((1s,4s)-4-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(4-fluorophenyl)methanone isobutylamide; N-((1s,4s)-4-((2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutylamide; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-(-4-methylpiperazin-1-yl)piperidin-1-yl)phenyl )amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (3,4-difluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)- 2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((1-cyclobutyl-1H-pyrazol-4-yl)amino)-4-((1s,4s)-4-( (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; -((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(2,4-difluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one -yl)prop-2-en-1-one; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)butanamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin- (2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone phenyl)methanone; (2-((4-(2-(dimethylamino)ethoxy)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; N-((1s, 4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)acetamide; N-( (1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(piperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propanamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propane-1-sulfonamide.

In certain embodiments, the compound of Formula I, its stereoisomers or pharmaceutically acceptable salts thereof are selected from the following compounds: (4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-fluorophenyl)(4-((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3 -d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(3-morpholinopropyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-((( ((1-(2-(diethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((1-(2-(diethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone.

The present invention provides a method for preparing a compound of formula I, including its isomers, hydrates, solvates, pharmaceutically acceptable salts and prodrugs thereof, comprising the following steps:

1) 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine The reaction obtains compound T1; preferably, the reaction is carried out in an organic solvent, and the organic solvent is a polar solvent; preferably, the organic solvent is dichloromethane or acetone.

2) Compound T1 reacts with 2-(trimethylsilyl)ethoxymethyl chloride (SEM-Cl) under heating conditions to obtain compound T2; preferably, the reaction temperature is 30-60° C.;

The reaction is carried out in an organic solvent, wherein the organic solvent is at least one of N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), N,N-diisopropylethylamine (DIPEA), N-methyl-2-pyrrolidone (NMP), ethylene glycol dimethyl ether, isopropanol, n-butanol, sec-butanol, and tert-butanol;

The reaction is carried out under alkaline conditions, and the base is an inorganic base or an organic base, wherein the inorganic base is selected from ammonia, potassium carbonate, sodium carbonate, and cesium carbonate, and the organic base is selected from at least one of sodium acetate, triethylamine, diisopropylethylamine, triethylenediamine, pyridine, 4-dimethylaminopyridine, 1,8-diazabicycloundec-7-ene, and N-methylmorpholine;

3) Compound T2 reacts with amine T3 under heating conditions to obtain compound T4;

Preferably, the reaction temperature is 80-100°C;

The reaction is carried out under the conditions of an organic solvent and a base, wherein the base and the organic solvent are independently selected from the relevant definitions in step 2);

4) Compound T4 reacts with amine T5 under heating conditions to obtain compound T6, preferably, the reaction temperature is 100-130° C.; the reaction is carried out under organic solvent and base conditions, and the base and organic solvent are independently selected from the relevant definitions in step 2;

The reaction is a Buchwald-Hartwig coupling reaction using a palladium catalyst and an organic phosphine ligand, wherein the palladium catalyst used is selected from palladium acetate, palladium dichloride, [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride, tris(dibenzylideneacetone)dipalladium, tetrakistriphenylphosphine palladium, and bistriphenylphosphine palladium dichloride, and the organic phosphine ligand used includes but is not limited to 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, 1,1'-binaphthyl-2,2'-bisdiphenylphosphine, and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl;

5) removing trimethylsilylethoxymethyl (SEM group) from compound T6 under acid conditions, wherein the acid is an inorganic acid or an organic acid, wherein the inorganic acid is selected from at least one of hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, nitric acid, nitrous acid, and metaphosphoric acid, and the organic acid is selected from at least one of acetic acid, trichloroacetic acid, trifluoroacetic acid, glycolic acid, sulfamic acid, p-toluenesulfonic acid, ethylenediaminetetraacetic acid (EDTA), oxalic acid, and citric acid;

6) The product obtained in step 5 is neutralized under alkaline conditions to obtain a compound of formula I;

The base is an inorganic base or an organic base, wherein the inorganic base is selected from ammonia, potassium carbonate, sodium carbonate, and cesium carbonate, and the organic base is selected from at least one of sodium acetate, triethylamine, diisopropylethylamine, triethylenediamine, pyridine, 4-dimethylaminopyridine, 1,8-diazabicycloundec-7-ene, and N-methylmorpholine.

Wherein, _R1 , _R2 , _R3 , _R4 , X, X1, X2, W, W2, W3, a, b, m, n, e are as defined above, and E is a protecting group selected from SEM group, benzyloxycarbonyl, p-toluenesulfonyl, benzenesulfonyl, acetyl, trifluoroacetyl, fluorenylmethoxycarbonyl, and benzyl.

The present invention provides a pharmaceutical composition comprising an effective dose of a compound of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable pharmaceutical excipients. The compound of the present invention can be prepared into different dosage forms alone or with one or more pharmaceutical carriers, such as tablets, pills, powders, capsules, granules, syrups, emulsions, microemulsions or injections, specifically such as intravenous infusion, subcutaneous infusion, intramuscular infusion, intraperitoneal infusion, transdermal infusion and direct infusion into tissues for clinical oral administration, injection or topical administration.

