CN112608318A

CN112608318A - Compound serving as protein kinase inhibitor and application thereof

Info

Publication number: CN112608318A
Application number: CN202011491197.5A
Authority: CN
Inventors: 刘冠锋; 岳春超; 李筛; 李英富; 原晨光
Original assignee: Chengdu Haibowei Pharmaceutical Co ltd
Current assignee: Chengdu Haibowei Pharmaceutical Co ltd
Priority date: 2019-12-16
Filing date: 2020-12-16
Publication date: 2021-04-06
Anticipated expiration: 2040-12-16
Also published as: CN112608318B

Abstract

The invention discloses a compound as a protein kinase inhibitor and application thereof, wherein the compound has obvious inhibition effect on the activity of protein kinase, can be used as a BTK inhibitor, is used for preparing medicines for treating BTK-mediated diseases such as malignant tumors, autoimmune diseases and the like, and has wide application prospect.

Description

Compound serving as protein kinase inhibitor and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a compound serving as a protein kinase inhibitor and application thereof.

Background

Bruton's tyrosine protein kinase (BTK) is a member of the non-receptor protein tyrosine kinase Tec family, and is predominantly expressed in a variety of hematopoietic cell lines. The Tec family is the second 2 large family of human non-receptor kinases to the Src family, the major members of which include BTK, BMX (etk), ITK, Tec, and txk (rlk). BTK was identified in 1993 as a defective protein in human X-linked agammaglobulinemia (XLA). BTK is a key regulator of a B Cell Receptor (BCR) signal transduction pathway, plays an important role in the processes of B cell activation, proliferation, differentiation and survival, and is closely related to various B cell tumors and autoimmune diseases.

BTK contains 5 major domains in its structure, the PH domain (Pleckstrin homology), the TH domain (Tec homology), the SH3 domain (Src homology 3), the SH2 domain (Src homology 2) and the SHl domain (Src homology1), respectively, and its activation (phosphorylation) occurs initially in the activation loop in the SHl domain, and further activation occurs in the SH2 and SH3 domains that contain major autophosphorylation sites. These SH domains also contain Nuclear Localization Signals (NLS) and Nuclear Export Sequences (NES) required for BTK to shuttle nuclear matter.

BTK plays an irreplaceable role in the generation process of B lymphocytes, controls the development and differentiation of B cells by activating cell cycle positive regulatory factors and differentiation factors, and also controls the survival and proliferation of the B cells by regulating the expression of pro-apoptotic and anti-apoptotic proteins. Sustained activation of BTK is a prerequisite for the development of Chronic Lymphocytic Leukemia (CLL). Aberrant BCR-BTK signaling promotes survival of activated B-cell subtypes in diffuse large B-cell lymphoma (DLBCL). Mutations of the BTK gain-of-function type have also been confirmed in colorectal cancer, Acute Lymphoblastic Leukemia (ALL), and Chronic Myelogenous Leukemia (CML). Thus, aberrant activation of BTK-dependent pathways has been shown to be closely associated with the development of multiple tumors.

The approved irreversible BTK inhibitors on the market, such as Ibrutinib (Ibrutinib), acarabtinib (acalaburtinib) and zanibrutinib (Zanburtinib), can selectively form irreversible covalent bond with cysteine residue (Cys-481) of BTK, and inhibit the activity of BTK to achieve the purpose of treating related diseases. However, a subset of cancer patients develop resistance to the first generation of BTK inhibitors, and thus an unmet new clinical need arises. Research evidence shows that the BTK-C481S mutation is one of the main drug resistance mechanisms related to the BTK-C481S mutation, so that a drug capable of targeting and inhibiting the BTK-C481S mutation is expected to provide a new treatment scheme.

Disclosure of Invention

The invention mainly solves the technical problem of providing a compound which can effectively inhibit protein kinase.

In order to solve the technical problems, the invention adopts a technical scheme that:

providing a compound having the structure of formula (I) or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, pharmaceutically acceptable hydrate, solvate, or salt thereof:

wherein:

A₁、A₂、A₃、A₄are each independently selected from CR₉、N；

Y is selected from CR₁₀ R₁₁、O、NR₁₀、S、S(O)、S(O)₂C-S-O, C-alkynyl;

m is selected from substituted or unsubstituted aryl or heteroaryl, substituted or unsubstituted cycloalkyl or heterocycloalkyl;

R₁、R₂、R₉、R₁₀、R₁₁each independently selected from hydrogen, halogen, substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, nitro, mercapto, amide, sulfonyl, phosphoryl, alkyl phosphoroxy, alkyl sulfone, alkyl sulfoxide; or R₁、R₂Together with the N atom to which they are attached, form a substituted or unsubstituted heterocycloalkyl;

the substituent groups are respectively and independently selected from halogen, alkyl or heteroalkyl, cycloalkyl or heterocycloalkyl, methylene, unsaturated cyclic or heterocyclic group, aryl, heteroaryl, phenoxy, hydroxyl, cyano, amino, ester group, nitro, sulfydryl, amido, sulfonyl, phosphoryl, alkyl oxyphosphoryl, alkyl sulfonyl and alkyl sulfoxide;

the cycloalkyl or cyclic group is selected from monocyclic, fused, bridged or spiro cycloalkyl or cyclic group;

R₁、R₂not hydrogen at the same time.

The heterocycloalkyl or heterocyclic group means a cycloalkyl or cyclic group containing a heteroatom, and thus the heterocycloalkyl or heterocyclic group is also selected from monocyclic, fused, bridged or spiro heterocycloalkyl or heterocyclic groups.

"Ring" refers to any covalently closed structure, including, for example, carbocycles (e.g., aryl or cycloalkyl), heterocycles (e.g., heteroaryl or heterocycloalkyl), aryls (e.g., aryl or heteroaryl), nonaromatic (e.g., cycloalkyl or heterocycloalkyl). The rings may be optionally substituted, and may be monocyclic or polycyclic. Typical polycyclic rings generally include bicyclic and tricyclic rings.

The "cycloalkyl" as used herein refers to a saturated cyclic structure (monocyclic, fused, bridged or spiro) alkyl group, and the "unsaturated cyclic group" refers to a cyclic structure (monocyclic, fused, bridged or spiro) hydrocarbon group containing an unsaturated bond (e.g., a carbon-carbon double bond) on the cyclic skeleton, and typical cycloalkyl or unsaturated cyclic groups include, but are not limited to:

similarly, the heterocycloalkyl or unsaturated heterocyclyl group may contain a heteroatom at any position on the above-described structural backbone, and typical heterocycloalkyl or heterocyclyl groups include, but are not limited to:

the methylene group is CH₂And a substituted carbon atom to form a double-bonded terminal alkene.

Further, A₁、A₂、A₃、A₄At least one of which is CR₉。

Further, the air conditioner is provided with a fan,

selected from substituted or unsubstituted indolyl, 7-azaindolyl, 5, 7-diazaindolyl and pyrazolopyrimidinyl, wherein the indolyl, 7-azaindolyl, 5, 7-diazaindolyl and pyrazolopyrimidinyl have the following structures:

further, said R₉Selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, hydroxyl, cyano, amino, ester, amide, sulfonyl, phosphoryl, alkylphosphoryl, alkylsulfone, and alkylsulfide;

further, said R₉Selected from the group consisting of hydrogen, halogen, methyl, trifluoromethyl, difluoromethyl, isopropyl, cyclopropyl, isobutyl, amide, dimethylphosphite, diethyloxyphosphite, cyano, hydroxy, amino, methoxy, ethoxy, cyclopropoxy, isobutoxy, methylamino, dimethylamino, ethylamino, diethylamino, cyclopropylamino, methylsulfoxide, methylsulfone;

further, said R₉Selected from hydrogen, halogen, C1-C3 alkyl, trifluoromethyl, difluoromethyl, methoxy, amido, dimethyl phosphoroso, cyano.

Further, the compound of the present invention has the structure shown in formula (II) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt thereof:

R₃selected from the group consisting of hydrogen, halogen, substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amide, substituted or unsubstituted aryl or heteroaryl, sulfonyl, phosphoryl, alkylphosphoryl, wherein the substituents are each independently selected from the group consisting of halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amide, aryl, heteroaryl, sulfonyl, phosphoryl, alkylphosphoryl, alkylsulfoxide, and alkylsulfoxide.

Further, A₁Or A₂Is CR₉，A₃Is N;

further, A₁Is CR₉，A₃Is N.

Further, R₁Selected from substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted aryl or heteroaryl; further, R₁Selected from substituted or unsubstituted alkyl, substituted or unsubstituted C3-15 cycloalkyl or heterocycloalkyl, substituted or unsubstituted C3-15 unsaturated heterocyclyl and substituted or unsubstituted heteroaryl, wherein hetero atoms in the heterocycloalkyl or heterocyclyl are selected from N, O, S;

R₂selected from hydrogen, halogen, substituted or unsubstituted alkyl or heteroalkyl; further, R₂Is H.

Further, R₁The compound is selected from substituted or unsubstituted alkyl, substituted or unsubstituted C3-15 cycloalkyl or oxygen-containing heterocycloalkyl, substituted or unsubstituted C3-15 unsaturated oxygen-containing heterocyclic group and substituted or unsubstituted nitrogen-containing heteroaryl, wherein the substituents are respectively and independently selected from halogen, C1-6 alkyl, C3-6 cycloalkyl, methylene and hydroxyl;

further, R₁Selected from substituted or unsubstituted alkyl, substituted or unsubstituted adamantyl, substituted or unsubstituted oxoheterocycloalkyl, substituted or unsubstituted dihydropyranSubstituted or unsubstituted nitrogen-containing heteroaryl, said oxygen-containing heterocycloalkyl being selected from tetrahydropyranyl, 6-oxaspiro [2.5]]Octyl, 3-oxabicyclo [4.1.0]]A heptylalkyl group, wherein the nitrogen-containing heteroaryl group is selected from pyridyl and pyrazinyl, wherein the substituents are independently selected from halogen, C1-C6 alkyl substituted by one or more hydroxyl groups, C1-C6 alkyl substituted by nitrogen-containing heterocycloalkyl groups, hydroxycyclopropyl, methylene and hydroxyl; further, the dihydropyran is 3, 6-dihydropyran.

The tetrahydropyran, 6-oxaspiro [2.5] octane, 3-oxabicyclo [4.1.0] heptane and 3, 6-dihydropyran have the following structures:

in the present invention

Tetrahydropyran substituted in position 4 by methylene, in R₁Falls within the scope of substituted tetrahydropyranyl.

Further, the compound of the present invention has the structure shown in formula (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt thereof:

e is selected from O, NR₁₂、S、S(O)、S(O)2、C＝O、C＝S；

n₁An integer selected from 0 to 8;

R₄selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, methylene, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, sulfonyl, phosphoryl, or two adjacent R₄To carbon atoms bound theretoThe two groups together form a substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl group, or R at two same sites₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄Together with the carbon atom to which they are attached form a carbon-carbon double bond, or two non-adjacent R₄Together form a bridged ring structure, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxy, cyano, amino, ester, amide, aryl, heteroaryl, sulfonyl;

"substituted or unsubstituted C1-6 alkyl or heteroalkyl" refers to: substituted or unsubstituted C1-6 alkyl, and substituted or unsubstituted C1-6 heteroalkyl, the same holds true for the rest.

Said "two adjacent R₄"means that the substituent R is at a carbon atom at two adjacent sites on the 6-membered ring backbone containing E₄Such as

The situation shown;

said "two non-adjacent R₄"means that the substituent R is at a carbon atom at two non-adjacent sites on the 6-membered ring backbone containing E₄Such as

Etc.;

said "R at two identical sites₄"means 2 substituents R on a carbon atom at a site on the 6-membered ring backbone containing E₄Such as

The situation shown;

said "two adjacent R₄The carbon atoms to which they are attached together form a carbon-carbon double bond "means that the substituents R on the carbon atoms at two adjacent sites on the 6-membered cyclic skeleton containing E₄Together with the two carbon atoms forming a carbon-carbon double bond, e.g.

Etc.;

when two adjacent R₄When the carbon atoms to which they are attached together form a carbon-carbon double bond, n is understood to mean₁Is 2.

R₃The aryl or heteroaryl substituted by C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, substituted or unsubstituted aryl or heteroaryl, sulfonyl, phosphoryl, and alkyl oxyphosphoryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl, sulfonyl, phosphoryl, alkyl oxyphosphoryl, alkyl sulfonyl, and alkyl sulfoxide;

R₁₂the aryl group is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, and substituted or unsubstituted aryl or heteroaryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl and heteroaryl.

