CN111763217B

CN111763217B - Thieno-nitrogen heterocyclic compounds, preparation method and application

Info

Publication number: CN111763217B
Application number: CN202010236388.0A
Authority: CN
Inventors: 万惠新; 沈竞康; 潘建峰; 马金贵; 查传涛
Original assignee: Shanghai Lingda Biomedical Co ltd
Current assignee: Shanghai Lingda Biomedical Co ltd
Priority date: 2019-03-30
Filing date: 2020-03-30
Publication date: 2022-06-28
Anticipated expiration: 2040-03-30
Also published as: WO2020200154A1; CN111763217A

Abstract

The invention discloses a thieno-nitrogen heterocyclic structure shown as a general formula I, or pharmaceutically acceptable salt thereof, or enantiomer, diastereoisomer, tautomer, solvate, polymorph or prodrug thereof, a preparation method and pharmaceutical application thereof.

Description

Thieno-nitrogen heterocyclic compounds, preparation method and application

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to thienoazacyclo JAK-STAT signal pathway inhibitor compounds, a preparation method and application thereof.

Background

The Signal Transducer and Activator of Transcription (STAT) family are potential cytoplasmic transcription factors coupled to tyrosine phosphorylation signaling pathways that serve the dual function of transmitting cytoplasmic signals and initiating transcription of nuclear genes in cells, particularly STAT3, which plays an important role in embryonic development and cell proliferative differentiation. Under normal physiological conditions, STAT3 activation is rapid and transient, lasting only minutes to hours and tightly regulated, whereas STAT3 is persistently activated in many tumor tissues, including leukemia, pancreatic cancer, multiple myeloma, head and neck squamous cell carcinoma, prostate cancer, breast cancer, lung cancer, ovarian cancer, osteosarcoma, and gastric cancer, among others. Meanwhile, STAT3 is a focus of convergence of multiple oncogenic tyrosine signal pathways such as EGFR, Src, IL-6/JAK and the like, and can be activated by various cytokines, hormones, growth factors and the like to generate phosphorylation (pSTAT3), thereby influencing proliferation, differentiation, apoptosis and the like of cells.

STAT3 forms a dimer after phosphorylation, migrates into cell nucleus, combines with STAT3 dependent gene promoter region specific DNA response element, starts target gene transcription, plays a role of oncogene in many tumors, and simultaneously STAT3 mediates the drug resistance generation of various chemotherapeutic drugs. Chemotherapeutic drugs generally have poor therapeutic effects on tumor drug-resistant cells abnormally activated by the STAT3 signaling pathway, and blocking the signaling pathway can improve or even reverse the tumor cell chemotherapy drug resistance. STAT3 is therefore an important potential target for tumor therapy. Therefore, the continuous activation state of STAT3 protein is widely existed in the process of tumorigenesis and development, and is a valve for continuous tumor replication and propagation.

Based on the signaling pathway of STAT3, researchers have attempted to design a series of inhibitors at different targets to close a "valve". The current research is mainly focused on the following four aspects: the method is characterized by searching molecules capable of inhibiting the phosphorylation of STAT3 protein, searching molecules capable of inhibiting the coupling of STAT3 protein, searching molecules capable of inhibiting the combination of STAT3 dimer and DNA, and searching molecules capable of directly inhibiting the expression of STAT3 protein. Some STAT3 inhibitors and research reports on the antitumor activity of the inhibitors have been reported at present, such as antisense nucleotide, JAK kinase inhibitors TG-101348, WP1066, INCB018424, S-Ruxolitinib, Static inhibiting STAT3 dimerization, and S3I-201. The above inhibitors have many problems: or the medicine effect is weak, the medicine property is poor, the target point is not clear, or the toxicity is large, so that the biological function research and the clinical application of the STAT3 inhibitor in the preclinical process are severely limited.

Therefore, the search for a novel micromolecular STAT3 inhibitor or a STAT3 protein degradation agent which has a novel structure, a definite mechanism, high efficiency, low toxicity and good drug formation property becomes one of the hotspots in the research and development field of the current antitumor drugs.

Disclosure of Invention

One of the technical problems to be solved by the invention is to provide a novel STAT3 inhibitor or degradation agent for preparing a therapeutic drug related to tumors or inflammatory diseases.

The scheme for solving the technical problems is as follows:

a thieno-nitrogen heterocyclic compound shown as a general formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, a diastereoisomer, a tautomer, a solvate, a polymorph or a prodrug thereof,

wherein R1 is hydrogen, halogen, C1-C6 alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-8 membered aryl, 5-8 membered heteroaryl, substituted or unsubstituted amino, -COOR, -CONHR, cyano, sulphonyl or sulphoxy; r is hydrogen, C1-C6 alkyl, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl;

r2 is a substituted or unsubstituted amino, amido, ester, 5-8 membered aryl or 5-8 membered heteroaryl;

n is an integer of 0 to 2;

m1, M2 and M3 are independently selected from N, O, S, NH, -CR3a ═ CR3 b-or-CR 3

Wherein R3, R3a, and R3b are independently hydrogen, halogen, C1-C10 alkyl, 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted amino, substituted or unsubstituted hydroxyl, cyano, sulfone, sulfoxide, acyl, sulfonyl, sulfinyl, ureido, sulfonylureido, alkenyl, alkynyl, mercapto, HC (═ O) NH-, or

Alternatively, R3a and R3b may form a 3 to 8 membered saturated, unsaturated or partially unsaturated ring system through a carbon chain or heteroatom;

alternatively, two adjacent R3 taken together with the carbon atom to which they are attached form a phenyl group; said phenyl being optionally substituted by one or more C1-C6 alkoxy groups;

one or more hydrogen atoms of any one of the groups R, R1, R2, R3, R3a, R3b, M1, M2, and M3 are optionally substituted with a substituted or unsubstituted group selected from the group consisting of: deuterium, halogen, hydroxyl, amino, cyclic amino, cyano, nitro, sulfone, sulfoxide, C1-C8 alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, C1-C8 alkoxy, C1-C8 alkylthio, C1-C18 alkylamino, alkenyl, alkynyl, acyl, sulfonyl, urea or sulfonylurea, 5-8 membered aryl, 5-8 membered heteroaryl, NH ₂S(＝O)₂-、NH₂C (═ O) -or Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O, P or S, the heterocycloalkyl group containing 1 to 3 heteroatoms selected from the group consisting of: n, O, P or S, said ring system including spiro, bridged, fused, etc. saturated or partially unsaturated ring systems;

or any adjacent two substituents on R3a, R3b together with the carbon atom to which they are attached form a 5-8 membered heterocycloalkenyl or 5-8 membered heteroaryl;

wherein said substituents are selected from the group consisting of 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, and 5-8 membered heteroaryl, optionally substituted with one or more substituents selected from the group consisting of: C1-C6 alkyl, C1-C6 alkyl substituted with one or more Rds, or 3-8 membered heterocycloalkyl;

rd is independently hydroxy, halo, C1-C6 alkoxy, 3-8 membered cycloalkyl,

Or (C1-C6 alkyl) -S (═ O)₂-；

Rd1 and Rd2 are independently hydrogen or C1-C6 alkyl;

among said substituents, said C1-C8 alkyl or C1-C8 alkoxy is optionally substituted with one or more substituents selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy, C1-C8 alkoxy substituted by one or more halogens,

NH₂C (═ O) -, (C1-C6 alkyl) -C (═ O) -, 3-8 membered heterocycloalkyl, 5-8 membered heteroaryl or (C1-C6 alkyl) -S (═ O) ₂-；

Re and Rf are independently hydrogen or C1-C6 alkyl;

m1 and m2 are independently 0 or 1;

L_a、L_band L_cIndependently becomeA bond, - (C1-C6 alkylene) -, - (C1-C6 alkylene) -O-, or-C (═ O) -;

ra is amino, 3-8 membered heterocycloalkyl, 6-10 membered aryl, 5-8 membered heteroaryl or

Said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino, halogen, C1-C6 alkyl, C1-C6 alkyl substituted by one or more halogens, C1-C6 alkoxy, C1-C6 alkoxy substituted by one or more halogens, NH₂-S(＝O)₂-or (C1-C6 alkyl) -S (═ O)₂-；

Ra1 and Ra2 are independently hydrogen or C1-C6 alkyl;

rb and Rc are independently 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl; the 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl is optionally substituted with one or more C1-C6 alkyl.

In some preferred embodiments of the present invention, in the thienonitrogen heterocycle compound of general formula I, or its pharmaceutically acceptable salt, or its enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug, the thienonitrogen heterocycle compound of general formula I is a compound of formula I-1, some groups are defined as follows (undefined groups are as defined in any of the embodiments of the present application),

In the formula: r1 is independently selected from hydrogen, halogen, C1-C6 alkyl, 3-8 membered cycloalkyl or heterocycloalkyl, 5-8 membered aryl or heteroaryl, substituted or unsubstituted amino, -COOR, -CONHR, cyano, sulfone, sulfoxide, and the like; r is selected from hydrogen, C1-C6 alkyl, 3-8 membered cycloalkyl or heterocycloalkyl, etc.;

r2 is independently selected from substituted or unsubstituted amino, amido, ester, 5-8 membered aryl or heteroaryl, and the like;

n is independently selected from an integer of 0-2;

m1, M2, M3 are each independently selected from N, O, S, -CR3a ═ CR3b-, -CR3, wherein R3, R3a, Rb are each independently selected from hydrogen, halogen, C1-C10 alkyl, 3-to 10-membered cycloalkyl or heterocycloalkyl, 5-to 10-membered aryl or heteroaryl, substituted or unsubstituted amino, substituted or unsubstituted hydroxyl, cyano, sulfone, sulfoxide, acyl, sulfonyl, sulfinyl, ureido, sulfonylureido, alkenyl, alkynyl and the like, and the above-mentioned R3a and R3b may form a 3-to 8-membered saturated, unsaturated or partially unsaturated ring system through a carbon chain or heteroatom;

one or more hydrogen atoms of any one of the groups R, R1, R2, R3, R3a, R3b, M1, M2, and M3 may be substituted with a substituted or unsubstituted group selected from the group consisting of: including but not limited to deuterium, halogen, hydroxy, amino or cyclic amino, cyano, nitro, sulfone or sulfoxide, C1-C8 alkyl, 3-8 membered cycloalkyl or heterocycloalkyl, C1-C8 alkoxy or alkylthio, C1-C8 alkylamino, alkenyl, alkynyl, acyl or sulfonyl, urea or sulfonylurea, 5-8 membered aryl or heteroaryl; wherein said heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O, P or S, the heterocycloalkyl group containing 1 to 3 heteroatoms selected from the group consisting of: n, O, P or S, said ring system including spiro, bridged, fused, etc. saturated or partially unsaturated ring systems.

Further preferred is a compound represented by the general formula (II), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof:

wherein R4 is independently selected from the group consisting of C1-C10 alkyl, 3-10 membered cycloalkyl or heterocycloalkyl, 5-12 membered aryl or heteroaryl, and the like, and one or more hydrogen atoms on the above R4 group may be substituted with a substituted or unsubstituted group selected from the group consisting of: including but not limited to deuterium, halogen, hydroxy, amino or cyclic amino, cyano, nitro, sulfone or sulfoxide, C1-C8 alkyl, 3-8 membered cycloalkyl or heterocycloalkyl, C1-C8 alkoxy or alkylthio, C1-C8 alkylamino, alkenyl, alkynyl, acyl or sulfonyl, urea or sulfonyl urea, 5-8 membered aryl or heteroaryl; n, M1, M2, M3 are as defined above.

Further preferred is a compound represented by the general formula (III), or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, wherein:

wherein R5 is independently selected from C1-C10 alkyl, 3-10 membered cycloalkyl or heterocycloalkyl, 5-12 membered aryl or heteroaryl, and the like; m is independently selected from an integer from 1 to 4, preferably from 1, 2; r3 and R4 are as defined above.

In some preferred embodiments of the present invention, in the thienonitrogen heterocycles of general formula I, or their pharmaceutically acceptable salts, or their enantiomers, diastereomers, tautomers, solvates, polymorphs, or prodrugs, the thienonitrogen heterocycles of general formula I are compounds of formula I-1, some of which are defined as follows (the undefined groups are as defined in any embodiment of the present application),

r1 is-CONH₂Or a cyano group;

r5 is 3-10 membered heterocycloalkyl or 5-12 membered heteroaryl; said 3-10 membered heterocycloalkyl is optionally substituted with one or more C1-C8 alkyl groups; said 5-12 membered heteroaryl is optionally substituted with one or more substituents independently selected from the group consisting of: C1-C6 alkyl, C1-C6 alkyl substituted with one or more Rds, and 3-8 membered heterocycloalkyl; in the 3-10 membered heterocycloalkyl and 5-12 membered heteroaryl, the heteroatoms are selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

rd is independently hydroxy, halo, C1-C6 alkoxy, 3-8 membered cycloalkyl,

Or (C1-C6 alkyl) -S (═ O)₂-；

Rd1 and Rd2 are independently hydrogen or C1-C6 alkyl;

M1, M2 and M3 are independently selected from N, O, S, NH, -CR3a ═ CR3 b-and-CR 3

R3 is independently hydrogen or halogen;

r3a and R3b are independently hydrogen, cyano, halogen, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted amino, substituted or unsubstituted hydroxy, mercapto, HC (═ O) NH-, or

In the 3-10 membered heterocycloalkyl and 5-10 membered heteroaryl, the heteroatoms are selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

alternatively, R3a and R3b form a phenyl or 5-8 membered heteroaryl group through a carbon chain atom; said phenyl is optionally substituted with one or more halogens;

in R3a and R3b, the 3-to 10-membered heterocycloalkyl group, the 5-to 10-membered aryl group, the 5-to 10-membered heteroaryl group, the substituted or unsubstituted hydroxyl group, the mercapto group, HC (═ O) NH-, or

One or more hydrogen atoms thereon are optionally substituted with a substituent selected from the group consisting of: hydroxy, halogen, cyano, C1-C8 alkyl, C1-C8 alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, NH₂S(＝O)₂-、NH₂C (═ O) -and Ra-L _a-(Rb)_m1-L_b-(Rc)_m2-L_c-; said C1-C8 alkyl is optionally substituted with one or more substituentsThe substituents of (a) are independently selected from: hydroxy, cyano, halogen, C1-C8 alkoxy,

NH₂C (═ O) -, (C1-C6 alkyl) -C (═ O) -, 3-8 membered heterocycloalkyl, 5-8 membered heteroaryl, and (C1-C6 alkyl) -S (═ O)₂-; said C1-C8 alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of: halogen and C1-C6 alkyl; in the 3-8 membered heterocycloalkyl, 3-10 membered heterocycloalkyl, 5-8 membered heteroaryl and 5-10 membered heteroaryl, the heteroatom is selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

or any adjacent two substituents on R3a, R3b together with the carbon atom to which they are attached form a 5-8 membered heterocycloalkenyl or 5-8 membered heteroaryl; in the 5-8 membered heteroaryl, the heteroatom is selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

re and Rf are independently hydrogen or C1-C6 alkyl;

L_a、L_band L_cIndependently a bond, - (C1-C6 alkylene) -, - (C1-C6 alkylene) -O-, or-C (═ O) -;

Said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino, halogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more halogens, C1-C6 alkoxy, C1-C6 alkoxy substituted with one or more halogens, NH₂-S(＝O)₂-and (C1-C6 alkyl) -S (═ O)₂-; in the 3-8 membered heterocycloalkyl and 5-8 membered heteroaryl, the heteroatoms are selected from N, O and S, and the number of heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

ra1 and Ra2 are independently hydrogen or C1-C6 alkyl;

rb and Rc are independently 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl; said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl is optionally substituted with one or more C1-C6 alkyl; in the 3-8 membered heterocycloalkyl and 5-8 membered heteroaryl, the heteroatoms are selected from N, O and S, and the number of heteroatoms is 1, 2 or 3; when the substituent is plural, the same or different.

In some preferred embodiments of the present invention, in the thienonitrogen heterocycles of general formula I, or their pharmaceutically acceptable salts, or their enantiomers, diastereomers, tautomers, solvates, polymorphs, or prodrugs, some groups are defined as follows (the undefined groups are as defined in any of the embodiments of the present application),

R5 is 5-12 membered heteroaryl; said 5-12 membered heteroaryl is optionally substituted with one or more substituents independently selected from the group consisting of: C1-C6 alkyl or C1-C6 alkyl substituted with one or more Rds.

r5 is 5-12 membered heteroaryl; the 5-12 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl.

rd is C1-C6 alkoxy.

