CN114853739A - Alkynyl pyrazine FGFR inhibitor and preparation method and application thereof - Google Patents

Abstract

The invention discloses an alkynylpyrazine compound as an FGFR inhibitor, a preparation method and medical application thereof. The invention relates to a compound shown in a general formula (I) and a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound and/or the pharmaceutically acceptable salt thereof, and application of the compound or the pharmaceutically acceptable salt thereof in medicaments for treating or preventing FGFR kinase related diseases, particularly tumors, and discloses a preparation method of the pharmaceutical composition containing the compound or the pharmaceutically acceptable salt thereof. Wherein each of the general formula (I)The substituents are as defined in the specification.

Description

Alkynyl pyrazine FGFR inhibitor and preparation method and application thereof

The technical field is as follows:

the invention relates to an alkynylpyrazine compound or pharmaceutically acceptable salt thereof used as an FGFR inhibitor; a pharmaceutical composition containing the alkynylpyrazine compound or pharmaceutically acceptable salt thereof; a preparation method of the alkynylpyrazine compound or the pharmaceutically acceptable salt thereof; the application of the alkynopyrazine compound or the pharmaceutically acceptable salt thereof or the pharmaceutical composition containing the alkynopyrazine compound or the pharmaceutically acceptable salt thereof in preparing a medicament for treating and/or preventing FGFR related diseases and tumors.

Background art:

fibroblast Growth Factor Receptors (FGFR) are a class of Receptor Tyrosine Kinases (RTKs), and the FGFR family mainly comprises four subtypes of FGFR1, FGFR2, FGFR3 and FGFR 4. FGFR1 is a transmembrane protein belonging to the receptor tyrosine kinase, consisting of three major components: namely an extracellular domain, a transmembrane domain and a cell domain, the extracellular domain is a binding domain of the ligand Fibroblast Growth Factors (FGFs). FGFs are also a polygenic family, and there are 19 members, namely FGF1 also known as acidic fibroblast growth factor (aFGF), FGF2 also known as basic fibroblast growth-related document factor (basic FGF, bFGF), which have biological activities of stimulating the growth of fibroblasts, vascular endothelial cells, smooth muscle cells and nerve cells, and FGFR1, which is a high affinity receptor. After FGF is combined with an extracellular segment of FGFR1, a tyrosine kinase active region of an inner segment of a receptor cell firstly generates autophosphorylation, then receptor target protein generates transphosphorylation, and a signal of a ligand is transmitted to a cell nucleus through a family structure and family relation of a protein cascade reaction, so that the effects of promoting damage repair, embryonic development and skeleton formation are achieved.

However, when FGFR is mutated or overexpressed, it causes excessive activation of the FGFR signaling pathway and further induces normal cell carcinogenesis. Wherein, over-activation of RAS-RAF-MAPK stimulates cell proliferation and differentiation; over-activation of PI3K-AKT results in inhibition of apoptosis; SATA is closely related to promoting tumor invasion and metastasis and enhancing tumor immune escape capacity; the PLC gamma signal channel is an important way for regulating and controlling the metastasis of tumor cells. Next Generation Sequencing (NGS) on 4853 solid tumor types showed, according to a study published in Clinical Cancer Research in 2015, FGFR aberrations (abortions) and abnormal activation were found in approximately 7.1% of cancers, mostly gene amplification (66%), followed by mutations (26%) and rearrangements (8%). FGFR distortion exists in almost all detected malignant tumors, and the cancers with high incidence rate include urothelial carcinoma, cholangiocarcinoma, breast cancer, endometrial carcinoma, squamous epithelial carcinoma and the like; meanwhile, the abnormal activation of FGFR is also found in tumors such as lung cancer, liver cancer, breast cancer and the like.

There are currently some non-FGFR specific drugs on the market, such as Sunitinib from pfizer, lentitini from Eisai, and nintedanib from Boehringer ingelheimer. Whereas the only FGFR inhibitors approved by the FDA to be marketed are balversa (erdafitinib) and pemazyre (pemigatinib). Small molecule inhibitors of FGFR1/2/3 entering the clinic are: infigrtinib (BGJ398) and AZD4547, fisogatinib (BLU-554), Roblitiniib (FGF401), H3B6527, lucitanib (E-3810), Futibatinib (TAS-120), RPN1371, ICP-192, derazatinib, 3D185, BPI-17509, HMPL-453.

Although the development of FGFR inhibitors has attracted the deployment of numerous companies both at home and abroad, and although 2 FGFR inhibitors are already on the market, there is still a need to develop new compounds due to the prospects they show in the treatment of various malignancies. Through continuous efforts, the invention designs an irreversible inhibitor which has proprietary intellectual property rights and shows excellent activity on FGFR-1-4 protein kinase.