When the pharmaceutical composition of the present invention is prepared in the form of an oral preparation, ingredients known in the art can be used as pharmaceutically acceptable carriers without restriction, as long as they do not interfere with the active expression of the active ingredient. The carrier may include, for example, excipients, diluents, disintegrants, binders, glidants, surfactants, emulsifiers, suspending agents, diluents, etc., but is not limited thereto.

When the pharmaceutical composition of the present invention is prepared in a parenteral dosage form, the parenteral dosage form is usually sterile or can be sterilized before being administered to a subject because its administration generally bypasses the subject's natural defenses against contaminants. Parenteral dosage forms include, but are not limited to, solutions prepared for injection, dry products prepared for dissolution or suspension in a pharmaceutically acceptable vehicle for injection, suspensions prepared for injection, and emulsions. Suitable carriers/vehicles that can be used to provide parenteral dosage forms are well known to those skilled in the art, including, but not limited to: Water for Injection USP; aqueous vehicles, such as, but not limited to, sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles, such as, but not limited to, ethanol, polyethylene glycol, and polypropylene glycol; and anhydrous vehicles, such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

In these different preparations, the content of the compound of the present invention can be 0.1%-99.9%. The dosage of the compound of the present invention can be 0.001-10000 mg/kg/0.3 day, which can be appropriately adjusted according to clinical needs.

The "pharmaceutically acceptable salt" described in the present invention refers to a pharmaceutically acceptable acid or base addition salt, or a solvate or hydrate thereof.

The present invention provides the use of the compound, its stereoisomer or its pharmaceutically acceptable salt and its pharmaceutical composition in the preparation of drugs for preventing and treating BTK-related diseases. The disease includes an autoimmune disease or a tumor. The autoimmune disease is selected from rheumatoid arthritis or lupus or relief of moderate to severe new coronary pneumonia, etc.; the tumor is a B-cell malignant tumor. The results of a series of tumor cell tests showed that the compounds of the present invention have good anti-BTK kinase activity, and their IC ₅₀ values are all in the nanomolar level, which is comparable to the activity of positive control drugs Fenebrutinib (GDC-0853), Nemtabrutinib (ARQ531), and Pirtobrutinib (Loxo-305), and some compounds are significantly better than positive drugs.

The present invention provides the use of the compound, its stereoisomer, its prodrug, its metabolite or its pharmaceutically acceptable salt and its pharmaceutical composition in the preparation of tumor drugs.

The tumor is a B-cell malignancy.

The tumor is selected from lymphoma, leukemia, prostate cancer, and gastric cancer.

Specifically, the tumor is selected from diffuse large B-cell lymphoma, chronic myeloid leukemia, mantle lymphoma, Burkitt’s lymphoma, prostate cancer, and gastric cancer.

The tumor is selected from hematopoietic or lymphoid tissue tumors.

The hematopoietic or lymphoid tissue tumor is selected from non-Hodgkin's lymphoma, Hodgkin's lymphoma, myeloproliferative neoplasms, myelodysplastic syndrome, precursor lymphocyte tumors or other malignant hematopoietic tissue tumors.

The non-Hodgkin's lymphoma is selected from mature B-cell lymphoma or precursor lymphocytic neoplasm.

The mature B cell neoplasm is selected from follicular lymphoma, diffuse large B cell lymphoma, mature B cell neoplasm with leukemic behavior, plasma cell neoplasm, other specified mature B cell neoplasms or lymphomas.

The diffuse large B-cell lymphoma is selected from primary mediastinal large B-cell lymphoma, intravascular large B-cell lymphoma, plasmablastic lymphoma, lymphomatoid granuloma, primary central nervous system diffuse large B-cell lymphoma, primary cutaneous diffuse large B-cell lymphoma, and primary effusion lymphoma.

The mature B cell tumor with leukemic behavior is selected from chronic lymphocytic leukemia or small lymphocytic lymphoma, B cell prolymphocytic leukemia, hairy cell leukemia, lymphoplasmacytic lymphoma.

The plasma cell tumor is selected from the group consisting of plasma cell myeloma, solitary plasmacytoma, extraosseous plasmacytoma, plasma cell leukemia, and monoclonal immunoglobulin deposition disease.

The other specific mature B cell tumors or lymphomas are selected from lymph node marginal zone lymphoma, extranodal marginal zone lymphoma of gastric mucosa-associated lymphoid tissue, extranodal marginal zone lymphoma outside the stomach or skin, lymphoplasmacytic lymphoma, mantle cell lymphoma, Burkitt's lymphoma including Burkitt's leukemia, splenic marginal zone B cell lymphoma, and vitreoretinal lymphoma.

The precursor lymphocytic tumor is selected from precursor B lymphoblastic tumors.

The precursor B lymphoblastic tumor is selected from B lymphoblastic leukemia or B lymphoblastic lymphoma.

The myeloproliferative neoplasm is selected from chronic myeloid leukemia.

The malignant hematopoietic tissue tumor is selected from acute leukemia, myeloid leukemia, chronic myeloid leukemia, and lymphocytic leukemia.

