CN109422760B - FGFR4 inhibitor and application thereof - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry, and particularly relates to a novel FGFR4 inhibitor, a pharmaceutical composition containing the inhibitor, and application of the inhibitor or the pharmaceutical composition as a cancer treatment drug.

Description

FGFR4 inhibitor and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and particularly relates to a novel FGFR4 inhibitor, a pharmaceutical composition containing the inhibitor, and application of the inhibitor or the pharmaceutical composition as a cancer treatment drug.

Background

The receptor tyrosine kinase plays a key role in various links such as tumorigenesis development, invasion and metastasis, drug resistance and the like due to abnormal expression activation or gene mutation, and becomes an important target for research and development of antitumor drugs. Fibroblast Growth Factor Receptors (FGFRs) are important members of the receptor tyrosine kinase family, mainly including four subtypes FGFR1, FGFR2, FGFR3 and FGFR 4. The ligand is Fibroblast Growth Factors (FGFs). These receptors form ternary complexes with FGFs and Heparan Sulfate Proteoglycans (HSPGs), which in turn trigger a series of signaling and participate in the regulation of various physiological and pathological processes in the organism.

The amino acid sequences between FGFR family members (FGFR1, FGFR2, FGFR3, and FGFR4) are highly conserved and show differences in ligand affinity, tissue distribution, and the like. Due to gene amplification, mutation, fusion or ligand induction and the like, all the FGFR members are continuously activated to induce the proliferation, invasion and migration of tumor cells, promote angiogenesis and promote the generation and development of tumors. FGFRs are highly expressed and abnormally activated in various tumors and are closely related to poor prognosis of tumor patients, such as non-small cell lung cancer, breast cancer, gastric cancer, bladder cancer, endometrial cancer, prostate cancer, cervical cancer, colon cancer, esophageal cancer, keratinocyte tumor, myeloma, rhabdomyosarcoma and the like. Fibroblast growth factor receptor 4(FGFR4) is a member of the fibroblast growth factor receptor family, encoded by the FGFR4 gene, and the FGFR4 genomic structure contains 18 exons. Ectopic mineralization was observed in rats treated with FGFR1 inhibitor, showing inappropriate calcium phosphorus deposition in soft tissue (Brown, AP et al (2005), Toxicol. Pathol, p. 449-455). This indicates that selective inhibition of FGFR4 is desirable to avoid certain toxicities. The research finds that FGFR4 is the only receptor with specificity shown by FGF19 (a physiological ligand of FGFR4), and the overexpression of FGF19 can cause the activation of FGF19-FGFR4 pathways, thereby causing cancers such as sarcoma, renal cell carcinoma, breast cancer, liver cancer and the like. FGFR4 inhibitor therapy is effective against tumors with FGF19 gene amplification. Ectopic mineralization was observed in rats treated with FGFR1 inhibitor and characterized by inappropriate calcium phosphorus deposition in soft tissues (Brown, AP et al (2005), toxicol. Pathol, p. 449-455). This indicates that selective inhibition of FGFR4, but not other subtypes of FGFR such as FGFR1, can avoid certain toxic side effects of the drug. Currently, a range of FGFR4 inhibitors are disclosed, including the compounds disclosed in WO2015/108992, WO2015/059668, WO2015061572 and the like. There is still a need to develop compounds with better efficacy, less toxicity and higher selectivity.

Disclosure of Invention

The invention aims to provide a compound with FGFR inhibitory activity shown in a general formula I or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof,

wherein:

warheads are moieties capable of forming covalent bonds with nucleophiles;

ring a is a saturated or unsaturated mono-, spiro-, fused or bridged heterocyclic group, wherein the mono-, spiro-, fused or bridged heterocyclic group is substituted with one or more optionally substituted aryl, heteroaryl, cycloalkyl or heterocyclic group, which is optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxyl, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkylacylalkyl, aryl, heteroaryl, cycloalkyl and heterocyclic groups;

each R₁、R₂、R₃Each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, cycloalkyl, heterocyclyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, amino, alkylamino, alkanoylamino, alkanoyl, aminoacyl, alkylaminoacyl, carboxy, nitro, cyano, aryl, and heteroaryl, wherein n, p, q are each independently selected from the group consisting of 1,2, 3, 4, and 5; and

R₄and R₅Each independently selected from H and optionally substituted alkyl.

It is another object of the present invention to provide a process for preparing the compound of formula I of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug.

It is a further object of the present invention to provide compositions comprising a compound of formula I of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug and a pharmaceutically acceptable carrier, and compositions comprising a compound of formula I of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug and another FGFR inhibitor or inhibitors.

Still another object of the present invention is to provide a method for treating cancer by the compound of formula I of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug, and the use of the compound of formula I of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug for the manufacture of a medicament for treating cancer.

Aiming at the above purpose, the invention provides the following technical scheme:

in a first aspect, the invention provides a fibroblast growth factor receptor inhibitor with a general formula I or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, which shows good FGFR inhibitory activity, especially selectively inhibits FGFR4,

wherein:

warheads are moieties capable of forming covalent bonds with nucleophiles;

ring a is a saturated or unsaturated mono-, spiro-, fused or bridged heterocyclic group, wherein the mono-, spiro-, fused or bridged heterocyclic group is substituted with one or more optionally substituted aryl, heteroaryl, cycloalkyl or heterocyclic group, and the spiro-, fused or bridged heterocyclic group is optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxyl, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkylacylylalkyl, aryl, heteroaryl, cycloalkyl and heterocyclic groups;

each R₁、R₂、R₃Each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, cycloalkyl, heterocyclyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, amino, alkylamino, alkylacylamino, alkanoyl, aminoacyl, alkylaminoacyl, carboxy, nitro, cyano, aryl and heteroaryl, wherein n, p, q are each independently selected from 1,2, 3, 4 and 5; and

R₄and R₅Each independently selected from H and optionally substituted alkyl.

In some preferred embodiments of the invention, a compound according to formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein:

ring A is a saturated or unsaturated 3-12 membered monocyclic azaheterocyclyl, monocyclic diazacyclo, monocyclic azaheterocyclyl, azaspiro cycloalkyl, diazaspiro cycloalkyl, azaspiro cycloalkyl, azafused ring alkyl, diazaspiro cycloalkyl, azabridged cycloalkyl or azabridged cycloalkyl group, wherein said 3-12 membered monocyclic azaheterocyclyl, monocyclic diazacyclo and monocyclic azaheterocyclyl are substituted with one or more C_6-12Aryl, 5-12 membered heteroaryl, C_3-12Cycloalkyl or 3-12 membered heterocyclyl, wherein said C is_6-12Aryl, 5-12 membered heteroaryl, C_3-12Cycloalkyl or 3-12 membered heterocyclyl optionally substituted by one or more halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, C_1-6Alkyl acyl radical C_1-6Alkyl, hydroxy C_1-6Alkyl, monoamino C_1-6Alkyl and diamino C_1-6Alkyl substitution; said 3-12 membered azaspiro cycloalkyl, diazaspiro cycloalkyl, oxaspiro cycloalkyl, aza-fused ring alkyl, diazepine-fused ring alkyl, oxafused cycloalkyl, aza-bridged cycloalkyl, diazepine-bridged cycloalkyl or oxabridged cycloalkyl group optionally substituted with one or more nitrogen atomsSelected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, aryl, heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl;