Further, A₁Is CR₉Or N, A₃Is N;

e is selected from O;

n₁an integer selected from 0 to 6;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or R at two same sites₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, naphthenic base and heteroalkylA group, heterocycloalkyl, hydroxy, cyano, amino, ester, amide, aryl, heteroaryl, sulfonyl;

R₃selected from the group consisting of hydrogen, halogen, methyl, trifluoromethyl, difluoromethyl, isopropyl, cyclopropyl, isobutyl, amide, dimethylphosphite, diethyloxyphosphite, cyano, hydroxy, amino, methoxy, ethoxy, cyclopropoxy, isobutoxy, methylamino, dimethylamino, ethylamino, diethylamino, cyclopropylamino, methylsulfoxide, methylsulfone;

further, n₁An integer selected from 0 to 4;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or R at two same sites₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

further, R₄Selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, alkyl of C1-6 substituted by hydroxyl, cycloalkyl of C3-6 substituted by hydroxyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or R at two same sites₄The substituent and the carbon atom connected with the substituent form a substituted or unsubstituted C3-10 cycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

further, M is selected from substituted or unsubstituted aryl or heteroaryl, wherein the substituent is selected from halogen, alkyl or heteroalkyl, C3-C6 cycloalkyl or heterocycloalkyl, unsaturated cyclic or heterocyclic group, aryl, heteroaryl, phenoxide, hydroxyl, cyano, amino, ester, amide, sulfonyl, phosphoryl, alkyl oxyphosphoryl;

further, M is selected from substituted or unsubstituted aryl, wherein the substituent is selected from halogen, alkyl or heteroalkyl, C3-C6 cycloalkyl or heterocycloalkyl, unsaturated cyclic or heterocyclic group, phenol oxy, amide group.

Further, the compound of the present invention has the structure shown in formula (IV) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture form, pharmaceutically acceptable hydrate, solvate or salt thereof:

b is selected from O,

Wherein R is₁₃、R₁₄、R₁₅Independently selected from hydrogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or with R₆And atoms which are connected together form a substituted or unsubstituted C5-8 cycloalkyl or heterocycloalkyl group, a substituted or unsubstituted C5-8 unsaturated cyclic or heterocyclic group, an aryl or heteroaryl group, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl and sulfonyl;

R₅the aryl-substituted aryl group is selected from hydrogen, halogen, hydroxyl, cyano, amino, substituted or unsubstituted C1-6 alkyl or heteroalkyl, and substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester group, amido, aryl, heteroaryl and sulfonyl; n is₂Selected from 0, 1,2, 3, 4;

R₆is selected from the group consisting ofSubstituted or unsubstituted monocyclic, bicyclic, tricyclic aryl or heteroaryl, wherein the substituents are selected from nitro, hydroxyl, amino, mercapto, halogen, cyano, ester, carboxyl, amido, phosphamido, alkyl phospho, C1-6 alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl, 5-6 membered aryl or heteroaryl.

Further, A₁Is CR₉；

n₁An integer selected from 0 to 4;

b is selected from O,

R₁₃Selected from hydrogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or with R₆And atoms connected together form substituted or unsubstituted C5-8 heterocycloalkyl, substituted or unsubstituted C5-8 unsaturated heterocyclyl, heteroaryl, R₁₄、R₁₅Selected from hydrogen, substituted or unsubstituted C1-6 alkyl orThe aryl group comprises a heteroalkyl group, a substituted or unsubstituted C3-6 cycloalkyl group or a heterocycloalkyl group, wherein the substituents are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester group, amido, aryl, heteroaryl and sulfonyl;

R₅the halogen-free substituted or unsubstituted C1-6 alkyl or heteroalkyl, wherein the substituents are independently selected from halogen and alkyl; n is selected from 0, 1 and 2;

R₆the aryl is selected from substituted or unsubstituted monocyclic or bicyclic aryl or heteroaryl, wherein the substituent is selected from hydroxyl, halogen, cyano, C1-6 alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl.

In a particular embodiment of the invention, B is selected from O,

R₁₄、R₁₅Selected from hydrogen, substituted or unsubstituted C1-6 alkyl, or B and R₆Are formed together

R₅Selected from hydrogen, halogen; n is₂Selected from 0, 1;

R₆the aryl is selected from substituted or unsubstituted phenyl, wherein the substituent is selected from halogen, cyano, C1-6 alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl.

Further, when R is₁In the case of substituted or unsubstituted tetrahydropyranyl, the compound of the invention has the structure shown in formula (V) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable hydrate, solvate or salt thereof:

n₁an integer selected from 0 to 7;

R₃the aryl-substituted aryl group is selected from hydrogen, halogen, hydroxyl, cyano, amino, substituted or unsubstituted C1-6 alkyl or heteroalkyl, and substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester group, amido, aryl, heteroaryl and sulfonyl;

R₈the compound is selected from hydrogen, nitro, hydroxyl, amino, sulfydryl, halogen, cyano, ester group, carboxyl, amido, phosphamido, alkyl oxyphosphoryl, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, and substituted or unsubstituted 5-6-membered aryl or heteroaryl, wherein the substituents are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester group, amido, aryl, heteroaryl and sulfonyl; n is₃An integer selected from 0 to 5;

R₇the aryl group is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, sulfonyl and phosphoryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl and sulfonyl;

further, A₁When is N, N₁+n₃≥2。

Further, A₁Is CR₉。

Further, n₁An integer selected from 0 to 3;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or R at two same sites₄To which the carbon atoms are attached to form substituted or unsubstituted C3E10 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

R₅selected from hydrogen, halogen;

R₈the aryl group is selected from hydrogen, nitro, hydroxyl, amino, halogen, cyano, amido, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl and sulfonyl;

R₇the aryl group is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amide, aryl, heteroaryl and sulfonyl.

Further, the air conditioner is provided with a fan,

n₁an integer selected from 0 to 2;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, C1-6 alkyl substituted by hydroxyl or halogen, C3-6 cycloalkyl substituted by hydroxyl or halogen, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-6 cycloalkyl or R at two same sites₄The substituent and the carbon atom connected with the substituent form a substituted or unsubstituted C3-6 cycloalkyl, or two adjacent R₄The carbon atoms connected with the carbon-carbon double bond,wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxy, cyano;

R₅selected from hydrogen, halogen;

R₈selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, hydroxy, cyano, wherein the substituents are selected from halogen, alkyl or heteroalkyl, cycloalkyl or heterocycloalkyl, hydroxy, cyano; n is₃Is selected from 0 or 1;

R₇the substituent is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, and hydroxyl, wherein the substituent is selected from halogen, alkyl or heteroalkyl, cycloalkyl or heterocycloalkyl, and hydroxyl.

In a particular embodiment of the present invention,

R₄selected from hydrogen, halogen, hydroxyl, cyano, methylene, C1-6 alkyl substituted by hydroxyl or halogen, C3-6 cycloalkyl substituted by hydroxyl or halogen, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-6 cycloalkyl or R at two same sites₄The substituent and the carbon atom connected with the substituent form a substituted or unsubstituted C3-6 cycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

R₅selected from hydrogen, halogen;

R₈selected from hydrogen, halogen, C1-6 alkyl, C3-6 cycloalkyl and cyano;

R₇selected from hydrogen, halogen, C1-6 alkyl substituted by hydroxyl or halogen, and C3-6 cycloalkyl substituted by hydroxyl or halogen; further, R₇Selected from hydroxymethyl and hydroxycyclopropyl.

Further, R₄Selected from hydrogen, halogen, hydroxy, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-6Cycloalkyl, or two adjacent R₄And the carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano.

Further, R₅Is halogen, R₈Is hydrogen.

In some embodiments of the invention, R₄Selected from hydrogen, F, hydroxy, or two adjacent R₄The carbon atom to which it is attached constituting a substituted or unsubstituted cyclopropyl group, or two adjacent R₄Together with the carbon atoms to which they are attached form a carbon-carbon double bond; further, R₅Is Cl, R₈Is hydrogen.

Specifically, the compound structure is selected from one of the following:

the invention also provides a medicinal composition, and the active ingredients of the medicinal composition are selected from one or more than two of the compounds or the stereoisomer, the solvate, the hydrate, the pharmaceutically acceptable salt or the eutectic combination thereof.

The invention also provides the use of the compound or the stereoisomer, the solvate, the hydrate, the pharmaceutically acceptable salt or the eutectic crystal thereof in the preparation of a protein kinase inhibitor; further, the kinase inhibitor is a BTK inhibitor.

The invention also provides the use of the above compound or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof in the manufacture of a medicament for the treatment of any one or more of an autoimmune disease, an inflammatory disease, a thromboembolic disease, an allergy, an infectious disease, a proliferative disorder and cancer.

Further, the disease is selected from: arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, crohn's disease, dermatitis, asthma, sjogren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, skin rash, anti-neutrophil cytoplasmic antibody vasculitis, pemphigus vulgaris, chronic obstructive pulmonary disease, psoriasis; breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular cancer, gastric cancer, glioma, endometrial cancer, melanoma, renal cancer, bladder cancer, melanoma, bladder cancer, biliary tract cancer, renal cancer, pancreatic cancer, lymphoma, hairy cell cancer, nasopharyngeal cancer, pharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, lung cancer, non-small cell lung cancer, small cell cancer, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular cancer, hodgkin's leukemia, bronchial cancer, thyroid cancer, uterine corpus cancer, cervical cancer, multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, chronic lymphoid leukemia, Myeloid leukemia, non-hodgkin's lymphoma, primary macroglobulinemia.

The invention also provides the use of the compound or the stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or eutectic crystal thereof in the preparation of medicaments for treating diseases causing over-expression of BTK kinase.

The invention also provides application of the compound or a stereoisomer, a solvate, a hydrate, a pharmaceutically acceptable salt or a co-crystal thereof in preparing a medicament for treating diseases caused by over-expression of BTK kinase. .

The pharmaceutical composition containing the compound of the invention or the stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or cocrystal thereof can contain pharmaceutically acceptable auxiliary materials.

As used herein, "pharmaceutically acceptable" is meant to include any material that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered.

The pharmaceutically acceptable auxiliary materials are general names of all the additional materials except the main medicine in the medicine, and the auxiliary materials have the following properties: (1) no toxic effect on human body and few side effects; (2) the chemical property is stable and is not easily influenced by temperature, pH, storage time and the like; (3) has no incompatibility with the main drug, and does not influence the curative effect and quality inspection of the main drug; (4) does not interact with the packaging material. The auxiliary materials in the invention include, but are not limited to, a filler (diluent), a lubricant (glidant or anti-adhesion agent), a dispersing agent, a wetting agent, an adhesive, a regulator, a solubilizer, an antioxidant, a bacteriostatic agent, an emulsifier, a disintegrating agent and the like. The binder comprises syrup, acacia, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethyl cellulose or hydroxypropyl methylcellulose), gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone; the filler comprises lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricant comprises superfine silica gel powder, magnesium stearate, talcum powder, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; the disintegrating agent comprises starch and its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; the wetting agent comprises sodium lauryl sulfate, water or alcohol, etc.; the antioxidant comprises sodium sulfite, sodium bisulfite, sodium pyrosulfite, dibutylbenzoic acid, etc.; the bacteriostatic agent comprises 0.5% of phenol, 0.3% of cresol, 0.5% of chlorobutanol and the like; the regulator comprises hydrochloric acid, citric acid, potassium (sodium) hydroxide, sodium citrate, and buffer (including sodium dihydrogen phosphate and disodium hydrogen phosphate); the emulsifier comprises polysorbate-80, sorbitan fatty acid, pluronic F-68, lecithin, soybean lecithin, etc.; the solubilizer comprises Tween-80, bile, glycerol, etc. The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. The acid base is a generalized Lewis acid base. Suitable acids for forming the salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils, or mixtures of such materials, and the like.

In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the invention can likewise be used in injectable preparations. Wherein the injection is selected from liquid injection (water injection), sterile powder for injection (powder injection) or tablet for injection (refers to impression tablet or machine pressing tablet prepared by aseptic operation method of medicine, and is dissolved with water for injection for subcutaneous or intramuscular injection when in use).

Wherein the powder for injection contains at least an excipient in addition to the above compound. The excipients, which are components intentionally added to a drug in the present invention, should not have pharmacological properties in the amounts used, however, the excipients may aid in the processing, dissolution or dissolution of the drug, delivery by a targeted route of administration, or stability.