A series of tumor cell test results show that the present invention provides a class of novel BTK inhibitory active compounds, which have lower tumor cell inhibition concentrations compared to existing compounds. The inhibitory activity of the compounds of the present invention on BTK kinase is at the nanomolar level, specifically less than 1nM, and the inhibitory activity of some compounds is in the range of 0.01-0.1nM.

The BTK inhibitor of the present invention has excellent drug activity and selectivity, lower toxic side effects and better physical and chemical properties. It is also found that it can respond to HCK kinase and synergistically treat tumor diseases such as lymphoma carrying C481S drug-resistant mutations.

The present invention provides a method for treating B-cell malignancies, comprising administering a therapeutically effective amount of the above-mentioned compound, its stereoisomer or pharmaceutically acceptable salt thereof and a pharmaceutical composition thereof to a subject or patient.

The compound provided by the present invention has excellent inhibitory activity on BTK kinase, has great industrialization and commercialization prospects and market value, and has significant economic benefits.

Although preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Now, those skilled in the art will appreciate that many changes, variations and substitutions without departing from the present invention. It should be understood that various substitutions to the embodiments of the present invention described herein may be used to implement the present invention. The appended claims are intended to define the scope of the invention, and thus cover methods and structures and their equivalents within the scope of these claims.

Some chemical terms

Unless otherwise defined, all technical terms herein have the same meaning as commonly understood by those skilled in the art to which the subject matter of the claims belongs. Unless otherwise indicated, all patents, patent applications, and public materials cited in the entirety of this article are incorporated herein by reference in their entirety.

It should be understood that the above brief description and the following detailed description are exemplary and are only used for explanation, and do not limit the subject matter of the present application. In this application, unless otherwise specifically stated, the use of the singular also includes the plural. It should also be noted that unless otherwise stated, the use of "or" and "or" means "and/or". In addition, the term "including" and other forms, such as "including", "containing" and "containing" are all non-restrictive descriptions.

Definitions of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4THED." Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise stated, conventional methods within the technical scope of the art, such as mass spectrometry, NMR and pharmacological methods, are used. Unless specific definitions are provided, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, the manufacturer's instructions for the use of the kit can be used, or the reaction and purification can be carried out in a manner known in the art or the description of this application. The above-mentioned techniques and methods can usually be implemented according to the descriptions in a plurality of summary and more specific documents cited and discussed in this specification, according to conventional methods well known in the art. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds.

When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, _CH2O is equivalent to _OCH2 .

The "compound" described in the present application is meant to include all stereoisomers, geometric isomers, tautomers and isotopes. The compounds of the present application may be asymmetric, for example, having one or more stereoisomers. Unless otherwise indicated, all stereoisomers are included, such as enantiomers and diastereomers. The compounds of the present application containing asymmetrically substituted carbon atoms can be separated in optically pure form or racemic form. Optically pure forms can be separated from racemic mixtures or synthesized by using chiral raw materials or chiral reagents. The compounds of the present application also include tautomeric forms. Tautomeric forms are derived from the exchange of a single bond with an adjacent double bond and are accompanied by the migration of a proton. The compounds of the present application also include atoms of all isotopes, whether in intermediates or final compounds. Isotopic atoms include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. That is, the compounds of the present application include compounds in which part or all of hydrogen (H) is replaced by tritium (T) and/or deuterium (D); compounds in which part or all of ¹² C is replaced by ¹³ C and/or ¹⁴ C; and compounds in which other isotopes (such as N, O, P, S) are replaced, such as ¹⁴ N and ¹⁵ N; ¹⁸ O and ¹⁷ O; ³¹ P and ³² P; ³⁵ S and ³⁶ S, etc. The compounds described herein may have one or more stereogenic centers, and each isomeric center may exist in the form of R or S configuration or a combination thereof. Similarly, the compounds described herein may have one or more double bonds, and each double bond may exist in the form of E (trans) or Z (cis) configuration or a combination thereof. A specific stereoisomer, structural isomer, diastereomer, enantiomer or epimer should be understood to include all possible isomers, such as stereoisomers, structural isomers, diastereomers, enantiomers or epimers and mixtures thereof. Therefore, the compounds described herein include all configurationally different stereoisomers, structural isomers, diastereomers, enantiomers or epimers and their corresponding mixtures. The techniques for converting a specific stereoisomer or keeping a specific stereoisomer intact, as well as the techniques for resolving a mixture of stereoisomers are well known in the art, and those skilled in the art can select the appropriate method for the specific situation.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence of said event or circumstance and the non-occurrence of said event or circumstance.

As used herein, _C1-3 or C1-3 means that there are 1-3 carbon atoms in the moiety, i.e., the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms. Thus, for example, "C1-3 alkyl" refers to an alkyl group having 1-3 carbon atoms, i.e., the alkyl group is selected from methyl, ethyl, propyl, isopropyl.

The term "alkyl" used herein alone or in combination refers to an optionally substituted straight chain or optionally substituted branched aliphatic hydrocarbon. The "alkyl" herein preferably has 1-10 carbon atoms, for example, 1-8 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms or 1-3 carbon atoms. Examples of alkyl groups herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, etc.