further preferably, ring A is a saturated or unsaturated 3-12 membered monocyclic azaheterocyclyl, monocyclic diazacyclo, monocyclic azaheterocyclyl, azaspirocycloalkyl, diazaspiro-cycloalkyl, azaspirocycloalkyl, azacondensed-ring alkyl, diazaspiro-ring alkyl, azacondensed-ring cycloalkyl, azabridged cycloalkyl, diazaspiro-bridged cycloalkyl or nitroxide-bridged cycloalkyl group, wherein said 3-12 membered monocyclic azaheterocyclyl, monocyclic diazacyclo and monocyclic azaheterocyclyl are substituted with C_6-8Aryl, 5-8 membered heteroaryl, C_3-8Cycloalkyl or 3-8 membered heterocyclyl, wherein said C is_6-8Aryl, 5-8 membered heteroaryl, C_3-8Cycloalkyl or 3-8 membered heterocyclyl optionally substituted by one or more halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, C_1-3Alkyl acyl radical C_1-3Alkyl, hydroxy C_1-3Alkyl, monoamino C_1-3Alkyl and diamino C_1-3Alkyl substitution; said 3-12 membered azaspiro cycloalkyl, diazaspiro cycloalkyl, oxaspiro cycloalkyl, aza-fused ring alkyl, diazepine-fused ring alkyl, oxafused cycloalkyl, aza-bridged ring alkyl, diazepine-bridged ring alkyl, and oxabridged cycloalkyl optionally substituted with one or more substituents selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl radical, C_1-3Alkyl acyl radical C_1-3Alkyl, 6-8 membered aryl, 5-8 membered heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl;

in some embodiments of the invention, a compound according to formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein:

ring A is

Wherein said radicals are optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, preferably these groups are optionally substituted by one or more substituents selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, C_1-3Alkyl acyl radical C_1-3Alkyl substituents.

In some preferred embodiments, the compounds of the present invention are of formula I or an isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug thereof, wherein:

each R₁、R₂、R₃Each independently selected from hydrogen, halogen, hydroxy, C_1-6Alkyl radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, amino, C_1-6Alkylamino radical, C_1-6Alkylacylamino group, C_1-6Alkyl acyl, amino acyl, C_1-6Alkyl amino acyl, carboxyl, nitro,Cyano radicals, C_6-12Aryl and 5-12 membered heteroaryl;

further preferably, each R₁、R₂、R₃Each independently selected from hydrogen, halogen, hydroxy, C_1-3Alkyl radical, C_3-10Cycloalkyl, 3-10 membered heterocyclyl, halo C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, amino, C_1-3Alkylamino radical, C_1-3Alkylacylamino group, C_1-3Alkyl acyl, amino acyl, C_1-3Alkylaminoacyl, carboxyl, nitro, cyano, C_6-10Aryl and 5-10 membered heteroaryl;

even more preferably, each R₁、R₂、R₃Each independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, iodo, hydroxy, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy, amino, methylamino, dimethylamino, ethylamino, diethylamino, methylethylamino, methylacylamino, ethylacylamino, vinylacylamino, methylacyl, ethylacoyl, vinylacyl, aminoacyl, methylaminoacyl, ethylaminoacyl, carboxy, nitro, cyano, phenyl and azetidinyl.

In other embodiments, a compound according to general formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein:

R₄and R₅Each independently selected from H and alkyl, wherein the alkyl is optionally substituted with a substituent selected from the group consisting of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkanoylalkyl, aryl, heteroaryl, cycloalkyl, and heterocyclyl;

further preferably, R₄And R₅Each independently selected from H and C_1-6Alkyl, wherein the alkyl is optionally halogenatedElement, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl radical, C_6-12Aryl, 5-12 membered heteroaryl, C_3-12Cycloalkyl or 3-12 membered heterocyclyl;

even more preferably, R₄And R₅Each independently selected from H and C_1-3Alkyl, wherein the alkyl is optionally substituted by halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl, C_1-3Alkyl acyl radical C_1-3Alkyl radical, C_6-10Aryl, 5-to 10-membered heteroaryl, C_3-10Cycloalkyl or 3-10 membered heterocyclyl.

In other embodiments, the compound according to formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein n, p, q are each independently selected from 1,2, 3 and 4.

In other further preferred embodiments of the present invention, a compound according to formula I, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein formula I has the structure shown in formula II below,

wherein

Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl, wherein said aryl, heteroaryl, cycloalkyl and heterocyclyl are optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkylacylalkyl, aryl, heteroaryl, cycloalkyl and heterocyclyl;

m is selected from 0, 1,2, 3, 4 and 5;

warhead, R₁、R₂、R₃、R₄、R₅N, p and q are defined as general formula I; .

In yet other preferred embodiments of the present invention, a compound according to formula II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring B is C_6-12Aryl, 5-12 membered heteroaryl, C_3-12Cycloalkyl or 3-12 membered heterocyclyl, said C_6-12Aryl, 5-12 membered heteroaryl, C_3-12Cycloalkyl or 3-12 membered heterocyclyl is optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, 6-12 membered aryl, 5-12 membered heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl.

In other preferred embodiments of the present invention, a compound according to formula II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring B is a 3-to 8-membered nitrogen monoheterocyclyl; still more preferably a 5-or 6-membered nitrogen heteromonocyclic group; non-limiting examples of ring B include substituted or unsubstituted pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyranyl, or tetrahydrofuranyl.

In further preferred embodiments of the present invention, the compound according to formula II or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring B is a 7-to 15-membered nitrogen polycyclic heterocyclic group, more preferably a 7-to 15-membered nitrogen spiro heterocyclic group, nitrogen fused heterocyclic group and nitrogen bridged heterocyclic group, still more preferably a 7-to 12-membered nitrogen mono spiro heterocyclic group and nitrogen bridged heterocyclic group, whereinSaid nitrogen polycyclic heterocyclic group, nitrogen spiro heterocyclic group, nitrogen fused heterocyclic group or nitrogen bridged heterocyclic group is optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, 6-12 membered aryl, 5-12 membered heteroaryl, C_3-8Cycloalkyl and C_3-8Heterocyclyl is substituted with a substituent; still more preferably, ring B is

Wherein said radicals are optionally substituted by one or more groups selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl radical, C_1-3Alkyl acyl radical C_1-3Alkyl substituents.

In other preferred embodiments of the present invention, a compound according to formula I, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein formula I as shown has the structure shown in formula III below:

wherein: warhead, R₁、R₂、R₃、R₄、R₅N, p and q are defined as general formula I;

m and d are each independently selected from 1,2, 3 and 4;

ring C is selected from cycloalkyl and heterocyclyl, wherein said cycloalkyl and heterocyclyl are optionally substituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkylacylalkyl, aryl, heteroaryl, cycloalkyl and heterocyclyl.

In yet further preferred embodiments of the present invention, a compound according to formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein ring C is selected from the group consisting of 3-8 membered cycloalkyl and heterocyclyl, preferably ring C is selected from the group consisting of C_3-8Cycloalkyl, 3-8 membered azaheterocyclyl, 3-8 membered oxaheterocyclyl and 3-8 membered thiaheterocyclyl wherein these groups are optionally substituted with one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, 6-12 membered aryl, 5-12 membered heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl. In some embodiments, non-limiting examples of ring C include cyclopentyl, cyclohexyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyranyl, or tetrahydrofuranyl, wherein these groups are optionally substituted with one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl substituents.