Disclosure of Invention

The invention provides an alkyne heterocyclic derivative compound shown in a general formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the compound contains an alkyne incense ring and an alpha, beta-unsaturated carboxylic acid amide structure, can be used as an irreversible FGFR inhibitor,

wherein:

ring B is a phenyl ring or a 5-6 membered heteroaromatic ring wherein the above phenyl and heteroaromatic rings are optionally substituted by one or more G ¹ Substituted;

R ¹ ，R ² independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -OR ⁴ 、-NR ⁴ R ⁵ 、-C(O)NR ⁴ R ⁵ Wherein said alkyl, cycloalkyl OR heterocycloalkyl is optionally substituted by cyano, halogen, -OR ⁶ 、-NR ⁶ R ⁷ 、C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocycloalkyl;

x is independently selected from-C _0-4 Alkyl-, -CR ⁸ R ⁹ -、-C _1-2 Alkyl (R) ⁸ )(OH)-、-C(O)-、-CR ⁸ R ⁹ O-、-OCR ⁸ R ⁹ -、-SCR ⁸ R ⁹ -、-CR ⁸ R ⁹ S-、-NR ⁸ -、-NR ⁸ C(O)-、-C(O)NR ⁸ -、-NR ⁸ C(O)NR ⁹ -、-CF ₂ -、-O-、-S-、-S(O) _m -、-NR ⁸ S(O) ₂ -、-S(O) ₂ NR ⁸ -；

Y is absent or C is selected _3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spiro cyclic group, 5-12 membered spiro heterocyclic group, aromatic group or heteroaromatic group, wherein said cycloalkyl, heterocycloalkyl, spiro cyclic group, fused heterocyclic group, spiro heterocyclic group, aromatic group or heteroaromatic group is optionally substituted with one or more G ² Substituted;

z is independently selected from cyano, -NR ¹⁰ CN、

Bond a is a double or triple bond;

when a is a double bond, R ^a 、R ^b And R ^c Each independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl. Wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ³ Substituted;

R ^a and R ^b Or R ^b And R ^c Optionally taken together with the carbon atom to which they are attached to form a 3-6 membered ring optionally containing heteroatoms;

when bond a is a triple bond, R ^a And R ^c Is absent, R ^b Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl by one or more G ⁴ Substituted;

R ¹⁰ independently selected from H, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl, wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ⁵ Substituted;

G ¹ 、G ² 、G ³ 、G ⁴ and G ⁵ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹¹ 、-OC(O)NR ¹¹ R ¹² 、-C(O)OR ¹¹ 、-C(O)NR ¹¹ R ¹² 、-C(O)R ¹¹ 、-NR ¹¹ R ¹² 、-NR ¹¹ C(O)R ¹² 、-NR ¹¹ C(O)NR ¹² R ¹³ 、-S(O) _m R ¹¹ or-NR ¹¹ S(O) _m R ¹² Wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by 1 or more cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹⁴ 、-OC(O)NR ¹⁴ R ¹⁵ 、-C(O)OR ¹⁴ 、-C(O)NR ¹⁴ R ¹⁵ 、-C(O)R ¹⁴ 、-NR ¹⁴ R ¹⁵ 、-NR ¹⁴ C(O)R ¹⁵ 、-NR ¹⁴ C(O)NR ¹⁵ R ¹⁶ 、-S(O) _m R ¹⁴ or-NR ¹⁴ S(O) _m R ¹⁵ Substituted with the substituent(s);

R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ 、R ⁹ 、R ¹¹ 、R ¹² 、R ¹³ 、R ¹⁴ 、R ¹⁵ and R ¹⁶ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl; and m is 1 or 2.

Another embodiment of the present invention relates to compounds of formula I as described above or prodrugs, stable isotopic derivatives, pharmaceutically acceptable salts, polymorphs, solvates, isomers and mixtures thereof.

Exemplary embodiments utilizing the principles of the present invention are set forth in the following detailed description of the invention. The features and advantages of the present invention may be better understood by reference to the following summary.

The compounds of the invention are capable of effectively inhibiting the activity of FGFR1, FGFR2, FGFR3 or FGFR4, which inhibit the IC of FGFR1, FGFR2, FGFR3 or FGFR4 ₅₀ Is 100 to 1000nM, more preferably IC ₅₀ Less than 100nM, optimal IC ₅₀ Less than 10 nM.

The compounds of the invention are useful for the treatment or prevention of FGFR-associated tumors, such as non-small cell lung cancer, esophageal cancer, melanoma rhabdomyosarcoma, cellular carcinoma, multiple myeloma, breast cancer, ovarian cancer, endometrial cancer, cervical cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, lung cancer, prostate cancer, and liver cancer (e.g., hepatocellular carcinoma), more particularly liver cancer, gastric cancer, and bladder cancer. Thus, in a further aspect, the present invention provides a method of treating or preventing FGFR-mediated diseases (e.g. of a neoplasm), which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of the present invention or a prodrug, stable isotope derivative, polymorph, solvate, pharmaceutically acceptable salt, isomer and mixtures thereof, or a pharmaceutical composition comprising the compound.

Another aspect of the present invention relates to a compound of formula I or a prodrug, stable isotope derivative, polymorph, solvate, pharmaceutically acceptable salt, isomer, and mixture thereof for pharmaceutical or medicinal use, for treating or preventing FGFR mediated diseases, such as tumor or inflammatory diseases, including but not limited to non-small cell lung cancer, esophageal cancer, melanin, rhabdomyosarcoma, wild cell cancer, multiple myeloma, breast cancer, ovarian cancer, endometrial cancer, uterine cancer, gastric cancer, diaphragm cancer, bladder cancer, pancreatic cancer, lung cancer, prostate cancer.