In certain preferred embodiments of the present invention, in the thienonitrogen heterocycles of general formula I, or their pharmaceutically acceptable salts, or their enantiomers, diastereomers, tautomers, solvates, polymorphs, or prodrugs, certain groups are defined as follows (the undefined groups are as defined in any embodiment of the present application),

M1, M2 and M3 are independently selected from N, O, S, -CR3a ═ CR3 b-or-CR 3.

r3 is hydrogen or halogen.

r3a and R3b are independently hydrogen, cyano, halogen, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted hydroxy, mercapto or HC (═ O) NH-.

R3a and R3b are independently hydrogen, cyano, halogen, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, or substituted or unsubstituted hydroxy.

in R3a and R3b, one or more hydrogen atoms of the 3-to 10-membered heterocycloalkyl, 5-to 10-membered aryl, 5-to 10-membered heteroaryl, substituted or unsubstituted hydroxyl, mercapto or HC (═ O) NH-are optionally substituted by a substituent selected from the group consisting of: hydroxy, halogen, cyano, C1-C8 alkyl, C1-C8 alkoxy, 3-8 membered heterocycloalkyl, NH₂C (═ O) -and Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-; said C1-C8 alkyl is optionally substituted with one or more substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy,

NH₂C (═ O) -, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said C1-C8 alkoxy group being optionally substituted with one or more halogens; the 3-8 membered heterocycloalkyl group is optionally substituted with one or more C1-C6 alkyl groups.

in R3a and R3b, one or more hydrogen atoms of the 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, or substituted or unsubstituted hydroxyl group are optionally substituted with a substituent selected from the group consisting of: hydroxy, halogen, C1-C8 alkyl, C1-C2 alkoxy, 3-8 membered heterocycloalkyl and Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-; said C1-C8 alkyl is optionally substituted with one or two substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy, NH₂C (═ O) -, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said C1-C8 alkoxy group being optionally substituted with one or more halogens; said 3-to 8-membered heterocycloalkyl group being optionally substituted by one or more C1-C6 alkyl groupsAnd (4) substituting.

L_a、L_bAnd L_cIndependently a bond, - (C1-C6 alkylene) -, - (C1-C6 alkylene) -O-or-O-.

Said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino, halogen, C1-C6 alkoxy, C1-C6 alkoxy substituted with one or more halogens, or (C1-C6 alkyl) -S (═ O)₂-。

ra is amino, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said 3-8 membered heterocycloalkyl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino and (C1-C6 alkyl) -S (═ O) ₂-。

rb and Rc are independently 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl; the 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl is optionally substituted with one or more C1-C6 alkyl groups.

when R5 is a 3-10 membered heterocycloalkyl, the 3-10 membered heterocycloalkyl is a 3-6 membered heterocycloalkyl wherein the heteroatom is selected from N, O and S, the number of heteroatoms is 1 or 2, for example

When R5 is 3-10 membered heterocycloalkyl, said 3-10 membered heterocycloalkyl is optionally substituted with one or more C1-C8 alkyl, said C1-C8 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

when R5 is a 5-12 membered heteroaryl, said 5-12 membered heteroaryl is a 5-6 membered heteroaryl wherein the heteroatoms are selected from N and O, the number of heteroatoms is 1, 2 or 3; for example

when R5 is a 5-12 membered heteroaryl, said 5-12 membered heteroaryl is optionally substituted by one or more substituents being C1-C6 alkyl or C1-C6 alkyl substituted by one or more Rd, said C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl or ethyl.

when R5 is a 5-12 membered heteroaryl, the 5-12 membered heteroaryl is optionally substituted with one or more substituents which are 3-8 membered heterocycloalkyl, the 3-8 membered heterocycloalkyl is a 3-6 membered heterocycloalkyl wherein the heteroatoms are selected from N, O and S, the number of heteroatoms is 1 or 2, for example

when Rd is independently halogen, said halogen is fluorine, chlorine, bromine or iodine, e.g. fluorine.

When Rd is independently C1-C6 alkoxy, said C1-C6 alkoxy is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, for example methoxy.

when Rd is independently a 3-8 membered cycloalkyl group, said 3-8 membered cycloalkyl group is a 3-6 membered cycloalkyl group, e.g.

when Rd is independently (C1-C6 alkyl) -S (═ O)₂-wherein C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, Such as methyl.

when Rd1 and Rd2 are independently C1-C6 alkyl, said C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

when R3 is independently halogen, the halogen is fluorine, chlorine, bromine or iodine, for example bromine.

When R3a and R3b are independently halogen, the halogen is fluorine, chlorine, bromine or iodine, for example bromine.

when R3a and R3b are independently 3-10 membered heterocycloalkyl, the heterocycloalkyl is 3-6 membered heterocycloalkyl wherein the heteroatoms are selected from N and O and the number of heteroatoms is 1 or 2Mono, preferably 5-6 membered heterocycloalkyl, e.g.

when R3a and R3b are independently 5-10 membered aryl, the 5-10 membered aryl is phenyl.

When R3a and R3b are independently 5-10 membered heteroaryl, the 5-10 membered heteroaryl is 5-6 membered heteroaryl, wherein the heteroatoms are selected from N and O and the number of heteroatoms is 1, 2 or 3, e.g. N and O

when R3a and R3b form a 5-8 membered heteroaryl group through a carbon chain atom, the 5-8 membered heteroaryl group is a 5-6 membered heteroaryl group, for example

when R3a and R3b form a phenyl group through a carbon chain atom, the phenyl group is optionally substituted with one or more halogens, the halogen is fluorine, chlorine, bromine or iodine, for example fluorine.

when R3a and R3b form a phenyl group through a carbon chain atom, the phenyl group being optionally substituted with one or more C1-C6 alkoxy groups, the C1-C6 alkoxy group is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, for example methoxy.

when two adjacent R3 groups together with the carbon atom to which they are attached form a phenyl group, which phenyl group is optionally substituted with one or more C1-C6 alkoxy groups, the C1-C6 alkoxy group is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, for example methoxy.

in R3a and R3b, when said 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted hydroxy, mercapto, HC (═ O) NH-, or

One or more of the hydrogen atoms in (a) are optionally substituted by a substituent, which when halogen is fluorine, chlorine, bromine or iodine, for example fluorine.

Wherein one or more hydrogen atoms are optionally substituted with a substituent, said substituent being a C1-C8 alkyl group, said C1-C8 alkyl group being a methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl group, such as methyl, ethyl, propyl, isopropyl or sec-butyl.

in R3a and R3b, when the 3-to 10-membered heterocycloalkyl group is present, the number is 5 to 10Aryl, 5-to 10-membered heteroaryl, substituted or unsubstituted hydroxy, mercapto, HC (═ O) NH-, or

Wherein one or more hydrogen atoms are optionally substituted with a substituent, said substituent being C1-C8 alkoxy, said C1-C8 alkoxy group being methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, for example methoxy, ethoxy, propoxy, isopropoxy or n-butoxy.

in R3a and R3b, when said 3-to 10-membered heterocycloalkyl group, 5-to 10-membered aryl group, 5-to 10-membered heteroaryl group, substituted or unsubstituted hydroxy group, mercapto group, HC (═ O) NH-, or

Wherein one or more hydrogen atoms are optionally substituted by a substituent which, when said substituent is a 3-to 8-membered cycloalkyl group, said 3-to 8-membered cycloalkyl group is a 3-to 6-membered cycloalkyl group, for example

Wherein one or more hydrogen atoms are optionally substituted by a substituent, and wherein when said substituent is a 3-8 membered heterocycloalkyl group, said 3-8 membered heterocycloalkyl group is a 3-6 membered heterocycloalkyl group, wherein the heteroatom is selected from N, O and S, and the number of heteroatoms is 1 or 2, e.g.

Wherein one or more hydrogen atoms are optionally substituted by a substituent selected from the group consisting of C1-C8 alkyl, and wherein when said C1-C8 alkyl is optionally substituted by one or more halogen, said halogen is fluorine, chlorine, bromine, or iodine, e.g., fluorine.

One or more hydrogen atoms of which are optionally substituted by a substituent,the substituent is C1-C8 alkyl, and when the C1-C8 alkyl is optionally substituted by one or more C1-C8 alkoxy groups, the C1-C8 alkoxy group is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, such as methoxy.

Wherein one or more hydrogen atoms are optionally substituted by a substituent selected from the group consisting of C1-C8 alkyl, C1-C8 alkyl optionally substituted by one or more (C1-C6 alkyl) -C (═ O) -or (C1-C6 alkyl) -S (═ O) ₂-when substituted, said (C1-C6 alkyl) -C (═ O) -or (C1-C6 alkyl) -S (═ O)₂C1-C6 alkyl in (A-C) is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

One or more hydrogen atoms of (A) are optionally substituted by substituentsSaid substituent is C1-C8 alkyl, said C1-C8 alkyl is optionally substituted with one or more 3-8 membered heterocycloalkyl, said 3-8 membered heterocycloalkyl is 3-6 membered heterocycloalkyl wherein the heteroatom is selected from N, O and S, the number of heteroatoms is 1 or 2, preferably 5-6 membered heterocycloalkyl, e.g. C1-C8 alkyl

Wherein one or more hydrogen atoms are optionally substituted by a substituent(s) selected from the group consisting of C1-C8 alkyl, C1-C8 alkyl when optionally substituted by one or more 5-8 membered heteroaryl, wherein 5-8 membered heteroaryl is 5-6 membered heteroaryl, wherein the heteroatoms are N and O, and the number of heteroatoms is 1, 2 or 3, e.g.

r3a and R3b when the 3-to 10-membered heterocycloalkyl group is present, the 5-to 10-membered heterocycleAryl, 5-to 10-membered heteroaryl, substituted or unsubstituted hydroxy, mercapto, HC (═ O) NH-, or

Optionally substituted with a substituent(s) which is C1-C8 alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl, said C1-C8 alkoxy, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl is optionally substituted with one or more halogen(s) which is fluorine, chlorine, bromine or iodine, e.g. fluorine.

Wherein one or more hydrogen atoms are optionally substituted by a substituent selected from the group consisting of C1-C8 alkoxy, 3-8 membered cycloalkyl and 3-8 membered heterocycloalkyl, wherein C1-C8 alkoxy, 3-8 membered cycloalkyl and 3-8 membered heterocycloalkyl are optionally substituted by one or more C1-C6 alkyl groups, and wherein C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl.

When any adjacent two substituents on R3a, R3b together with the carbon atom to which they are attached form a 5-8 membered heterocycloalkenyl,said 5-8 membered heterocycloalkenyl is 5-6 membered heterocycloalkenyl, e.g.

when any adjacent two substituents on R3a, R3b taken together with the carbon atom to which they are attached form a 5-8 membered heteroaryl, said 5-8 membered heteroaryl is a 5-6 membered heteroaryl wherein the heteroatoms are selected from N and O and the heteroatoms are 1 or 2, e.g. N and O

when Re and Rf are independently C1-C6 alkyl, the C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, such as methyl.

when L is_a、L_bAnd L_cIndependently is- (C1-C6 alkylene) -or- (C1-C6 alkylene) -O-, C1-C6 alkylene in said- (C1-C6 alkylene) -or- (C1-C6 alkylene) -O-is methylene,Ethylene, propylene, isopropylene, n-butylene, isobutylene, sec-butylene or tert-butylene, e.g. -CH₂-or

when Ra is 3-8 membered heterocycloalkyl, said 3-8 membered heterocycloalkyl is 3-6 membered heterocycloalkyl wherein the heteroatom is selected from N, O and S, the number of heteroatoms is 1 or 2, preferably 5-6 membered heterocycloalkyl, e.g.

when Ra is 6-10 membered aryl, said 6-10 membered aryl is phenyl.

when Ra is 5-8 membered heteroaryl, said 5-8 membered heteroaryl is 5-6 membered heteroaryl, for example

When Ra is 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl, said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted by one or more substituents which are halogen or C1-C6 alkoxy substituted by one or more halogens, wherein said halogen is fluorine, chlorine, bromine or iodine, e.g. fluorine.

when Ra is 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl, said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted by one or more substituents C1-C6 alkoxy or C1-C6 alkoxy substituted by one or more halogens, wherein said C1-C6 alkoxy is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, for example methoxy.

when Ra is 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl, said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted by one or more substituents which are (C1-C6 alkyl) -S (═ O)₂-wherein said C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

when Ra1 and Ra2 are independently C1-C6 alkyl, said C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

when Rb and Rc are independently 3-8 membered heterocycloalkyl, said 3-8 membered heterocycloalkyl is 3-6 membered heterocycloalkyl wherein the heteroatoms are selected from N, O and S, the number of heteroatoms is 1 or 2, preferably 5-6 membered heterocycloalkyl, e.g.

when Rb and Rc are independently 6-10 membered aryl, the 6-10 membered aryl is phenyl.

When Rb and Rc are independently 5-10 membered heteroaryl, the 5-10 membered heteroaryl is 5-6 membered monocyclic heteroaryl or 8-10 membered bicyclic heteroaryl, the 5-6 membered monocyclic heteroaryl for example

The 8-to 10-membered bicyclic heteroaryl group is, for example, indolyl

when Rb and Rc are independently 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl, said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl being optionally substituted by one or more C1-C6 alkyl groups, said C1-C6 alkyl group is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

One or more hydrogen atoms of (a) are optionally substituted by a substituent of Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_cWhen said Ra-L is_a-(Rb)_m1-L_b-(Rc)_m2-L_c-is of

(6-10-membered aryl) - (C1-C6 alkylene) -, (5-8-membered heteroaryl) - (C1-C6 alkylene) -, (3-8-membered heterocycloalkyl) - (C1-C6 alkylene) - (5-10-membered heteroaryl) -, (3-8-membered heterocycloalkyl) -C (═ O) - (5-10-membered heteroaryl) -, (3-8-membered heterocycloalkyl) - (C1-C6 alkylene) - (6-10-membered aryl) - (C1-C6 alkylene) -, (3-8-membered heterocycloalkyl) -O- (6-10-membered aryl) - (C1-C6 alkylene) -, (3-8-membered heterocycloalkyl) - (C1-C6 alkylene) -O- (6-10-membered aryl) -, (3-8-membered heterocycloalkyl) - (C1-C6 alkylene) -, (6-10-membered aryl) -, or (C), (3-8-membered heterocycloalkyl) - (6-10-membered aryl) - (C1-C6 alkylene) -, (3-8-membered heterocycloalkyl) - (6-10-membered aryl) -O-, (3-8-membered heterocycloalkyl) -C (═ O) - (5-10-membered heteroaryl) -O-, (3-8-membered heterocycloalkyl) - (5-10-membered heteroaryl) -O-, (6-10-membered aryl) - (C1-C6 alkylene) - (3-8-membered heterocycloalkyl) -C (═ O) - (5-10-membered heteroaryl), (6-10-membered aryl) - (3-8-membered heterocycloalkyl) -C (═ O) - (5-10-membered heteroaryl) — or a pharmaceutically acceptable salt thereof, (6-10-membered aryl) - (C1-C6 alkylene) - (3-8-membered heterocycloalkyl) -C (═ O) -, (6-10-membered aryl) - (C1-C6 alkylene) - (3-8-membered heterocycloalkyl) -, (6-10-membered aryl) - (3-8-membered heterocycloalkyl) -C (═ O) -, (6-10-membered aryl) - (C1-C6 alkylene) - (3-8-membered heterocycloalkyl) -C (═ O) - (5-10-membered heteroaryl) -O-, (6-10-membered aryl) - (C1-C6 alkylene) - (3-8-membered heterocycloalkyl) -C (═ O) -, (6-10-membered heterocycloalkyl) - (3-8-membered heterocycloalkyl) -C ) -C (═ O) - (6-10 membered aryl) -O-or (6-10 membered aryl) - (3-8 membered heterocycloalkyl) -C (═ O) - (6-10 membered aryl) -O-.

rd is hydroxy, fluoro, methoxy, cyclopropyl,

is composed of

Preferably is

r3 is H or Br.