"alkyl" refers to an aliphatic hydrocarbon group and to a saturated hydrocarbon group. The alkyl moiety may be a straight chain or branched chain alkyl.

The C1-n used in the invention comprises C1-2 and C1-3 … … C1-n. n is an integer greater than one. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, and the like.

"heteroalkyl" refers to an alkyl group containing a heteroatom, including, but not limited to O, S, N, P, and the like; alkoxy, thioalkyl, aminoalkyl and the like are all intended to be heteroalkyl.

In the present invention, the term "C1-n" represents a heteroalkyl group, where n is an integer greater than one and refers to the number of carbon atoms in the heteroalkyl group.

In the present invention, in the "cycloalkyl group or heterocycloalkyl group having C3-6", when the definition of the heterocycloalkyl group is defined by "C3-6", the definition of the heterocycloalkyl group is to be understood as 3-6 membered heterocycloalkyl, and the similar definitions of the heterocycloalkyl group are the same.

"amido" is a chemical structure having the formula-C (O) NHR or-NHC (O) R, wherein R is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl.

"ester group" means a chemical structure having the formula-COOR or-OC (O) R, wherein R is selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl.

"Sulfonyl" is a compound having the formula-S (═ O)₂The chemical structure of R, including sulfonamide, wherein R can be selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, amino, and the like;

"phosphoryl" is a chemical structure having the formula-P (═ O) RR ', where R, R' may each be independently selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, hydroxy, amino, and the like;

"alkynyl" refers to a group having a carbon-carbon triple bond.

"element" means the number of skeleton atoms constituting a ring. Typical 5-membered rings may include cyclopentyl, pyrrole, imidazole, thiazole, furan, thiophene, and the like; typical 6-membered rings include cyclohexyl, pyridine, pyran, pyrazine, thiopyran, pyridazine, pyrimidine, benzene, and the like. Wherein, the skeleton atom contains the ring of heteroatom, namely is the heterocycle; aryl consisting of a heterocycle is heteroaryl; the nonaromatic group consisting of a heterocycle is a heterocycloalkyl group.

Heteroatoms include, but are not limited to O, S, N, P and the like.

Typical heteroaryl or heteroaryl groups include, but are not limited to:

"aryl" means a planar ring having a delocalized pi-electron system and containing 4n +2 pi electrons, where n is an integer. An aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. Aromatic groups include, but are not limited to, phenyl, naphthyl, phenanthryl, anthracyl, fluorenyl, indenyl, and the like.

"halogen" or "halo" refers to fluorine, chlorine, bromine or iodine.

The alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl, and the like, as described herein, may be unsubstituted alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl, or may be substituted alkyl, heteroalkyl, cyclic, heterocyclic, amino, ester, carbonyl, amide, sulfonyl, phosphoryl.

Hereinbefore, except where already indicated, "substituted" in said "substituted or unsubstituted" means that the mentioned groups may be substituted by one or more additional groups each and independently selected from alkyl, cycloalkyl, aryl, carboxy, heteroaryl, heterocycloalkyl, hydroxy, alkoxy, alkylthio, aryloxy, nitro, acyl, halogen, haloalkyl, amino and the like.

"inhibitor" refers to a substance that decreases the activity of an enzyme.

The invention has the beneficial effects that: the invention provides a series of compounds with BTK activity inhibition, tests show that the compounds have obvious inhibition effect on BTK, a new scheme is provided for treating diseases taking BTK as a treatment target, such as malignant tumor diseases or autoimmune diseases, the compounds can be used for preparing medicines for treating related diseases, and the compounds have wide application prospects.

Detailed Description

The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In the present invention, the structure of the compound is determined by Mass Spectrometry (MS) and/or nuclear magnetic resonance (1HNMR) equipment. The abbreviations have the following meanings:

DMF: n, N-dimethylformamide

n-BuLi: n-butyl lithium

THF: tetrahydrofuran (THF)

The DIAD: diisopropyl azodicarboxylate

DIEA: n, N-diisopropylethylamine

PE: petroleum ether

EA: ethyl acetate

DCM: methylene dichloride

TSOH: p-toluenesulfonic acid

HATU: o- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea

Pd₂(DBA)₃: tris (dibenzylideneacetone) dipalladium

BINAP: 1,1 '-binaphthyl-2, 2' -bis (diphenylphosphines)

DMSO, DMSO: dimethyl sulfoxide

m-CPBA: meta-chloroperoxybenzoic acid

Dess-Martin: dess-martin oxidizer

TBAF: tetrabutylammonium fluoride

DAST: diethylaminosulfur trifluoride

DIBAL-H: diisobutylaluminum hydride

Example 1

Step 1: synthesis of Compounds 1-3

Compound 1-1(11.232g, 72.2mmol), potassium carbonate (14.967g, 108.3mmol), 1-2(8.154g, 86.6mmol), DMF (60mL) were added to the flask and the reaction was stirred at room temperature overnight with TLC indicating completion of the reaction. Pouring the reaction system into water, extracting twice with ethyl acetate, and extracting with ethyl acetateThe ester layers were combined, washed twice with water, washed with brine and anhydrous Na₂SO₄Drying and concentrating under reduced pressure to obtain 16.5g of product 1-3, yield: 99 percent. The product was used in the next step without purification.

Step 2: synthesis of Compounds 1-4

To a reaction flask were added compound 1-3(16.5g, 72mmol), aqueous KOH (220mL, 5M), and ethanol (45mL), and the reaction was allowed to warm to 100 ℃ overnight with TLC indicating complete reaction of the starting material. Cooling the reaction system to 0 ℃, adding concentrated hydrochloric acid to adjust the reaction system to be acidic, separating out solids, filtering, washing with water, and drying a filter cake to obtain 17.3g of products 1-4 with yield: 96 percent. The product was used in the next step without purification.

And step 3: synthesis of Compounds 1-5

The compounds 1-4(17.3g, 69.6mmol), potassium carbonate (14.4g, 76.6mmol) and DMF (50mL) were added to a reaction flask, methyl iodide (4.8mL, 76.6mmol) was added dropwise to the reaction solution under ice bath, after the addition, the reaction solution was warmed to room temperature and stirred for 5 hours, and TLC showed completion of the reaction. Adding water into the reaction solution, extracting with ethyl acetate, washing the organic phase with saturated brine, drying with anhydrous sodium sulfate, and concentrating under reduced pressure to obtain 18g of product 1-5, yield: 98 percent. The product was used in the next step without purification.

And 4, step 4: synthesis of Compounds 1-7

The compound 1-6(1.00g,3.8mmol) was added to a three-necked reaction flask, the mixture was purged with nitrogen, tetrahydrofuran (35mL) was added, the reaction mixture was cooled to-78 ℃, n-BuLi tetrahydrofuran solution (3.2mL, 2.5M) was added dropwise, the mixture was stirred for 30 minutes, and after reaction, 1-5(887mg,3.8mmol) tetrahydrofuran solution was added dropwise to the reaction mixture, and the reaction was continued for 2 hours. Heating the reaction solution to room temperature, adding saturated ammonium chloride solution to quench the reaction, adding ethyl acetate to extract twice, drying the obtained organic phase with anhydrous sodium sulfate, concentrating under reduced pressure, and purifying the crude product by silica gel column chromatography to obtain 850mg of products 1-7 with the yield: 58 percent.

And 5: synthesis of Compounds 1-9

To a reaction flask were added compound 1-8(2.5g, 10.361mmol), tetrahydrofuran (12.5mL), cooled to 0 deg.C, borane dimethylsulfide solution (10mL, 20.723mmol) was added slowly, the reaction mixture was stirred at room temperature for 16 h, and TLC indicated completion of the reaction. The reaction was quenched with methanol, water was added, extracted with ethyl acetate (2 x 50mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to give 2.1g of product 1-9, yield: 91 percent. The product was used in the next step without purification.

Step 6: synthesis of Compounds 1-10

To a reaction flask were added compounds 1-9(1.8g, 7.930mmol), triphenylphosphine (3.1g, 11.894mmol), phthalimide (1.2g, 7.930mmol), tetrahydrofuran (25mL), cooled to 0 deg.C and slowly added dropwise DIAD (2.4g, 11.894mmol), and the reaction mixture stirred at room temperature for 0.5 h, TLC indicated completion of the reaction. The reaction system was extracted with water and ethyl acetate (3 × 50mL), the organic phases were combined, washed with brine, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE/EA ═ 10/1) to give 2.6g of product 1-10, yield: 93 percent.

And 7: synthesis of Compounds 1-11

To the reaction flask were added compound 1-10(2.6g, 7.303mmol), ethanol (25mL), hydrazine hydrate (8.7mL) slowly dropwise, and the reaction mixture was stirred at room temperature for 4 hours, TLC indicated completion of the reaction. Adding water into a reaction system, extracting with ethyl acetate (3 × 50mL), combining organic phases, washing with saturated saline solution, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain 1.2g of a product 1-11, wherein the yield is as follows: 75 percent. The product was used in the next step without purification.

And 8: synthesis of Compounds 1-12

To a reaction flask were added compounds 1-11(30mg, 0.133mmol), 1-7(50mg, 0.133mmol), n-butanol (2mL) and DIEA (51mg, 0.3982mmol), and the reaction mixture was heated to 100 deg.C and stirred for 1 hour, TLC indicated completion of the reaction. The reaction system is directly decompressed and concentrated after being cooled. The crude product was purified on preparative silica gel plate (DCM/MeOH ═ 20/1) to give 55mg of the product 1-12, yield: 72 percent.

And step 9: synthesis of Compound 1

The above-mentioned compounds 1 to 12(55mg, 0.096mmol) and a hydrogen chloride dioxane solution (2mL) were added to a reaction flask, and the reaction mixture was stirred at room temperature for 3 hours, and TLC showed that the starting material had reacted completely. The reaction was directly evaporated in vacuo and lyophilized to give 55mg of product 1 (hydrochloride salt) in yield: 100 percent.

LC/MS:m/z＝474.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.38-1.45(2H,m),1.49-1.57(2H,m),1.65-1.71(2H,m),1.93-2.01(2H,m),3.63-3.72(3H,m),7.04(1H,dd,J＝8.5,2.4Hz),7.18-7.32(5H,m),7.48-7.51(2H,m),7.59-7.64(2H,m),7.78-7.81(1H,m),8.25(1H,s),8.81-8.83(1H,m),12.79(1H,brs).

Example 2

Step 1: synthesis of Compound 2-2

Compound 2-1(200mg, 1.31mmol) and THF (6mL) were added to the reaction flask, lithium aluminum hydride (100mg, 2.63mmol) was added to the reaction in portions, the reaction was stirred overnight at room temperature, and TLC indicated completion of the reaction. The reaction solution was cooled in an ice-water bath and quenched slowly with 200mg of water. And (3) carrying out suction filtration on the obtained suspension, washing a filter cake by using THF, and carrying out reduced pressure concentration on the filtrate to obtain 120mg of a product 2-2, wherein the yield is as follows: 74 percent. The product was used in the next step without purification.

Step 2: synthesis of Compound 2

To a reaction flask were added compound 1-7(50mg, 0.13mmol), 2-2(32mg, 0.26mmol), TsOH (25mg, 0.13mmol) and n-butanol (2ml), and the reaction solution was reacted at 120 ℃ for 2 hours and TLC showed completion of the reaction. Water was added to the reaction system, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by preparative silica gel plate (DCM/MeOH ═ 20/1) to give 30mg of product 2, yield: 49 percent.

LC/MS:m/z＝472.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ4.66(2H,d,J＝5.5Hz),5.44(1H,t,J＝5.5Hz),7.05(1H,dd,J＝8.5,2.4Hz),7.19-7.29(4H,m),7.48-7.52(2H,m),7.68(1H,d,J＝8.5Hz),7.90(1H,s),8.24-8.38(2H,m),8.53(1H,s),8.94(1H,d,J＝2.2Hz),11.25(1H,s),13.10(1H,brs).

Example 3

To a reaction flask was added compound 1(60mg, 0.126mmol), potassium carbonate (53mg, 0.381mmol), anhydrous ethanol (1mL) and bromoethanol (16mg, 0.126mmol), and the reaction mixture was heated to reflux and stirred for 1 hour, TLC showed completion of the reaction. The reaction was cooled to room temperature and concentrated directly to dryness under reduced pressure to afford 10mg of product 3 by purification on preparative silica gel plates (DCM/MeOH ═ 10/1) yield: 16 percent.