In some embodiments of the invention, a compound according to formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein:

radical (I)

Is selected from

Wherein these radicals are optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, preferably these groups are optionally substituted by one or more substituents selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl radical, C_1-3Alkoxy radical C_1-3Alkyl radical, C_1-3Alkyl acyl radical, C_1-3Alkyl acyl radical C_1-3Alkyl substituents.

In some embodiments of the invention, a compound according to formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein

Is selected from the group

Wherein said groups are optionally substituted by one or more substituentsSubstituted with one or more substituents selected from halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkanoyl, alkanoylalkyl, aryl, heteroaryl, cycloalkyl and heterocyclyl, preferably said groups are optionally substituted with one or more substituents selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, 6-12 membered aryl, 5-12 membered heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl.

In some particular embodiments of the invention, a compound according to formula I or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein

Is selected from the group

In other preferred embodiments of the present invention, the compound according to formula I, formula II or formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein the warhead is selected from the group consisting of:

wherein R is_a、R_b、R_cEach independently selected from H, alkyl, cycloalkyl, cyano, wherein the alkyl or cycloalkyl is optionally substituted with one or more substituents selected from halogen, alkyl, haloAlkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkylacylalkyl, Xa is selected from halogen and triflate; preferably, R_a、R_b、R_cEach independently selected from H, C_1-6Alkyl radical, C_3-8Cycloalkyl, cyano, wherein the alkyl or cycloalkyl is optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl, alkoxy C_1-6Alkyl radical, C_1-6Alkyl acyl radical C_1-6Radical substitution of alkyl; further preferably, R_a、R_b、R_cEach independently selected from H, C_1-3Alkyl radical, C_3-6Cycloalkyl, cyano, wherein the alkyl or cycloalkyl is optionally substituted by one or more groups selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl, alkoxy C_1-3Alkyl radical, C_1-3Alkyl acyl radical C_1-3Alkyl groups.

In yet other preferred embodiments of the present invention, the compound according to formula I, formula II or formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein the warhead is selected from the group consisting of acryloyl, methacryloyl and propargyl acyl.

In other preferred embodiments of the present invention, a compound according to formula I, formula II or formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein each R is₁、R₂Each independently selected from hydrogen and C_1-6Alkyl radical, C_1-6An alkoxy group; preferably hydrogen, C_1-3Alkyl radical, C_1-3An alkoxy group; more preferably selected from hydrogen, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxyI-propoxy, wherein n is selected from 1,2 and 3; and p is selected from 1 and 2.

In other preferred embodiments, a compound according to formula I, formula II, or formula III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein each R is₃Each independently selected from halogen, C_1-6An alkoxy group; preferably selected from halogen, C_1-3An alkoxy group; each R₃Non-limiting examples of each independently include fluorine, chlorine, bromine, methoxy, ethoxy, propoxy, isopropoxy, wherein q is selected from 1,2, 3, 4 and 5; in some specific embodiments, each R is₃Each independently selected from fluorine, chlorine, bromine, methoxy, ethoxy, propoxy, isopropoxy and q is 4.

In other preferred embodiments, a compound according to formula I, formula II or formula III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₄Selected from H and C_1-6An alkyl group.

In other preferred embodiments, a compound according to formula I, formula II or formula III, or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein R is₅Selected from H and C_1-6Alkyl, preferably C_1-3Alkyl, more preferably selected from methyl, ethyl, propyl and isopropyl.

In some embodiments of the present invention, non-limiting examples of the compounds of formula I, formula II, or formula III, or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, include:

in other preferred embodiments, the compound according to formula I, formula II or formula III or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, wherein the salt comprises a conventional non-toxic salt of the compound according to formula I, formula II or formula III with an inorganic or organic acid. Conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and the like, as well as salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid and the like. Pharmaceutically acceptable salts of the compounds of the present invention can be synthesized by conventional chemical methods from compounds of the present invention that contain an acidic or basic moiety, and salts of the basic compounds of the present invention can be prepared by, for example, exchange chromatography, or by reacting the free base with a stoichiometric amount or an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents.

In another aspect, the present invention provides a process for the preparation of a compound of formula I according to the invention, comprising the steps of:

(1) reacting the compound of the formula 1 to prepare a compound of a formula 2;

(2) reacting the compound of formula 2 with the compound of formula 3 to produce a compound of formula 4;

(3) the compound of formula 4 and the compound of formula 5 are reacted via palladium mediated coupling to form the compound of formula 6;

(4) reducing the nitro group on the compound of formula 6 to an amino group to obtain a compound of formula 7;

(5) the compound of the formula 7 and warhead-LG 3 are subjected to amide coupling reaction to obtain a compound of the formula I;

wherein, warhead, R₁、R₂、R₃、R₄、R₅N, p, q and ring A have the meanings given in formula I; LG1, LG2, LG3 represent leaving groups, which may be the same or different, preferably halogen or sulfonyloxy, more preferably Cl, Br, I;

the nitro compound is reduced to prepare the aromatic primary amine by adopting the conventional reduction reaction, and the common reagents comprise metal and acid, wherein the metal uses iron, zinc or tin, and the acid uses hydrochloric acid, sulfuric acid or acetic acid and the like;

the nitro group can also be reduced by a catalytic hydrogenation method, common catalysts comprise Ni, Pt, Pd and the like, and the reaction can be carried out under neutral conditions.

On the basis of the preparation method of the compound of the general formula I or the pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof, the compound of the general formula II or the general formula III or the pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof can be obtained by selecting conventional raw materials by a person skilled in the art.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, a pharmaceutically acceptable salt, a crystal, a solvate or a prodrug thereof.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention, or an isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug thereof, further comprising one or more selected from the group consisting of: FGFR inhibitors, IDH inhibitors, tyrosine protease inhibitors, EGFR inhibitors, VEGFR inhibitors, Bcr-Abl inhibitors, c-kit inhibitors, c-Met inhibitors, Raf inhibitors, MEK inhibitors, histone deacetylase inhibitors, VEGF antibodies, EGF antibodies, HIV protein kinase inhibitors, HMG-CoA reductase inhibitors, and the like.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention, or an isomer, a pharmaceutically acceptable salt, a crystal, a solvate or a prodrug thereof, and a pharmaceutically acceptable carrier.

The compound of the present invention or its isomer, pharmaceutically acceptable salt, crystal, solvate or prodrug can be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation suitable for oral or parenteral administration. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulations may be administered by any route, for example by infusion or bolus injection, by a route of absorption through epithelial or cutaneous mucosa (e.g. oral mucosa or rectum, etc.). Administration may be systemic or local. Examples of the formulation for oral administration include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions and the like. The formulations may be prepared by methods known in the art and include carriers, diluents or excipients conventionally used in the art of pharmaceutical formulation.

In a fourth aspect, the invention provides a method for treating and/or preventing tumors by using a compound shown in a general formula I, a general formula II or a general formula III, or a pharmaceutically acceptable salt, an isomer, a solvate, a crystal or a prodrug thereof, or a pharmaceutical composition of the invention, and an application of the method in preparing a medicament for treating and/or preventing tumors.