[0046] "Alkyl" used in combination herein includes alkyl groups in combination with other groups, such as an alkyl group in an alkoxy group.

The term "5-12 membered heteroaromatic ring" refers to a 5- to 12-membered conjugated ring system group having 4 to 11 carbon atoms and 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur in the ring. Unless otherwise specifically indicated in this specification, the heteroaryl group may be a monocyclic, bicyclic, tricyclic or more ring system, and may also be fused with a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is connected to the rest of the molecule by a single bond via an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group may be optionally oxidized; the nitrogen atom may be optionally quaternized. For the purposes of the present invention, the heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur.

The term "halogen" as used herein, alone or in combination, is selected from F, Cl, Br, I.

As used herein, the term "treat" and other similar synonyms include alleviating, relieving or ameliorating symptoms of a disease or condition, inhibiting a disease or condition, such as preventing the development of a disease or condition, alleviating a disease or condition, making a disease or condition better, alleviating symptoms caused by a disease or condition, or stopping symptoms of a disease or condition, preventing other symptoms, improving or preventing the underlying metabolic causes of symptoms, and in addition, the term includes the purpose of prevention. The term also includes obtaining a therapeutic effect and/or a prophylactic effect. The therapeutic effect refers to the cure or improvement of the underlying disease being treated. In addition, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, for example, although the patient may still be affected by the underlying disease, the patient's condition is observed to improve. In terms of prophylactic effects, the composition can be administered to a patient at risk for a particular disease, or even if a disease diagnosis has not yet been made, the composition can be administered to a patient who has one or more physiological symptoms of the disease.

As used herein, the term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" refers to an amount of at least one active substance (such as a compound of the present application) sufficient to relieve one or more symptoms of the disease or condition being treated to some extent after administration. The result can be the reduction and/or alleviation of signs, symptoms or causes of disease, or any other desired changes in biological systems. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein required to provide a significant symptom alleviation effect clinically. Techniques such as dose escalation trials can be used to determine the effective amount suitable for any individual case.

The term "acceptable" with respect to a formulation, composition or ingredient, as used herein, means having no long-term detrimental effect on the general health of the subject being treated.

The term "pharmaceutically acceptable" as used herein refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application and is relatively non-toxic, that is, the substance can be administered to a subject without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.

The term "pharmaceutical composition" as used herein refers to a mixture of the compound of the present application and at least one pharmaceutically acceptable substance. The pharmaceutically acceptable substance includes, but is not limited to, a carrier, a stabilizer, a diluent, a dispersant, a suspending agent, a thickener and/or an excipient.

As used herein, the term "carrier" refers to a relatively nontoxic substance that facilitates the introduction of a compound of the present application into cells or tissues.

The term "pharmaceutically acceptable salt" as used herein refers to a salt that retains the biological effectiveness of the free acid and free base of the specified compound and has no adverse effects in biology or other aspects. The compounds of the present application also include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to the form of a basic group in a parent compound converted into a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amine (amino) groups. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound, that is, the basic group in the parent compound is reacted with 1-4 equivalents of acid in a solvent system. Suitable salts are listed in Remingtong's Pharmaceutical Scicences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p.1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

Unless otherwise indicated, the salts in the present application refer to acid salts formed with organic acids/inorganic acids, and basic salts formed with organic bases/inorganic bases.

"BTK" refers to Bruton's tyrosine kinase.

Herein, unless otherwise indicated, the dosages and ranges provided herein are calculated based on the molecular weight of the free base form of the compounds of the invention.

Unless otherwise specified, expressions such as R1, NH2, and CH3 in the text are equivalent to R ₁ , NH ₂ , and CH ₃ .

DETAILED DESCRIPTION

In order to more clearly illustrate the present invention, a series of embodiments are listed here. These embodiments are illustrative and should not be understood as limiting the present invention.

The present invention provides methods for preparing corresponding compounds. A variety of synthetic methods can be used to prepare the compounds described herein, including the methods involved in the following embodiments. The compounds of the present invention or pharmaceutically acceptable salts, isomers or hydrates thereof can be synthesized using the following methods and synthetic methods known in the field of organic chemical synthesis, or by variations of these methods understood by those skilled in the art. Preferred methods include but are not limited to the following methods.

Example 1

(4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone (DSB-001)

2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (5 g) and dichloromethane (150 mL) were added to a 250 mL round-bottom flask, and aluminum chloride (14.15 g) was added under stirring in an ice-water bath. After the addition was completed, the temperature was raised to room temperature and stirred for 0.5 h, and benzoyl chloride (9.2 mL) was added, and the temperature was raised to 40°C and stirred for 24 h. The reaction solution was poured into ice water, the aqueous phase was extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried by spin drying. 1.99 g of light yellow solid (2,4-dichloro-5-benzoyl-7H-pyrrolo[2,3-d]pyrimidine, intermediate 1a) was separated by silica gel column chromatography (petroleum ether: ethyl acetate 8:1-3:1).