One or more hydrogen atoms of (a) are optionally substituted by a substituent selected from the group consisting of hydroxy, halo, cyano, C1-C8 alkyl, C1-C8 alkoxy, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, NH₂S(＝O)₂-or NH₂When C (═ O) -, the substituent is methoxy,

Trifluoromethyl, hydroxy, butoxy, methyl, NH₂S(＝O)₂-, chloro, ethoxy, isopropoxy, ethyl, cyano,

N-propyl, sec-butyl, isopropyl,

R5 is

Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-is of

r3a or R3b are independently H, Br, F, CN, NH₂、

when R3a and R3b form a phenyl or 5-8 membered heteroaryl group through a carbon chain atom,

Is composed of

when two adjacent R3 taken together with the carbon atom to which they are attached form a phenyl group,

is composed of

r1 is-CONH₂Or cyano;

r5 is 5-12 membered heteroaryl; said 5-12 membered heteroaryl is optionally substituted with one or more substituents independently selected from the group consisting of: C1-C6 alkyl or C1-C6 alkyl substituted with one or more Rds;

rd is C1-C6 alkoxy;

m1, M2 and M3 are independently selected from N, O, S, NH, -CR3a ═ CR3 b-or-CR 3;

r3 is independently hydrogen or halogen;

R3a and R3b are independently hydrogen, cyano, halogen, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted hydroxy, mercapto or HC (═ O) NH —;

in R3a and R3b, one or more hydrogen atoms of the 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, substituted or unsubstituted hydroxyl, mercapto or HC (═ O) NH "are optionally substituted by a substituent selected from the group consisting of: hydroxy, halogen, cyano, C1-C8 alkyl, C1-C8 alkoxy, 3-8 membered heterocycloalkyl, NH₂C (═ O) -and Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-; said C1-C8 alkyl is optionally substituted with one or more substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy,

NH₂C (═ O) -, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said C1-C8 alkoxy group being optionally substituted with one or more halogens; said 3-8 membered heterocycloalkyl is optionally substituted with one or more C1-C6 alkyl groups;

Re and Rf are independently hydrogen or C1-C6 alkyl;

Said 3-8 membered heterocycloalkyl, 6-10 membered aryl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino, halogen, C1-C6 alkoxy, C1-C6 alkoxy substituted with one or more halogens, or (C1-C6 alkyl) -S (═ O)₂-；

Ra1 and Ra2 are independently hydrogen or C1-C6 alkyl;

r1 is-CONH₂Or cyano;

r5 is 5-12 membered heteroaryl; said 5-12 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl;

Rd is C1-C6 alkoxy;

m1, M2 and M3 are independently selected from N, O, S, -CR3a ═ CR3 b-or-CR 3;

r3 is independently hydrogen or halogen;

r3a and R3b are independently hydrogen, cyano, halogen, 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, or substituted or unsubstituted hydroxy;

in R3a and R3b, one or more hydrogen atoms of the 3-10 membered heterocycloalkyl, 5-10 membered aryl, 5-10 membered heteroaryl, or substituted or unsubstituted hydroxyl group are optionally substituted with a substituent selected from the group consisting of: hydroxy, halogen, C1-C8 alkyl, C1-C2 alkoxy, 3-8 membered heterocycloalkyl, and Ra-L_a-(Rb)_m1-L_b-(Rc)_m2-L_c-; said C1-C8 alkyl is optionally substituted with one or two substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy, NH₂C (═ O) -, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said C1-C8 alkoxy group being optionally substituted with one or more halogens; said 3-8 membered heterocycloalkyl is optionally substituted with one or more C1-C6 alkyl groups;

Re and Rf are independently hydrogen or C1-C6 alkyl;

L_a、L_band L_cIndependently a bond, - (C1-C6 alkylene) -, - (C1-C6 alkylene) -O-or-O-;

ra is amino, 3-8 membered heterocycloalkyl, or 5-8 membered heteroaryl; said 3-8 membered heterocycloalkyl or 5-8 membered heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: amino and (C1-C6 alkyl) -S (═ O)₂-；

Ra1 and Ra2 are independently hydrogen or C1-C6 alkyl;

In some preferred embodiments of the present invention, in the thienonitrogen heterocycle compound shown in formula I, or its pharmaceutically acceptable salt, or its enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug, the thienonitrogen heterocycle compound shown in formula I is a compound shown in formula I-2 or formula I-3, some groups are defined as follows (undefined groups are as described in any embodiment of the present application),

wherein R1, M1, M2, M3 and R5 are as defined in any of the previous schemes;

R5a is C1-C6 alkyl, C1-C6 alkyl substituted with one or more Rd, or 3-8 membered heterocycloalkyl;

rd is independently hydroxy, halo, C1-C6 alkoxy, 3-8 membered cycloalkyl,

Or (C1-C6 alkyl) -S (═ O)₂-。

In some preferred embodiments of the present invention, the thienonitrogen heterocycle compound of formula I, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, is a thienonitrogen heterocycle compound of formula I, wherein the thienonitrogen heterocycle compound of formula I is a compound of formula I-5, I-6, I-7, I-8, I-9, I-10, I-11 or I-12, and some groups are defined as follows (undefined groups are defined as in any embodiment of the present application),

wherein R1, R5, R3a, R3b and R3 are as defined in any of the previous embodiments.

In some preferred embodiments of the invention, the thieno-nitrogen heterocycle compound shown in the general formula I can be any one of the following structures,

a process for the preparation of a compound of formula (II), characterized in that it comprises the steps a-b:

a) reacting a compound of formula (A1) with R under condensation reagent conditions₄NH₂Reacting the compound to obtain a compound with a general formula (B1); or converting the compound of the general formula (A2) into an intermediate (B2) by functional groups such as alkylation, acylation, reductive amination and the like; and

b) Carrying out substitution reaction or transition metal catalyzed coupling reaction on the compound with the general formula (B1) or (B2) and the compound with the general formula (C) under alkaline conditions to obtain a compound with a general formula (II); the definition of each group is as described above;

preferably, said steps a), b) are each carried out in a solvent and said solvent is selected from the group consisting of: water, methanol, ethanol, isopropanol, butanol, ethylene glycol methyl ether, N-methyl pyrrolidone, dimethyl sulfoxide, tetrahydrofuran, toluene, dichloromethane, 1, 2-dichloroethane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, dioxane, or a combination thereof.

Preferably, the transition metal catalyst is selected from the group consisting of: tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) Tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Palladium acetate, palladium chloride, dichlorobis (triphenylphosphine) palladium, palladium trifluoroacetate, triphenylphosphine palladium acetate, [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride, bis (tri-o-phenylphosphino) palladium dichloride, 1, 2-bis (diphenylphosphino) ethane palladium dichloride, or a combination thereof; the catalyst ligand is selected from the group consisting of: tri-tert-butylphosphine, tri-tert-butylphosphine tetrafluoroborate, tri-n-butylphosphine, triphenylphosphine, tri-p-benzylphosphine, tricyclohexylphosphine, tri-o-phenylphosphine, or a combination thereof.

Preferably, the inorganic base is selected from the group consisting of: sodium hydride, potassium hydroxide, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, or combinations thereof; the organic base is selected from the group consisting of: pyridine, triethylamine, N, N-diisopropylethylamine, 1, 8-diazabicyclo [5.4.0] undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide, lutidine, or a combination thereof.

The invention provides a preparation method of a thieno-nitrogen heterocyclic compound shown as a general formula I, which comprises the following steps: in a solvent, in the presence of alkali, carrying out substitution reaction on a compound shown as a formula B and a compound shown as a formula C as shown in the specification;

wherein n, R1, R2, M1, M2, and M3 are as defined in any of the previous versions;

x is chlorine, bromine, iodine or methylsulfonyl.

In the preparation method, the solvent may be one or more of water, an alcohol solvent (e.g., one or more of methanol, ethanol, isopropanol, butanol and ethylene glycol), an ether solvent (e.g., one or more of ethylene glycol methyl ether, tetrahydrofuran and dioxane), a ketone solvent (e.g., N-methylpyrrolidone), a sulfoxide solvent (e.g., dimethyl sulfoxide), an aromatic hydrocarbon solvent (e.g., toluene), a halogenated hydrocarbon solvent (e.g., dichloromethane and/or 1, 2-dichloroethane), a nitrile solvent (e.g., acetonitrile), an amide solvent (e.g., N-dimethylformamide and/or N, N-dimethylacetamide), preferably an ether solvent and/or an amide solvent.

In the preparation method, the solvent can be inorganic base and/or organic base. The inorganic base can be one or more of sodium hydride, potassium hydroxide, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, potassium fluoride, cesium fluoride, potassium phosphate, potassium carbonate, potassium bicarbonate, sodium carbonate and sodium bicarbonate. The organic base may be one or more of pyridine, triethylamine, N, N-diisopropylethylamine, 1, 8-diazabicyclo [5.4.0] undec-7-ene (DBU), lithium hexamethyldisilazide, sodium hexamethyldisilazide and lutidine, preferably N-diisopropylethylamine.

Another object of the present invention is to provide a medicament for treating or preventing tumor or inflammatory diseases and a composition thereof. The technical scheme for realizing the purpose is as follows:

a pharmaceutical composition for treating tumor or inflammatory diseases, which comprises the thienoazacyclo-compound shown in the general formula (I), or pharmaceutically acceptable salt thereof, or enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof and pharmaceutically acceptable carrier.

Another object of the present invention is to provide a use of the above compound. The technical scheme for realizing the purpose is as follows:

The thieno-nitrogen heterocyclic compound shown in the general formula (I) or pharmaceutically acceptable salt thereof, or enantiomer, diastereoisomer, tautomer, solvate, polymorph or prodrug thereof is used for preparing medicaments for treating diseases related to STAT protein activity or expression quantity, in particular to medicaments for treating tumors, immunological diseases and inflammatory diseases. The tumor is independently selected from non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostatic cancer, liver cancer, skin cancer, gastric cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymph cancer, fibroma, sarcoma, basal cell carcinoma, glioma, renal carcinoma, melanoma, bone cancer, thyroid cancer, nasopharyngeal carcinoma, pancreatic cancer and the like. The immune disease and the inflammatory disease are independently selected from the group consisting of rejection of transplanted organs, gout, rhinitis, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, arthritis, allergic dermatitis, Behcet's disease, bullous skin disease, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease, liver cirrhosis, degenerative joint disease, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, gastritis, nephritis, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease, irritable bowel syndrome, Kawasaki disease, meningococcus, multiple sclerosis, myocarditis, myasthenia gravis, mycosis fungoides, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, psoriasis, and chronic obstructive pulmonary disease, Fibrosis, lupus erythematosus, tissue graft rejection, thyroiditis, type I diabetes, urethritis, uveitis, vasculitis, vitiligo, Waldenstrom's macroglobulinemia, and the like.

The invention relates to a thieno-nitrogen heterocyclic compound with the structural characteristics of a general formula (I), which can inhibit various tumor cells, particularly can efficiently kill tumors related to STAT3 signal channel abnormity, such as leukemia, prostatic cancer, pancreatic cancer, breast cancer and the like, and is a therapeutic drug with a brand new action mechanism.

The invention provides a pharmaceutical composition, which comprises the thienonitrogen heterocyclic compound shown as the general formula I, or pharmaceutically acceptable salt thereof, or enantiomer, diastereoisomer, tautomer, solvate, polymorph or prodrug thereof, and a pharmaceutically acceptable carrier. The thienonitrogen heterocycles of formula I or IV, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph or prodrug thereof, may be in a therapeutically effective amount.

The invention also provides an application of the thieno nitrogen heterocyclic compound shown as the general formula I, or pharmaceutically acceptable salt thereof, or an enantiomer, a diastereoisomer, a tautomer, a solvate, a polymorph or a prodrug thereof, or the pharmaceutical composition in preparing STAT inhibitors. The STAT inhibitor is preferably a STAT3 inhibitor.

The invention also provides an application of the thieno-nitrogen heterocyclic compound shown as the general formula I, or pharmaceutically acceptable salt thereof, or enantiomer, diastereoisomer, tautomer, solvate, polymorph or prodrug thereof, or the pharmaceutical composition in preparing medicines for preventing or treating diseases related to STAT protein activity or expression quantity. The disease related to the STAT protein activity or expression quantity is preferably one or more of tumor, immune disease and inflammatory disease.

The invention also provides the application of the thieno-nitrogen heterocyclic compound shown as the general formula I, or pharmaceutically acceptable salt thereof, or enantiomer, diastereoisomer, tautomer, solvate, polymorph or prodrug thereof, or the pharmaceutical composition in preparing medicines. The medicament is preferably a medicament for preventing or treating one or more of tumors, immune diseases and inflammatory diseases.

The present invention also provides a method of treating one or more of a tumor, an immune disease, and an inflammatory disease, comprising administering to a patient a therapeutically effective amount of a thienonitrogen heterocycle as shown in formula I above, or a pharmaceutically acceptable salt thereof, or an enantiomer, diastereomer, tautomer, solvate, polymorph, or prodrug thereof, or a pharmaceutical composition thereof.

The tumor is independently selected from non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostatic cancer, liver cancer, skin cancer, gastric cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymph cancer, fibroma, sarcoma, basal cell carcinoma, glioma, renal carcinoma, melanoma, bone cancer, thyroid cancer, nasopharyngeal carcinoma, pancreatic cancer and the like.

The immune disease and the inflammatory disease are independently selected from the group consisting of rejection of transplanted organs, gout, rhinitis, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, arthritis, allergic dermatitis, Behcet's disease, bullous skin disease, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease, liver cirrhosis, degenerative joint disease, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, gastritis, nephritis, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease, irritable bowel syndrome, Kawasaki disease, meningococcus, multiple sclerosis, myocarditis, myasthenia gravis, mycosis fungoides, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, psoriasis, and chronic obstructive pulmonary disease, Fibrosis, lupus erythematosus, tissue graft rejection, thyroiditis, type I diabetes, urethritis, uveitis, vasculitis, vitiligo, Waldenstrom's macroglobulinemia, and the like.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. The space is not described herein in a repeated fashion.

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the thieno-nitrogen heterocyclic compound shown as the general formula I or IV has a novel structure, has good inhibitory activity and selectivity on STAT, particularly STAT3, has good metabolic stability and pharmacokinetic properties, and is good in oral bioavailability, high in safety and good in inhibitory effect on various tumors.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

Example 1 the inventors have made extensive and intensive studies for a long time to prepare a compound having a novel structure represented by formula I or formula II, and found that the compound has a good activity of inhibiting phosphorylation of STAT3 or degrading STAT3 protein, and that the compound has a specific inhibition or degradation effect on STAT3 at a very low concentration (which may be as low as ≤ 100nmol/L), i.e., on phosphorylation of STAT3, and has a very excellent inhibitory activity on cell proliferation associated with positive STAT3, and thus, the compound can be used for treating diseases associated with abnormal expression of STAT3, such as tumors. Based on the above findings, the inventors have completed the present invention.

Term(s)

Unless defined otherwise, all technical and scientific terms herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter claimed. In this application, the use of the singular also includes the plural unless specifically stated otherwise. It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are not limiting.

Definitions for the terms of the standardization sector can be found in the literature references including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols.A (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art are employed, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods. Unless a specific definition is set forth, the terms used herein in the pertinent description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the instructions of the kit from the manufacturer, or according to the methods known in the art or the instructions of the present invention. The techniques and methods described above can generally be practiced according to conventional methods well known in the art, as described in various general and more specific documents referred to and discussed in this specification. In the present specification, groups and substituents thereof may be selected by one skilled in the art to provide stable moieties and compounds.

When a substituent is described by a general formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the formula is written from right to left. For example, -CH₂O-is equivalent to-OCH₂-。

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and treatises, are hereby incorporated by reference in their entirety.

Certain chemical groups defined herein are preceded by a shorthand notation to indicate the total number of carbon atoms present in the group. For example, C1-6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the shorthand notation excludes carbons that may be present in a substituent of the group.