The invention provides a method for treating and/or preventing diseases or symptoms mediated by FGFR-4 or FGF19 by using a compound shown in a general formula I, a general formula II or a general formula III or a pharmaceutically acceptable salt, an isomer, a solvate, a crystal or a prodrug thereof or a pharmaceutical composition of the invention, and application of the compound in preparing a medicament for treating and/or preventing diseases or symptoms mediated by FGFR-4 or FGF19, wherein the compound is characterized in that FGFR-4 or FGF19 is overexpressed, FGFR4 or FGF19 is amplified in an individual. In a preferred embodiment, the invention relates to a method for treating and/or preventing tumors by using a compound shown in a general formula I, a general formula II or a general formula III or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof or the pharmaceutical composition disclosed by the invention, and application of the compound in preparing a medicament for treating and/or preventing tumors, wherein the tumors are mediated by FGFR 4. In a preferred embodiment, a method for treating and/or preventing tumor and application of the compound with the general formula I, the general formula II or the general formula III or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof or the pharmaceutical composition of the invention in preparing a medicament for treating and/or preventing tumor are provided, wherein the tumor is selected from breast cancer, ovarian cancer, lung cancer, liver cancer and sarcoma. In a specific embodiment, the liver cancer is hepatocellular carcinoma. The reduction of FGF19 levels can promote bile acid synthesis, and therefore compounds that reduce FGF19 levels are useful for the treatment of hyperlipidemia. In a specific embodiment, the invention provides a method for treating and/or preventing hyperlipidemia by using the compound shown in the general formula I, the general formula II or the general formula III or pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof or the pharmaceutical composition of the invention, and application of the compound in preparing a medicament for treating and/or preventing hyperlipidemia.

Description of the terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

"isomers" in the present invention include cis-trans isomers in either the cis or trans configuration, as well as enantiomers and diastereomers arising from chiral carbons.

The "pharmaceutically acceptable salt" of the present invention refers to a pharmaceutically acceptable salt of the compound of the present invention with an acid, which may be selected from: phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, citric acid, maleic acid, malonic acid, mandelic acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-toluenesulfonic acid, malic acid, methanesulfonic acid, and the like.

"solvate" of the present invention refers to a form of the compound of the present invention which forms a solid or liquid complex by coordination with a solvent molecule. Hydrates are a special form of solvates in which coordination occurs with water. Within the scope of the present invention, the solvate is preferably a hydrate.

The term "crystalline" as used herein refers to the various solid forms formed by the compounds of the present invention, including crystalline forms and amorphous forms.

The "prodrug" of the present invention refers to a compound which is converted into the compound of the present invention by reacting with an enzyme, gastric acid or the like under physiological conditions of an organism, that is, a compound which is converted into the compound of the present invention by oxidation, reduction, hydrolysis or the like by an enzyme and/or a compound which is converted into the compound of the present invention by hydrolysis or the like by gastric acid or the like.

The "pharmaceutical composition" of the present invention is intended to include a mixture of any one of the compounds described herein, including isomers, prodrugs, solvates, pharmaceutically acceptable salts, or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers.

The "warhead" of the invention is the moiety that forms a covalent bond with the nucleophile between the warhead of the inhibitor and the cysteine residue of FGFR 4. The warhead includes, but is not limited to, alkyl halides, alkyl sulfonates, heteroaryl halides, epoxides, haloacetamides, maleimides, sulfonates, α - β unsaturated ketones, α - β unsaturated esters, vinyl sulfones, propargyl acyl, acryloyl. The structure of an exemplary warhead is as follows:

wherein Xa represents a leaving group which may be halogen or an activated hydroxy moiety (e.g. triflate); r_a、R_b、R_cEach independently selected from H, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted cycloalkyl, cyano, and the like, wherein the substituents are selected from one or more of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, nitro, cyano, hydroxy, amino, carboxy, aminoalkyl, hydroxyalkyl, alkoxyalkyl, alkylacylalkyl.

"cycloalkyl" in the context of the present invention means a saturated or partially unsaturated mono-or polycyclic substituent comprising from 3 to 20 carbon atoms, preferably from 4 to 13 carbon atoms. Non-limiting examples of monocycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like, with cyclopentyl, cyclohexyl being preferred. Polycyclic alkyl groups include spiro, fused and bridged cycloalkyl groups.

"spirocycloalkyl" as used herein refers to a 5-20 membered polycyclic group having a single ring with a common carbon atom (referred to as a spiro atom) between the rings, which spirocycloalkyl group may contain one or more double bonds, preferably 5-14 membered, more preferably 7-10 membered. Spirocycloalkyl groups are classified into mono-spirocycloalkyl groups, di-spirocycloalkyl groups or multi-spirocycloalkyl groups, preferably mono-spirocycloalkyl groups or di-spiroalkyl groups, according to the number of spiro atoms shared between rings. More preferred is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/6-membered single spirocycloalkyl group. Non-limiting examples of spiro rings include

"fused cycloalkyl" as used herein refers to a 5-20 membered multicyclic alkyl group in which each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system, wherein one or more of the rings may contain one or more double bonds, preferably 6-14 membered, more preferably 7-10 membered, and may be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicycloalkyl groups, depending on the number of constituent rings. Non-limiting examples of fused ring alkyl groups are:

"bridged cycloalkyl" in the context of the present invention refers to an all-carbon polycyclic group of 5 to 20 members, any two rings sharing two carbon atoms not directly attached, which may contain one or more double bonds. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic cycloalkyl groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl radicals include

The cycloalkyl groups of the present invention may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy.

"Heterocyclyl" in accordance with the present invention means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, wherein one or more ring atoms are selected from the group consisting of N, O and S (O)_e(wherein e is an integer of 0 to 2) and the remaining ring atoms are carbon. Wherein heterocyclyl preferably includes 3 to 12 ring atoms containing 1 to 4 heteroatoms; more preferably, heterocyclyl contains 5 to 11 ring atoms of which 1 to 2 are heteroatoms.

"Heterocyclyl" as used herein refers to a saturated or unsaturated, non-aromatic, monocyclic, fused, spiro or bridged ring containing 3 to 20 carbon atoms and containing heteroatoms such as N, O or S in place of one or more C atoms. Examples of heterocyclyl groups include tetrahydrofuranyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, indolinyl, isoindolinyl, morpholinyl, thiomorpholinyl, homomorpholinyl, homopiperidinyl, homopiperazinyl, homothiomorpholinyl, thiomorpholinyl-S-oxide, thiomorpholinyl-S, S-dioxide, pyranyl, tetrahydropyranyl, tetrahydrothienyl, homothiomorpholinyl-S, S-dioxide, oxazolidinyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidyl, dihydrofuranyl, dihydropyranyl, tetrahydrothienyl-S-oxide, tetrahydrothienyl-S, S-dioxide, homothiomorpholinyl-S-oxide, pyrazolinyl-S-oxide, pyrazolidinyl, pyrazolinyl-S-oxide, thiomorpholinyl-S-oxide, and the like, 2-oxa-5-azabicyclo [2.2.1] heptane, 8-oxa-3-azabicyclo [3.2.1] octane, 3, 8-diazabicyclo [3.2.1] octane, 2, 5-diazabicyclo [2.2.1] heptane, 3, 8-diazabicyclo [3.2.1] octane, 3, 9-diazabicyclo [4.2.1] nonane and 2, 6-diazabicyclo [3.2.2] nonane.