LC/MS:m/z＝518.2[M+H]+.

1H NMR(400MHz,d6-DMSO)δ1.33-1.38(2H,m),1.46-1.53(2H,m),1.61-1.70(2H,m),1.92-1.99(2H,m),3.54-3.67(3H,m),3.78-3.82(2H,m),3.88-3.93(1H,m),4.15-4.16(1H,m),4.53(1H,t,J＝5.7Hz),7.03(1H,dd,J＝8.5,2.5Hz),7.19-7.26(4H,m),7.47-7.51(2H,m),7.59-7.64(2H,m),8.25(1H,s),8.80-8.83(1H,m),12.67(1H,brs).

Example 4

To a reaction flask were added compound 1(50mg, 0.110mmol), glycolic acid (8mg, 0.110mmol), DIEA (57mg, 0.44mmol) and DCM (5ml), followed by HATU (63mg, 0.165mmol) in one portion, and the reaction was reacted at room temperature for 2 hours, TLC showed completion of the reaction. Water was added to the reaction system, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified on preparative silica gel plate (DCM/MeOH ═ 20/1) to give 38mg of product 4, yield: 65.5 percent.

LC/MS:m/z＝532.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.34-1.39(2H,m),1.48-1.55(2H,m),1.63-1.71(2H,m),1.92-1.99(2H,m),3.57-3.66(3H,m),3.90-3.96(1H,m),4.16-4.19(1H,m),4.57(1H,t,J＝5.7Hz),7.04(1H,dd,J＝8.5,2.5Hz),7.19-7.26(4H,m),7.47-7.51(2H,m),7.59-7.64(2H,m),8.25(1H,s),8.80-8.83(1H,m),12.76(1H,brs).

Example 5

Compound 1(60mg, 0.126mmol), DIEA (18mg, 0.126mmol), absolute ethanol (1mL) and glycidol (48mg, 0.634mmol) were added to a reaction flask, the reaction mixture was heated to reflux and stirred for 4 hours, TLC showed completion of the reaction. The reaction was cooled to room temperature and concentrated directly to dryness under reduced pressure to afford 10mg of product 5 by purification on preparative silica gel plates (DCM/MeOH ═ 10/1) yield: 14 percent.

LC/MS:m/z＝548.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.20-1.35(2H,m),1.53-1.62(1H,m),1.74-1.86(2H,m),1.96-2.02(1H,m),2.16-2.34(2H,m),3.37-3.44(4H,m),3.52-3.72(4H,m),4.46-4.48(1H,m),4.59-4.62(1H,m),7.04(1H,dd,J＝8.4,2.3Hz),7.19-7.20(3H,m),7.26(1H,t,J＝7.4Hz),7.47-7.51(2H,m),7.59(1H,d,J＝8.5Hz),7.62(1H,brs),8.24(1H,s),8.80-8.84(1H,m),12.74(1H,brs).

Example 6

To a reaction flask were added compound 1-7(20mg, 0.052mmol), 3-amino-1, 2-propanediol (6mg, 0.066mmol), n-butanol (1mL) and DIEA (20mg, 0.156mmol), and the reaction mixture was heated to 100 ℃ and stirred for 1 hour, TLC indicated completion of the reaction. The reaction system is directly decompressed and concentrated after being cooled. The crude product was purified on preparative silica gel plate (DCM/MeOH ═ 10/1) to give 15mg of product 6, yield: 66 percent.

LC/MS:m/z＝439.1[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ2.82-2.86(1H,m),3.28-3.32(1H,m),3.45-3.56(3H,m),4.84(1H,t,J＝5.8Hz),4.93-4.95(1H,m),7.03(1H,dd,J＝8.5,2.4Hz),7.18-7.20(3H,m),7.26(1H,t,J＝7.4Hz),7.49(2H,t,J＝8.0Hz),7.59-7.63(2H,m),8.25(1H,s),8.33(1H,d,J＝9.1Hz),12.73(1H,brs).

Examples 7 to 9

The preparation method is implemented by using commercially available raw materials according to the detailed operation procedures of the above example 2 and example 6, and the detailed operation procedures are shown in the following table 1:

TABLE 1

Example 10

Step 1: synthesis of Compound 10-2

Compound 10-1(2.0g, 8.65mmol, prepared in reference Eur. J. org. chem.2003, 2418-2427) and hydrogen chloride dioxane solution (20mL) were added to the reaction flask, the reaction was stirred at room temperature for 3 hours, and TLC showed complete reaction of the starting materials. The reaction solution was directly evaporated to dryness in vacuo to give 1.5g of product 10-2, yield: 100 percent. The product was used in the next step without purification.

Step 2: synthesis of Compound 10-4

To a reaction flask were added compound 10-3(5.0g, 20.3mmol) and methanol (50mL), concentrated sulfuric acid (5mL) and warmed to 100 deg.C overnight. TLC showed the reaction was complete and cooled. Pouring the reaction solution into water, extracting with ethyl acetate for 3 times, combining organic phases, washing with water, washing with saturated saline solution, and anhydrous Na₂SO₄Drying and vacuum evaporation to dryness gave 5.3g of product 10-4, yield: 100 percent. The product was used in the next step without purification.

And step 3: synthesis of Compound 10-5

To a three-necked flask, compound 10-4(2.0g, 7.72mmol), tetrahydrofuran (20mL) was added, replaced with nitrogen, cooled to-70 ℃ in an ethanol dry ice bath, and ethyl magnesium bromide (28mL, 1mol/L) was added dropwise and reacted at this temperature for 1 hour. The reaction was poured into saturated ammonium chloride solution, extracted three times with ethyl acetate, the organic phases combined, added to silica gel and stirred, then purified through silica gel column to give 680mg of product 10-5, yield: 45 percent.

And 4, step 4: synthesis of Compound 10-7

The compound 10-5(200mg,0.93mmol) and DCM (10mL) were added to the reaction flask, cooled to 0 ℃ in ice bath, trifluoromethanesulfonic acid (1410mg, 9.4mmol) was added dropwise, reaction was carried out for 30min, the compound 10-6(330mg,1.22mmol, prepared by refluxing 1-4 with thionyl chloride for 2 hours and then concentrating to dryness) was added to the reaction solution, the reaction solution was slowly warmed to room temperature and stirred for 1 hour, TLC showed completion of the reaction. The reaction was quenched by addition of 1mL methanol, poured into saturated sodium bicarbonate solution, extracted 4 times with dichloromethane, the organic layers combined, evaporated in vacuo and the crude purified by silica gel column to give 240mg of product 10-7, yield: 60 percent.

¹H NMR(400MHz,CDCl₃)δ4.02(3H,s),6.93(1H,dd,J＝8.5,2.4Hz),7.07(1H,d,J＝2.4Hz),7.10-7.13(2H,m),7.22-7.25(1H,m),7.42-7.45(2H,m),7.56(2H,dd,J＝8.3,4.3Hz),7.63(1H,d,J＝2.9Hz),7.82(1H,d,J＝8.1Hz),10.51(1H,s).

And 5: synthesis of Compound 10-8

To a reaction flask were added compound 10-7(100mg,0.21mmol), 10-2(35mg,0.21mmol), Pd₂(DBA)₃(10mg,0.01mmol), BINAP (6mg,0.01mmol), cesium carbonate (134mg,0.41mmol) and dioxane (4mL), purged with nitrogen, and stirred overnight at 100 ℃. TLC showed the reaction was complete, cooled, poured into water, extracted 3 times with ethyl acetate, dried over anhydrous sodium sulfate, evaporated in vacuo and purified by preparative silica gel plate (DCM/MeOH ═ 10/1) to give 20mg of compound 10-8. Yield: 19 percent.

Step 6: synthesis of Compound 10-9

To a reaction flask, compound 10-8(20mg,0.041mmol), lithium hydroxide monohydrate (9mg,0.21mmol), tetrahydrofuran (5mL) and water (1mL) were added, and the mixture was heated to 60 ℃ for reaction overnight. TLC showed the reaction was complete and cooled. The reaction solution was poured into water, 200mg of glacial acetic acid was added, followed by extraction with ethyl acetate 3 times and evaporation in vacuo to give 15mg of compound 10-9. Yield: 70 percent. The product was used in the next step without purification.

And 7: synthesis of Compound 10

To a reaction flask were added compound 10-9(15mg,0.029mmol), DIEA (16mg,0.12mmol), NH₄Cl (8mg,0.15mmol) and DMF (2mL) were added in one portion to HATU (17mg,0.045mmol) and reacted at room temperature overnight. The reaction was poured into water, extracted 3 times with ethyl acetate, the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and purified by preparative silica gel plate (DCM/MeOH ═ 10/1) after evaporation in vacuo to give 8mg of product 10. Yield: 53 percent.

LC/MS:m/z＝520.1[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.34-1.43(1H,m),1.53-1.62(1H,m),1.76-1.80(1H,m),2.17-2.22(1H,m),3.12(1H,t,J＝11.2Hz),3.40-3.46(3H,m),4.11-4.18(2H,m),4.68(1H,t,J＝5.4Hz,1H),6.88(1H,dd,J＝8.5,2.4Hz),7.02(1H,d,J＝2.4Hz),7.08-7.11(2H,m),7.20-7.22(1H,m),7.40-7.43(2H,m),7.54(2H,dd,J＝8.3,4.3Hz),7.61-7.65(3H,m),7.72(1H,d,J＝8.3Hz),10.50(1H,s).

Example 11

Synthesis of Compound 11-1 reference was made to the synthesis of Compound 10-7.

Step 1: synthesis of Compound 11-2

To a reaction flask were added compound 11-1(70mg,0.17mmol), compound 10-2(35mg,0.21mmol), DIEA (66mg,0.51mmol) and DMSO (5mL), and the temperature was raised to 120 ℃ for 12 hours. TLC showed the reaction was complete, cooled, poured into water, extracted 3 times with ethyl acetate, the organic phases combined, washed with brine, dried over anhydrous sodium sulfate, evaporated in vacuo and purified by preparative silica gel plate (DCM/MeOH ═ 10/1) to give 50mg of compound 11-2. Yield: 53.3 percent.

Step 2: synthesis of Compound 11-3

Compound 11-2(50mg,0.091mmol), lithium hydroxide (19mg,0.453mmol), tetrahydrofuran (5mL) and water (1mL) were added to the reaction flask and allowed to warm to 60 ℃ for reaction overnight. TLC showed the reaction was complete and cooled. The reaction solution was poured into water, 200mg of glacial acetic acid was added, followed by extraction with ethyl acetate 3 times and evaporation in vacuo to give 48mg of Compound 11-3. Yield: 98 percent. The product was used in the next step without purification.

And step 3: synthesis of Compound 11

To a reaction flask was added compound 11-3(48mg,0.089mmol), DIEA (46mg,0356mmol), NH₄Cl (14mg,0.267mmol) and DMF (2mL) were added in one portion to HATU (51mg,0.134mmol) and reacted at room temperature overnight. The reaction was poured into water, extracted 3 times with ethyl acetate, the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and purified by preparative silica gel plate (DCM/MeOH ═ 10/1) after evaporation in vacuo to give 11mg of product 11. Yield: 23 percent.

LC/MS:m/z＝538.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.34-1.44(1H,m),1.53-1.63(1H,m),1.76-1.80(1H,m),2.18-2.23(1H,m),3.14(1H,t,J＝10.8Hz),3.40-3.47(3H,m),4.11-4.18(2H,m),4.69(1H,t,J＝5.4Hz,1H),7.03(1H,d,J＝2.4Hz),7.08-7.11(2H,m),7.20-7.22(1H,m),7.40-7.43(2H,m),7.54(2H,dd,J＝8.3,4.3Hz),7.60-7.66(3H,m),7.70(1H,m),10.52(1H,s).

Example 12

Step 1: synthesis of Compound 12-2

To a reaction flask were added compound 12-1(500mg, 2.124mmol), potassium carbonate (439mg, 3.177mmol) and DMF (5 mL). After cooling in an ice bath, methyl iodide (331mg,2.332mmol) was added dropwise, and after the addition was complete, the reaction solution was warmed to room temperature and stirred for 3 hours, and TLC showed completion of the reaction. Water was added to the reaction mixture, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 520mg of compound 12-2, yield: 98 percent. The product was used in the next step without purification.