The invention further relates to a pharmaceutical composition which comprises the compound or the prodrug, the stable isotope derivative, the pharmaceutically acceptable salt isomer and the mixture thereof, and pharmaceutically acceptable carriers, diluents and excipients.

Another aspect of the present invention relates to the use of the compounds represented by the general formula I or their prodrug stable isotope derivatives, pharmaceutically acceptable salts, isomers and mixtures thereof, or pharmaceutical compositions thereof for the preparation of medicaments for the treatment or prevention of FGFR mediated diseases such as tumors and inflammatory diseases.

According to the present invention, the drug may be in any pharmaceutical dosage form including, but not limited to, tablets, sachets, solutions, lyophilized formulations, injections.

Certain chemical terms

Unless stated to the contrary, the following terms are used in the specification and claims.

Has the following meanings and is used herein in the manner of _x-y "denotes the range of the number of carbon atoms, wherein x and y are each an integer, e.g. C _3-8 Cycloalkyl denotes cycloalkyl having 3 to 8 carbon atoms, i.e. cycloalkyl having 3,4, 5, 6, 7 or 8 carbon atoms. It is also understood that "C" is _3-8 "also includes any subrange therein, e.g. C _3-7 、C _3-6 、C _4-7 、C _4-6 、C _5-6 And the like.

"alkyl" refers to a straight or branched chain hydrocarbyl group containing 1 to 20 carbon atoms, for example 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples of alkyl groups include methyl, 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, and 2-ethylbutyl. The alkyl group may be substituted or unsubstituted.

"alkenyl" refers to a straight or branched chain hydrocarbyl group containing at least one carbon-carbon double bond and typically 2 to 20 carbon atoms, e.g., 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1, 4-pentadienyl, and 1, 4-butadienyl. The alkenyl group may be substituted or unsubstituted.

"alkynyl" refers to a straight or branched chain hydrocarbyl group containing at least one carbon-carbon triple bond and typically 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl. The alkynyl group may be substituted or unsubstituted.

"cycloalkyl" refers to a saturated cyclic hydrocarbyl substituent containing from 3 to 14 carbon ring atoms. Cycloalkyl groups may be monocyclic, typically containing from 3 to 7 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Cycloalkyl groups may alternatively be bi-or tricyclic fused together, such as decahydronaphthyl, which may be substituted or unsubstituted.

"Heterocyclyl", "heterocycloalkyl", "heterocycle" means a stable 3-to 18-membered monovalent non-aromatic ring comprising 2 to 12 carbon atoms, 1 to 6 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise specified, a heterocyclyl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may contain fused, spiro or bridged ring systems, the nitrogen, carbon or sulfur of the heterocyclyl group may optionally be oxidized, the nitrogen atom may optionally be quaternized, and the heterocyclyl group may be partially or fully saturated. The heterocyclic group may be attached to the rest of the molecule through a single bond via a carbon or heteroatom in the ring. The heterocyclic group containing fused rings may contain one or more aromatic or heteroaromatic rings, provided that the atoms on the non-aromatic ring are attached to the rest of the molecule. For purposes of this application, a heterocyclyl group is preferably a stable 4-11 membered monovalent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably a stable 4-8 membered monovalent non-aromatic monocyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclyl groups include azepanyl, azetidinyl, decahydroisoquinolinyl, dihydrofuranyl, indolinyl, dioxolanyl, 1-dioxo-thiomorpholinyl, imidazolidinyl, imidazolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinonyl, pyranyl, pyrazolidinyl, pyrrolidinyl, quinolizinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydropyranyl and the like.

"spiroheterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group which shares one atom (called the spiro atom) between single rings, wherein one or more ring atoms are selected from nitrogen, oxygen or a heteroatom of S (0) whose m is an integer of 0 to 2, and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings have a fully conjugated electronic system, preferably 6 to 14, more preferably 7 to 10. The spirocycloalkyl group is classified into a single spiroheterocyclyl group, a double spiroheterocyclyl group or a multiple spiroheterocyclyl group, preferably a single spirocycloalkyl group and a double spirocycloalkyl group, according to the number of spiro atoms shared between rings. More preferably a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monocyclic group. Non-limiting examples of spirocycloalkyl 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, one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system in which one or more ring atoms are selected from nitrogen, oxygen or S (O) _m (wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

"aryl" or "aryl" refers to an aromatic ring or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 members, such as phenyl and naphthyl, most preferably the aryl ring of the phenyl group may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the rings attached to the parent structure are aryl rings, non-limiting examples of which include:

"heteroaryl" or "heteroaryl" refers to a 5-16 membered ring system containing 1-15 carbon atoms, preferably 1-10 carbon atoms, 1-4 heteroatoms selected from nitrogen, oxygen and sulfur, at least one aromatic ring. Unless otherwise specified, heteroaryl groups may be monocyclic, bicyclic, tricyclic or tetracyclic ring systems, which may contain fused or bridged ring systems, provided that the point of attachment to the rest of the molecule is an aromatic ring atom, which may be selectively oxidized at nitrogen, carbon and sulfur atoms, and which may optionally be quaternized. For the purposes of the present invention, heteroaryl groups are preferably stable 4-11 membered monocyclic aromatic rings containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably stable 5-8 membered monocyclic aromatic rings containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heteroaryl groups include acridinyl, azepinyl, benzimidazolyl, benzindolyl, benzodioxinyl, benzodioxolyl, benzofuranonyl, benzofuranyl, benzonaphthofuranyl, benzopyranonyl, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazolyl, furanyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quininyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, and the like. In the present application, heteroaryl is preferably 5-8 membered heteroaryl comprising 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably pyridyl, pyrimidinyl, thiazolyl. The heteroaryl group may be substituted or unsubstituted.

"halogen" means fluorine, chlorine, bromine or iodine.

"hydroxy" means-OH, and "amino" means-NH ₂ "amido" means-NHCO-, "cyano" means-CN, and "nitro" means-NO ₂ "isocyano" means-NC and "trifluoromethyl" means-CF ₃ 。

The term "heteroatom" or "hetero", as used herein alone or as part of another ingredient, refers to atoms other than carbon and hydrogen, and is independently selected from, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin, and in embodiments where two or more heteroatoms are present, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different.

The terms "fused" or "fused ring" as used herein, alone or in combination, refer to a cyclic structure in which two or more rings share one or more bonds.

The term "spiro" or "spirocyclic" as used herein, alone or in combination, refers to a cyclic structure in which two or more rings share one or more atoms.

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

"substituted" means that one or more atoms, preferably 5, more preferably 1 to 3 atoms, in the group are independently substituted with a corresponding number of substituents. It goes without saying that the skilled person in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort, when the substituents are in their possible chemical positions. For example, having a free amine or hydroxyl group may be unstable in combination with a carbon atom having an unsaturated (e.g., olefinic) bond. Such substituents include, but are not limited to, hydroxy, amine, halogen, cyano, C _1-6 Alkyl radical, C _1-6 Alkoxy radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl groups, and the like.

"pharmaceutical composition" refers to a composition containing one or more compounds described herein, or a pharmaceutically acceptable salt or prodrug thereof, and other ingredients such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote administration to the organism, facilitate absorption of the active ingredient and further exert biological activity.

"isomers" refer to compounds having the same molecular formula but differing in the nature or order of their bonding of atoms or the spatial arrangement of their atoms, referred to as "isomers", and isomers differing in the spatial arrangement of their atoms, referred to as "stereoisomers". Stereoisomers include optical isomers, geometric isomers and conformational isomers. The compounds of the present invention may exist in the form of optical isomers. Depending on the configuration of the substituents around the chiral carbon atom, these optical isomers are either in the "R" or "S" configuration. Optical isomers, including enantiomers and diastereomers, and methods of preparing and separating optical isomers are known in the art.

Geometric isomers may also exist for the compounds of the present invention. The present invention contemplates various geometric isomers and mixtures thereof resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as either the Z or E configuration, substituents around cycloalkyl or heterocyclic rings are designated as either the cis or trans configuration.

The compounds of the invention may also exhibit tautomerism, such as keto-enol tautomerism.

It is to be understood that the present invention includes any tautomeric or stereoisomeric form and mixtures thereof, and is not to be limited solely to any one tautomeric or stereoisomeric form employed in the nomenclature or chemical structure of the compounds.

"isotopes" are all isotopes of atoms occurring in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into compounds of the invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as but not limited to ² H、 ³ H、 ¹³ C、 ¹⁴ C、 ¹⁵ N、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F and ³⁶ and (4) Cl. Isotopically-labelled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples using appropriate isotopically-labelled reagentsPreparing a non-isotopic labeled preparation. Such compounds have a variety of potential uses, for example, as standards and reagents in the determination of biological activity. In the case of stable isotopes, such compounds have the potential to favorably alter biological, pharmacological or pharmacokinetic properties.

By "prodrug" is meant that the compounds of the present invention can be administered in the form of a prodrug. Prodrugs refer to derivatives that are converted to the biologically active compounds of the present invention under physiological conditions in vivo, e.g., by oxidation, reduction, hydrolysis, and the like, each of which utilizes or proceeds without the participation of an enzyme. Examples of prodrugs are the following compounds: compounds in which the amine group in the compounds of the invention is acylated, alkylated or phosphorylated, for example eicosanoylamino, propylaminoylamino, pivaloyloxymethylamino, or in which the hydroxyl group is acylated, alkylated, phosphorylated or converted to a borate, for example acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaroyloxy, propylaminoyloxy, or in which the carboxyl group is esterified or amidated, or in which the sulfhydryl group forms a disulfide bridge with a carrier molecule, for example a peptide, which selectively delivers a drug to the target and/or to the cytosol of the cell, can be prepared from the compounds of the invention according to well-known methods.