"spiroheterocyclyl" means a 5-20 membered polycyclic heterocyclic group having a single atom (the spiro atom) between monocyclic rings, wherein one or more of the ring atoms is selected from the group consisting of N, O and S (O)_e(wherein e is an integer from 0 to 2) and the remaining ring atoms are carbon, the spiroheterocyclyl group may contain one or more double bonds, preferably 6-14, more preferably 7-12. When a spiro heterocyclic group contains one or more nitrogen atoms, it is simply referred to as "nitrogen spiro heterocyclic group". The spirocycloalkyl group is classified into a single spiroheterocyclic group, a double spiroheterocyclic group or a multiple spiroheterocyclic group, preferably a single spiroheterocyclic group or a double spiroheterocyclic group, according to the number of spiro atoms shared between rings. More preferred are 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/6-membered mono spiroheterocyclic groups. Non-limiting examples of spiroheterocyclic groups include

"fused heterocyclyl" means a 5-to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, and in which one or more of the rings may contain one or more double bonds, and in which one or more of the ring atoms is selected from N, O or S (O)_e(wherein e is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 12. When a fused heterocyclic group containing one or more nitrogen atoms is used, the term "nitrogen fused heterocyclic group" is used for short. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicycloalkyl groups. Non-limiting examples of fused ring heterocyclic groups include

"bridged heterocyclyl" according to this invention means 5-14 membered polycyclic heterocyclic groups in which any two rings share two atoms not directly attached, which may contain one or more double bonds, one or more of which are further atoms selected from N, O or S (O)_e(wherein e is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 12. When a bridged heterocyclic group containing one or more nitrogen atoms is used, it is simply referred to as "nitrogen-bridged heterocyclic group". According to the number of constituent rings, which may be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged ring groups, preferably bicyclic, tricyclic or tetracyclic, non-limiting examples of bridged heterocyclic groups include

The heterocyclyl group may be substituted or unsubstituted and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy.

According to the invention

Refers to heterocyclic groups containing one or more nitrogen atoms, abbreviated as "nitrogen heterocyclic groups". The heterocyclyl radical may also contain one or more further heteroatoms, e.g. O or S (O)_e(wherein e is an integer of 0 to 2). When ring a is a mono-heterocyclic ring, it is referred to herein as a "nitrogen mono-heterocyclic group"; when ring a is a polyheterocycle, it is referred to herein as a "nitrogen polyheterocycle group"; in turn, when ring a is a single spiroheterocyclyl, it is referred to herein as a "nitrogen single spiroheterocyclyl".

The "aryl" of the present invention refers to an aromatic system which may comprise a single or multiple condensed rings such as bicyclic or tricyclic aromatic rings, wherein at least a portion of the condensed rings form a conjugated aromatic system containing 5 to 50 carbon atoms, preferably about 6 to about 14 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, anthracenyl, tetrahydronaphthyl, fluorenyl, indanyl, biphenylenyl, and acenaphthenyl.

Aryl groups may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy.

The term "heteroaryl" as used herein refers to an aromatic group having at least one carbon atom of an aromatic monocyclic or polycyclic ring such as bicyclic or tricyclic ring replaced by a heteroatom, said heteroatom being O, S, N. Suitable heteroaryl groups include, but are not limited to, imidazolyl, benzimidazolyl, imidazopyridinyl, quinazolinyl, pyrrolyl, imidazolonyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, and the like.

Heteroaryl groups may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy.

"halogen" in the context of the present invention means fluorine, chlorine, bromine or iodine.

"oxo" in the context of the present invention means O ═ e.g. formed by substitution of a carbon atom with an oxo group

The sulfur atom being substituted by an oxo group to form

The sulfur atom being substituted by two oxo groups

"alkyl" in the context of the present invention means a straight or branched chain saturated hydrocarbon radical, preferably C_1-8Alkyl, more preferably C_1-6An alkyl group. Non-limiting examples of alkyl groups include, but are not limited to, phenyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl.

The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, hydroxy, mercapto, cyano, nitro, azido, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylthio, aryl, aryloxy, arylthio, heteroaryl, heteroaryloxy.

"haloalkyl" in the context of the present invention means an alkyl group substituted with at least one halogen atom.

"alkoxy" in the context of the present invention means-O-alkyl.

"Haloalkoxy" of the invention "Means alkoxy substituted by at least one halogen, preferably C substituted by at least one halogen_1-6Alkoxy, more preferably C substituted by at least one halogen_1-3Alkoxy, suitably halo C_1-3Alkoxy is chloromethoxy, fluoromethoxy, dichloromethoxy, difluoromethoxy, trichloromethoxy or trifluoromethoxy; dichloroethoxy, difluoroethoxy, trichloroethoxy, trifluoroethoxy.

The "nitro group" in the present invention means-NO₂。

The "cyano group" of the present invention means-CN.

The "hydroxyl group" in the present invention means-OH.

"hydroxyalkyl" in the context of the present invention means OH-alkyl-.

"Alkylacylalkyl" according to the invention means alkyl-C (O) -alkyl-

"amino" in the context of the present invention means-NH₂-NH- (alkyl) or-N (alkyl).

"aminoalkyl" according to the invention means NH₂-alkyl-, (alkyl) NH-alkyl-, or (alkyl) N-alkyl-.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl" means that an alkyl may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl and the heterocyclic group is not substituted with an alkyl.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.

"Hydrogen" and "carbon" packages in the compounds of the inventionIncluding all isotopes thereof. Isotopes are understood to include atoms having the same number of atoms but different mass numbers, e.g. isotopes of hydrogen including tritium and deuterium, and isotopes of carbon including¹³C and¹⁴c, isotopes of oxygen including¹⁸O。

Detailed Description

The present invention will be further described and explained with reference to the following examples, which are not intended to limit the scope of the present invention. The materials used in the following examples are all commercially available unless otherwise specified.

EXAMPLE 1N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) acrylamide

Step 12, 4-dichloro-3-isocyanato-1, 5-dimethoxybenzene preparation

In a 500mL single-necked flask, 1, 4-dioxane (200mL) was added, followed by the sequential addition of 2,6-dichloro-3, 5-dimethoxyaniline (11g), triphosgene (8.86g) and triethylamine (4.02g), and the reaction was carried out at 130 ℃ for 2 hours. TLC shows that the reaction is finished, the solvent is removed by rotary evaporation to obtain a crude product, and the crude product is separated and purified by column chromatography to obtain the title compound.¹H-NMR(400MHz,DMSO-d₆):δ3.82(s,6H),6.79(s,1H).ESI-MS m/z:[M+H]⁺＝248.0.

Step preparation of 21- (6-Chloropyrimidin-4-yl) -3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylurea

6-chloro-N-methylpyrimidin-4-amine (4g) was dissolved in 25mL of N, N-dimethylformamide and hydrogen was added with stirring at-5 deg.CSodium (2g) is dissolved, stirring is continued for 0.5h, then 2, 4-dichloro-3-isocyanato-1, 5-dimethoxybenzene (9g) is added, the mixture is stirred for 1h at room temperature, LC-MS monitors that the reaction is finished, and the reaction solution is directly put into the next step. ESI-MS M/z [ M + H ]]⁺＝391.0.

Step preparation of 31- (6-Chloropyrimidin-4-yl) -3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) urea

Slowly adding 2- (trimethylsilyl) ethoxymethyl chloride (10mL) into the reaction liquid in the step 2, continuously stirring at room temperature for 1h, monitoring the reaction completion by LC-MS, filtering to remove a white solid, adding a saturated ammonium chloride solution into the filtrate for quenching, extracting by dichloromethane, concentrating, and carrying out column chromatography separation and purification to obtain the title compound. ESI-MS M/z [ M + H ]]⁺＝521.1.