Step 2: synthesis of Compound 12-4

To a reaction flask were added compound 12-2(460mg, 1.844mmol), 12-3(220mg,1.844mmol), Pd₂(DBA)₃(34mg,0.037mmol), X-Phos (35mg,0.073mmol), cesium carbonate (1.212g,3.72mmol) and dioxane (10mL), the reaction was replaced with liquid nitrogen and reacted at 100 ℃ for 5 hours, TLC indicated complete reaction of starting material. After the reaction system is cooled, silica gel is directly added for sample mixing, and then the product 12-4 of 310mg is obtained by silica gel column purification, and the yield is as follows: 70 percent.

And step 3: synthesis of Compounds 12-5

The compound 1-6(100mg,0.43mmol) was added to a three-necked reaction flask, nitrogen gas was substituted, tetrahydrofuran (4mL) was added, the reaction solution was cooled to-72 ℃, an n-BuLi tetrahydrofuran solution (0.36mL, 2.5M) was added dropwise, after stirring and reacting for 40 minutes, 12-4(123mg,0.43mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction solution, and the reaction was continued for 2 hours. Heating the reaction solution to room temperature, adding saturated ammonium chloride solution to quench the reaction, adding ethyl acetate to extract twice, combining organic phases, concentrating under reduced pressure, and purifying by silica gel column chromatography to obtain 26mg of a product 12-5 with yield: 15 percent.

And 4, step 4: synthesis of example 12

To a reaction flask were added compound 12-5(26mg, 0.063mmol), 10-2(10mg,0.063mmol), DIEA (32mg,0.252mmol) and n-butanol (1mL), heated to 100 ℃ for 2 hours and TLC indicated complete reaction of starting materials. Evaporation in vacuo followed by purification by preparative silica gel plate (DCM/MeOH ═ 15/1) afforded 12mg of product 12, yield: 38 percent.

LC/MS:m/z＝504.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.37-1.44(1H,m),1.55-1.62(1H,m),1.77-1.81(1H,m),2.16-2.23(1H,m),2.49-2.50(1H,m),3.12-3.17(2H,m),3.27-3.33(1H,m),3.34-3.48(3H,m),4.03(1H,t,J＝8.5Hz),4.13-4.20(2H,m),4.65-4.70(1H,m),6.78-6.86(1H,m),7.14-7.21(1H,m),7.25-7.32(2H,m),7.53-7.59(1H,m),7.68-7.86(3H,m),8.25(1H,d,J＝8.4Hz),8.64(1H,dd,J＝16.3,7.4Hz),12.52(1H,brs).

Example 13

Step 1: synthesis of Compound 13-2

Compound 13-1(11.0g, 32mmol, prepared by reference Eur. J. org. chem.2003, 2418-2427) and methylene chloride (110mL) were added to the reaction flask, the reaction was replaced with gaseous nitrogen, the ice-water bath was cooled, then m-CPBA (11.1g, 64mmol) was added in portions, after the addition was complete, the ice was removed, the reaction mixture was stirred overnight at room temperature, and TLC indicated completion of the reaction. Filtering the reaction system, washing with dichloromethane, adding sodium sulfite solution into the filtrate, stirring for 10min, extracting with dichloromethane for 2 times, combining the organic phases, and extracting with NaHCO₃The aqueous solution was washed 2 times with brine, directly evaporated in vacuo and purified by silica gel column to give 8.6g of 13-2 as a product in yield: 75 percent.

Step 2: synthesis of Compounds 13-3a and 13-3b

LAH (1363mg, 35.9mmol) and tetrahydrofuran (50mL) were added to the reaction flask, the reaction was replaced with liquid nitrogen, cooled in an ice-salt bath, and a solution of compound 13-2(8.6g, 23.9mmol) in tetrahydrofuran (50mL) was added dropwise, after dropping, the ice was removed after stirring for 10min, and stirred at room temperature overnight, and TLC indicated complete reaction. The reaction solution was cooled in an ice-water bath, and then water (2.8mL) was slowly added dropwise to the reaction solution to quench the reaction. The suspension is filtered, the filter cake is washed with ethyl acetate, the filtrate is concentrated in vacuum and purified by a silica gel column to obtain 300mg of product 13-3a and 3237mg of product 13-3b, the yield is as follows: 41 percent.

And step 3: synthesis of Compound 13-4

13-4(1500mg, 4.15mmol) and dichloromethane (30mL) were added to the flask, the reaction was purged with nitrogen, Dess-Martin oxidant (3519mg, 8.30mmol) was added to the flask under ice-water bath, the flask was stirred overnight at room temperature, and TLC indicated completion of the reaction. Silica gel was added directly to the reaction solution to mix the sample, which was then purified through silica gel column to give 1158mg of product 13-4, yield: 78 percent.

And 4, step 4: synthesis of Compound 13-5

To a reaction flask were added compound 13-4(550mg, 1.53mmol), methyl triphenyl phosphonium bromide (2186mg, 6.12mmol) and tetrahydrofuran solution (25mL), the reaction was replaced with nitrogen gas, cooled in an ice-water bath, and then potassium tert-butoxide (1373mg, 12.2mmol) was added in one portion. After stirring for 30min, the temperature was slowly raised to room temperature and stirring was continued for 1.5 h. TLC showed the starting material reaction was complete. The reaction solution was quenched with aqueous ammonium chloride solution, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure, and purified by silica gel column to give 302mg of product 13-5, yield: and 55 percent.

And 5: synthesis of Compound 13-6

Diiodomethane (375mg, 1.40mmol) and dichloromethane (6mL) were added to the reaction flask, the reaction liquid was replaced with gaseous nitrogen, diethylzinc (1.4mL, 1M) was added dropwise to the reaction liquid, and the mixture was stirred for 5 minutes and then cooled in an ice-water bath. After dropwise addition of a solution of 13-5(100mg, 0.28mmol) in dichloromethane (1mL) to the reaction mixture, the reaction mixture was slowly warmed to room temperature and stirred for 3 hours, TLC showed completion of the reaction, diluted hydrochloric acid (0.1M, 10mL) was added to quench the reaction, after stirring for 5 minutes, the reaction was extracted twice with dichloromethane, the organic phases were combined, concentrated under reduced pressure and purified by silica gel column to give 64mg of 13-6, yield: 62 percent.

Step 6: synthesis of Compound 13-7

To a reaction flask were added compound 13-6(40mg, 0.108mmol) and tetrahydrofuran (2mL), and TBAF (0.22mL, 1M in THF) was added to the reaction solution, which was stirred at room temperature for 2.5 h. TLC showed the reaction of the starting materials was complete, water was added to the reaction mixture, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. Adding hydrogen chloride dioxane solution (2mL) into the residue, stirring the reaction liquid at room temperature for reacting for 3 hours, directly evaporating the reaction liquid in vacuum to dry to obtain a product 13-7, and directly using the product in the next step without purification.

And 7: synthesis of Compound 13

To a reaction flask were added the above-mentioned compounds 13-7(0.108mmol, in terms of theory), 1-7(41mg, 0.108mmol), n-BuoH (2mL) and DIPEA (70mg,0.54mmol), and the reaction was heated to 120 ℃ and stirred for 2 hours, TLC showed the formation of the product. After cooling the reaction system and evaporation in vacuo, purification by preparative silica gel plate (DCM/MeOH ═ 10/1) afforded 18mg of product 13, yield: 33% in two steps.

LC/MS:m/z＝505.1[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ0.26-0.37(2H,m),0.46-0.50(1H,m),0.77-0.81(1H,m),1.80-1.85(1H,m),1.97-2.03(1H,m),3.25(1H,t,J＝10.4Hz),3.42-3.52(3H,m),3.99(1H,dd,J＝10.4,4.4Hz),4.62-4.70(2H,m),7.02(1H,dd,J＝8.5,2.4Hz),7.18-7.20(3H,m),7.26(1H,t,J＝7.4Hz),7.49(2H,t,J＝8.0Hz),7.59(1H,d,J＝8.5Hz),7.65(1H,s),8.25(1H,s),8.33(1H,d,J＝9.1Hz),12.63(1H,brs).

Examples 14 to 15

Synthesis of Compound 14-1:

compound 13-3a (200mg, 0.554mmol), dichloromethane (2mL) was added to the reaction flask, the temperature was reduced to 0 deg.C, DAST (94mg, 0.582mmol) was added, the reaction mixture was stirred at 0 deg.C for 0.5 h, and TLC indicated completion of the reaction. The reaction was purified directly by silica gel column chromatography (PE/EA ═ 20/1) to give 75mg of the product 14-1, yield: 38 percent.

Synthesis of Compound 15-1: refer to the detailed procedure for synthesis 14-1.

Synthesis of compounds 14, 15:

compounds 14 and 15 were synthesized from intermediates 14-1 and 15-1 by the method described in example 13, and the structures of the compounds are shown in Table 2.

Example 16

To a reaction flask were added compound 13-4(120mg,0.334mmol), DCE (4mL) and DAST (119mg,0.735mmol), the reaction mixture was warmed to 60 deg.C and stirred for 1.5h, TLC showed a new spot. The reaction was cooled and purified directly by silica gel column chromatography (PE/EA ═ 10/1) to give 10mg of product 16-1, yield: 8 percent. Compound 16 was synthesized via intermediate 16-1, according to the method of example 13, and the specific structure of the compound is shown in Table 2.

Examples 17 to 20

Compound 17: compound 17 was synthesized via intermediate 13-1, according to the method of example 13.

Compound 18: compound 18 was synthesized via intermediate 13-3a, according to the procedure of example 13.

Compound 19: compound 19 was synthesized via intermediate 13-3b, according to the procedure of example 13.

Compound 20: compound 20 was synthesized via intermediate 13-5, according to the procedure of example 13.

The specific structures of compounds 17-20 are shown in Table 2

Example 21

Diiodomethane (312mg, 1.165mmol) and dichloromethane (6mL) were added to the reaction flask, the reaction liquid was replaced with nitrogen gas, diethyl zinc (1.2mL,1M) was added dropwise to the reaction liquid, the reaction liquid was stirred at room temperature for 5 minutes and then cooled in an ice water bath, then a solution of compound 13-1(100mg, 0.291mmol) in dichloromethane (1mL) was added dropwise, after dropping, the reaction mixture was slowly warmed to room temperature and stirred for 3 hours, and TLC showed substantial completion of the reaction. The reaction was quenched by addition of dilute hydrochloric acid (0.1M, 10mL), extracted twice with dichloromethane, combined the organic phases, concentrated under reduced pressure and purified by silica gel column to give 33mg of product 21-1, yield: 32 percent. Compound 21 was synthesized from intermediate 21-1 by the method described in example 13, and the specific structure of the compound is shown in Table 2.

Example 22

Step 1: synthesis of Compound 22-1

Oxalyl chloride (110mg, 0.866mmol) and dichloromethane (2mL) were added to a reaction flask, the temperature was reduced to-72 ℃ and replaced with nitrogen, DMSO (101mg, 1.30mmol) was slowly dropped and stirred at the temperature for 30min after dropping, compound 10-1(100mg, 0.433mmol) was dissolved in dichloromethane (0.5mL) and slowly dropped and stirred at the temperature for 30min after dropping, triethylamine (220mg, 2.17mmol) was slowly added and stirred at-72 ℃ for 0.5 h after adding, and TLC showed completion of the reaction. Adding water into a reaction system, extracting with dichloromethane for 3 times, combining organic phases, washing with saturated saline, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain 90mg of a product 22-1, wherein the yield is as follows: 91 percent. The product was used in the next step without purification.

Step 2: synthesis of Compound 22-2

Compound 22-1(90mg, 0.393mmol) and dichloromethane (2mL) were added to the reaction flask, the temperature was reduced to-72 deg.C and DAST (127mg, 0.786mmol) was added and the reaction mixture was stirred at-72 deg.C for 0.5 h, TLC showed substantial completion of the reaction. Adding water into a reaction system, extracting with dichloromethane for 3 times, combining organic phases, washing with saturated saline, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure to obtain 90mg of a product 22-2, wherein the yield is as follows: 91 percent. The product was used in the next step without purification. Compound 22 was synthesized via intermediate 22-2, again according to the procedure of example 13, and the specific structure of the compound is shown in Table 2.

Example 23

Step 1: synthesis of Compound 23-3

To a reaction flask were added compound 23-1(200mg, 1.06mmol), 23-2(131mg, 1.17mmol), potassium carbonate (220mg, 1.59mmol) and DMF (3 mL). The reaction was heated to 80 ℃ and stirred overnight, TLC showed the reaction was complete. Adding water to the reaction solution, extracting twice with ethyl acetate, combining the organic phases, washing with saturated brine, drying over anhydrous sodium sulfate, concentrating under reduced pressure, and purifying with silica gel column to obtain 232mg of compound 23-3 with yield: 78 percent.