"pharmaceutically acceptable salt" or "pharmaceutically acceptable" refers to those made from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. Where the compounds of the invention contain one or more acidic or basic groups, the invention also includes their corresponding pharmaceutically acceptable salts. Thus, the compounds of the invention containing acidic groups can be present in the form of salts and can be used according to the invention, for example as alkali metal salts, alkaline earth metal salts or as ammonium salts. More specific examples of such salts include sodium, potassium, calcium, magnesium or salts with amines or organic amines, such as primary, secondary, tertiary, cyclic amines, and the like, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purines, piperazine, piperidine, choline, caffeine, and the like, with particularly preferred organic bases being isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. The compounds of the invention containing basic groups can be present in the form of salts and can be used according to the invention in the form of their addition to inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. If the compounds of the invention contain both acidic and basic groups in the molecule, the invention also includes inner salts or betaine salts in addition to the salt forms mentioned. The salts are obtained by conventional methods known to the person skilled in the art, for example by contacting these with organic or inorganic acids or bases in solvents or dispersants or by anion exchange or cation exchange with other salts.

Thus, when reference is made in this application to "a compound", "a compound of the invention" or "a compound of the invention", all said compound forms are included, such as prodrugs, stable isotopic derivatives, pharmaceutically acceptable salts, isomers, meso-forms, racemates, enantiomers, diastereomers and mixtures thereof.

In this context, the term "tumor" includes both benign tumors and malignant tumors (e.g., cancers).

The term "cancer" as used herein includes various malignancies in which Bruton's tyrosine kinase is involved, including, but not limited to, non-small cell lung cancer, esophageal cancer, melanoma, striated muscle garnet, cell carcinoma, multiple myeloma, breast cancer ovarian cancer, endometrial cancer, cervical cancer, gastric cancer, colon cancer, bladder cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer and liver cancer (e.g., hepatocellular carcinoma), more specifically liver cancer, gastric cancer and bladder cancer.

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 term "polymorph" or "polymorph" as used herein means that the compounds of the present invention have multiple crystal lattice forms, some of the compounds of the present invention may have more than one crystal form, and the present invention encompasses all polymorphic forms or mixtures thereof.

Intermediate compounds of the present invention and polymorphs thereof are also within the scope of the present invention.

Crystallization often results in a solvate of a compound of the present invention, and the term "solvate" as used herein refers to an association of one or more molecules of a compound of the present invention and one or more molecules of a solvent.

The solvent may be water, in which case the solvate is a hydrate. In addition, an organic solvent may be used. Thus, the compounds of the present invention may exist as hydrates, including monohydrate, dihydrate, hemihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compounds of the invention may be true solvates, but in other cases the compounds of the invention may also be present only occasionally as water or as a mixture of water with some other solvent the compounds of the invention may be reacted in a solvent or precipitated or crystallized in a solvent. Solvates of the compounds of the invention are also included within the scope of the invention.

As used herein, the term "acceptable" in reference to a formulation, composition or ingredient means that there is no lasting deleterious effect on the overall health of the subject being treated.

The term "pharmaceutically acceptable" as used herein refers to a substance (e.g., carrier or diluent) that does not affect 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 adverse biological response or interacting in an adverse manner with any of the components contained in the composition.

"pharmaceutically acceptable carriers" include, but are not limited to, adjuvants, carriers, excipients, adjuvants, deodorants, diluents, preservatives, dyes/colorants, flavor enhancers, surfactants and wetting agents, dispersants, suspending agents, stabilizers, isotonizing agents, solvents, or emulsifiers that have been approved by the relevant governmental authorities for use in humans and domestic animals.

As used herein, the term "subject", "patient", "subject" or "individual" refers to an individual suffering from a disease, disorder or condition, and the like, including mammals and non-mammals, examples of which include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment related to the methods and compositions provided herein, the mammal is a human.

The term "treatment" as used herein refers to the treatment of a disease condition associated with a mammal, particularly a human, and includes

(i) Preventing the development of a disease or condition in a mammal, particularly a mammal that has been previously exposed to the disease or condition but has not been diagnosed as having the disease or condition;

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

(iii) relieving the disease or condition, i.e., slowing the regression of the disease or condition;

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

The terms "disease" and "condition" as used herein may be used interchangeably and may have different meanings, as certain specific diseases or conditions have no known causative agent (and therefore the cause of the disease is not yet clear) and therefore are not considered as a disease but can be considered as an unwanted condition or syndrome, with more or less specific symptoms being confirmed by clinical researchers.

The terms "administering," "administration," "administering," and the like as used herein refer to methods that are capable of delivering a compound or composition to a desired site for biological action. Including, but not limited to, oral, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

Synthesis method

The invention also provides a method for preparing the compound. The preparation of the compounds of the general formula (I) according to the invention can be carried out by the following exemplary methods and examples, which should not be construed as limiting the scope of the invention in any way. The compounds of the invention may also be synthesized by synthetic techniques known to those skilled in the art, or a combination of methods known in the art and those described herein may be used. The product of each step is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatography, and the like. The starting materials and chemical reagents required for the synthesis can be routinely synthesized or purchased according to the literature (reaxys).