Step 4 preparation of bis (tert-butyloxycarbonyl) -4-bromo-2-nitroaniline

In a 500mL single-necked flask, methylene chloride (200mL) was added, followed by addition of 4-bromo-2-nitroaniline (25g), di-tert-butyl dicarbonate (50g), triethylamine (11.66g) and 4-dimethylaminopyridine (7.04g) in that order, and stirring was carried out at room temperature for 1 h. TLC shows that the reaction is finished, suction filtration and drying are carried out to obtain a crude product, and the crude product is separated and purified by column chromatography to obtain the title compound.¹H-NMR(400MHz,DMSO-d₆):δ1.41(s,18H), 7.22(d,1H),7.77(dd,1H),8.21(d,1H).

Step 5 preparation of bis (tert-butyloxycarbonyl) -2-nitro-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) aniline

Bis (tert-butyloxycarbonyl) -4-bromo-2-nitroaniline (2.5g), 8-oxa-2-aza-amineSpiro [4.5]]Decane hydrochloride (2g), tris (dibenzylideneacetone) dipalladium (0.72g), 2-dicyclohexylphosphorus-2 ',6' -diisopropoxy-1, 1' -biphenyl (0.734g) and cesium carbonate (7.68g) were added in this order to 1, 4-dioxane (50mL), and reacted at 100 ℃ for 18 hours under nitrogen protection, the reaction solution was filtered, the filtrate was concentrated, and the title compound was obtained by column chromatography separation and purification. ESI-MS M/z [ M + H ]]⁺＝478.3.

Step preparation of 62-Nitro-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) aniline

The bis (tert-butyloxycarbonyl) -2-nitro-4- (8-oxa-2-azaspiro [4.5] prepared in the step 5]Decan-2-yl) aniline (1.8g) was dissolved in dichloromethane (30mL), 40 times the molar amount of trifluoroacetic acid was added, the mixture was stirred at room temperature for 2 hours, after completion of the reaction, saturated sodium bicarbonate was added to quench, and the mixture was extracted with dichloromethane and concentrated to give the title compound. ESI-MS M/z [ M + H ]]⁺＝278.1.

Step preparation of 71- (2,6-dichloro-3,5-dimethoxyphenyl) -3-methyl-3- (6- ((2-nitro-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) amino) pyrimidin-4-yl) -1- ((2- (trimethylsilyl) ethoxy) methyl) urea

2-nitro-4- (8-oxa-2-azaspiro [4.5]]Decan-2-yl) aniline (1.6g), 1- (6-chloropyrimidin-4-yl) -3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) urea (2.7g), tris (dibenzylideneacetone) dipalladium (0.474g), 2-dicyclohexylphosphonium-2 ',6' -diisopropoxy-1, 1' -biphenyl (0.483g), cesium carbonate (5.06g), successively added to 1, 4-dioxane (50mL), reacted at 100 ℃ for 18 hours under nitrogen protection, the reaction solution was filtered by column chromatography, the filtrate was concentrated, and purified by separation to give the title compound. ESI-MS M/z [ M + H ]]⁺＝762.2.

Step 8 preparation of tert-butyl (6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) ureido) pyrimidin-4-yl) (2-nitro-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) carbamate

1- (2,6-dichloro-3,5-dimethoxyphenyl) -3-methyl-3- (6- ((2-nitro-4- (8-oxa-2-azaspiro [4.5 ]) prepared in the step 7]Decan-2-yl) phenyl) amino) pyrimidin-4-yl) -1- ((2- (trimethylsilyl) ethoxy) methyl) urea (1.8g) was dissolved in tetrahydrofuran (70mL), a catalytic amount of 4-dimethylaminopyridine and di-tert-butyl dicarbonate (0.644g) were added, and the reaction was carried out at 80 ℃ for 2 hours, followed by water washing, extraction with dichloromethane, concentration, and purification by column chromatography to give the title compound. ESI-MS M/z [ M + H ]]⁺＝862.6.

Step 9 preparation of tert-butyl (2-amino-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) (6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) ureido) pyrimidin-4-yl) carbamate

1- (6- ((4- ((1R,4R) -2-oxa-5-azabicyclo [ 2.2.1) prepared in the step 8]Dissolving 1g of hept-5-yl) -2-nitrophenyl) amino) pyrimidin-4-yl) -3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylurea in 10mL of anhydrous tetrahydrofuran and 10mL of methanol, adding 2-3 mL of Raney-Ni aqueous solution under stirring at room temperature, introducing hydrogen, stirring for 3 hours, filtering after the reaction is finished, washing the filtrate with water, extracting with dichloromethane, concentrating, and purifying by column chromatography to obtain the title compound. ESI-MS M/z [ M + H ]]⁺＝832.3.

Step 10 preparation of tert-butyl (2-acrylamido-4- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) (6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) ureido) pyrimidin-4-yl) carbamate

The (2-amino-4- (8-oxa-2-azaspiro [4.5 ]) prepared in the step 9]Decan-2-yl) phenyl) (6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) ureido) pyrimidin-4-yl) carbamic acid tert-butyl ester (200mg) was dissolved in dichloromethane (30mL), N-diisopropylethylamine (125mg) was added, acryloyl chloride (23uL) was added at 10 ℃ and stirring was continued for 30min, after the reaction was complete, saturated sodium bicarbonate was quenched, extracted with dichloromethane, concentrated and purified by column chromatography to give the title compound. ESI-MS M/z [ M + H ]]⁺＝886.7.

Step 11 preparation of N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- (8-oxa-2-azaspiro [4.5] decan-2-yl) phenyl) acrylamide

The (2-acrylamido-4- (8-oxa-2-azaspiro [4.5] obtained in the step 10]Decan-2-yl) phenyl) (6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methyl-3- ((2- (trimethylsilyl) ethoxy) methyl) ureido) pyrimidin-4-yl) carbamic acid (200mg) was dissolved in dichloromethane (30mL), 40 equivalents of trifluoroacetic acid was added, stirring was carried out at room temperature for 30min, after the reaction was completed, quenching was carried out with saturated sodium bicarbonate, extraction was carried out with dichloromethane, concentration and purification was carried out by column chromatography to give the title compound.¹H-NMR(400MHz,DMSO-d₆):δ1.54-1.58(m, 4H),1.90(t,2H),3.16(s,2H),3.20(s,3H),3.31(t,2H),3.59-3.64(m,4H),3.94(s,6H),5.7(dd,1H), 6.10(brs,1H),6.24(dd,1H),6.40-6.53(m,2H),6.89(s,1H),6.93(s,1H),7.20(d,1H),8.30(s,1H),8.61(s, 1H),9.57(s,1H),12.14(s,1H).ESI-MS m/z:[M+H]⁺＝656.4.