Step 2: synthesis of Compound 23-4

The compound 1-6(192mg,0.83mmol) was added to a three-necked reaction flask, nitrogen gas was used for replacement, tetrahydrofuran (5mL) was added, the reaction solution was cooled to-72 ℃, an n-BuLi tetrahydrofuran solution (0.73mL,1.83mmol,2.5M) was added dropwise, after stirring for 40 minutes for reaction, 23-3(232mg,0.83mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction solution, and the reaction was continued for 2 hours. Heating the reaction solution to room temperature, adding saturated ammonium chloride solution to quench the reaction, adding ethyl acetate to extract twice, combining organic phases, concentrating under reduced pressure, and purifying by silica gel column chromatography to obtain 140mg of a product 23-4 with yield: 42 percent.

And step 3: synthesis of Compound 23

To a reaction flask were added compound 23-4(50mg, 0.124mmol), 10-2(21mg,0.124mmol), DIEA (64mg,0.496mmol) and n-butanol (2mL), heated to 120 ℃ for 2 hours and TLC showed the starting material was essentially reacted. The reaction was cooled, evaporated in vacuo and purified by preparative silica gel plate (DCM/MeOH ═ 15/1) to give 15mg of product 23, yield: 24 percent.

LC/MS:m/z＝497.1[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.33-1.43(1H,m),1.52-1.62(1H,m),1.76-1.80(1H,m),2.17-2.23(1H,m),3.12(1H,t,J＝11.2Hz),3.38-3.46(3H,m),4.10-4.18(2H,m),4.68(1H,t,J＝5.4Hz,1H),7.03(1H,dd,J＝8.5,2.3Hz),7.22-7.28(2H,m),7.38(1H,m),7.51-7.56(2H,m),7.61(1H,d,J＝8.5Hz),7.69(1H,s),8.28(1H,s),8.83(1H,d,J＝7.4Hz),12.79(1H,brs).

Examples 24 to 26

Compounds 24-26 were synthesized by reference to intermediates prepared above and to the procedure of example 23, and the specific structures of the compounds are shown in Table 2.

Example 27

Step 1: synthesis of Compound 27-1

To the reaction flask were added 22-1(600mg,2.64mmol), tert-butanol (7.5mL), water (3mL) and 2-methyl-2-butene (926mg,13.2 mmol). To the reaction mixture were added an aqueous solution (3mL) of sodium dihydrogenphosphate (1584mg,13.2mmol) and sodium chlorite (955mg,10.6 mmol). The reaction was stirred at room temperature for 1.5h and TLC showed the reaction was complete. Quenching the reaction system by using sodium sulfite aqueous solution, then adjusting acid by using dilute hydrochloric acid, extracting for 2 times by using ethyl acetate, combining organic phases, washing by using saturated saline solution, drying by using anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain 500mg of a product 27-1, wherein the yield is as follows: 78 percent. The product was used in the next step without purification.

Step 2: synthesis of Compound 27-2

To a reaction flask were added compound 27-1(500mg, 2.06mmol), potassium carbonate (426mg, 3.08mmol) and DMF (6mL), followed by methyl iodide (437mg,3.08 mmol). The reaction solution was stirred overnight at room temperature and TLC showed the reaction was complete. Adding water to the reaction solution, extracting twice with ethyl acetate, combining the organic phases, washing with saturated brine, drying over anhydrous sodium sulfate, concentrating under reduced pressure, and purifying with silica gel column to obtain 310mg of compound 27-2, yield: 58 percent.

And step 3: synthesis of Compound 27-3

Compound 27-2(310mg, 1.2mmol) and DMF (3mL) were added to the reaction flask, replaced with nitrogen, and cooled in an ice-water bath. Tetraisopropyl titanate (136mg, 0.48mmol) was added to the reaction solution, followed by slow dropwise addition of ethyl magnesium bromide (3mL, 3.0mmol, 1M). After the addition, the reaction solution was warmed to room temperature and stirred for 2 hours, and TLC showed completion of the reaction. The reaction solution was quenched by pouring into aqueous ammonium chloride solution, extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified by silica gel column to obtain 112mg of compound 27-3, yield: 37 percent. Compound 27 was synthesized via intermediate 27-3, again following the procedure of example 13, and the detailed structure is shown in Table 2.

TABLE 2

Example 28

Step 1: synthesis of Compound 28-1

Compound 1-3(2.0g,8.7mmol) and THF (10mL) were added to a three-necked flask, the reaction was replaced with nitrogen, cooled in an ice-water bath, DIBAL-H (13.1mL,1M in toluene) was added dropwise, after the addition was complete, the ice was removed, the reaction was stirred at room temperature for 5 hours, and TLC indicated completion of the reaction. The reaction solution was introduced into dilute hydrochloric acid, stirred for 3 hours, extracted 2 times with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure, and purified by silica gel column to give 1.7g of 28-1, yield: 84 percent.

Step 2: synthesis of Compound 28-3

To a reaction flask were added compound 28-2(500mg,3.28mmol), compound 28-1(763mg,3.28mmol) and methanol (10ml), followed by potassium tert-butoxide (1840mg,16.4mmol), and the reaction was stirred at room temperature overnight with TLC showing the formation of the product. The reaction solution was poured into water, extracted three times with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified by preparative silica gel column to give 435mg of the product 28-3, yield: 28 percent.

And step 3: synthesis of Compound 28-4

Compound 28-3(40mg,0.104mmol) and dichloromethane (2mL) were added to the flask, the reaction was replaced with liquid nitrogen, Dess-Martin oxidant (88mg, 0.208mmol) was added to the flask under ice water bath, the reaction was stirred at room temperature for 1 hour, and TLC indicated completion of the reaction. The reaction solution was poured into water, extracted three times with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by preparative silica gel plates to give 16mg of 28-4, yield: 40 percent.

And 4, step 4: synthesis of example 28

Compound 28-4(16mg,0.042mmol), 10-2(21mg, 0.126mmol), NMP (1mL) and DIPEA (33mg,0.252mmol) were added to the reaction flask, the reaction was heated to 180 ℃ and stirred for 8 hours, TLC showed the formation of the product. After the reaction system was cooled, poured into water, extracted three times with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by preparative silica gel plates to give 9mg of product 28, yield: 45 percent.

LC/MS:m/z＝478.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.41-1.56(2H,m),1.77-1.80(1H,m),2.21-2.26(1H,m),3.12(1H,t,J＝10.4Hz),3.34-3.45(3H,m),3.55-3.62(1H,m),4.11-4.14(1H,m),4.68(1H,t,J＝5.7Hz),6.41(1H,d,J＝5.7Hz),7.02(1H,dd,J＝8.4,2.3Hz),7.18-7.20(3H,m),7.26(1H,t,J＝7.4Hz),7.46-7.50(2H,m),7.53-7.55(2H,m),7.93(1H,d,J＝5.5Hz),8.39-8.43(1H,m),12.44(1H,brs).

Example 29

29-2 reference the procedure for the synthesis of 1-7, the yield of this step is very low.

Synthesis of 29 reference was made to the method of synthesis 23.

LC/MS:m/z＝480.1[M+H]⁺.

Example 30

Step 1: synthesis of Compound 30-2

Compound 30-1(2.5g,12.7mmol) and THF (25mL) were added to a three-necked flask, the reaction was replaced with nitrogen, cooled in an ice-water bath, sodium hydride (60%, 761mg,19.0mmol) was added in portions, after the addition, the ice was removed, and the reaction was stirred at room temperature for 0.5 hour. The reaction solution was cooled again in an ice-water bath, TIPSCl (3.66g,19.0mmol) was added dropwise thereto, ice was removed after the addition, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was slowly poured into ice water to quench, extracted 2 times with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified by silica gel column to give 3.92g of product 30-2, yield: 88 percent.

Step 2: synthesis of Compound 30-3

Compound 30-2(3.92g,11.1mmol) and THF (25mL) were added to the reaction flask, the reaction was replaced with liquid nitrogen, the bath was cooled to-72 ℃ in a dry ice/ethanol bath, n-butyllithium (2.5M,8.9mL,22.2mmol) was added dropwise, and after the addition, stirring was carried out for 0.5 hour under constant temperature. To the reaction mixture was added dropwise a solution of NFSI (4.2g,13.3mmol) in THF (10mL), and after the addition, the reaction mixture was slowly warmed to room temperature and stirred for 1 hour. The reaction solution was slowly poured into ice water to quench, extracted 2 times with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified by silica gel column to give 1.69g of product 30-3, yield: 52 percent.

And step 3: synthesis of Compound 30-4

The reaction flask was charged with compound 30-3(400mg,1.37mmol) and THF (5mL), the reaction was replaced with liquid nitrogen, the dry ice/ethanol bath was cooled to-72 deg.C, sec-butyllithium (1.3M,2.1mL,2.74mmol) was added dropwise, and after addition, stirring was maintained for 0.5 h. To the reaction mixture was added dropwise a solution of NFSI (518mg,1.64mmol) in THF (3mL), and after the addition, the reaction mixture was slowly warmed to room temperature and stirred for 1 hour. The reaction solution was slowly poured into ice water to quench, extracted 2 times with ethyl acetate, the organic phases were combined, washed with saturated brine, and concentrated under reduced pressure to give 800mg of crude product 30-4, which was used in the next step without purification.

And 4, step 4: synthesis of Compound 30-5

To the reaction flask were added compound 30-4(800mg,2.58mmol), THF (8mL) and TBAF (1M,3.2mL), and the reaction was stirred at room temperature for 0.5 h. The reaction solution is directly decompressed and concentrated and then purified by a silica gel column to obtain 30-5 of 150mg of product, and the yield of the two steps is as follows: 71 percent.

And 5: synthesis of Compound 30-6

Compound 30-5(95mg,0.62mmol) and DMF (4mL) were added to the reaction flask, the reaction was cooled in an ice-water bath, NBS (110mg,0.62mmol) was added, and the reaction was allowed to warm to room temperature slowly and stirred for 0.5 h. The reaction was poured into water, extracted 2 times with ethyl acetate, the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified on silica gel plates to give 120mg of product 30-6, yield: 83 percent.

30-7 reference is made to the methods of syntheses 1-7.

Synthesis of 30 reference was made to the method of synthesis 28.

LC/MS:m/z＝496.1[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ1.35-1.56(2H,m),1.75-1.78(1H,m),2.22-2.25(1H,m),3.14(1H,t,J＝10.7Hz),3.30-3.35(3H,m),3.78-3.83(1H,m),4.10-4.12(1H,m),7.02(1H,dd,J＝8.5,2.3Hz),7.18-7.20(2H,m),7.26(1H,t,J＝7.4Hz),7.48(2H,t,J＝8.0Hz),7.56(1H,d,J＝8.5Hz),7.66(1H,s),8.04(1H,d,J＝6.6Hz),8.43(1H,d,J＝8.4Hz),12.66(1H,brs).

Example 31

31-1 reference is made to the procedure for the synthesis of 30-4, replacing NFSI with NCS.

31 reference is made to the method for the synthesis of 30 from 30-4.

LC/MS:m/z＝512.2[M+H]⁺.

Example 32

32-1 Synthesis reference is made to the procedure for the synthesis of 30-4, replacing NFSI with carbon tetrabromide.

32-2 reference is made to the method of synthesis 30-5.

32-3 Synthesis reference is made to the method of synthesis 30-6, replacing NBS with NIS.

32 refers to the method of synthesis of 30 from 30-6.

LC/MS:m/z＝556.1[M+H]⁺.

Example 33

Adding into a reaction flaskCompound 32(10mg,0.018mmol), Zinc cyanide (4.2mg,0.036mmol), DMA (0.5mL), Zinc powder (1.0mg), dppf (2.0mg) and Pd₂(dba)₃(2.0mg), the reaction was replaced with liquid nitrogen gas, heated to 100 ℃ and stirred for 2 hours. The reaction was extracted 2 times with ethyl acetate and the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified through silica gel prep plates to give 3mg of product 33, yield: 33 percent.

LC/MS:m/z＝503.1[M+H]⁺.

Example 34

34-1 reference is made to the procedure for the synthesis of 13-7.

34 reference is made to the method of synthesis 30.

LC/MS:m/z＝508.2[M+H]⁺.