The alkynylpyrazine heterocyclic compound shown in the general formula I can be synthesized according to the following route

The method comprises the following steps: 1. starting material a1 was coupled by sonogashira to afford a 2; 2. a2 and precursor H-X-Y-Boc are subjected to aromatic nucleophilic substitution reaction under the action of alkali to generate A3; 3. deprotection of the amine group in A3 provides A4; 4. the amine group in A4 is derivatized with a chemical reagent (e.g., acryloyl chloride, etc.) containing a functional group that reacts with the cysteine residue in the kinase ligand binding domain to provide the compound of formula I.

The second method comprises the following steps: 1. the initiator A1 and the precursor H-X-Y-Boc are subjected to aromatic nucleophilic substitution reaction under the action of alkali to obtain A2'; 2. a 2' was coupled by sonogashira to yield A3; 3. and the subsequent 2 steps are the same as the first method to obtain the compound shown in the general formula I.

Unless otherwise indicated, temperatures are in degrees celsius. Reagents were purchased from commercial suppliers such as Combi-blocks Inc, Astatech Inc or mcalin, and these reagents were used directly without further purification unless otherwise stated.

Unless otherwise stated, the following reactions are carried out at room temperature, in anhydrous solvents, under positive pressure of nitrogen or argon, or using a drying tube; glassware was dried and/or heat dried.

Unless otherwise stated, column chromatography purification was performed using 200-300 mesh silica gel from Qingdao oceanic plants; preparation of thin-layer chromatography silica gel precast slab (HSGF254) produced by Nicotiana chemical industry research institute; MS was measured using a Thermo Fisher LCQ fly model (ESI) liquid chromatography-mass spectrometer.

Nuclear magnetic data ( ¹ H NMR) Using a Bruker Avance-400MHz or Varian Oxford-400Hz Nuclear magnetic Analyzer, the Nuclear magnetic data were obtained using CDCl as the solvent ₃ 、CD ₃ OD、D ₂ O、DMSO-d ₆ Etc., based on tetramethylsilane (0.000ppm) or based on residual solvent (CDCl) ₃ :7.26ppm；CD ₃ OD:3.31ppm；D ₂ O:4.79ppm；DMSO-d ₆ 2.50ppm) when indicating the diversity of the peak shapes, the following abbreviations indicate the different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (doublet of doublets), dt (doublet of triplets). If the coupling constant is given, it is given in Hertz (Hz).

Example 1: preparation of (S) -2- (1-acryloyl-pyrrolidin-3-amino) -6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 1)

Step 1: synthesis of Compound 1b

A reaction flask was charged with compound 1a (1.49g,10.0mmol), 3, 5-dimethoxyphenylacetylene (1.62g,10.0mmol), bis (triphenylphosphine) palladium dichloride (702mg,1.0mmol), cuprous iodide (190mg,1.0mmol), triethylamine (5.06g,50.0mmol) and 40ml of N, N-dimethylformamide. The mixture was purged with nitrogen 3 times, and reacted at 100 ℃ overnight with stirring. Cooled to room temperature, the reaction solution was diluted with ethyl acetate and water, and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 1b (1.90g, yield 69%) as a pale yellow solid. LC/MS (ESI) M/z 275.1[ M + H ]] ⁺ .

Step 2: synthesis of Compound 1c

To a reaction flask were added compound 1b (0.82g,3.0mmol), (S) -1-tert-butoxycarbonyl-3-aminopyrrolidine (0.67g,3.6mmol), potassium carbonate (0.83g,6.0mmol) and 12ml of N, N-dimethylformamide. The reaction was carried out at 90 ℃ for 6 hours with stirring. Cooled to room temperature, the reaction solution was diluted with ethyl acetate and water, and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 1c (0.93g, yield 73%) as a pale yellow solid. LC/MS (ESI) M/z 325.2[ M + H ]] ⁺ .

And step 3: synthesis of Compound 1d

Intermediate 1c (0.85g,2.0mmol), 4ml ethyl acetate, 4ml HCl in 1, 4-dioxane were added to the reaction flask. After stirring at room temperature for 2 hours, the reaction solution was neutralized with 1N sodium hydroxide solution and extracted with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and the organic phase was evaporated to dryness under reduced pressure. Compound 1d (0.60g, 92% yield) was obtained as a yellow oil and used directly in the next step. LC/MS (ESI) M/z 325.2[ M + H ]] ⁺ .

And 4, step 4: synthesis of Compound 1

A reaction flask was charged with compound 1d (324mg,1.0mmol), triethylamine (152mg,1.5mmol), and 4ml of dichloromethaneAlkane, cooled in an ice-water bath and a solution of acryloyl chloride (136mg,1.5mmol) in 0.5ml of dichloromethane was slowly added dropwise. After the addition was complete, stirring was continued for 3 hours. The reaction solution was quenched with methanol and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 1(148mg, yield 39%) as a pale yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:7.95(s,1H),7.31(s,1H),6.74(d,2H),6.45(t,1H),6.23(dd,1H),6.01(dd,1H),5.75(s,1H),5.32(dd,1H),3.81(s,6H),3.69-3.28(m,4H),2.83-2.76(m,1H),1.91-1.65(m,2H)；LC/MS(ESI):m/z＝379.2[M+H] ⁺ .