Example 2N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- ((1R,4R) -5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) phenyl) acrylamide

The preparation method is the same as that of example1, except that the starting material 8-oxa-2-azaspiro [4.5]]Replacement of decane hydrochloride by (1R,4R) -2-methyl-2, 5-diazabicyclo [2.2.1]Heptane dihydrochloride to yield the title compound.¹H-NMR(400MHz,DMSO-d₆): δ1.87(d,1H),1.89(d,1H),2.28(s,3H),2.79(d,1H),3.15(d,1H),3.22(s,3H),3.31(s,2H),3.44(s,1H), 3.94(s,6H),4.24(s,1H),5.70(d,1H),6.10(brs,1H),6.24(d,1H),6.44-6.53(m,2H),6.90(s,1H),6.96(s, 1H),7.20(d,1H),8.31(s,1H),8.61(s,1H),9.57(s,1H),12.14(s,1H).ESI-MS m/z:[M+H]⁺＝627.2.

Example 3N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- ((1S,4S) -5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) phenyl) acrylamide

The preparation method is the same as that of example1, except that the starting material 8-oxa-2-azaspiro [4.5]]Replacement of decane hydrochloride by (1S,4S) -2-methyl-2, 5-diazabicyclo [2.2.1]Heptane dihydrobromide salt produced the title compound.¹H-NMR(400MHz,

DMSO-d₆):δ1.78(d,1H),1.88(d,1H),2.28(s,3H),2.79(d,1H),3.15(d,1H),3.21(s,3H),3.30(s,2H), 3.44(s,1H),3.94(s,6H),4.24(s,1H),5.70(d,1H),6.14(brs,1H),6.23(d,1H),6.46-6.52(m,2H),6.90(s, 1H),6.96(s,1H),7.20(d,1H),8.31(s,1H),8.61(s,1H),9.56(s,1H),12.13(s,1H).ESI-MS m/z:[M+H]⁺＝ 627.2.

Example 4N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- (4-morpholinylpiperidin-1-yl) phenyl) acrylamide

The preparation method is the same as that of example1, except that the starting material 8-oxa-2-azaspiro [4.5]]Replacement of decane hydrochloride with 4- (piperidin-4-yl) morpholine gave the title compound.¹H-NMR(400MHz,DMSO-d₆):δ1.48(q,2H),1.87(q,2H), 2.24-2.34(m,1H),2.69(t,2H),3.23(s,3H),3.31-3.33(m,4H),3.58(s,4H),3.67-3.70(m,2H),3.94(s,6H), 5.71(d,1H),6.21(s,1H),6.25(s,1H),6.46-6.55(m,1H),6.82(d,1H),6.90(s,1H),7.26-7.32(m,2H), 8.32(s,1H),8.72(s,1H),9.58(s,1H),12.08(s,1H).ESI-MS m/z:[M+H]⁺＝685.3.

Example 5N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- (4- (4-ethylpiperazin-1-yl) piperidin-1-yl) phenyl) acrylamide

The preparation method is the same as that of example1, except that the starting material 8-oxa-2-azaspiro [4.5]]Replacement of decane hydrochloride with 1-ethyl-4- (piperidin-4-yl) piperazine hydrochloride gave the title compound.¹H-NMR(400MHz,DMSO-d₆):δ0.98(t,3H), 1.45-1.54(m,2H),1.84-1.87(m,2H),2.26-2.50(m,10H),2.68(t,3H),3.23(s,3H),3.67-3.70(m,2H), 3.94(s,6H),5.72(d,1H),6.21(s,1H),6.26(s,1H),6.46-6.55(m,1H),6.82(d,1H),6.90(s,1H),7.27(d,1H), 7.31(s,1H),8.32(s,1H),8.74(s,1H),9.59(s,1H),12.10(s,1H).ESI-MS m/z:[M+H]⁺＝712.3.

Example 6N- (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) -5- (2-oxa-6-azaspiro [3.4] octan-6-yl) phenyl) acrylamide

The preparation was carried out in the same manner as in example1 except that the starting material, 8-oxa-2-azaspiro was used[4.5]Replacement of decane hydrochloride by 2-oxa-6-azaspiro [3.4]]Octane hemioxalate gave the title compound.¹H-NMR(400MHz,DMSO-d₆):δ2.27(t,2H), 3.20(s,3H),3.26(t,2H),3.50(s,2H),3.94(s,6H),4.53(d,2H),4.60(d,2H),5.72(d,1H),6.10(brs,1H), 6.24(d,1H),6.41(d,1H),6.46-6.53(m,1H),6.90(s,1H),6.96(s,1H),7.20(d,1H),8.30(s,1H),8.64(s, 1H),9.57(s,1H),12.13(s,1H).ESI-MS m/z:[M+H]⁺＝628.2.

Example 7N-5- ((1S,4S) -2-oxa-5-azabicyclo [2.2.1] heptan-5-yl) - (2- ((6- (3- (2,6-dichloro-3,5-dimethoxyphenyl) -1-methylureido) pyrimidin-4-yl) amino) phenyl) acrylamide

The preparation method is the same as that of example1, except that the starting material 8-oxa-2-azaspiro [4.5]]Replacement of decane hydrochloride by (1S,4S) -2-oxa-5-azabicyclo [2.2.1]Heptane hydrochloride gave the title compound.¹H-NMR(400MHz,DMSO-d₆):δ1.85(d,1H),1.95(d,1H),2.97(d,1H),3.22(s,3H),3.50(d,1H),3.70(d,1H),3.76(d,1H),3.93(s,6H), 4.50(s,1H),4.62(s,1H),5.71(d,1H),6.15(brs,1H),6.24(d,1H),6.46-6.53(m,2H),6.89(s,1H),6.99(s, 1H),7.23(d,1H),8.31(s,1H),8.64(s,1H),9.59(s,1H),12.14(s,1H).ESI-MS m/z:[M+H]⁺＝614.4.

Examples 8-17 were synthesized according to the synthesis method of examples 1-7 of the present invention using different starting materials, and the characterization parameters of examples 8-17 are shown in table 1:

table 1:

comparative example1

The Compound name N- ((3S,4S) -3- ((6- (2,6-dichloro-3, 5-dichlorphenyl) quinazolin-2-yl) amino) tetrahydrogen-2H-pyran-4-yl) acrylamide (BLU-554) was prepared according to the synthetic method of Compound 40 in WO2015061572 and identified by hydrogen spectroscopy and mass spectrometry.

Comparative example 2

Name of compound

N- (2- ((6- (3- (2, 6-dichoro-3, 5-dimethoxyphenyl) -1-methyllureido) pyrimidin-4-yl) amino) -5- (4-ethylpipera zin-1-yl) phenyl) acrylamide (H3B-6527) was prepared according to the synthesis method of the sample 108 compound in WO2015057938 and identified by hydrogen spectroscopy and mass spectrometry.

The inhibitory activity of compound a and compound B against FGFR4 kinase and FGFR1 kinase, and the inhibitory activity against human hepatoma cell line Hep3B cells were tested using the methods of experimental example1 and experimental example 2 below. The experimental results show that the selectivity of the compound A and the compound B on FGFR4 kinase and FGFR1 kinase is obviously lower than that of the compound of the invention, and the inhibitory activity on human liver cancer cell strain Hep3B cells is obviously weaker than that of the compound of the invention.

Experimental example1 in vitro kinase Activity evaluation

1 test materials

FGFR1, available from Carna under catalog No. 08-133;

FGFR4, available from Carna under catalog No. 08-136;

p22 peptide, available from GL Biochem under catalog number 112393;

staurosporine9, available from Sigma under catalog number S4400-1 MG;

2 method of experiment

1) Preparation of 1 Xkinase base buffer and stop buffer

A.1 × kinase base buffer: 20mM HEPES、pH 7.5，0.01％Triton X-100，10mM MgCl₂，2mM DTT。

B. Stop buffer: 100mM HEPES, pH7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50mM EDTA.

2) Preparation of the Compounds

A. 10mM stock solutions of the compounds of the examples of the invention and of the reference compounds A and B were prepared.