Example 35

35 reference is made to the method of synthesis 28.

LC/MS:m/z＝490.2[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ0.71-0.75(1H,m),0.81-0.86(1H,m),1.26-1.34(1H,m),1.50-1.56(1H,m),2.70-2.75(1H,m),3.48-3.61(3H,m),3.88(1H,dd,J＝11.6,6.0Hz),4.13-4.23(1H,m),4.84(1H,t,J＝5.6Hz),6.47(1H,d,J＝5.8Hz),6.99-7.06(2H,m),7.18-7.20(2H,m),7.25(1H,t,J＝7.4Hz),7.40-7.56(4H,m),7.92(1H,d,J＝5.6Hz),8.54(1H,d,J＝7.7Hz),12.40(1H,brs).

Example 36

Synthesis of 36-1 reference was made to the method of Synthesis 13-7.

Synthesis of 36 reference the method of synthesis 28.

LC/MS:m/z＝476.1[M+H]⁺.

Example 37

Synthesis step 1: synthesis of Compound 37-2

To a reaction flask, compound 37-1(652-67-5,200mg,1.37mmol), phthalimide (201mg,1.37mmol), triphenylphosphine (718mg,2.74mmol) and THF (5mL) were added, DIAD (553mg,2.74mmol) was added dropwise to the reaction solution, and after completion of addition, the reaction solution was warmed to a high temperature and stirred at room temperature for 2 hours. The reaction mixture was directly concentrated under reduced pressure and purified by a silica gel column to obtain 351mg of 37-2 (containing triphenylphosphine oxide) as a product, yield: 93 percent.

Step 2: synthesis of Compound 37-3

To a reaction flask were added compound 37-2(200mg,0.73mmol), ethanol (4ml) and hydrazine hydrate (80%, 91mg,1.45mmol), and the reaction solution was stirred at room temperature overnight. The reaction solution is filtered, washed by a small amount of ethanol, and the filtrate is concentrated under reduced pressure to obtain a crude product 37-3 without further purification.

37 methods of synthesis reference is made to the methods of synthesis 1-12.

LC/MS:m/z＝492.9[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ3.69(1H,d,J＝9.6Hz),3.80-3.85(2H,m),3.93(1H,dd,J＝9.5,4.2Hz),4.14(1H,s),4.45(1H,d,J＝3.6Hz),4.60(2H,d,J＝3.5Hz),5.24(1H,d,J＝3.8Hz),7.02(1H,dd,J＝8.5,2.4Hz),7.17-7.20(3H,m),7.23-7.27(1H,m),7.45-7.51(2H,m),7.59(1H,d,J＝8.5Hz),7.68(1H,s),8.30(1H,s),8.87(1H,d,J＝7.1Hz),12.82(1H,brs).

Example 38

Synthesis step 1: synthesis of Compound 38-2

Addition of Compound 38 to the reaction flask1(60mg,0.215mmol), phenylacetylene (44mg,0.429mmol), DMF (1mL), TEA (43mg,0.429mmol), CuI (10mg) and PdCl₂(PPh)₃(20mg), the reaction solution was replaced with liquid nitrogen, and the mixture was heated to 120 ℃ and stirred for 5 hours. The reaction was extracted 2 times with ethyl acetate and the organic phases were combined, washed with saturated brine, concentrated under reduced pressure and purified on silica gel plates to give 46mg of product 38-2, yield: 84 percent.

38 with reference to methods of synthesis 1-12.

LC/MS:m/z＝349.1[M+H]⁺.

Example 39

39 synthetic method referring to the synthesis of 37 from 37-1, the starting material was replaced with 39-1 (641-74-7).

LC/MS:m/z＝493.0[M+H]⁺.

¹H NMR(400MHz,d6-DMSO)δ3.44(1H,d,J＝8.0Hz),3.78(1H,dd,J＝8.4,6.6Hz),3.93(1H,d,J＝9.3Hz),4.02(1H,dd,J＝9.5,4.3Hz),4.14-4.20(1H,m),4.46-4.51(2H,m),4.58-4.61(1H,m),4.94(1H,d,J＝6.3Hz),7.02(1H,dd,J＝8.5,2.4Hz),7.17-7.21(3H,m),7.26(1H,t,J＝7.4Hz),7.46-7.50(2H,m),7.58(1H,d,J＝8.5Hz),7.66(1H,s),8.30(1H,s),8.89(1H,d,J＝6.9Hz),12.81(1H,brs).

Example 40

The synthesis method of 40 refers to the synthesis method of 38, and the raw material phenylacetylene is replaced by 2-chlorophenylacetylene.

LC/MS:m/z＝383.0[M+H]⁺.

EXAMPLE 41

41-1, Organic Letters,2015, vol.17, #18, p.4640-4643.

41 Synthesis method with reference to the method for synthesizing 38, the starting material phenylacetylene was replaced with 41-1.

LC/MS:m/z＝441.0[M+H]⁺.

Test of drug efficacy

Test example 1: in vitro BTK inhibition kinase activity assay

1: compound preparation

Compound powders were dissolved in 100% DMSO to make 10mM stock solutions. And (4) freezing and storing at-20 ℃ in a dark place.

2: kinase reaction process

(1) Preparing 1 XKinase buffer;

(2) preparation of compound concentration gradient: test compounds were tested at 1 μ M concentration, diluted to 100-fold final concentration in 100% DMSO solutions in 384source plates, 3-fold compound dilutions, 10 concentrations. Using the dispenser Echo 550 to the target plate OptiPlate-384F transfer 250nL 100 times the final concentration of compounds;

(3) preparing a Kinase solution with 2.5 times of final concentration by using 1 XKinase buffer;

(4) add 10. mu.L of 2.5 fold final concentration kinase solution to the compound well and positive control well, respectively; add 10. mu.L of 1 XKinase buffer to the negative control wells;

(5) centrifuging at 1000rpm for 30 s, oscillating and mixing the reaction plate, and incubating at room temperature for 10 min;

(6) preparing a mixed solution of ATP and Kinase substrate2 with a final concentration of 5/3 times by using 1 XKinase buffer;

(7) adding 15 μ L of a mixed solution of ATP and substrate at 5/3 times final concentration to initiate the reaction;

(8) centrifuging a 384-hole plate at 1000rpm for 30 seconds, shaking and uniformly mixing, and incubating for 10 minutes at room temperature;

(9) adding 30 mu L of termination detection solution to stop kinase reaction, centrifuging at 1000rpm for 30 seconds, and uniformly mixing by oscillation;

(10) the conversion was read using a Caliper EZ Reader.

3: data analysis

Calculating the formula:

wherein: conversion% _ sample is the Conversion reading for the sample; conversion% _ min: negative control well mean, representing conversion readings without enzyme live wells; conversion% _ max: positive control wells are averaged for conversion readings in wells without compound inhibition.

Fitting dose-effect curve

The log values of the concentrations were taken as the X-axis and the percent inhibition as the Y-axis, and the log (inhibitor) vs. response-Variable slope of the analytical software GraphPad Prism5 was used to fit the dose-effect curves to obtain IC50 values for the enzyme activities of the respective compounds.

The formula is calculated as that Y ═ Bottom + (Top-Bottom)/(1+10^ ((Logic 50-X). HillSlope)) compounds have inhibitory activity on BTK wild type and BTK mutant C481S kinase as shown in Table 3.

Table 3: compound has activity of inhibiting BTK and BTK-C481S kinase

Test example 2: liver microsome stability test

1: adding 10 μ L of a test or control working solution and 80 μ L of a microsome working solution (the concentration of liver microsome protein is 0.5mg/mL) to the well site of the T0, T5, T10, T20, T30, T60, and NCF60 samples, adding only the microsome working solution to the well site of Blank60, and then placing the samples Blank60, T5, T10, T20, T30, and T60 except for T0 and NCF60 in a water bath at 37 ℃ for pre-incubation for about 10 minutes;

2: adding 300 mu L of termination solution (acetonitrile solution of conjugation 200ng/mL tolbutamide and 200ng/mL labetalol) into a T0 sample, and then adding 10ul of NADPH regeneration system working solution;

3: after the preincubation of the incubation plates Blank60, T5, T10, T20, T30 and T60 is finished, 10uL of NADPH regeneration system working solution is added into each sample well to start the reaction, and 10uL of 100mM potassium phosphate buffer solution is added into the NCF60 sample well;

4: after incubation for an appropriate time (e.g., 5, 10, 20, 30, and 60 minutes), 300. mu.L of stop solution was added to each of the test sample wells and the control sample wells of Blank60, T5, T10, T20, T30, T60, and NCF60 plates, respectively, to stop the reaction.

5: all sample plates were shaken and centrifuged at 4000rpm for 20 minutes, 100. mu.L of test or control supernatant, respectively, was diluted into 300. mu.L of pure water for LC-MS/MS analysis

6: data analysis, calculation of T1/2 and CL based on first order elimination kinetics_int(mic)(μ L/min/mg) value, first order elimination kinetics equation:

the results of the human and rat liver microsome stability tests are shown in table 4.

Table 4 results of liver microsome stability test of the compounds of the present invention

Test example 3: compounds for in vitro cell proliferation (TMD8) inhibition activity assay

1: cell lines

Culturing at 37 deg.C under 5% CO2 and 95% humidity.

2: compound configuration

The test compounds were diluted with DMSO to give a final concentration of 10mM stock solution.

3: cell culture and inoculation

(1) Cells in the logarithmic growth phase were harvested and counted using a platelet counter. Detecting the cell viability by using a trypan blue exclusion method to ensure that the cell viability is over 90 percent;

(2) adjusting the cell concentration; add 90 μ Ι _ of cell suspension to 96-well plates, respectively;

(3) cells in 96-well plates were incubated overnight at 37 ℃ with 5% CO2 and 95% humidity.

4: drug dilution and dosing

(1) Preparing 10 times of drug solution, wherein the highest concentration is 10 mu M, the concentration is 9, the dilution is 3.16 times, 10 mu L of drug solution is added into each hole of a 96-hole plate inoculated with cells, three multiple holes are arranged for each drug concentration, the final acting concentration of the compound is 1 mu M, the concentration is 9, the dilution is 3.16 times, and the final acting concentration of DMSO is 0.1%;

(2) the cells in the dosed 96-well plate were incubated for a further 72 hours at 37 ℃ under 5% CO2 and 95% humidity, after which they were subjected to CTG analysis.

5: terminal reading board

(1) Melt CTG reagents and equilibrate cell plates to room temperature for 30 minutes;

(2) adding equal volume of CTG solution into each well;

(3) the cells were lysed by shaking on an orbital shaker for 5 minutes;

(4) the cell plate was left at room temperature for 20 minutes to stabilize the luminescence signal;

(5) and reading the cold light value.

6: data processing

Data were analyzed using GraphPad Prism 7.0 software, fitted to the data using non-linear S-curve regression to derive a dose-effect curve, and IC50 values were calculated therefrom.

The cell survival rate (%) × (Lum test drug-Lum culture solution control)/(Lum cell control-Lum culture solution control) × 100%.

The results of the in vitro TMD8 cell proliferation inhibitory activity are shown in Table 5.

Table 5: inhibitory Activity of some Compounds of the present invention on TMD8 cell proliferation

Examples	TMD8 IC50(nm)
		17	36
21	24
		34	61
35	71
		ARQ-531	35

From the above examples, it can be seen that the compound of the present invention, which is an inhibitor of BTK protein kinase, has strong inhibitory effect on both wild-type BTK and mutant BTK (C481S) kinase, has good stability of liver microsome and good cell activity, and can be used for preparing drugs for treating diseases caused by over-expression of BTK kinase.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. A compound having the structure of formula I or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, pharmaceutically acceptable hydrate, solvate, or salt thereof:

wherein:

A₁、A₂、A₃、A₄are each independently selected from CR₉、N；

Y is selected from CR₁₀R₁₁、O、NR₁₀、S、S(O)、S(O)₂C-S-O, C-alkynyl;

R₁、R₂、R₉、R₁₀、R₁₁each independently selected from hydrogen, halogen, substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, nitro, mercapto, amidoSulfonyl, phosphoryl, alkylphosphoryl, alkylsulfonyl, alkylsulfoxide; or R₁、R₂Together with the N atom to which they are attached, form a substituted or unsubstituted heterocycloalkyl;

R₁、R₂not hydrogen at the same time.