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

In the present application, the term "halogen" means fluorine, chlorine, bromine or iodine; "hydroxy" means an-OH group; "hydroxyalkyl" refers to an alkyl group as defined below substituted with a hydroxyl (-OH) group; "carbonyl" refers to a-C (═ O) -group; "nitro" means-NO ₂(ii) a "cyano" means-CN; "amino" means-NH₂(ii) a "substituted amino" means substituted with oneOr two alkyl, alkylcarbonyl, aralkyl, heteroaralkyl substituted amino groups as defined below, for example, monoalkylamino, dialkylamino, alkylamido, aralkylamino, heteroaralkylamino; "carboxyl" means-COOH.

In the present application, the term "alkyl", as a group or as part of another group (e.g. as used in groups such as halogen-substituted alkyl), means a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing no unsaturated bonds, having, for example, from 1 to 12 (preferably from 1 to 8, more preferably from 1 to 6) carbon atoms and being attached to the rest of the molecule by single bonds. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2-dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl, decyl, and the like.

In the present application, the term "alkenyl" as a group or part of another group means a straight or branched hydrocarbon chain group consisting of only carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms, and being connected to the rest of the molecule by a single bond, such as, but not limited to, vinyl, propenyl, allyl, but-1-enyl, but-2-enyl, pent-1, 4-dienyl, and the like.

In the present application, the term "alkynyl" as a group or part of another group means a straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, containing at least one triple bond and optionally one or more double bonds, having for example 2 to 14 (preferably 2 to 10, more preferably 2 to 6) carbon atoms and being connected to the rest of the molecule by single bonds, such as but not limited to ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-en-4-ynyl and the like.

In the present application, the term "cycloalkyl" as a group or as part of another group means a stable saturated or partially unsaturated monocyclic or polycyclic hydrocarbon group consisting of only carbon atoms and hydrogen atoms, which may include fused, bridged or spiro ring systems, having from 3 to 15 carbon atoms, preferably from 3 to 10 carbon atoms, more preferably from 3 to 8 carbon atoms, and which is saturated or unsaturated and may be attached to the rest of the molecule by a single bond via any suitable carbon atom. Unless otherwise specifically indicated in the specification, carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indenyl, 2, 3-indanyl, 1,2,3, 4-tetrahydro-naphthyl, 5,6,7, 8-tetrahydro-naphthyl, 8, 9-dihydro-7H-benzocyclohepten-6-yl, 6,7,8, 9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9, 10-hexahydro-benzocyclooctenyl, fluorenyl, bicyclo [2.2.1] heptyl, 7-dimethyl-bicyclo [2.2.1] heptyl, bicyclo [2.2.1] heptenyl, bicyclo [2.2.2] octyl, bicyclo [3.1.1] heptyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octenyl, Bicyclo [3.2.1] octenyl, adamantyl, octahydro-4, 7-methylene-1H-indenyl, octahydro-2, 5-methylene-pentalenyl and the like. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, cyclooctyl, 1,2,3, 4-tetrahydro-naphthyl, 5,6,7, 8-tetrahydro-naphthyl, and the like.

In this application, the term "heterocyclyl" as a group or part of another group means a stable 3-to 20-membered non-aromatic cyclic group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen, and sulfur. Unless otherwise specifically indicated in the specification, a heterocyclic group may be a monocyclic, bicyclic, tricyclic or higher ring system, which may include fused ring systems, bridged ring systems or spiro ring systems; wherein the nitrogen, carbon or sulfur atom in the heterocyclic group thereof may be optionally oxidized; the nitrogen atoms may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule via a carbon atom or a heteroatom and by a single bond. In heterocyclic groups containing fused rings, one or more of the rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purposes of the present invention, heterocyclyl is preferably a stable 4-to 11-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 4-to 8-membered non-aromatic monocyclic, bicyclic, bridged or spiro group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclyl groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2, 7-diaza-spiro [3.5] nonan-7-yl, 2-oxa-6-aza-spiro [3.3] heptan-6-yl, 2, 5-diaza-bicyclo [2.2.1] heptan-2-yl, azetidinyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxolanyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, quinolizinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indolinyl, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl, phthalimidyl, and the like.

In this application, the term "aryl" as a group or as part of another group means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms, preferably having 6 to 10 carbon atoms. For the purposes of the present invention, an aryl group may be a monocyclic, bicyclic, tricyclic or higher polycyclic ring system and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the aryl group is attached to the remainder of the molecule by a single bond via an atom on the aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthryl, phenanthryl, fluorenyl, 2, 3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazin-3 (4H) -on-7-yl, and the like.

In the present application, the term "arylalkyl" refers to an alkyl group as defined above substituted with an aryl group as defined above.

In this application, the term "heteroaryl" as a group or part of another group means a 5-to 16-membered conjugated ring system group having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur in the ring. Unless otherwise specifically indicated in the specification, a heteroaryl group may be a monocyclic, bicyclic, tricyclic or higher ring system, and may also be fused to a cycloalkyl or heterocyclic group as defined above, provided that the heteroaryl group is attached to the rest of the molecule by a single bond via an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group may be optionally oxidized; the nitrogen atoms may optionally be quaternized. For the purposes of the present invention, heteroaryl is preferably a stable 5-to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a stable 5-to 10-membered aromatic group containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur or a 5-to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoindolyl, indazolyl, isoindolyl, purinyl, quinolyl, isoquinolyl, diazonaphthyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxazolyl, cinnolinyl, quinazolinyl, thiophenyl, indolizinyl, orthophenanthrolidinyl, isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6, 7-tetrahydrobenzo [ b ] thienyl, naphthopyridyl, pyridinyl, and the like, [1,2,4] triazolo [4,3-b ] pyridazine, [1,2,4] triazolo [4,3-a ] pyrazine, [1,2,4] triazolo [4,3-c ] pyrimidine, [1,2,4] triazolo [4,3-a ] pyridine, imidazo [1,2-b ] pyridazine, imidazo [1,2-a ] pyrazine and the like.

In the present application, the term "heteroarylalkyl" refers to an alkyl group as defined above substituted with a heteroaryl group as defined above.

In this application, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups.

The terms "moiety," "structural moiety," "chemical moiety," "group," "chemical group" as used herein refer to a specific fragment or functional group in a molecule. Chemical moieties are generally considered to be chemical entities that are embedded in or attached to a molecule.

"stereoisomers" refers to compounds that consist of the same atoms, are bonded by the same bonds, but have different three-dimensional structures. The present invention is intended to cover various stereoisomers and mixtures thereof.

When the compounds of the present invention contain olefinic double bonds, the compounds of the present invention are intended to include both E-and Z-geometric isomers unless otherwise specified.

"tautomer" refers to an isomer formed by the transfer of a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the invention are also intended to be included within the scope of the invention.

The compounds of the present invention or pharmaceutically acceptable salts thereof may contain one or more chiral carbon atoms and may therefore give rise to enantiomers, diastereomers and other stereoisomeric forms. Each chiral carbon atom may be defined as (R) -or (S) -, based on stereochemistry. The present invention is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the invention may be prepared by selecting as starting materials or intermediates racemates, diastereomers or enantiomers. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, e.g., crystallization and chiral chromatography.

Conventional techniques for the preparation/separation of individual isomers include Chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, Chiral high performance liquid chromatography, as described, for example, in Gerald Gubitz and Martin G.Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol.243, 2004; m. Stalcup, Chiral Separations, Annu. Rev. anal. chem.3:341-63, 2010; fumiss et al (eds.), VOGEL' S ENCYCOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5. TH ED., Longman Scientific and Technical Ltd., Essex,1991, 809-816; heller, acc, chem, res, 1990,23,128.

In the present application, the term "pharmaceutically acceptable salts" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"pharmaceutically acceptable acid addition salts" refers to salts with inorganic or organic acids which retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, and the like; organic acid salts include, but are not limited to, formates, acetates, 2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, caprates, undecenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbates, salicylates, 4-aminosalicylates, napadisylates, and the like. These salts can be prepared by methods known in the art.

"pharmaceutically acceptable base addition salts" refers to salts with inorganic or organic bases which maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

"polymorphs" refer to the distinct solid crystalline phases of certain compounds of the present invention in the solid state resulting from the presence of two or more distinct molecular arrangements. Certain compounds of the present invention may exist in more than one crystalline form, and the present invention is intended to include the various crystalline forms and mixtures thereof.

Typically, crystallization will result in solvates of the compounds of the invention. The term "solvate" as used herein refers to an aggregate comprising one or more molecules of the compound of the present invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the compounds of the present invention may exist as hydrates, including monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compounds of the invention may form true solvates, but in some cases it is also possible to retain only adventitious water or a mixture of water plus some adventitious solvent. The compounds of the invention may be reacted in a solvent or precipitated or crystallized from a solvent. Solvates of the compounds of the invention are also included within the scope of the invention.

The invention also includes prodrugs of the above compounds. In the present application, the term "prodrug" denotes a compound that can be converted under physiological conditions or by solvolysis to the biologically active compound of the invention. Thus, the term "prodrug" refers to a pharmaceutically acceptable metabolic precursor of a compound of the invention. Prodrugs may not be active when administered to a subject in need thereof, but are converted in vivo to the active compounds of the invention. Prodrugs are generally rapidly converted in vivo to yield the parent compound of the invention, for example, by hydrolysis in blood. Prodrug compounds generally provide solubility, histocompatibility, or sustained release advantages in mammalian organisms. Prodrugs include known amino protecting groups and carboxyl protecting groups. Specific methods for preparing prodrugs can be found in Saulnier, M.G., et al, bioorg.Med.chem.Lett.1994,4, 1985-1990; greenwald, r.b., et al, j.med.chem.2000,43,475.

In the present application, a "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for delivery of biologically active compounds to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of active ingredients and exert biological activity.

The term "pharmaceutically acceptable" as used herein refers to a substance (e.g., carrier or diluent) that does not interfere with the biological activity or properties of the compounds of the present invention, and is relatively non-toxic, i.e., the substance can be administered to an individual without causing an undesirable biological response or interacting in an undesirable manner with any of the components contained in the composition.

As used herein, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonizing agent, solvent, or emulsifier that is approved by the relevant governmental regulatory agency for human or livestock use.

The "tumor" and "diseases related to abnormal cell proliferation" include, but are not limited to, leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, pancreatic cancer, squamous cell lung cancer, lung adenocarcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, intestinal cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, renal cancer, oral cancer, and the like.

The terms "preventing," "prevention," and "prevention" as used herein include reducing the likelihood of occurrence or worsening of a disease or disorder in a patient.

The term "treatment" and other similar synonyms as used herein include the following meanings:

(i) preventing the occurrence of a disease or condition in a mammal, particularly when such mammal is susceptible to the disease or condition, but has not been diagnosed as having the disease or condition;

(ii) inhibiting the disease or disorder, i.e., arresting its development;

(iii) alleviating the disease or condition, i.e., causing regression of the state of the disease or condition; or

(iv) Alleviating the symptoms caused by the disease or disorder.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The terms "administering," "administration," "administering," and the like, as used herein, refer to a method of delivering a compound or composition to a desired site for a biological effect. These methods include, but are not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Administration techniques useful for The compounds and methods described herein are well known to those skilled in The art, for example, in Goodman and Gilman, The pharmaceutical Basis of Therapeutics, current ed.; pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

The terms "drug combination", "administering other treatment", "administering other therapeutic agent" and the like as used herein refer to a drug treatment obtained by mixing or combining more than one active ingredient, including fixed and unfixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one co-agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, concomitant administration, or sequential administration at variable intervals of at least one compound described herein and at least one synergistic formulation to a patient as separate entities. These also apply to cocktail therapy, for example the administration of three or more active ingredients.

It will also be appreciated by those skilled in the art that in the processes described below, the intermediate compound functional groups may need to be protected by appropriate protecting groups. Such functional groups include hydroxyl, amino, mercapto, and carboxylic acid. Suitable hydroxyl protecting groups include trialkylsilyl or diarylalkylsilyl groups (e.g.t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino include t-butyloxycarbonyl, benzyloxycarbonyl and the like. Suitable thiol protecting groups include-C (O) -R "(where R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl, and the like. Suitable carboxyl protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups may be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting Groups is described in detail in Greene, T.W. and P.G.M.Wuts, Protective Groups in organic Synthesis, (1999),4th Ed., Wiley. The protecting group may also be a polymeric resin.

The invention will be further illustrated with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally according to conventional conditions, or according to conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise indicated.

Preparation of intermediate A

An intermediate A: n- (3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: N-Boc-4-piperidone (5.0g, 25.1mmol) was dissolved in ethanol (EtOH) (75mL), malononitrile (1.8g,27.3mmol), sublimed sulfur (0.88g,27.5mmol) and morpholine (4.4g,50.6mmol) were added under nitrogen, heated to 88 deg.C and stirred for 2.5 h. Cooling the reaction liquid to room temperature, filtering, washing filter cakes with ethanol, and drying to obtain 2-amino-3-cyano-4, 7-dihydrothieno [2,3-c ]]Pyridine-6 (5H) -carboxylic acid tert-butyl ester (3.3g, yellow solid). LC-MS M/z 278.0[ M-H ]]⁺。

The second step is that: 2-amino-3-cyano-4, 7-dihydrothieno [2,3-c ]]Tert-butyl pyridine-6 (5H) -carboxylate (22.2g, 79.6mmol) and 1-methyl-1H-pyrazole-4-carboxylate (13.0g, 103.2mmol) were dissolved in Dichloromethane (DCM) (330mL), 2-chloro-1-methylpyridine iodide (26.5g, 103.9mmol), N-Dimethylaminopyridine (DMAP) (2.1g,17.2mmol), Triethylamine (TEA) (49mL,350.2mmol) were added under nitrogen, and the mixture was heated to 50 ℃ and stirred overnight. The reaction solution is cooled to room temperature and saturated sodium carbonate Na₂CO₃Washing the solution for 2 times, then washing with water for 2 times, concentrating under reduced pressure, pulping ethyl acetate and petroleum ether (petroleum ether PE: ethyl acetate EA ═ 5: 1) and purifying to obtain 3-cyano-2- (1-methyl-1H-pyrazole-4-amido) -4, 7-dihydrothieno [2, 3-c) ]Pyridine-6 (5H) -carboxylic acid tert-butyl ester (17.6g, yellow solid). LC-MS M/z 410.0[ M + Na ]]⁺；¹H-NMR(400MHz,DMSO-d6):11.53(s,1H),8.47(s,1H),8.07(s,1H),4.46(s,2H),3.91(s,3H),3.61-3.64(m,2H),2.60(t,J＝5.6Hz,2H),1.43(s,9H)。

The third step: reacting 3-cyano-2- (1-methyl-1H-pyrazole-4-amido) -4, 7-dihydrothieno [2,3-c]Pyridine-6 (5H) -carboxylic acid tert-butyl ester (7.6g, 19.6mmol) was dissolved in anhydrous dichloromethane (57mL), trifluoroacetic acid (TFA) (28mL) was added dropwise, reacted at room temperature for 1H, and concentrated to give intermediate A (5.6g, yellow solid) which was used directly in the next reaction. LC-MS M/z 288.0[ M + H ]]⁺。

Examples preparation

EXAMPLE 1N- (6- (5-Bromopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-carboxamide

Intermediate A (5.6g, 19.5mmol) was dissolved in N, N-Dimethylformamide (DMF) (80mL), 5-bromo-2-chloropyrimidine (3.2g, 16.7mmol) and N, N-Diisopropylethylamine (DIEA) (10.6g, 82.2mmol) were added, and the reaction was allowed to proceed overnight at 90 degrees. The reaction solution was cooled to room temperature, a solid precipitated, petroleum ether was added, stirred for 10 minutes, filtered, and the filter cake was purified by slurrying with methanol to obtain the objective compound (3.2g, yellow solid). MS (M + H) 444.0/446.1;¹H-NMR(400MHz,DMSO-d6):8.53(s,2H),8.46(s,1H),8.06(s,1H),4.82(s,2H),4.06(t,J＝5.6Hz,2H),3.91(s,3H),2.68(t,J＝5.2Hz,2H)。

example 2: n- (3-cyano-6- (5-cyanopyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The target compound was prepared using intermediate a and 5-cyano-2-chloropyrimidine as starting materials according to the synthetic method of example 1. MS (M + H) 391.2; ¹H-NMR(400MHz,DMSO-d6)：11.53(s,1H),8.83(s,2H),8.46(s,1H),8.07(s,1H),4.95(s,2H),4.18(t,J＝5.6Hz,2H),3.93(s,3H),2.71(t,J＝5.6Hz,2H)。