Add intermediate 1a (0.82 g), potassium carbonate (1.2 g) and DMF (20 mL) to a 100 mL flask, add SEM-Cl (0.75 mL) diluted with DMF (20 mL) dropwise under stirring in an oil bath at 40°C, and maintain stirring at 40°C for 2 h. Most of the DMF is removed by rotary evaporation, and the residue is suspended in water, stirred and filtered, and the filter cake is rinsed with water and dried to obtain 0.77 g of a white solid (2,4-dichloro-5-benzoyl-7-trimethylsilylethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine, intermediate 2a).

Add intermediate 2a (0.47 g), DMF (20 mL), potassium carbonate (0.62 g) and cis-4-aminocyclohexane methanol hydrochloride (0.28 g) to a 250 mL eggplant-shaped bottle and stir in an oil bath at 90°C for 6 h. Remove most of the DMF by rotary evaporation, add water to the system, extract the aqueous phase with ethyl acetate (50 mL × 2), wash the organic phase with saturated brine, dry it with anhydrous sodium sulfate, and spin dry it. Separate it by silica gel column chromatography (petroleum ether: ethyl acetate 10:1-5:1) to obtain 0.60 g of a white solid (2-chloro-4-(4-(1s,4s)-4-hydroxymethylcyclohexane-1-amino)-5-benzoyl-7-trimethylsilylethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine, intermediate 3a).

Add intermediate 3a (200 mg), 3-amino-1-methyl-pyrazole (50 μL), Pd2(dba)3 (50 mg), X-phos (80 mg), and potassium carbonate (200 mg) to a 250 mL eggplant-shaped bottle, and stir in an oil bath at 110°C for 5 h under nitrogen protection. The reaction solution was spin-dried and separated by silica gel column chromatography (dichloromethane: methanol 50:1) to obtain 350 mg of a yellow solid (2-(1-methyl-3-pyrazolylamino)-4-(4-(1s,4s)-4-hydroxymethylcyclohexane-1-amino)-5-benzoyl-7-trimethylsilylethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine, intermediate 4a).

Add intermediate 4a (350 mg), dichloromethane (10 mL) and trifluoroacetic acid (3 mL) to a 100 mL eggplant-shaped bottle and stir at room temperature for 8 h. The reaction solution was dried by spin drying, methanol (10 mL) and ammonia water (3 mL) were added, and stirred at room temperature for 12 h. The system was poured into 100 mL of water, the aqueous phase was extracted with dichloromethane (50 mL), the dichloromethane phase was dried by spin drying, and separated by silica gel column chromatography (dichloromethane: methanol 30:1) to obtain 160 mg of yellow solid (4-(((1s, 4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone. ¹ H NMR (600 MHz, DMSO-d ₆ )δ12.23(s,1H),9.42–8.83(m,2H),7.78–7.73(m,2H),7.65–7.59(m,1H),7.57–7.50(m,3H),7.43(d,J＝2.3Hz,1H),6.66(d,J＝2.1Hz,1H),4.44(t,J ＝5.3Hz,1H),4.42–4.36(m,1H),3.74(s,3H),3.31–3.27(m,2H),1.89–1.80(m,2H),1.72–1.61(m,4H),1.58–1.48(m,1H),1.43–1.34(m,2H).MS/[M +H]+:446.2.

Examples 2-47 were carried out using the synthesis method of general formula I in combination with Example 1.

Example 48 BTK in vitro enzymatic assay

1. Preparation of 1X Kinase Buffer

One volume of 5-fold concentrated enzyme buffer was diluted with 4 volumes of distilled water; 5 mM magnesium chloride; 1 mM DTT; 1 mM MnCl ₂ .

2. Compound activity test method

a) Using Echo 655, transfer compound dilutions to each well of the assay plate (784075, Greiner);

b) Seal the assay plate and centrifuge it at 1000 g for 1 minute;

c) Prepare 2X BTK in 1X kinase buffer;

d) Add 5 μl of 2X BTK into the 384-well assay plate;

e) The test plate was centrifuged at 1000 g for 30 seconds and incubated at room temperature for 15 minutes;

f) preparing a 2X mixture of polyE4Y1 and ATP in 1X kinase buffer;

g) Add 2X 2X polyE4Y1 and ATP mixture to start the reaction;

h) Centrifuge the plate at 1000 g for 30 seconds. Seal the plate and incubate at room temperature for 1 hour;

i) Add ADP-Glo reagent and incubate at room temperature for 1 hour;

j) adding kinase detection reagent and incubating at room temperature for 1 hour;

k) The test plate was centrifuged at 1000 g for 30 seconds and incubated at room temperature for 1 hour;

l) Read the fluorescence signal on an Envision 2104 plate reader.

3. Data Analysis

3.1 Detect the signal of each well.

3.2 The inhibition rate is calculated as follows:

% inhibition rate=100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100.

3.3 Calculate the _IC50 value of the compound and draw the dose-effect curve of the compound:

GraphPad 6.0 was used to plot the logarithm of compound concentration against inhibition rate to calculate the _IC50 value.

Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC ₅₀ -X) * slope))

X: logarithm of inhibitor concentration; Y: inhibition rate.

3.4 Reporting Double Inspection

3.4.1 After one inspector completes the report, another inspector will review the report again to ensure that the data analysis is correct.

3.4.2 Data were exported from the software and analyzed manually.

3.4.2.1 Conversion of ratios to inhibition rates All _IC50 values were calculated using inhibition rates using Prism GraphPad 6.0.

3.4.2.2 The ratio was then used to recalculate the _IC50 value to check the accuracy of the analysis results.

3.4.3 Using the above method, the activity of the compounds of the present invention on BTK kinase was determined.

The results of the determination of the kinase activity of the compounds of the present invention on BTK are shown in Table 1.

Table 1 Kinase inhibitory activity IC50 (nM) of compounds against BTK

+ means less than or equal to 100 nM; ++ means less than or equal to 10 nM; +++ means less than or equal to 1 nM; nd means not tested.

The compounds of the present invention have good inhibitory activity on BTK kinase, wherein the IC50 values of the compounds listed in Table 1 are all less than 1 nM, which is equivalent to that of positive drugs.

Example 49 HCK in vitro enzymatic assay

Preparation of 1x kinase buffer

One volume of 5-fold concentrated enzyme buffer was diluted with 4 volumes of distilled water; 5 mM magnesium chloride; 1 mM DTT; 1 mM MnCl ₂ .

2. Compound activity test method

a) Using Echo 550, transfer compound dilutions to each well of the assay plate (784075, Greiner);

b) Seal the assay plate and centrifuge it at 1000 g for 1 minute;

c) Prepare 2X HCK in 1X kinase buffer;

d) Add 5 μl of HCK kinase into a 384-well assay plate (784075, Greiner);

e) The test plate was centrifuged at 1000 g for 30 seconds and incubated at room temperature for 10 minutes;

f) Add 2x TK-substrate-biotin and ATP to 1x kinase buffer and mix;

g) Add 5 μl of TK-substrate-biotin and ATP mixture to start the reaction;

h) Centrifuge the plate at 1000 g for 30 seconds. Seal the plate and incubate at room temperature for 40 minutes;

i) Prepare 4X Sa-XL 665 in HTRF detection buffer;

j) Add 5 μl Sa-XL 665 and 5 μl TK-antibody-Cryptate to each well of the detection plate;

k) The test plate was centrifuged at 1000 g for 30 seconds and incubated at room temperature for 1 hour;

l) Fluorescence signals at 615 nm (Cryptate) and 665 nm (XL665) were read on an Envision 2104 plate reader.

3. Data Analysis

3.1 Calculate the ratio (665/615 nm) for each well.

3.2 The inhibition rate is calculated as follows:

% inhibition rate=100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100.

3.3 Calculate the IC50 value of the compound and draw the dose-effect curve of the compound:

GraphPad 6.0 was used to plot the logarithm of compound concentration against inhibition rate to calculate the IC50 value.

Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50-X) * slope))

X: logarithm of inhibitor concentration; Y: inhibition rate.

3.4 Reporting Double Inspection

3.4.1 After one inspector completes the report, another inspector will review the report again to ensure that the data analysis is correct.

3.4.2 Data were exported from the software and analyzed manually.

3.4.2.1 Conversion of ratios to inhibition rates All IC50 values were first calculated using inhibition rates using Prism GraphPad 6.0.

3.4.2.2 The IC50 value was then recalculated using the ratio to check the accuracy of the analysis results.

3.4.3 Using the above method, the activity of the compounds of the present invention on HCK kinase was determined.

The results of the determination of the kinase activity of the compounds of the present invention on HCK are shown in Table 2.

Table 2 Kinase inhibitory activity IC50 (nM) of compounds against HCK

+ means less than or equal to 100 nM; ++ means less than or equal to 10 nM; +++ means less than or equal to 1 nM; nd means not tested.

The compounds of the present invention have good inhibitory activity on HCK kinase, wherein the IC50 values of the compounds listed in Table 2 are all less than 1 nM, which is better than positive drugs.

Example 50 BTK-C481S in vitro enzymatic assay

The method was the same as the BTK kinase activity screening experiment in Example 48, except that BTK-C481S kinase was used to replace BTK kinase. The results are shown in Table 3.

Table 3 Kinase inhibitory activity of compounds against BTK-C481S (IC50, nM)

+ means less than or equal to 100 nM; ++ means less than or equal to 10 nM; +++ means less than or equal to 1 nM; ++++ means less than or equal to 0.1 nM; nd means not tested.

The compounds of the present invention have good inhibitory activity against BTK-C481S kinase, wherein the IC50 values of the compounds listed in Table 3 are mostly less than 1 nM, and the inhibitory activity of some compounds is in the range of 0.01-0.1 nM, which is better than the positive drugs Ibrutinib and Fenebrutinib (GDC-0853).