2. The compound of claim 1, wherein a is₁、A₂、A₃、A₄At least one of which is CR₉。

3. The compound of claim 1,

selected from the group consisting of substituted or unsubstituted indolyl, 7-azaindolyl, 5, 7-diazaindolyl, pyrazolopyrimidinyl;

further, said R₉Selected from the group consisting of hydrogen, halogen, methyl, trifluoromethyl, difluoromethyl, isopropyl, cyclopropyl, isobutyl, amido, dimethylphosphite, diethylphosphophite, cyano, hydroxy, amino, methoxy, ethoxy, cyclopropoxy, isobutoxy, methylamino, dimethylaminoEthylamino, diethylamino, cyclopropylamino, methylsulfonyl and methylsulfonyl;

further, said R₉Selected from hydrogen, halogen, C1-3 alkyl, trifluoromethyl, difluoromethyl, methoxy, amido, dimethylphosphite and cyano.

4. A compound according to any one of claims 1 to 3, having a structure according to formula (II) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

5. A compound according to claim 4, wherein A is₁Or A₂Is CR₉，A₃Is N;

further, A₁Is CR₉，A₃Is N.

6. A compound of claim 1, wherein R is₁Selected from substituted or unsubstituted alkyl or heteroalkyl, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted unsaturated cyclic or heterocyclic, substituted or unsubstituted arylOr a heteroaryl group; further, R₁Selected from substituted or unsubstituted alkyl, substituted or unsubstituted C3-15 cycloalkyl or heterocycloalkyl, substituted or unsubstituted C3-15 unsaturated heterocyclyl and substituted or unsubstituted heteroaryl, wherein hetero atoms in the heterocycloalkyl or heterocyclyl are selected from N, O, S;

7. A compound of claim 6, wherein R is₁The compound is selected from substituted or unsubstituted alkyl, substituted or unsubstituted C3-15 cycloalkyl or oxygen-containing heterocycloalkyl, substituted or unsubstituted C3-C15 unsaturated oxygen-containing heterocyclic group and substituted or unsubstituted nitrogen-containing heteroaryl, wherein the substituents are respectively and independently selected from halogen, C1-C6 alkyl, C3-6 cycloalkyl, methylene and hydroxyl.

8. A compound of claim 7, wherein R is₁Selected from substituted or unsubstituted alkyl, substituted or unsubstituted adamantyl, substituted or unsubstituted oxygen-containing heterocycloalkyl, substituted or unsubstituted dihydropyran, substituted or unsubstituted nitrogen-containing heteroaryl, said oxygen-containing heterocycloalkyl being selected from tetrahydropyranyl, 6-oxaspiro [2.5] O]Octyl, 3-oxabicyclo [4.1.0]]A heptylalkyl group, wherein the nitrogen-containing heteroaryl group is selected from pyridyl and pyrazinyl, wherein the substituents are respectively and independently selected from halogen, C1-6 alkyl substituted by one or more hydroxyl, C1-6 alkyl substituted by nitrogen-containing heterocycloalkyl, hydroxycyclopropyl, methylene and hydroxyl; further, the dihydropyran is 3, 6-dihydropyran.

9. The compound according to any one of claims 1 to 6, having a structure represented by formula (III) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

e is selected from O, NR₁₂、S、S(O)、S(O)₂、C＝O、C＝S；

n₁An integer selected from 0 to 8;

R₄selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, methylene, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, sulfonyl, phosphoryl, or two adjacent R₄The two carbon atoms are connected together to form a substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or R at two same sites₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄Together with the carbon atom to which they are attached form a carbon-carbon double bond, or two non-adjacent R₄Together form a bridged ring structure, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxy, cyano, amino, ester, amide, aryl, heteroaryl, sulfonyl;

R₃selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, substituted or unsubstituted aryl or heteroaryl, sulfonyl, phosphoryl, and alkylphosphoryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl, sulfonyl, phosphoryl, alkylphosphoryl, alkylsulfone, and alkylsulfoxide;

R₁₂is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycleAlkyl, hydroxyl, cyano, amino, ester, amide, aryl, heteroaryl.

10. The compound of claim 9,

A₁is CR₉Or N, A₃Is N;

e is selected from O;

n₁an integer selected from 0 to 6;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or R at two same sites₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester group, amido, aryl, heteroaryl and sulfonyl;

R₃selected from the group consisting of hydrogen, halogen, methyl, trifluoromethyl, difluoromethyl, isopropyl, cyclopropyl, isobutyl, amide, dimethylphosphite, diethyloxyphosphite, cyano, hydroxy, amino, methoxy, ethoxy, cyclopropoxy, isobutoxy, methylamino, dimethylamino, ethylamino, diethylamino, cyclopropylamino, methylsulfoxide, methylsulfone.

11. A compound according to claim 10, wherein n is₁An integer selected from 0 to 4;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or two adjacent R₄The carbon atoms connected with the aryl form substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl or twoR at the same position₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl, or two adjacent R₄And the carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano.

12. The compound of claim 11, wherein R is₄Selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, alkyl of C1-6 substituted by hydroxyl, cycloalkyl of C3-6 substituted by hydroxyl, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-10 cycloalkyl or R at two same sites₄The substituent and the carbon atom connected with the substituent form a substituted or unsubstituted C3-10 cycloalkyl, or two adjacent R₄And the carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano.

13. The compound of claim 1, wherein M is selected from substituted or unsubstituted aryl or heteroaryl, wherein said substituents are selected from halogen, alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl, unsaturated cyclic or heterocyclic, aryl, heteroaryl, phenoxy, hydroxy, cyano, amino, ester, amide, sulfonyl, phosphoryl, alkylphosphoroxy;

further, M is selected from substituted or unsubstituted aryl, wherein the substituent is selected from halogen, alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl, unsaturated cyclic or heterocyclic group, phenoloxy, and amide.

14. A compound according to any one of claims 9 to 13, having the structure of formula (IV) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

b is selected from O,

R₆the aryl or heteroaryl group is selected from substituted or unsubstituted monocyclic, bicyclic, tricyclic aryl or heteroaryl, wherein the substituent is selected from nitro, hydroxyl, amino, sulfydryl, halogen, cyano, ester group, carboxyl, amido, phosphamido, alkyl phospho, C1-6 alkyl or heteroalkyl, C3-6 cycloalkyl or heterocycloalkyl, and 5-6-membered aryl or heteroaryl.

15. The compound of claim 14,

A₁is CR₉；

n₁An integer selected from 0 to 4;

b is selected from O,

R₁₃Selected from hydrogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, or with R₆And atoms connected together form substituted or unsubstituted C5-8 heterocycloalkyl, substituted or unsubstituted C5-8 unsaturated heterocyclyl, heteroaryl, R₁₄、R₁₅The aryl-substituted aryl group is selected from hydrogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, and substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, wherein the substituents are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl and sulfonyl;

16. A compound according to claim 8,

b is selected from O,

R₅Selected from hydrogen, halogen; n is₂Selected from 0, 1;

17. A compound according to any one of claims 14 to 16, having a structure according to formula (V) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, pharmaceutically acceptable hydrate, solvate or salt thereof:

n₁an integer selected from 0 to 7;

R₈selected from hydrogen, nitro, hydroxyl, amino, sulfydryl, halogen, cyano, ester group, carboxyl, amido, phosphamido, alkyl oxygen phosphorus group, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted 5-6 membered aryl or heteroaryl, and the likeWherein the substituents are each independently selected from the group consisting of halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxy, cyano, amino, ester, amide, aryl, heteroaryl, sulfonyl; n is₃An integer selected from 0 to 5;

R₇the aryl group is selected from hydrogen, halogen, substituted or unsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, substituted or unsubstituted aryl or heteroaryl, hydroxyl, cyano, amino, ester, amido, sulfonyl and phosphoryl, wherein the substituents are independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl, cyano, amino, ester, amido, aryl, heteroaryl and sulfonyl.

18. The compound of claim 17, wherein a is₁Is N, N₁+n₃≥2。

19. The compound of claim 17, wherein a is₁Is CR₉。

20. A compound according to any one of claims 17 to 19,

n₁an integer selected from 0 to 3;

R₃selected from hydrogen, halogen, methyl, trifluoromethyl, difluoromethyl, isopropylA group, cyclopropyl group, isobutyl group, amide group, dimethylphosphide group, diethyloxyphosphide group, cyano group, hydroxyl group, amino group, methoxy group, ethoxy group, cyclopropoxy group, isobutoxy group, methylamino group, dimethylamino group, ethylamino group, diethylamino group, cyclopropylamino group, methylsulfoxide group, methylsulfone group;

R₅selected from hydrogen, halogen;

21. The compound of claim 20,

n₁an integer selected from 0 to 2;

R₄selected from hydrogen, halogen, hydroxyl, cyano, amino, methylene, C1-6 alkyl substituted by hydroxyl or halogen, C3-6 cycloalkyl substituted by hydroxyl or halogen, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-6 cycloalkyl or R at two same sites₄The substituent and the carbon atom connected with the substituent form a substituted or unsubstituted C3-6 cycloalkyl, or two adjacent R₄The carbon atoms connected with the substituent groups form a carbon-carbon double bond together, wherein the substituent groups are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

R₅selected from hydrogen, halogen;

R₈selected from hydrogen, halogen, substituted orUnsubstituted C1-6 alkyl or heteroalkyl, substituted or unsubstituted C3-6 cycloalkyl or heterocycloalkyl, hydroxy, cyano, wherein the substituents are selected from the group consisting of halogen, alkyl or heteroalkyl, cycloalkyl or heterocycloalkyl, hydroxy, cyano; n is₃Is selected from 0 or 1;

22. The compound of claim 21,

R₅selected from hydrogen, halogen;

R₈selected from hydrogen, halogen, C1-6 alkyl, C3-6 cycloalkyl and cyano;

23. The compound of claim 22, wherein R is₄Selected from hydrogen, halogen, hydroxy, or two adjacent R₄The carbon atoms connected with the substituted or unsubstituted C3-6 cycloalkyl or two adjacent R₄Together with the carbon atom to which they are attached form a carbon-carbon double bond, whereinThe substituents are respectively and independently selected from halogen, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, hydroxyl and cyano;

further, R₅Is halogen, R₈Is hydrogen.

24. The compound of claim 23, wherein R is₄Selected from hydrogen, F, hydroxy, or two adjacent R₄The carbon atom to which it is attached constituting a substituted or unsubstituted cyclopropyl group, or two adjacent R₄Together with the carbon atoms to which they are attached form a carbon-carbon double bond;

further, R₅Is Cl, R₈Is hydrogen.

25. The compound of claim 1, wherein the compound structure is selected from one of the following:

26. a pharmaceutical composition, wherein the active ingredient of the pharmaceutical composition is selected from the compounds of any one of claims 1 to 25, or one or more of stereoisomers, solvates, hydrates, pharmaceutically acceptable salts and co-crystals thereof.

27. Use of a compound of any one of claims 1 to 25, or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof, for the preparation of a protein kinase inhibitor; further, the kinase inhibitor is a BTK inhibitor.

28. Use of a compound according to any one of claims 1 to 25, or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof, in the manufacture of a medicament for use in the treatment of any one or more of an autoimmune disease, an inflammatory disease, a thromboembolic disease, an allergy, an infectious disease, a proliferative disorder and cancer.

29. The use according to claim 28, wherein the disease is selected from: arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, crohn's disease, dermatitis, asthma, sjogren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, skin rash, anti-neutrophil cytoplasmic antibody vasculitis, pemphigus vulgaris, chronic obstructive pulmonary disease, psoriasis; breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular cancer, gastric cancer, glioma, endometrial cancer, melanoma, renal cancer, bladder cancer, melanoma, bladder cancer, biliary tract cancer, renal cancer, pancreatic cancer, lymphoma, hairy cell cancer, nasopharyngeal cancer, pharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, lung cancer, non-small cell lung cancer, small cell cancer, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular cancer, hodgkin's leukemia, bronchial cancer, thyroid cancer, uterine corpus cancer, cervical cancer, multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, chronic lymphoid leukemia, Myeloid leukemia, non-hodgkin's lymphoma, primary macroglobulinemia.

30. Use of a compound of any one of claims 1 to 25, or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof, in the manufacture of a medicament for the treatment of a disease that causes overexpression of a BTK kinase.

31. Use of a compound of any one of claims 1 to 25, or a stereoisomer, solvate, hydrate, pharmaceutically acceptable salt or co-crystal thereof, for the manufacture of a medicament for the treatment of a disease caused by overexpression of a BTK kinase.