Example 3: n- (3-cyano-6- (5-fluoropyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The target compound was prepared using intermediate a and 5-fluoro-2-chloropyrimidine as starting materials with reference to the synthesis method of example 1. MS (M + H) 384.2;¹H-NMR(400MHz,DMSO-d6)：11.52(s,1H),8.54(s,2H),8.49(s,1H),8.06(s,1H),4.83(s,2H),4.10(t,J＝5.6Hz,2H),3.91(s,3H),2.71(t,J＝5.6Hz,2H)。

example 4: n- (6- (5-bromopiperazin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The intermediate a and 2, 5-dibromopyrazine are used as raw materials, and the target compound is prepared according to the synthesis method of the example 1. MS (M + H): 444.2;¹H-NMR(400MHz,CD₃OD)：11.52(s,1H),8.46(s,1H),8.29(s,1H),8.27(s,1H),8.06(s,1H),4.71(s,2H),3.95(t,J＝4.8Hz,2H),3.91(s,3H),2.72(t,J＝4.8Hz,2H)。

example 5: n- (6- (3-bromopyridin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The target compound was prepared from intermediate a and 2-chloro-3-bromopyridine by the synthesis method of example 1. MS (M + H): 443.2;¹H-NMR(400MHz,DMSO-d6)：11.50(s,1H),8.44(s,1H),8.26(dd,J＝1.6,4.8Hz,1H),8.05(s,1H),8.01(dd,J＝1.6,8.0Hz,1H),6.96(dd,J＝4.8,8.0Hz,2H),4.39(s,2H),3.91(s,3H),3.60(t,J＝5.6Hz,2H),2.80(s,2H)。

example 6: n- (6- (5- (benzyloxy) pyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The target compound was prepared using intermediate a and 2-chloro-5- (benzyloxy) pyrimidine as starting materials, according to the synthetic method of example 1. MS (M + H): 472.1.¹H-NMR(400MHz,DMSO-d6):11.50(br s,1H),8.47(s,1H),8.31(s,2H),8.06(s,1H),7.35-7.43(m,5H),5.12(s,2H),4.77(s,2H),4.01(br s,2H),3.91(s,3H),2.65(br s,2H)。

Example 7: n- (6- (4-bromothiazol-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Dissolving intermediate A (120mg, 0.42mmol), 2, 4-dibromothiazole (100mg, 0.41mmol), 1 '-binaphthyl-2, 2' -bis-diphenylphosphine (BINAP) (50mg, 0.080mmol) and sodium tert-butoxide (80mg, 0.83mmol) in 1, 4-dioxane (10mL), and adding tris (dibenzylideneacetone) dipalladium [ Pd ] under nitrogen protection₂(dba)₃](50mg, 0.080mmol), heated to 80 ℃ and stirred overnight. Cooling the reaction solution to room temperature, filtering by using diatomite, concentrating under reduced pressure, and purifying by using column chromatography to obtain the target compound. MS (M + H): 449.2;¹H-NMR(400MHz,DMSO-d6)：8.29(s,1H),7.93(s,1H),6.92(s,1H),4.52(s,2H),3.88(s,3H),3.77(t,J＝5.6Hz,2H),2.72(t,J＝5.6Hz,2H)。

example 8: n- (3-cyano-6- (5- (3-methoxyphenyl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (110mg, 0.25mmol) and 3-methoxyphenylboronic acid (57mg,0.38mmol) were dissolved in 1, 4-dioxane/water (5mL/1mL) and sodium carbonate (Na) was added under nitrogen (Na-N-O-N-O-N-O-N-O-N-O-N-O-N-O-C-N-2-N-C-O-N-O-N-N₂CO₃) (53mg,0.50mmol) and [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride Dichloromethane Complex [ Pd (dppf) Cl₂](12.4mg, 0.017mmol) and heated to 90 deg.C for reaction overnight. The reaction was cooled to room temperature, concentrated under reduced pressure, and purified by preparative chromatography to give the objective compound (9.8mg, pale yellow solid). MS (M + H): 472.4; ¹H-NMR(400MHz,DMSO-d6)：11.52(s,1H),8.78(s,2H),8.46(s,1H),8.06(s,1H),7.35(t,J＝8.0Hz,1H),7.21-7.23(m,2H),6.92(d,J＝8.0Hz,1H),4.91(s,2H),4.15(t,J＝5.6Hz,2H),3.91(s,3H),3.81(s,3H),2.71(brs,2H)。

The following compounds of examples were prepared according to the synthetic method of example 8 using the compound of example 1 and commercial reagents as starting materials.

Example 43: n- (3-cyano-6- (5- (1-isobutyl-1H-pyrazol-5-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: 1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazole-5-boronic acid pinacol ester (3.5g,12.6mmol) and 5-bromo-2- (methylthio) pyrimidine (1.7g,8.3mmol) are dissolved in 1, 4-dioxane/water (60mL/6mL) and potassium carbonate (K) is added under nitrogen protection₂CO₃) (2.7g,19.6mmol) and Pd (dppf) Cl₂(310mg,0.42mmol), replaced three more times with nitrogen, and heated to 90 degrees for reaction overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to give the objective compound (1.1g, white solid). MS (M + H): 277.0.

The second step is that: 2- (methylthio) -5- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) pyrimidine (1.2g,4.35mmol) was dissolved in methanol (30mL), and a methanol solution of Hydrogen Chloride (HC) was addedL/MeOH, 0.5mL, 4mol/L), under nitrogen at room temperature for 4.5 h, and concentrated under reduced pressure to give the title compound (956mg, white solid). MS (M + H) 193.1;¹H-NMR(400MHz,DMSO-d6)：9.04(s,2H),7.84(s,1H),6.86(s,1H),2.55(s,3H)。

the third step: 2- (methylthio) -5- (1H-pyrazol-5-yl) pyrimidine (200mg,1.04mmol) and bromoisobutane (285mg,2.09mmol) were dissolved in DMF (10mL), and K was added ₂CO₃(575mg,4.17mmol) was heated to 65 ℃ under nitrogen for reaction overnight. Cooling the reaction liquid to room temperature, and adding saturated ammonium chloride NH₄The Cl solution was extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound (223mg, pale yellow solid). MS (M + H) 249.1;¹H-NMR(400MHz,DMSO-d6)：9.00(s,2H),7.84(d,J＝2.0Hz,1H),6.84(d,J＝2.0Hz,1H),3.96-3.98(m,2H),2.57(s,3H),2.14-2.17(m,1H),0.85-0.87(m,6H)。

the fourth step: 5- (1-isobutyl-1H-pyrazol-5-yl) -2- (methylthio) pyrimidine (223mg,0.90mmol) was dissolved in a tetrahydrofuran/water mixed solvent (THF/H)₂O, 6mL/6mL), Oxone (Oxone) (1.1g,1.8mmol) was added and the reaction was allowed to proceed overnight at room temperature under nitrogen. THF was removed by concentration under reduced pressure, water was added to precipitate a white solid, which was filtered, and the cake was dried to give the objective compound (144mg, white solid). MS (M + H): 281.1;¹H-NMR(400MHz,DMSO-d6)：9.41(s,2H),7.94(d,J＝1.6Hz,1H),7.07(d,J＝1.6Hz,1H),4.04(d,J＝7.2Hz,2H),3.42(s,3H),2.17-2.20(m,1H),0.88(d,J＝6.4Hz,6H)。

the fifth step: 5- (1-isobutyl-1H-pyrazol-5-yl) -2- (methylsulfonyl) pyrimidine (144mg,0.51mmol) and N- (3-cyano-4, 5,6, 7-tetrahydrothiophene [2, 3-c)]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (177mg,0.62mmol) was dissolved in 1, 4-dioxane (5mL), DIEA (198mg,1.53mmol) was added, the mixture was heated to 100 ℃ under nitrogen for reaction overnight, DIEA (198mg,1.53mmol) was added, and the mixture was heated to 120 ℃ for reaction overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to give the objective compound (32mg, pale yellow solid). MS (M + H): 488.1; ¹H-NMR(400MHz,DMSO-d6)：11.51(s,1H),8.78(s,2H),8.47(s,1H),8.07(s,1H),7.74-7.76(m,1H),6.66(s,1H),4.87(s,2H),4.12(brs,2H),3.91-3.94(m,5H),2.70(brs,2H),2.12-2.15(m,1H),0.85(d,J＝6.8Hz,6H)。

The following compounds of examples were prepared according to the synthetic method of example 43 using commercially available reagents as starting materials.

Example 48: n- (3-cyano-6- (5- (2-isopropoxypyridin-4-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-carboxamide (example 1) (300mg,0.68mmol) and 2-fluoropyridineboronic acid (140mg,0.99mmol) were dissolved in 1, 4-dioxane/water (10mL/2mL) and Na was added under nitrogen₂CO₃(144mg,1.36mmol) and Pd (dppf) Cl₂(50mg, 0.068mmol), heated to 85 ℃ and reacted overnight. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and purified by preparative chromatography to give the objective compound (310 mg). MS (M + H): 461.5;¹H-NMR(400MHz,DMSO-d6)：11.53(s,1H),8.99(s,2H),8.47(s,1H),8.27(d,J＝5.2Hz,1H),8.07(s,1H),7.74(d,J＝5.2Hz,1H),7.59(s,1H),4.96(s,2H),4.19(t,J＝5.6Hz,2H),3.91(s,3H),2.73(t,J＝5.6Hz,2H)。

the second step is that: mixing N- (3-cyano-6- (5- (2-fluoropyridin-4-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (60mg, 0.13mmol) was dissolved in isopropanol (3mL), NaH (15mg, 60% in mineral oil, 0.38mmol) was added under nitrogen and heated to 65 ℃ for 3 days. Cooling the reaction liquid to room temperature, decompressing and concentrating to prepareAnd (5) carrying out chromatographic purification to obtain the target compound. MS (M + H) 501.5; ¹H-NMR(400MHz,CD₃OD)：12.34(brs,1H),9.71(s,2H),9.29(s,1H),8.98(d,J＝5.6Hz,1H),8.88(s,1H),8.10(dd,J＝1.2,5.6Hz,1H),7.90(s,1H),6.08-6.12(m,1H),5.75(s,2H),4.98(t,J＝5.6Hz,2H),4.72(s,3H),3.52(brs,2H),2.12(d,J＝6.0Hz,6H)。

Example 49: n- (3-cyano-6- (5- (2-morpholinopyridin-4-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The title compound was prepared according to the synthesis method of example 48. MS (M + H) 528.5;¹H-NMR(400MHz,CD₃OD)：8.77(s,2H),8.28(s,1H),8.13(d,J＝7.2Hz,1H),8.07(s,1H),6.95-7.02(m,2H),5.20(s,2H),4.24(t,J＝5.6Hz,2H),3.87(s,3H),3.77-3.82(m,4H),3.50-3.54(m,4H),2.79(brs,2H)。

example 50: n- (3-cyano-6- (5- (1- (piperidin-4-yl) -1H-pyrazol-4-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (150mg,0.34mmol) and 1- (N-Boc-piperidin-4-yl) -4-pyrazole boronic acid pinacol ester (192mg,0.51mmol) were dissolved in 1, 4-dioxane/water (10mL/2mL) and Na was added under nitrogen₂CO₃(72mg,0.68mmol) and Pd (dppf) Cl₂(25mg, 0.034mmol), heated to 85 ℃ and reacted overnight. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and purified by preparative chromatography to give the objective compound (150 mg). MS (M + H) 615.0.

The second step is that: 4- (4- (2- (3-cyano-2- (1-methyl-1H-pyrazole-4-carboxamide) -4, 7-dihydrothiophene [2, 3-c)]Pyridin-6 (5H) -yl) pyrimidin-5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylic acid tert-butyl ester(150mg, 0.24mmol) was dissolved in methanol (5mL), and 1M hydrogen chloride solution in methanol (5mL) was added to react at room temperature for 2 hours. Concentration under reduced pressure and purification by preparative chromatography gave the title compound (63 mg). MS (M + H) 515.0; ¹H-NMR(400MHz,DMSO-d6)：11.52(s,1H),9.11(brs,1H),8.85(brs,1H),8.70(s,2H),8.47(s,1H),8.21(s,1H),8.07(s,1H),7.93(s,1H),4.87(s,2H),4.46-4.53(m,1H),4.11(t,J＝5.6Hz,2H),3.91(s,3H),3.37-3.40(m,2H),3.04-3.09(m,2H),2.70(brs,2H),2.10-2.24(m,4H)。

Example 51: n- (3-cyano-6- (5- (4-methylpiperazin-1-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Reacting N- (6- (5-bromopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (110mg, 0.25mmol), 1-methylpiperazine (35mg, 0.35mmol), BINAP (35mg, 0.06mmol) and sodium tert-butoxide (50mg, 0.52mmol) were dissolved in toluene (10mL), the mixture was replaced three times with nitrogen and Pd was added rapidly₂(dba)₃(25mg, 0.03mmol), heated to 100 ℃ under nitrogen and stirred overnight. The reaction solution was cooled to room temperature, filtered through celite, washed with ethyl acetate, and the filtrate was washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by preparative chromatography to give the objective compound (20mg, pale yellow solid). MS (M + H): 464.2;¹H-NMR(400MHz,DMSO-d6):11.51(s,1H),8.47(s,1H),8.31(s,2H),8.06(s,1H),4.79(s,2H),4.01-4.08(m,2H),3.91(s,3H),3.64-3.67(m,2H),3.49-3.52(m,2H),3.16-3.19(m,2H),2.90-2.96(m,2H),2.85(s,3H),2.55(t,J＝5.2Hz,2H)。

example 52: n- (3-cyano-6- (5- (2-methylpyridin-4-yl) piperazin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Prepared by the synthesis method of reference example 8 using the compound of example 4 as a starting materialAnd preparing the target compound. MS (M + H) 457.2;¹H-NMR(400MHz,DMSO-d6)：11.54(s,1H),8.85(d,J＝1.2Hz,1H),8.56(d,J＝1.2Hz,1H),8.48(d,J＝5.6Hz,2H),8.07(s,1H),7.83(s,1H),7.74(d,J＝5.6Hz,1H),4.84(s,2H),4.07(t,J＝5.6Hz,2H),3.91(s,3H),2.78(t,J＝5.6Hz,2H),2.49(s,3H)。

example 53: n- (3-cyano-6- (5- (2- (4- (methylsulfonyl) piperidine-1-carbonyl) -1H-indol-5-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (1.0g,2.26mmol) and 2-methoxycarbonylindole-5-boronic acid pinacol ester (1.0g,3.32mmol) were dissolved in 1, 4-dioxane/water (20mL/4mL) and Na was added under nitrogen₂CO₃(478mg,4.51mmol) and Pd (dppf) Cl₂(165mg, 0.23mmol), heated to 90 ℃ and reacted overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by beating to give the objective compound (650mg, yellow solid). MS (M + H) 539.5;¹H-NMR(400MHz,DMSO-d6)：12.02(brs,1H),11.52(brs,1H),8.76(s,2H),8.46(s,1H),8.06(s,1H),7.91(s,1H),7.54(brs,2H),7.18(s,1H),4.90(s,2H),4.14(brs,2H),3.90(s,3H),3.88(s,3H),2.71(brs,2H)。

the second step is that: reacting 5- (2- (3-cyano-2- (1-methyl-1H-pyrazole-4-amido) -4, 7-dihydrothieno [2, 3-c)]Pyridin-6 (5H) -yl) pyrimidin-5-yl) -1H-indole-2-carboxylic acid methyl ester (600mg, 1.12mmol) in MeOH/H₂To O (5/1, 15mL), sodium hydroxide (NaOH) (440mg,11.0mmol) was added, and the reaction was carried out at room temperature for 2 hours. The mixture was diluted with water, concentrated under reduced pressure to remove most of the organic solvent, and 1M diluted hydrochloric acid was concentrated to pH 3 to precipitate a solid, which was filtered and dried to obtain the objective compound (400mg, yellow solid). MS (M + H) 525.5;¹H-NMR(400MHz,DMSO-d6)：11.85(brs,1H),11.52(brs,1H),8.76(s,2H),8.47(s,1H),8.07(s,1H),7.90(s,1H),7.52(brs,2H),7.12(d,J＝1.2Hz,1H),4.91(s,2H),4.13(brs,2H),3.91(s,3H),2.72(brs,2H)。

the third step: reacting 5- (2- (3-cyano-2- (1-methyl-1H-pyrazol-4-amido) -4, 7-dihydrothieno [2, 3-c)]Pyridin-6 (5H) -yl) pyrimidin-5-yl) -1H-indole-2-carboxylic acid (80mg, 0.15mmol), 4- (methylsulfonyl) piperidine (32mg, 0.20mmol) and DIPEA (59mg, 0.46mmol) were dissolved in DMF (2mL), HATU (77mg, 0.20mmol) was added under nitrogen and stirred at room temperature overnight. Concentration under reduced pressure and purification by preparative chromatography gave the title compound (30mg, yellow solid). MS (M + H): 670.6; ¹H-NMR(400MHz,DMSO-d6)：11.69(brs,1H),11.52(brs,1H),8.76(s,2H),8.47(s,1H),8.07(s,1H),7.86(s,1H),7.49(brs,2H),6.85(d,J＝1.6Hz,1H),4.92(s,2H),4.56-4.59(m,2H),3.85-4.19(m,7H),3.40-3.50(m,1H),2.97(s,3H),2.72(brs,2H),2.12-2.16(m,2H),1.62-1.65(m,2H)。