Example 51 Cell proliferation inhibition experiment

1. Experimental Grouping

Experimental group (As): cells + culture medium + drugs + cck-8; control group (A ₀ ): cells + culture medium + cck-8, no drugs; blank group (Ac): culture medium + cck-8, no cells and drugs.

2. Experimental cell lines

TMD8, human diffuse large B-lymphoma cells;

K562, human chronic myeloid leukemia cells;

Jeko-1, human mantle lymphoma cells;

Raji, human Burkitt’s lymphoma cells;

CEM/C1, human acute lymphoblastic leukemia cells;

PC-3, human prostate cancer cells;

DU145, human prostate cancer cells;

LNCaP, human prostate cancer cells;

BGC-283, human gastric cancer cells.

3. Cell Culture

3.1 Cell recovery

Take out the cryopreservation tube of the above cells from liquid nitrogen, immediately put it into 37°C warm water and shake it quickly until the cryopreservation solution is completely dissolved; transfer the cell cryopreservation solution to a centrifuge tube, add about 3 ml of culture solution, gently blow to mix and centrifuge at 1000r/min for 5 minutes; discard the supernatant, add an appropriate amount of culture solution and transfer to a culture bottle for culture.

3.2 Cell passaging

When the cell coverage rate in the culture bottle reaches 80%-90%, the original culture medium is sucked off and an appropriate amount of trypsin is added; after the cells become round, the trypsin is discarded, an appropriate amount of culture medium is added and mixed with a pipette; an appropriate amount of cell suspension is discarded and an equal amount of culture medium is added, and the cells are placed in an incubator for continued culture.

4. Effects of drugs on cell lines

Take the above-mentioned cells in logarithmic growth phase, adjust the density to 1500-20000 cells/ml with culture medium, 100μl per well, inoculate in 96-well plate, and add 200μl physiological saline to each well at the outermost periphery. After 6-30 hours of culture in the incubator, add different concentrations of compounds, 3 replicates for each drug concentration, and administer 100μl per well. Culture for 72h, then add CCK-8 solution, 15μl/well. After continuing to culture for 2h, detect the absorbance value at a wavelength of 450nm with an enzyme marker. Inhibition rate % = [(A0-AS)/(A0-AC)]×100%. Use Prism GraphPad 5.0 software to calculate the IC ₅₀ value of the compound on cells.

Table 4 Antiproliferative activity of compounds against TMD8 (IC50, nM)

+ means less than or equal to 100 nM; ++ means less than or equal to 10 nM; +++ means less than or equal to 1 nM; nd means not tested.

The compounds of the present invention have good antiproliferative activity against TMD8, wherein the IC50 values of the compounds listed in Table 4 are less than 100 nM, and the antiproliferative activity of most of the compounds is in the range of 0.1-10 nM, which is better than that of positive drugs.

Table 5 In vitro anti-cell proliferation activity (IC50, nM) of DSB-013, DSB-016 and DSB-026

nd, means not tested.

Claims (10)