Example 2: preparation of (S) -2- (1-acryloylpyrrolidin-3-amino) -5-methyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 2)

In a similar manner to example 1 (starting material was changed to 2-methyl-3, 5-dichloropyrazine), compound 2(162mg, yield 41%, which is the final yield, the same applies hereinafter) was obtained as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:7.25(s,1H),6.76(d,2H),6.47(t,1H),6.24(dd,1H),5.99(dd,1H),5.82(s,1H),5.36(dd,1H),3.81(s,6H),3.72-3.31(m,4H),2.81-2.73(m,1H),2.39(s,3H),1.93-1.68(m,2H)；LC/MS(ESI):m/z＝393.2[M+H] ⁺ .

Example 3: preparation of (S) -2- (1-acryloylpyrrolidin-3-amino) -5-fluoro-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 3)

In a similar manner to example 1 (starting material was changed to 2-fluoro-3, 5-dichloropyrazine) was used to give compound 3(147mg, yield 38%) as a pale yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:7.62(s,1H),6.74(d,2H),6.44(t,1H),6.28(dd,1H),6.06(dd,1H),5.81(s,1H),5.40(dd,1H),3.80(s,6H),3.68-3.28(m,4H),2.85-2.73(m,1H),1.98-1.67(m,2H)；LC/MS(ESI):m/z＝397.2[M+H] ⁺ .

Example 4: preparation of (S) -2- (1-acryloylpyrrolidin-3-amino) -5-cyano-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 4)

In a similar manner to example 1 (starting material was changed to 2-cyano-3, 5-dichloropyrazine) was used to give compound 4(129mg, yield 32%) as a pale yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.06(s,1H),6.75(d,2H),6.43(t,1H),6.25(dd,1H),6.05(dd,1H),5.63(s,1H),5.35(dd,1H),3.81(s,6H),3.72-3.34(m,4H),2.86-2.77(m,1H),1.92-1.65(m,2H)；LC/MS(ESI):m/z＝404.2[M+H] ⁺ .

Example 5: preparation of (S) -2- (1-acryloylpyrrolidin-3-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 5)

In a similar manner to example 1 (starting material was changed to 3, 5-dichloropyrazine-2-carboxamide) gave compound 5(136mg, yield 37%) as a yellow solid. ¹ HNMR(400MHz,DMSO-d ₆ )δ:8.12(s,1H),8.06(s,1H),7.98(s,1H),6.82(d,2H),6.45(t,1H),6.21(dd,1H),6.02(dd,1H),5.83(s,1H),5.41(dd,1H),3.81(s,6H),3.69-3.31(m,4H),2.91-2.78(m,1H),1.90-1.62(m,2H)；LC/MS(ESI):m/z＝422.2[M+H] ⁺ .

Example 6: preparation of (S) -2- (1-acryloylpiperidin-3-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 6)

Using the intermediates 5-chloro-3- (3, 5-dimethoxyphenylethynyl) pyrazine-2-carboxamide of example 5 and (S) -1-tert-butoxycarbonyl-3-aminopiperidine, the subsequent 2 steps were similar to those of example 1 to obtainTo compound 6(168mg, 47% yield) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.05(s,1H),7.96(s,1H),6.79(d,2H),6.43(t,1H),6.17(dd,1H),5.98(dd,1H),5.76(s,1H),5.39(dd,1H),3.81(s,6H),3.78-3.31(m,4H),2.72-2.65(m,1H),1.91-1.43(m,4H)；LC/MS(ESI):m/z＝436.2[M+H] ⁺ .

Example 7: preparation of (S) -2- (1-acryloylpiperidin-4-ylamino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 7)

Using the intermediates 5-chloro-3- (3, 5-dimethoxyphenylethynyl) pyrazine-2-carboxamide in example 5 and (S) -1-tert-butoxycarbonyl-4-aminopiperidine, followed by 2 steps similar to example 1, compound 7(146mg, yield 41%) was obtained as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.05(s,1H),7.97(s,1H),6.80(d,2H),6.45(t,1H),6.21(dd,1H),6.02(dd,1H),5.81(s,1H),5.43(dd,1H),3.81(s,6H),3.25-2.95(m,4H),2.71-2.64(m,1H),1.84-1.45(m,4H)；LC/MS(ESI):m/z＝436.2[M+H] ⁺ .

Example 8: preparation of (S) -2- (but-2-ynoylpyrrolidin-3-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 8)

Reaction of the intermediate (S) -2- (3-aminopyrrolidine) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine of example 5 with 2-butynoyl chloride gave compound 8(134mg, 33% yield) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.12(s,1H),8.06(s,1H),7.98(s,1H),6.82(d,2H),6.45(t,1H),5.91(s,1H),3.81(s,6H),3.75-3.31(m,7H),2.94-2.85(m,1H),1.91-1.58(m,2H)；LC/MS(ESI):m/z＝434.2[M+H] ⁺ .