B. Preparation of 50 × compound solution: the compounds of the above inventive examples and the reference compounds were diluted sequentially 3-fold in DMSO starting from 500 μ M for 10 concentrations.

C. 100 μ L of 100% DMSO was added to each of 2 empty wells of the same 96-well plate as a no-compound and no-kinase control. Label the 96-well plate as the source plate.

D. Preparing a middle plate: transfer 10 μ L of compound from source plate to a new 96-well plate as an intermediate plate; add 90. mu.L of 1 Xkinase buffer to each well of the intermediate plate; shaking and mixing for 10 min.

3) Preparing an experimental plate: transfer 5 μ L per well from 96 well intermediate plates to 384 well plates, 2 wells.

4) Kinase reaction

A. Preparation of 2.5 × kinase solution: the kinases were added separately to 1 × base buffer.

B. Preparation of 2.5 × polypeptide solution: FAM-labeled polypeptide and ATP were added to 1 × base buffer.

C. The assay plates already contained 5. mu.L of compound (10% DMSO).

D. Transfer 2.5 × kinase solution to assay plate: add 10. mu.L of 2.5 Xkinase solution to each well of a 384 well assay plate.

E. Incubate at room temperature for 10 min.

F. Transfer 2.5 × polypeptide solution to assay plate: add 10. mu.L of 2.5 Xpolypeptide solution to each well of a 384 well assay plate.

G. Kinase reaction and termination: incubating at 28 ℃ for a certain period of time; the reaction was stopped by adding 25. mu.L of stop buffer.

5) Caliper instrument reading: data were read on the Caliper instrument.

6) Fitting curve

A. Translation value data is obtained from the Caliper program.

B. The conversion value was converted into the inhibition rate.

Inhibition [% ], [% inhibition [% ], [% inhibition [% ] represents the DMSO vehicle control with or without kinase compound, and "[% ] represents the control without kinase compound.

C. Computing IC by Xlfit excel add-in 4.3.1 data processing software₅₀Values were obtained and the selectivity (IC for FGFR1 kinase) for each compound was calculated₅₀(nM) value/IC against FGFR4 kinase₅₀(nM) values), the results are shown in tables 2-3:

TABLE 2

TABLE 3

Experimental results show that the compound has strong inhibitory activity on FGFR4 kinase, low inhibitory activity on FGFR1 kinase and obviously better selectivity on FGFR4 kinase and FGFR1 kinase than a reference compound. Therefore, the compounds of the invention are active and advantageous kinase inhibitors selective for FGFR4, useful for the treatment of FGFR4 kinase-associated diseases and possibly reducing side effects caused by inhibition of FGFR1 kinase.

Experimental example 2 cell proliferation inhibition experiment

1 materials of the experiment

1.1 Compounds: this experiment was carried out using the compound of the present invention example, compound B.

1.2 cells: hep3B cells, provided by Shanghai Mingkude New drug development Co., Ltd.

1.3 reagent: FBS and DMEM, and the culture medium is purchased from GIBCO company;

CellTiter Glo, available from Promega corporation.

1.4 Instrument Tecan D300e quick pipettor; biotek fluorescence detector

2 method of experiment

All compounds were dissolved in DMSO and stored in a-20 ℃ freezer.

Day-1: according to 3X 10³Density of individual cells/well cells were added to 96 wells, 100 μ l per well;

blank control was added to 100. mu.l of medium per well;

the remaining untested edge wells were loaded with 100 μ l PBS;

day 0: setting a compound loading program, and automatically loading by using Tecan D300 e;

the initial concentration of test compound was 10 μ M, 3-fold dilution, 9 concentration, duplicate wells;

the initial concentration of positive reference Staurosporine is 1 mu M, 3 times of dilution, 9 concentrations and multiple wells;

day 3. equilibrate experimental 96-well plates for 30min at room temperature;

compound solubility and cell status were observed prior to addition of CTG;

add 50. mu.l CellTiter Glo reagent to each well and detect the signal 10 minutes later with BioTec (luminescence).

3, data analysis:

XL-fit software analysis data (supplier: ID Business Solution Ltd., version: XL fit 5.0)

Calculating the formula: r (%) {1-RLU_{Compound (I)}-RLU_{Blank space}}/{RLU_Control-RLU_{Blank space}} × 100%, the results are shown in Table 4:

TABLE 4

The experimental result shows that the compound has very good inhibitory activity on proliferation of human hepatoma cell line Hep3B cells amplified by FGFR4DNA, the activity of part of the compound is far superior to that of a reference compound, and the compound is an effective FGFR4 selective inhibitor.

Although the present invention has been described in detail above, those skilled in the art will appreciate that various modifications and changes can be made to the present invention without departing from the spirit and scope of the invention. The scope of the invention is not to be limited by the above detailed description but is only limited by the claims.

Claims (7)

1. A compound shown in a general formula I or a pharmaceutically acceptable salt thereof,

wherein: the warhead is selected from the following groups:

wherein R is_a、R_b、R_cEach independently selected from H, C_1-6Alkyl radical, C_3-8Cycloalkyl, cyano, wherein the alkyl or cycloalkyl is optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl, alkoxy C_1-6Alkyl radical, C_1-6Alkyl acyl radical C_1-6Radical substitution of alkyl;

ring A

Is selected from the group

Wherein said radicals are optionally substituted by one or more groups selected from halogen, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl acyl radical, C_1-6Alkyl acyl radical C_1-6Alkyl, 6-12 membered aryl, 5-12 membered heteroaryl, C_3-8Cycloalkyl and 3-8 membered heterocyclyl;

each R₁、R₂、R₃Each independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethyl, trifluoroethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, methoxy, ethoxy, propoxy, isopropoxy, amino, methylamino, dimethylamino, ethylamino, diethylamino, methylethylamino, methylacylamino, ethylacylamino, vinylacylamino, methylacyl, ethylacoyl, vinylacyl, aminoacyl, methylaminoacyl, ethylaminoacyl, carboxyl, nitro, cyano, phenyl and azetidinyl, wherein n, p, q are each independently selected from the group consisting of 1,2, 3, 4 and 5; and

R₄and R₅Each independently selected from H and C_1-6An alkyl group.

2. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R_a、R_b、R_cEach independently selected from H, C_1-3Alkyl radical, C_3-6Cycloalkyl, cyano, wherein the alkyl or cycloalkyl is optionally substituted by one or more groups selected from halogen, C_1-3Alkyl, halo C_1-3Alkyl radical, C_1-3Alkoxy, halo C_1-3Alkoxy, nitro, cyano, hydroxy, amino, carboxyl, amino C_1-3Alkyl, hydroxy C_1-3Alkyl, alkoxy C_1-3Alkyl radical, C_1-3Alkyl acyl radical C_1-3Alkyl groups.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

4. a pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof or a composition according to claim 4 in the manufacture of a medicament for the prophylaxis and/or treatment of an anti-tumour agent.

6. Use according to claim 5, wherein the tumour is selected from the group consisting of liver cancer, breast cancer, ovarian cancer, lung cancer and sarcoma.

7. The use according to claim 5, wherein the tumour is mediated by FGFR4 kinase.