1. A compound as shown in formula I, its stereoisomer or a pharmaceutically acceptable salt thereof: X is NH, C or O; X1, X2, W, W2, W3 are independently selected from C, N, O; a, b, and m represent the number of W, W2, and W3, respectively, and are independently selected from 0, 1, 2, or 3; n is 0 or 1; e is a chemical bond, representing any reasonable conformation; R1 is selected from H, halogen, C1-3 alkyl; the number of R1 is 1, 2 or 3, and R1 is located at any reasonable position of the benzene ring; R2 is selected from substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C1-6 alkoxy, substituted or unsubstituted C3-8 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group; the substituent is selected from C1-6 alkyl, C1-6 alkoxy, 5-8 membered heterocyclic group, wherein the 5-8 membered heterocyclic group may be further substituted by C1-6 alkyl; Said Ra and Rb are independently selected from C1-6 alkyl; R3 is selected from H, substituted or unsubstituted C1-6 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the substituent of the C1-6 alkyl is selected from OH, NH2, -OC(=O)Rc; the Rc is selected from C1-6 alkyl, C2-6 alkenyl, C3-8 cycloalkyl; R4 is selected from H, halogen, C1-6 alkyl or C1-6 alkoxy; The heterocyclic group contains 1-3 heteroatoms, and the heteroatoms are selected from N, O, and S. 2. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that W is C, a is 1, as shown in Formula II, Wherein, X is NH or O, b is 1, m is 0 or 1, W2 and W3 are independently selected from C, N, and O; R1 is selected from H or halogen, the number of R1 is 1 or 2, and R1 is located at any reasonable position of the benzene ring; X1 and X2 are independently selected from C or N; n is 0 or 1; R2 is selected from substituted or unsubstituted C1-6 alkyl, C3-6 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group. The substituent of the C1-6 alkyl is selected from C1-4 alkoxy, 5-8 membered heterocyclic group, The substituent of the 5-8 membered heterocyclic group is a C1-6 alkyl group; Said Ra and Rb are independently selected from C1-4 alkyl; R3 is selected from H, substituted or unsubstituted C1-4 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the Rc is selected from C1-4 alkyl, C2-4 alkenyl, C3-6 cycloalkyl, and the substituent of the C1-6 alkyl is selected from OH, NH2; R4 is selected from H, halogen or C1-3 alkoxy. 3. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that W is C, a is 1, and X is NH, as shown in Formula III, wherein b is 1, m is 0 or 1, W2 and W3 are independently selected from C, N, and O; R1 is selected from H or halogen, the number of R1 is 1 or 2, and R1 is located at any reasonable position of the benzene ring; X1 and X2 are independently selected from C or N; n is 0 or 1; R2 is selected from substituted or unsubstituted C1-6 alkyl, C3-6 cycloalkyl, substituted or unsubstituted 5-8 membered heterocyclic group. The substituent of the C1-6 alkyl is selected from C1-4 alkoxy, 5-8 membered heterocyclic group, The substituent of the 5-8 membered heterocyclic group is a C1-6 alkyl group; Said Ra and Rb are independently selected from C1-4 alkyl; R3 is selected from H, substituted or unsubstituted C1-4 alkyl, -NHCORc, -C(=O)Rc, -NHS(O)2Rc; the Rc is selected from C1-4 alkyl, C2-4 alkenyl, C3-6 cycloalkyl, and the substituent of the C1-6 alkyl is selected from OH, NH2; R4 is selected from H, halogen or C1-3 alkoxy. 4. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that R4 is H, n is 0, W is C, a is 1, and X is N, as shown in Formula IV, in, R1 is selected from H, halogen, C1-3 alkyl; R2 is selected from substituted or unsubstituted C1-4 alkyl, substituted or unsubstituted C1-4 alkoxy; the substituent is selected from C1-3 alkyl, C1-3 alkoxy, 5-8 membered heterocyclic group; the heterocyclic group contains 1-2 heteroatoms selected from N, O, and S; the Ra and Rb are independently selected from C1-3 alkyl; X1, X2, W2, W3 are independently selected from C, N, O; b and m are independently 1 or 2; R3 is selected from substituted or unsubstituted C1-3 alkyl, and the substituent is OH, NH2. 5. The compound according to claim 4, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that X1 and X2 are N, as shown in Formula V, Wherein, R1 is selected from H, halogen, R1 is located in the para position of the benzene ring; R2 is selected from substituted or unsubstituted C1-4 alkyl, substituted or unsubstituted C1-4 alkoxy; the substituent is selected from C1-3 alkyl, 5-6 membered heterocyclic group; the heterocyclic group contains 1-2 heteroatoms selected from N, O, and S; the Ra and Rb are independently selected from C1-3 alkyl; X1 and X2 are N; W2 and W3 are C; b and m are independently 1 or 2. 6. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, selected from the following compounds: (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-(3-morpholinopropyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (2-((1-(2-(diethylamino)ethyl)-1H-pyrazol-4-yl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-(dimethylamino)piperidin-1-yl)phenyl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; 2-((4-(4-(dimethylamino)piperidin-1-yl)-2-methoxyphenyl)amino)-4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; N-((1s,4s)-4-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; N-((1s,4s)-4-((2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-(-4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-fluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (3,4-difluorophenyl)(4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((1-cyclobutyl-1H-pyrazol-4-yl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(4-fluorophenyl)methanone; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(4-fluorobenzoyl)-2-((1-(2-morpholinoethyl)-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((2-((1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; (R)-1-(3-((5-(2,4-difluorobenzoyl)-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)pyrrolidin-1-yl)prop-2-en-1-one; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)butanamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)cyclopropanecarboxamide; (2-Fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(2-(dimethylamino)ethoxy)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)methanesulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)acetamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(piperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propanamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propane-1-sulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)cyclopropanesulfonamide; N-((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((5-methyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((5-isopropyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (2-((5-cyclopropyl-1H-pyrazol-3-yl)amino)-4-((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; N-((1r,4r)-4-((2-((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; N-((1r,4r)-4-((2-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-5-(4-fluorobenzoyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; (3-Fluorophenyl)(4-(((1r,4r)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; N-((1r,4r)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyramide; (4-Fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydro-2H-pyran-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (R)-(4-Fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydrofuran-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (S)-(4-Fluorophenyl)(2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4-((tetrahydrofuran-3-yl)oxy)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)methanone; (1s,4s)-methyl 4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)isobutyrate; (1s,4s)-methyl 4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)propanoate; Methyl ((1s,4s)-4-((5-(4-fluorobenzoyl)-2-((4-(4-methylpiperazin-1-yl)phenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclohexyl)acetate; (4-(((1s,4s)-4-(hydroxymethyl)cyclohexyl)amino)-2-((4-morpholinophenyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone; (4-(((1s,4s)-4-(Hydroxymethyl)cyclohexyl)amino)-2-((1-methyl-1H-pyrazol-3-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)(phenyl)methanone. 7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients. 8. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7 in the preparation of a drug for preventing and treating BTK-related diseases. 9. The use according to claim 8, characterized in that the disease is an autoimmune disease or a tumor. 10. The use according to claim 9, characterized in that the tumor is selected from hematopoietic or lymphoid tissue tumors, prostate cancer, and gastric cancer.