Example 54: 5- (2- (3-cyano-2- (1-methyl-1H-pyrazol-4-ylamino) -4, 7-dihydrothieno [2,3-c ] pyridin-6 (5H) -yl) pyrimidin-5-yl) -N- (2- (dimethylamino) ethyl) -1H-indole-2-amide

The title compound was prepared according to the synthesis method of example 53. MS (M + H) 595.6;¹H-NMR(400MHz,DMSO-d6)：11.73(brs,1H),11.53(s,1H),9.47(brs,1H),8.74-8.78(m,2H),8.47(s,1H),8.07(s,1H),7.92(s,1H),7.48-7.54(m,2H),7.16(d,J＝1.2Hz,1H),4.92(s,2H),4.15(t,J＝5.6Hz,2H),3.92(s,3H),3.63-3.67(m,2H),3.28-3.30(m,2H),2.88(d,J＝4.0Hz,6H),2.72(brs,2H)。

example 55: 5- (2- (3-cyano-2- (1-methyl-1H-pyrazol-4-ylamino) -4, 7-dihydrothieno [2,3-c ] pyridin-6 (5H) -yl) pyrimidin-5-yl) -1H-indole-2-amide

The title compound was prepared according to the synthesis method of example 53. MS (M + H) 524.5;¹H-NMR(400MHz,DMSO-d6)：11.60(brs,1H),8.75(s,2H),8.00(brs,2H),7.86(s,1H),7.72(brs,1H),7.38-7.50(m,3H),7.16(s,1H),4.76(s,2H),4.12(t,J＝5.6Hz,2H),3.85(s,3H),2.62(brs,2H)。

example 56: n- (3-cyano-6- (5- (pyridin-3-ylmethoxy) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: 2-Chloropyrimidin-5-ol (390mg,3.0mmol), 3- (chloromethyl) pyridine hydrochloride (538mg,3.3mmol) were dissolved in DMF (15mL) and K was added under nitrogen protection₂CO₃(621mg,4.5mmol), and stirred at room temperature overnight. Pouring the reaction solution into water, extracting with ethyl acetate for three times, washing the extract with water and saturated saline respectively, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, and purifying by column chromatography to obtain the target compound. MS (M + H): 222.1.

The second step is that: intermediate A (170mg, 0.59mmol) was dissolved in DMF (10mL), and 2-chloro-5- (pyridin-3-ylmethoxy) pyrimidine (120mg, 0.54mmol) and DIEA (270mg, 2.1mmol) were added and reacted overnight at 90 degrees. Cooling the reaction liquid to room temperature, pouring into water, separating out solids, filtering, and pulping and purifying a filter cake by using methanol to obtain the target compound. MS (M + H): 473.2; ¹H-NMR(400MHz,DMSO-d6):8.60-8.65(m,3H),8.16(s,2H),8.00(s,1H),7.95(s,1H),7.76(d,J＝7.2Hz,1H),7.33-7.36(m,1H),5.17(s,2H),4.83(s,2H),4.09(t,J＝5.6Hz,2H),3.99(s,3H),2.78(t,t＝5.6Hz,2H)。

Example 57: n- (2- (3-cyano-2- (1-methyl-1H-pyrazole-4-amide) -4, 7-dihydrothieno [2,3-c ] pyridin-6 (5H) -yl) pyrimidin-5-yl) -1-methylpiperidine-4-amide

The first step is as follows: intermediate A (500mg, 1.74mmol) was dissolved in DMF (20mL) and 5-nitro-2-chloropyrimidine (270mg, 1.70mmol) and DIEA (877mg, 6.8mmol) were added and reacted at 50 ℃ overnight. Cooling the reaction liquid to room temperature, separating out solids, adding petroleum ether, stirring for 10 minutes, filtering, and pulping and purifying a filter cake by using methanol to obtain the target compound. MS (M + H) 411.2.

The second step is that: n- (3-cyano-6- (5-nitropyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-C ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (450mg, 1.1mmol) was dissolved in THF/DMF (20mL/4mL), 10% Pd/C (45mg, 10% wt) was added, and the reaction was carried out overnight at room temperature under a 15psi hydrogen atmosphere. Filtering the reaction solution by using diatomite, concentrating the filtrate under reduced pressure, pulping by using methanol and purifying to obtain the target compound. MS (M + H) 381.4.

The third step: mixing N- (6- (5-aminopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (120mg, 0.32mmol), 1-methylpiperidine-4-carboxylic acid (50mg, 0.35mmol) and DIPEA (62mg, 0.48mmol) were dissolved in DMF (10mL), 2- (7-oxybenzotriazole) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU) (160mg, 0.42mmol) was added under nitrogen protection, and stirred at room temperature overnight. Pouring the reaction solution into water, extracting with ethyl acetate for three times, washing the extract with water and saturated saline respectively, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, and separating and purifying by column chromatography to obtain the target compound. MS (M + H): 506.2. ¹H-NMR(400MHz,DMSO-d6):11.35(br s,1H),9.83(s,1H),8.59(s,2H),8.39(s,1H),8.01(s,1H),4.78(s,2H),4.04(t,J＝5.6Hz,2H),3.90(s,3H),2.87(d,J＝11.6Hz,2H),2.65(brs,2H),2.22-2.29(m,4H),1.95-2.00(m,2H),1.60-1.78(m,4H)。

Example 58: n- (3-cyano-6- (5- (4- ((1- (methylsulfonyl) piperidin-4-yl) methoxy) phenyl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: 4-bromophenol (300mg,1.7mmol), 4- (bromomethyl) -1- (methylsulfonyl) piperidine (434mg,1.7mmol) were dissolved in DMF (15mL) and K was added under nitrogen protection₂CO₃(704mg,5.1mmol) and stirred at room temperature overnight. Pouring the reaction solution into water, extracting with ethyl acetate for three times, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, and purifying by column chromatography to obtain the target compound. MS (M + H): 348.1/350.1.

The second step is that: 4- ((4-bromophenoxy) methyl) -1- (methylsulfonyl) piperidine (350mg, 1.0mmol), diboron pinacol ester (305mg, 1.2mmol) and dppf (111mg, 0.2mmol) were dissolved in 1, 4-dioxane (15mL), and potassium acetate (200mg, 2.0mmol) and Pd (dppf) Cl were added under nitrogen protection₂(73mg, 0.1mmol), 80 ℃ overnight. Cooling the reaction liquid to room temperature, concentrating under reduced pressure, and purifying by column chromatography to obtain the target compound. MS (M + H) 396.2.

The third step: reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (110mg, 0.25mmol) and 1- (methylsulfonyl) -4- ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) methyl) piperidine (120mg,0.30mmol) were dissolved in 1, 4-dioxane/water (10mL/2mL) and Na was added under nitrogen ₂CO₃(53mg,0.50mmol) and Pd (dppf) Cl₂(12.4mg, 0.017mmol) and heated to 90 degrees for reaction overnight. And cooling the reaction liquid to room temperature, concentrating under reduced pressure, and purifying by preparative chromatography to obtain the target compound. MS (M + H) 633.2;¹H-NMR(400MHz,DMSO-d6)：11.50(s,1H),8.71(s,2H),8.37(s,1H),8.05(s,1H),7.57(d,J＝8.4Hz,2H),7.02(d,J＝8.0Hz,2H),4.90(s,2H),4.07-4.12(m,2H),3.84-3.90(m,5H),3.55-3.58(m,2H),2.85(s,3H),2.63-2.75(m,4H),1.83-1.89(m,3H),1.36-1.39(m,2H)。

example 59: n- (6- (5-bromopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -2-methylnicotinamide

The target compound was prepared according to the synthesis method of intermediate a and example 1. MS (M + H): 455.2;¹H-NMR(400MHz,DMSO-d6)：12.32(s,1H),8.59(dd,J＝1.6,4.8Hz,1H),8.54(s,2H),7.92(dd,J＝1.6,8.0Hz,1H),7.37(dd,J＝4.8,8.0Hz,1H),4.85(d,2H),4.07(t,J＝5.6Hz,2H),2.70(t,J＝5.6Hz,2H),2.54(s,3H)。

example 60: n- (6- (5-bromopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) nicotinamide

The target compound was prepared according to the synthesis method of intermediate a and example 1. MS (M + H): 441.2;¹H-NMR(400MHz,DMSO-d6)：12.18(s,1H),9.08(s,1H),8.80(d,J＝5.6Hz,1H),8.53(s,2H),8.28-8.31(m,1H),7.58-7.62(m,1H),4.86(s,2H),4.08(t,J＝5.6Hz,2H),2.70(t,J＝5.6Hz,2H)。

example 61: n- (6- (5-bromopyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1- (2-methoxyethyl) -1H-pyrazole-4-amide

The target compound was prepared according to the synthesis method of intermediate a and example 1. MS (M + H): 488.4;¹H-NMR(400MHz,CD₃OD)：11.52(s,1H),8.53(s,2H),8.48(s,1H),8.08(s,1H),4.82(s,2H),4.34(t,J＝4.4Hz,2H),4.06(t,J＝4.4Hz,2H),3.71(t,J＝4.4Hz,2H),3.24(s,3H),2.68(t,J＝4.4Hz,2H)。

example 62: n- (6- (5- ((2-amino-3-chloropyridin-4-yl) thio) pyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (example 1) (300mg, 0.68mmol), tetramethylphenanthroline (47mg,0.20mmol), potassium carbonate (376mg,2.72mmol) and CuI (38mg,0.20mmol) were added to a DMSO solution (3mL) of 2-amino-3-chloropyridine-4-thiophenol sodium salt (2.03mmol) and stirred at 120 ℃ under nitrogen for 16H. The reaction was cooled to room temperature, taken up in water, extracted 3 times with dichloromethane, the organic phases combined, concentrated under reduced pressure, purified by column chromatography to give a crude product as a yellow oil, and further purified by preparative chromatography to give the title compound (10.1mg, yellow solid). MS (M + H) 524.4; ¹H-NMR(400MHz,DMSO-d6):11.53(s,1H),8.57(s,2H),8.47(s,1H),8.07(s,1H),7.69(d,J＝5.6Hz,1H),6.84(brs,2H),5.95(d,J＝6.0Hz,1H),4.95(s,2H),4.18(t,J＝5.2Hz,2H),3.91(s,3H),2.74(t,J＝5.2Hz,2H)。

Example 63: n- (3-cyano-6- (4-methoxy-1H-benzo [ d ] imidazol-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: 2-amino-3-methoxybenzoic acid (1.0g,6.0mmol) was dissolved in 1,4-dioxane (20mL), DPPA (3.29g,12.0mmol) and TEA (2.42g,24.0mmol) were added under nitrogen, and the mixture was heated to 110 ℃ for reaction overnight. The reaction was cooled to room temperature, concentrated under reduced pressure, and 50mL of water was added followed by saturated sodium bicarbonate (NaHCO)₃) The aqueous solution was adjusted to pH 9, and 15mL of ethyl acetate was added, followed by stirring for 30 minutes to precipitate a solid, which was filtered and dried to obtain the objective compound (250mg, white solid). MS (M + H) 165.4.

The second step is that: reacting 4-methoxy-1H-benzo [ d ]]Imidazole-2-ol (250mg,1.52mmol) dissolved in phosphorus oxychloride POCl₃In (5mL), 1 drop of N, N-dimethylaniline was added under nitrogen and the mixture was heated to 110 ℃ for overnight reaction. The reaction was cooled to room temperature, concentrated under reduced pressure, 50mL of water was added, and saturated NaHCO was used again₃The aqueous solution was adjusted to pH 6 to precipitate a solid, which was filtered and dried to give the title compound (202mg, white solid). MS (M + H): 183.2.

The third step: mixing N- (3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-carboxamide (150mg, 0.52mmol) was dissolved in 1,4-dioxane (10mL) and 2-chloro-4-methoxy-1H-benzo [ d ] was added ]Imidazole (185mg, 1.02mmol) and DIEA (336mg, 2.60mmol), heated to 90 degrees for 2 days. And cooling the reaction solution to room temperature, pouring the reaction solution into water (50mL), separating out a solid, filtering, drying a filter cake, and performing preparative chromatography purification to obtain the target compound. MS (M + H) 434.1;¹H-NMR(400MHz,DMSO-d6)：11.54(brs,1H),8.31(s,1H),8.26(s,1H),7.95(s,1H),6.87(brs,2H),6.58(d,J＝5.6Hz,1H),4.64(s,2H),3.87-3.89(m,8H),2.69(brs,2H)。

example 64: n- (6- (5- (1- (3-bromobenzyl) -5-methyl-1H-1, 2, 4-triazol-3-yl) pyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: 2-Chloropyrimidine-5-carbonitrile (3.0g,21.6mmol) was dissolved in MeOH (30mL), MeONa (3.5g,64.8mmol) was added, and the reaction was allowed to react at room temperature for 1 hour. AcOH was added to adjust pH 7 and concentrated under reduced pressure to give the title compound (8.7g, crude, white solid). MS (M + H) 168.4.

The second step is that: methyl 2-methoxypyrimidine-5-imidate (1.5g, crude) and acethydrazide (318mg,4.3mmol) were dissolved in DMF (6mL) and heated to 130 ℃ for reaction overnight. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated and purified by column chromatography to give the objective compound (560mg, yellow solid). MS (M + H) 192.4.

The third step: 2-methoxy-5- (5-methyl-1H-1, 2, 4-triazol-3-yl) pyrimidine (2.0g,10.5mmol) was dissolved in DMF (80mL), NaH (60% in mineral oil, 0.52g,12.9mmol) was added in portions, reaction was carried out at room temperature for 1 hour, 1-bromo-3- (chloromethyl) benzene (2.4g,11.8mmol) was added, and reaction was continued at room temperature for 2 hours. The reaction solution was added with 400mL of water, filtered, and the filter cake was dried to give the objective compound (2.2g, pale yellow solid). MS (M + H): 359.9.

The fourth step: 5- (1- (3-bromobenzyl) -5-methyl-1H-1, 2, 4-triazol-3-yl) -2-methoxypyrimidine (2.0g,5.6mmol) was dissolved in MeOH (5mL), concentrated hydrochloric acid (15mL) was added, and the reaction was heated to 80 ℃ for 2 hours. The reaction was cooled to room temperature and concentrated under reduced pressure to give the title compound (2.8g, crude, pale yellow solid). MS (M + H) 345.9/347.9.

The fifth step: dissolving 5- (1- (3-bromobenzyl) -5-methyl-1H-1, 2, 4-triazol-3-yl) pyrimidin-2-ol (2.8g, crude product) in POCl₃(10mL), the mixture was heated to 120 ℃ and reacted for 3 hours. The reaction was cooled to room temperature and concentrated under reduced pressure to give the title compound (3.4g, crude, pale yellow solid). MS (M + H): 364.2.

And a sixth step: 5- (1- (3-bromobenzyl) -5-methyl-1H-1, 2, 4-triazol-3-yl) -2-chloropyrimidine (340mg, crude), N- (3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (200mg, 0.7mmol) and DIEA (452mg,3.5mmol) were dissolved in DMF (5mL) and heated to 95 ℃ for reaction overnight. The reaction solution was cooled to room temperature, diluted with ethyl acetate (50mL), washed with water (50mL) 3 times, and the organic phase was dried, filtered, concentrated under reduced pressure, and purified by column chromatography to give the objective compound (55mg, yellow solid). MS (M + H): 615.3/617.3.

Example 65: n- (3-cyano-6- (5- ((5- (4- (4- (difluoromethoxy) benzyl) piperazin-1-yl) pyridin-2-yl) oxo) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothiophene [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The first step is as follows: methyl 6-fluoronicotinate (2.3g, 14.8mmol) was dissolved in DMF (50mL), 2-chloropyrimidin-5-ol (2.85g,21.9mmol) and cesium carbonate (9.6g, 29.4mmol) were added, and the mixture was heated to 60 ℃ under nitrogen and stirred overnight. The reaction solution was cooled to room temperature, and a saturated aqueous solution of ammonium chloride was added, followed by extraction with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give the objective compound (2.6g, white solid). MS (M + H): 266.0;¹H-NMR(400MHz,DMSO-d6)：8.88(s,2H),8.70(d,J＝2.0Hz,1H),8.40(dd,J＝2.4,8.8Hz,1H),7.38(d,J＝8.4Hz,1H),3.87(s,3H)。

the second step is that: mixing N- (3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (1.0g, 3.5mmol) and methyl 6- ((2-chloropyrimidin-5-yl) oxo) nicotinate (1.0g, 3.8mmol) were dissolved in DMF (30mL) and DIEA (2.2g,17.1mmol) was added under nitrogen and heated to 90 deg.C and stirred overnight. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography to give the objective compound (0.91g, yellow solid). MS (M + H): 517.0;¹H-NMR(400MHz,DMSO-d6)：11.53(brs,1H),8.67(d,J＝2.0Hz,1H),8.31-8.44(m,4H),8.02(s,1H),7.25(d,J＝8.8Hz,1H),4.85(s,2H),4.10(t,J＝5.6Hz,2H),3.90(s,3H),3.85(s,3H),2.70(brs,2H)。

the third step: reacting 6- ((2- (3-cyano-2- (1-methyl-1H-pyrazole) -4-acyl) -4, 7-dihydrothiophene [2,3-c]Pyridin-6 (5H) -yl) pyrimidin-5-yl) oxo) nicotinic acid methyl ester (700mg, 1.36mmol) was dissolved in MeOH (40.5mL), added NaOH (13.5mL, 1mol/L) aqueous solution, and reacted at room temperature for 3.5 hours. The pH was adjusted to 5 to 6 with 1mol/L HCl solution, a large amount of solid was precipitated, filtered, and the filter cake was washed with water and dried to obtain the objective compound (638mg, yellow solid). MS (M + H) 502.9;¹H-NMR(400MHz,DMSO-d6)：11.56(brs,1H),8.64(d,J＝2.0Hz,1H),8.29-8.48(m,4H),8.08(s,1H),7.22(d,J＝8.8Hz,1H),4.87(s,2H),4.11(t,J＝5.6Hz,2H),3.92(s,3H),2.72(brs,2H)。

the fourth step: mixing 6- ((2- (3-cyano-2- (1-methyl-1H-pyrazol-4-acyl) -4, 7-dihydrothiophene [2, 3-c)]Pyridin-6 (5H) -yl) pyrimidin-5-yl) oxo) nicotinic acid (638mg, 1.3mmol) was dissolved in DMF (30mL), 1-Boc-piperazine (236mg, 1.3mmol), HATU (964mg, 2.5mmol) and DIEA (672mg, 5.2mmol) were added, and the mixture was stirred at room temperature for 3 hours. Pouring saturated NH₄The Cl solution was filtered, and the filter cake was washed with water and dried to give the title compound (580mg, yellow solid). MS (M + H) 671.0;¹H-NMR(400MHz,DMSO-d6)：11.51(brs,1H),8.42-8.47(m,3H),8.19(d,J＝1.6Hz,1H),8.05(s,1H),7.93(dd,J＝2.0,8.4Hz,1H),7.20(d,J＝8.0Hz,1H),4.86(s,2H),4.10(t,J＝5.2Hz,2H),3.91(s,3H),3.36-3.54(m,8H),2.71(brs,2H),1.40(s,9H)。

the fifth step: 4- (6- ((2- (3-cyano-2- (1-methyl-1H-pyrazol-4-acyl) -4, 7-dihydrothiophene [2, 3-c)]Pyridin-6 (5H) -yl) pyrimidin-5-yl) oxo) nicotinoyl) piperazine-1-carboxylic acid tert-butyl ester (530mg, 0.79mmol) was dissolved in anhydrous dichloromethane (10mL), TFA (5mL) was added dropwise, and the reaction was carried out at room temperature for 2 hours. Concentrating to obtain crude product, and adding saturated NaHCO ₃The solution was slurried twice with deionized water, filtered, and the filter cake was washed with water and dried to give the title compound (270mg, yellow solid). MS (M + H) 571.2;¹H-NMR(400MHz,DMSO-d6)：8.38-8.42(m,3H),8.17(s,1H),8.00(s,1H),7.90-7.93(m,1H),7.18(d,J＝8.4Hz,1H),4.84(s,2H),4.08(t,J＝5.6Hz,2H),3.90(s,3H),3.36-3.38(m,4H),2.69-2.77(m,6H)。

and a sixth step: reacting N- (3-cyano-6- (5- ((5- (piperazine-1-acyl) pyridine-2-yl) oxo) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothiophene [2,3-c ]]Pyridin-2-yl) -1-methyl-1H-pyrazol-4-ylAmine (100mg, 0.17mmol), 4- (difluoromethoxy) benzaldehyde (45mg, 0.26mmol) and NaBH₃CN (11mg, 0.17mmol) was dissolved in anhydrous methanol (5mL), and AcOH (0.5mL) was added to the solution to react at room temperature for 4 hours. Concentrating to obtain crude product, and adding saturated NaHCO₃The solution was slurried twice with deionized water, filtered, and the filter cake was purified by preparative chromatography to give the title compound (25mg, yellow solid). MS (M + H) 727.1;¹H-NMR(400MHz,DMSO-d6)：11.53(brs,1H),8.41-8.47(m,3H),8.16(d,J＝2.0Hz,1H),8.05(s,1H),7.89-7.92(m,1H),7.34-7.40(m,2H),7.12-7.21(m,4H),4.83(s,2H),4.10(t,J＝5.6Hz,2H),3.91(s,3H),3.27-3.63(m,6H),2.70(brs,2H),2.30(brs,4H)。

example 66: n- (3-cyano-6- (5- ((5- (4- (4-methoxybenzyl) piperazin-1-yl) pyridin-2-yl) oxo) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

The title compound was prepared according to the synthesis method of example 65. MS (M + H) 691.2;¹H-NMR(400MHz,DMSO-d6)：11.53(brs,1H),8.41(s,3H),8.15(d,J＝2.0Hz,1H),8.05(s,1H),7.88-7.91(m,1H),7.16-7.21(m,3H),6.86-6.89(m,2H),4.85(s,2H),4.10(t,J＝4.8Hz,2H),3.91(s,3H),3.73(s,3H),3.29-3.58(m,6H),2.70(brs,2H),2.33(brs,4H)。

example 67: n- (3-cyano-6- (5- (1- (2-hydroxyethyl) -1H-pyrazol-5-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole e-4-amide

Example 67, MS (M + H): 476.5;¹H-NMR(400MHz,DMSO-d6)：11.51(brs,1H),8.80(s,2H),8.47(s,1H),8.07(s,1H),7.75(d,J＝2.0Hz,1H),6.66(d,J＝2.4Hz,1H),4.89-4.92(m,3H),4.12-4.18(m,4H),3.91(s,3H),3.74-3.78(m,2H),2.67(brs,2H)。

example 68: n- (3-cyano-6- (5- (1- (2-fluoroethyl) -1H-pyrazol-5-yl) pyrimidin-2-yl-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Example 68 was obtained in the same manner and with success as example 43, MS (M + H): 478.2;¹H-NMR(400MHz,DMSO-d6)：11.51(s,1H),8.81(s,2H),8.47(s,1H),8.07(s,1H),7.82(d,J＝2.4Hz,1H),6.71(d,J＝2.4Hz,1H),4.89(s,2H),4.72-4.86(m,2H),4.43-4.51(m,2H),4.13(t,J＝5.6Hz,2H),3.91(s,3H),2.71(t,J＝5.6Hz,2H)。

example 69: n- (3-cyano-6- (5- (5-methyl-1- (3- (4-methylpiperazin-1-yl) benzyl) -1H-1,2, 4-triazol-3-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Using example 64(40mg,0.065mmol) as a starting material, N-methylpiperazine (10mg,0.1mmol) and Cs₂CO₃(46mg,0.14mmol), X-Phos (3.3mg,0.007mmol), X-Phos-Pd-G3(5.5mg,0.007mmol) and Pd₂(dba)₃(6.4mg,0.014mmol) was dissolved in DMSO (3mL) and heated to 100 ℃ under nitrogen for overnight reaction. Ethyl acetate (20mL) was added for dilution and washed with water (10mL)3 times. The organic phase was dried, concentrated and purified by preparative chromatography to give the title compound (10mg, yellow solid). MS (M + H): 635.8;¹H-NMR(400MHz,DMSO-d6)：11.46(brs,1H),8.89(s,2H),8.46(s,1H),8.06(s,1H),7.52(d,J＝8.0Hz,1H),7.48(s,1H),7.34(t,J＝8.0Hz,1H),7.23(d,J＝8.0Hz,1H),5.43(s,2H),4.91(s,2H),4.15(t,J＝5.6Hz,2H),3.91(s,3H),2.70-2.73(m,2H)，2.47(s,3H)。

example 70: 6- (5-bromopyrimidin-2-yl) -2- (1-methyl-1H-pyrazol-4-ylamino) -4,5,6, 7-tetrahydrothiophene [2,3-c ] pyridine-3-carboxamide

Reacting N- (6- (5-bromopyrimidine-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothiophene [2,3-c ] ]Pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide (100mg,0.22mmol) was dissolved in concentrated hydrochloric acid (3mL), reacted overnight at room temperature under nitrogen, water was added to precipitate a solid, and filtered to give the title compound (45mg, black solid). MS (M + H) 461.9;¹H-NMR(400MHz,DMSO-d6):12.35(s,1H),8.53(s,2H),8.32(s,1H),7.83(s,1H),4.84(s,2H),4.01(t,J＝5.6Hz,2H),3.92(s,3H),2.91(brs,2H)。

example 71: n- (6- (5- (1- (2-amino-2-oxoethyl) -1H-pyrazol-5-yl) pyrimidin-2-yl) -3-cyano-4, 5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Using example 1 and (1- (2-amino-2-oxoethyl) -1H-pyrazole-5 boronic acid as starting materials, example 71(23mg, pale yellow solid). MS (M + H): 489.1; 1H-NMR (400MHz, DMSO-d 6): 11.51(s,1H),8.80(s,2H),8.47(s,1H),8.07(s,1H),7.75(d, J ═ 2.4Hz,1H),7.51(s,1H),7.27(s,1H),6.70(d, J ═ 2.0Hz,1H),4.89(s,2H),4.79(s,2H),4.13(t, J ═ 5.6Hz,2H),3.91(s,3H),2.71(brs,2H) was prepared in the same manner as in example 50.

Example 72: n- (3-cyano-6- (5- (1- (cyanomethyl) -1H-pyrazol-5-yl) pyrimidin-2-yl) -4,5,6, 7-tetrahydrothieno [2,3-c ] pyridin-2-yl) -1-methyl-1H-pyrazole-4-amide

Example 72(35mg, yellow solid) was prepared using example 1 and (1- (cyanoethyl) -1H-pyrazol-5-yl) boronic acid as starting materials in the same manner as in example 50. . MS (M + H) 471.1; 1H-NMR (400MHz, DMSO-d 6): 11.50(s,1H),8.78(s,2H),8.49(s,1H),8.05(s,1H),7.73(d, J ═ 2.4Hz,1H),7.47(s,1H),5.09(s,2H),4.68(s,2H),4.15(t, J ═ 5.6Hz,2H),3.90(s,3H),2.73(brs, 2H).

Test example 1: DU145 cells were tested for inhibition of pY705-STAT3 by Western bolt.

And (3) testing: 1. paving a plate and adding medicine: DU145 cells were cultured at 5X 10⁵Cell amount per well was seeded into six well plates; culturing at 37 deg.C for 24 hr, adding compound, and culturing for 24 hr. 2. Total protein extraction and quantification of DU145 cells: taking out the six-hole plate, completely sucking culture solution, and washing with PBS for three times; adding 200 μ L of 2% SDS cell lysate to each well of cells; carrying out ultrasonic crushing, and centrifuging at 14000Xg for 10min at 4 ℃; the supernatant was transferred and the protein concentration was determined by the Bradford method. 3. Loading: add 15. mu.g of protein per well. 4. Western Blot analysis: performing constant voltage electrophoresis at 80V for 30min, and performing constant voltage electrophoresis at 120V for 65min after the bromophenol blue migrates to the separation gel; rotating the membrane at constant pressure of 100V for 90 min; after the membrane is converted, closing the membrane and cutting the membrane according to the molecular weight of the target protein; after incubation with primary and secondary antibodies, respectively, ECL was imaged and analyzed using a Bio-Rad chemiluminescent imaging system.

The results show that most of the compounds in the examples have stronger inhibition effect on the phosphorylation of the pY705-STAT3 protein of DU145 cells, for example, the inhibition effect of the compounds in the examples 1, 10, 12, 21, 28, 33, 35, 38, 39, 40, 68 and the like at the concentration of 1uM is stronger than the inhibition effect of the compounds in the positive control compound C188-9 (at the concentration of 30 uM) on the phosphorylation of the Y705-STAT3 protein.

Test example 2: EXAMPLES Effect of Compounds on the proliferation potency of STAT 3-positive cells, such as DU145, PANC-1, BxPC-3, and MiaPaca-2

1. The Cell counting kit-8(CCK-8) is used for respectively detecting the growth influence of the compounds of the examples on DU145, PANC-1, BxPC-3 and MiaPaca-2, Cells are inoculated in a 96-well plate, 3000Cells/well are added after 24h according to the preset concentration, 10uL CCK-8/well is added after 24h, the temperature is 37 ℃, the luminescence is measured by using Top Count 384. The inhibition rate (%) of the compound on the growth of tumor cells was calculated by the following formula (OD negative control well-OD administration well)/OD negative control well × 100%. IC (integrated circuit)₅₀The values were determined by regression with a four parameter method using a microplate reader random plus software. (A represents IC)₅₀<Or 100nM, B stands for 100nM<IC₅₀<Or ═500nM, C500 nM<IC₅₀<Or 1000nM, D stands for IC₅₀>1000nM)

2. As a result, the compounds of the examples provided by the present invention have proliferation inhibitory activity on cells such as DU145, PANC-1, BxPC-3, and MiaPaca-2, and most of the IC₅₀Values less than 20000 nM; proliferation inhibitory Activity of most of the example Compounds on the above cells IC₅₀Values less than 1000 nM; inhibitory Activity IC of some of the example Compounds, e.g., examples 1, 10, 12, 13, 14, 21, 28, 33, 35, 38, 39, 40, etc ₅₀Values even less than 100nM show strong cell proliferation inhibitory activity.

Test example 3: test of the Compounds of the examples for the inhibitory Activity on different kinases

The compounds of the invention were also tested for their inhibitory activity against different kinases such as the CDK family, the EGFR family, the JAK family, the ROCK family such as CDK1, CDK2, CDK9, EGFR, VEGFR, PDGFR, FGFR, RET, MET, Src, Lyn, Syk, MEK, RAF, ROCK1, ROCK 2, etc., and some of the example compounds such as 1, 10, 12, 21, 28, 35, 40, 68, etc., showed good target selectivity, greater than 100-fold.

Test example 4: example compounds assay for STAT3 non-activated cell proliferation inhibitory Activity

The cell proliferation inhibition activity of STAT3 non-activated cells NIH-3T3 is tested by using a CCK8 cell proliferation inhibition experiment method, and part of examples of the invention, such as 1, 10, 12, 21, 28, 35, 40, 68, and the like, have weak cell proliferation inhibition activity on NIH-3T3, and the IC50 is more than 1uM, so that the cell proliferation inhibition selectivity is better.

Test example 5: ADMET testing of the example Compounds

(1) Metabolic stability test: the system is 150 mu L liver microsome (final concentration is 0.5mg/mL) for metabolic stability incubation, the system contains NADPH (final concentration is 1mM), 1 mu M test compound and positive control midazolam or negative control atenolol, the reaction is stopped by acetonitrile containing tinidazole at 0min, 5min, 10min and 30min respectively, vortex for 10min, centrifuge for 10min at 15000rmp, and 50 mu L supernatant is taken to be injected into a 96-well plate. The metabolic stability of the compounds was calculated by determining the relative decrease of the bulk drug.

As a result: the compound of the embodiment of the invention has high stability to liver microsomes of various species (rats, mice, dogs and humans), and the half-life period is more than 30min, such as the compounds 1, 10, 12, 21 and the like of the embodiment.

(2) Direct inhibition assay (DI assay): the incubation was directly inhibited with 100. mu.L of human liver microsomes (final concentration 0.2mg/mL), which contained NADPH (final concentration 1mM), 10. mu.M of compound, cococktail (ketoconazole 10. mu.M, quinidine 10. mu.M, sulfaphenazole 100. mu.M, alpha-naphthoflavone 10. mu.M, tranylcypromine 1000. mu.M), negative control (BPS with 0.1% DMSO), and mixed probe substrate (midazolam 10. mu.M, testosterone 100. mu.M, dextromethorphan 10. mu.M, diclofenac 20. mu.M, phenacetin 100. mu.M, and mefenton 100. mu.M), and the reaction was terminated after incubation for 20 min. The relative activity of the enzyme was calculated by measuring the relative production of metabolites.

As a result: the compound pairs of the examples of the invention, such as the compounds 1, 10, 12, 21, 28, 33, 38 and the like of the examples, have no obvious inhibition on main metabolic enzyme subtypes (CYP1A2, 2C8, 2C19 and 3A4), and IC50 is more than 10 uM.

(3) hERG inhibition assay: compound stock solution of 20mM was diluted with DMSO, 10 μ L of 20mM compound stock solution was added to 20 μ L DMSO solution, and 3-fold serial dilutions were made to 6 DMSO concentrations; respectively taking 4 mu L of compounds with 6 DMSO concentrations, adding the compounds into 396 mu L of extracellular fluid, diluting the compounds to 6 intermediate concentrations by 100 times, then respectively taking 80 mu L of compounds with 6 intermediate concentrations, adding the compounds into 320 mu L of extracellular fluid, and diluting the compounds to a final concentration to be tested by 5 times; the highest test concentration is 40 mu M, and the highest test concentration is respectively 40, 13.3, 4.4, 1.48, 0.494 and 0.165 mu M for 6 concentrations; the DMSO content in the final assay concentration did not exceed 0.2%, and DMSO at this concentration had no effect on the hERG potassium channel; compounds were prepared for the entire dilution process by Bravo instrument; the current and time course of compound to hERG potassium channel were read and the inhibition curve of compound to hERG was made by fitting the curve.

As a result: the compounds of the embodiment of the invention such as 1, 7, 10, 12, 21, 28, 33, 35, 38, 40 and the like have no obvious inhibition effect on hERG potassium channel, IC₅₀Greater than 20 uM.

Test example 6: in vivo pharmacokinetic parameter testing of example Compounds in rats, mice

Dividing 6 male SPF SD rats or Balb c mice (Shanghai Spire-BiKa experimental animals) into two groups, and preparing the tested compound into appropriate solution or suspension; one group was administered intravenously (1mg/kg) and one group was administered orally (5 mg/kg). Blood is collected by jugular venipuncture, about 0.2 mL/time point of each sample is collected, heparin sodium is anticoagulated, and the blood collection time points are as follows: pre-dose and 5, 15 and 30min, 1, 2, 4, 6, 8 and 24h after dose administration; blood samples were collected and placed on ice, plasma was centrifuged (centrifugation conditions: 8000 rpm, 6 min, 2-8 ℃) and collected plasma was stored at-80 ℃ before analysis. Plasma samples were analyzed by LC-MS/MS.

According to the data of the blood concentration of the drug, pharmacokinetic calculation software WinNonlin5.2 non-atrioventricular model is used for respectively calculating the pharmacokinetic parameters AUC of the test sample_0-t、AUC_0-∞、MRT_0-∞、C_max、T_max、T_1/2And V_dIsoparametric and their mean and standard deviation. In addition, the bioavailability (F) will be calculated by the following formula.

For samples with concentrations below the lower limit of quantitation, C is reached when pharmacokinetic parameter calculations are performed_maxThe previously sampled samples should be counted as zero values,in the process of reaching C_maxSamples from later sampling points should be calculated as not quantifiable (BLQ).

As a result: some of the compounds 1, 10, 12, 21, etc. of the examples of the present invention have good pharmacokinetic properties in mice, and have oral bioavailability greater than 30%, half-life greater than 2 hours, Cmax greater than 250nM, AUC (hr × nM) greater than 2000, even greater than 25000, and bioavailability greater than 30%.

Test example 7: EXAMPLES test of Compounds for growth inhibition of PANC-1, MiaPaca-2, BxPC-3 nude mouse transplanted tumors

Cutting tumor tissue in vigorous growth stage into 1.5mm³And left and right, under aseptic conditions, inoculated subcutaneously in the right axilla of nude mice. Measuring the diameter of the transplanted tumor by using a vernier caliper in the nude mouse subcutaneous transplanted tumor until the average tumor volume reaches 100mm³Animals were randomized into groups. The compound of the example (prepared to the required concentration with water for injection containing 1% Tween 80) was administered orally at the given dose daily for three weeks with the solvent control group given an equal amount of solvent. Throughout the experiment, the diameter of the transplanted tumor was measured 2 times per week, while the body weight of the mice was weighed. The formula for Tumor Volume (TV) is: TV 1/2 × a × b ²Wherein a and b represent length and width, respectively. Calculating Relative Tumor Volume (RTV) according to the measurement result, wherein the calculation formula is as follows: RTV is Vt/V0. Where V0 is the tumor volume measured at the time of caged administration (i.e., d0) and Vt is the tumor volume at each measurement. The evaluation index of the antitumor activity is 1) the relative tumor proliferation rate T/C (%), and the calculation formula is as follows: T/C (%) (TRTV/CRTV) × 100%, TRTV: treatment group RTV; CRTV: negative control group RTV; 2) the tumor volume increase inhibition rate GI% is calculated according to the following formula: GI% ═ 1- (TVt-TV0)/(CVt-CT0)]×100％，TVt is the tumor volume measured for each treatment group; TV0 is the tumor volume obtained when therapeutic components were administered in cages; CVt is the tumor volume measured in each time in the control group; CV0 is the tumor volume obtained when the control component was administered in cages; 3) the tumor weight inhibition rate is calculated according to the following formula: tumor weight inhibition ratio [ (% Wc-W) ]_T) Wc × 100%, Wc: tumor weight of control group, W_T: the treated group had heavy tumor.

As a result: the compounds of some examples of the invention, such as examples 1, 10, 12, 39, etc., show better in vivo tumor inhibition effect after being orally administered once a day for 28 days under the dosage of 10 mg/kg-100 mg/kg, the tumor inhibition rate is more than 50%, and the tumor inhibition rate of some compounds, such as examples 1, 12, etc., is more than 70%.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. A thieno-nitrogen heterocyclic compound shown as a general formula I or a pharmaceutically acceptable salt thereof is characterized in that the thieno-nitrogen heterocyclic compound shown as the general formula I is a compound shown as a formula I-1,

wherein R1 is-CONHR or cyano; r is hydrogen or C1-C6 alkyl;

r5 is 5-6 membered heteroaryl; said 5-6 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl; r5 wherein the heteroaryl group contains 1 to 3 heteroatoms selected from the group consisting of: n, O and S;

is composed of

Wherein R3 is hydrogen or halogen;

r3a is independently hydrogen;

r3b is independently cyano, halogen, phenyl, 5-10 membered heteroaryl or

When R3b is independently a 5-10 membered heteroaryl, the 5-10 membered heteroaryl is

In R3b, one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent selected from the group consisting of: hydroxy, halogen, C1-C8 alkyl, and C1-C8 alkoxy; said C1-C8 alkyl is optionally substituted with one or more substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy, NH ₂C (═ O) -and 5-8 membered heteroaryl;

r3b wherein in the 5-8 membered heteroaryl, the heteroatom is selected from N, O and S;

when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent which is a C1-C8 alkoxy group, the C1-C8 alkoxy group is methoxy or ethoxy.

2. The thieno-nitrogen heterocycle compound of formula I or its pharmaceutically acceptable salt according to claim 1,

r1 is-CONH₂Or a cyano group;

in R5, in the 5-6 membered heteroaryl, the heteroatom is selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituents are plural, the same or different;

r3a is independently hydrogen;

in R3b, in the 5-8 membered heteroaryl, the heteroatom is selected from N, O and S, and the number of the heteroatoms is 1, 2 or 3; when the substituent is plural, the same or different.

3. The thienoazacycles compound of general formula I or a pharmaceutically acceptable salt thereof as claimed in claim 1,

when R5 is a 5-6 membered heteroaryl group, wherein the heteroatoms are selected from N and O, the number of heteroatoms is 1, 2 or 3; and/or, when R5 is 5-6 membered heteroaryl, said 5-6 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl, said C1-C6 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl;

And/or, when R3 is independently halogen, the halogen is fluorine, chlorine, bromine or iodine;

and/or, when R3b is independently halogen, the halogen is fluorine, chlorine, bromine or iodine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl is optionally substituted by a substituent which is halogen, the halogen is fluorine, chlorine, bromine or iodine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-to 10-membered heteroaryl is optionally substituted by a substituent, which is C1-C8 alkyl, the C1-C8 alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-to 10-membered heteroaryl are optionally substituted by a substituent, said substituent is C1-C8 alkyl, said C1-C8 alkyl is optionally substituted by one or more halogens, said halogen is fluorine, chlorine, bromine or iodine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-to 10-membered heteroaryl are optionally substituted by a substituent, the substituent is C1-C8 alkyl, the C1-C8 alkyl is optionally substituted by one or more C1-C8 alkoxy, the C1-C8 alkoxy is methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy;

And/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent, the substituent is C1-C8 alkyl, the C1-C8 alkyl is optionally substituted by one or more 5-8 membered heteroaryl, the 5-8 membered heteroaryl is 5-6 membered heteroaryl, wherein the heteroatoms are selected from N and O, and the number of heteroatoms is 1, 2 or 3.

4. The thieno-nitrogen heterocycle compound of formula I or its pharmaceutically acceptable salt according to claim 3,

when R5 is a 5-6 membered heteroaryl, the 5-6 membered heteroaryl is

And/or, when R5 is a 5-6 membered heteroaryl, said 5-6 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl, said C1-C6 alkyl is methyl or ethyl;

and/or, when R3 is independently halogen, the halogen is bromine;

and/or, when R3b is independently halogen, said halogen is bromine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl is optionally substituted by a substituent which is halogen, the halogen is fluorine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent which is C1-C8 alkyl, the C1-C8 alkyl is methyl, ethyl, propyl, isopropyl, or sec-butyl;

And/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent, said substituent is C1-C8 alkyl, said C1-C8 alkyl is optionally substituted by one or more halogens, said halogen is fluorine;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent, said substituent is C1-C8 alkyl, said C1-C8 alkyl is optionally substituted by one or more C1-C8 alkoxy, said C1-C8 alkoxy is methoxy;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent, said substituent is C1-C8 alkyl, said C1-C8 alkyl is optionally substituted by one or more 5-8 membered heteroaryl, said 5-8 membered heteroaryl is

5. The thienoazacycles compound of general formula I or its pharmaceutically acceptable salt according to claim 3 or 4,

r3 is H or Br;

and/or, in R3b, when one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent selected from the group consisting of hydroxy, halogen, cyano, C1-C8 alkyl or C1-C8 alkoxy, the substituent is methoxy, trifluoromethyl, hydroxy, methyl, chloro, ethoxy, ethyl, cyano, nitro, or a pharmaceutically acceptable salt thereof,

N-propyl, sec-butyl, isopropyl,

6. The thieno-nitrogen heterocycle compound of formula I or its pharmaceutically acceptable salt according to claim 5,

r5 is

And/or, R3b is independently Br, F, CN,

7. The thieno-nitrogen heterocyclic compound represented by general formula I or the pharmaceutically acceptable salt thereof according to claim 1, wherein the thieno-nitrogen heterocyclic compound represented by general formula I is scheme two, scheme three or scheme four;

scheme two is as follows:

r1 is-CONH₂Or cyano;

r5 is 5-6 membered heteroaryl; said 5-6 membered heteroaryl is optionally substituted with one or more substituents which are C1-C6 alkyl;

is composed of

R3 is independently hydrogen or halogen;

r3a is independently H;

r3b is independently cyano, halogen, phenyl, 5-10 membered heteroaryl or

In R3b, one or more hydrogen atoms on the phenyl or 5-10 membered heteroaryl are optionally substituted by a substituent selected from the group consisting of: hydroxy, halogen, C1-C8 alkyl, and C1-C2 alkoxy; said C1-C8 alkyl is optionally substituted with one or two substituents independently selected from the group consisting of: hydroxy, cyano, halogen, C1-C8 alkoxy, NH ₂C (═ O) -and 5-8 membered heteroaryl;

and a third scheme is as follows:

the thieno-nitrogen heterocyclic compound shown as the general formula I is a compound shown as a general formula I-2 or a general formula I-3,

wherein R1, M1, M2, M3 and R5 are as defined in claim 1;

r5a is C1-C6 alkyl;

and the scheme is as follows:

the thieno-nitrogen heterocyclic compound shown as the general formula I is a compound shown as a general formula I-7,

wherein R1, R5, R3a, R3b and R3 are as defined in claim 1.

8. A thieno-nitrogen heterocyclic compound shown as a general formula I or a pharmaceutically acceptable salt thereof is characterized in that the thieno-nitrogen heterocyclic compound shown as the general formula I is of any one of the following structures,

9. a process for the preparation of thienoazacycles of general formula I as defined in any of claims 1 to 8 comprising the following steps: in a solvent, in the presence of alkali, carrying out substitution reaction on a compound shown as a formula B and a compound shown as a formula C as shown in the specification;

wherein R1, R5, M1, M2 and M3 are as defined in any one of claims 1 to 8, n is 1;

x is chlorine, bromine, iodine or methylsulfonyl.

10. A pharmaceutical composition comprising the thienoazacycles of formula I or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 8 and a pharmaceutically acceptable carrier.

11. Use of the thienonitrogen heterocycle of general formula I or a pharmaceutically acceptable salt thereof as described in any of claims 1-8 or of the pharmaceutical composition of claim 10 for the preparation of a STAT inhibitor, a medicament for the prevention or treatment of a disease associated with STAT protein activity or expression, or a medicament for the prevention or treatment of one or more of a tumor, an immune disease and an inflammatory disease.

12. The use according to claim 11,

the tumor is independently selected from non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, breast cancer, prostate cancer, liver cancer, skin cancer, stomach cancer, intestinal cancer, cholangiocarcinoma, brain cancer, leukemia, lymph cancer, fibroma, sarcoma, basal cell carcinoma, glioma, kidney cancer, melanoma, bone cancer, thyroid cancer, nasopharyngeal cancer and pancreatic cancer.

13. The use according to claim 11,

the immune disease and inflammatory disease are independently selected from rejection of transplanted organs, gout, rhinitis, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, arthritis, allergic dermatitis, Behcet's disease, bullous skin disease, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease, liver cirrhosis, degenerative joint disease, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, gastritis, nephritis, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease, irritable bowel syndrome, Kawasaki disease, meningitis, multiple sclerosis, myocarditis, myasthenia gravis, mycosis fungoides, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, psoriasis, and the like, Fibrosis, lupus erythematosus, tissue transplant rejection, thyroiditis, type I diabetes, urethritis, uveitis, vasculitis, vitiligo, and Waldenstrom's macroglobulinemia.