Example 9: preparation of (S) -2- (1-dimethylaminopropylenepyrrolidin-3-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 9)

Intermediate (S) -2- (3-aminopyrrolidine) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine from example 5 and (E) -3- (dimethylamine) -acryloyl chloride gave compound 9(136mg, 32% yield) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.12(s,1H),8.06(s,1H),7.98(s,1H),6.82(d,2H),6.45(m,2H),5.86(s,1H),5.05(d,1H),3.80(s,6H),3.73(s,6H),3.75-3.28(m,4H),2.70-2.64(m,1H),1.87-1.54(m,2H)；LC/MS(ESI):m/z＝465.2[M+H] ⁺ .

Example 10: preparation of (S) -2- (1-acrylamido-3-pyrrolidinyl) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 10)

Using the intermediates 5-chloro-3- (3, 5-dimethoxyphenylethynyl) pyrazine-2-carboxamide in example 5 and (S) -3-tert-butoxycarbonylaminopyrrolidine, the subsequent 2 steps were similar to example 1 to give compound 10(158mg, yield 45%) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.12(s,1H),8.06(s,1H),7.98(s,1H),6.82(d,2H),6.45(t,1H),6.21(dd,1H),6.09(dd,1H),5.77(s,1H),5.48(dd,1H),3.81(s,6H),3.74-3.64(m,1H),2.93-2.65(m,4H),1.95-1.70(m,2H)；LC/MS(ESI):m/z＝436.2[M+H] ⁺ .

Example 11: preparation of 2- (2-acryloyl-2-azaspiro [3,3] heptan-6-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 11)

A similar procedure was used as in example 10 (intermediate exchanged for 6-amino-2-azaspiro [3 ].3]Heptane-2-carboxylic acid tert-butyl ester) gave compound 11(130mg, yield 29%) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.07(s,1H),7.95(s,1H),6.80(d,2H),6.47(t,1H),6.23(dd,1H),6.04(dd,1H),5.74(s,1H),5.35(dd,1H),3.81(s,6H),3.72-3.53(m,4H),3.15-3.04(m,1H),2.16-1.87(m,4H)；LC/MS(ESI):m/z＝448.2[M+H] ⁺ .

Example 12: preparation of 2- (2-acryloyl-2-azaspiro [3,4] octane-7-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 12)

Using a method similar to example 10 (intermediate exchanged for 2-amino-6-azaspiro [3.4 ]]Octane-6-carboxylic acid tert-butyl ester) gave compound 12(148mg, yield 35%) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.08(s,1H),7.96(s,1H),6.77(d,2H),6.46(t,1H),6.28(dd,1H),6.09(dd,1H),5.82(s,1H),5.37(dd,1H),3.80(s,6H),3.55-3.21(m,4H),3.12-3.04(m,1H),2.23-1.90(m,4H),1.63-1.49(m,2H)；LC/MS(ESI):m/z＝462.2[M+H] ⁺ .

Example 13: preparation of 2- (6-acryloyl-6-azaspiro [3,5] nonan-2-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 13)

Using a method similar to example 10 (intermediate exchanged for 2-amino-7-azaspiro [3.5 ]]Nonane-7-carboxylic acid tert-butyl ester) gave compound 13(171mg, 43% yield) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.06(s,1H),7.96(s,1H),6.77(d,2H),6.45(t,1H),6.25(dd,1H),6.06(dd,1H),5.75(s,1H),5.41(dd,1H),3.81(s,6H),3.56-3.28(m,4H),3.12-3.06(m,1H),2.21-1.92(m,4H),1.58-1.39(m,4H)；LC/MS(ESI):m/z＝476.2[M+H] ⁺ .

Example 14: preparation of 2- (7-acryloyl-7-azaspiro [3,5] nonan-2-amino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 14)

Using a method similar to example 10 (intermediate exchanged for 2-amino-7-azaspiro [3.5 ]]Nonane-7-carboxylic acid tert-butyl ester) gave compound 13(182mg, yield 48%) as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.06(s,1H),7.95(s,1H),6.77(d,2H),6.45(t,1H),6.21(dd,1H),5.98(dd,1H),5.79(s,1H),5.39(dd,1H),3.81(s,6H),3.65-3.32(m,4H),3.16-2.90(m,4H),2.72-2.65(m,1H),1.83-1.49(m,4H)；LC/MS(ESI):m/z＝476.2[M+H] ⁺ .

Example 15: preparation of 2- (1-acryloylpiperidine-4-methylamino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 15)

Using a method similar to example 10 (intermediate was changed to 1-tert-butoxycarbonyl-4-aminomethylpiperidine), compound 15(149mg, yield 40%) was obtained as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.07(s,1H),7.97(s,1H),6.78(d,2H),6.45(t,1H),6.19(dd,1H),6.01(dd,1H),5.81(s,1H),5.35(dd,1H),3.81(s,6H),3.41-3.22(m,4H),3.06-2.91(m,2H),2.03-1.72(m,5H)；LC/MS(ESI):m/z＝450.2[M+H] ⁺ .

Example 16: preparation of 2- (1-acryloylpiperidine-3-methylamino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 16)

Using a method similar to example 10 (intermediate was changed to 1-tert-butoxycarbonyl-3-aminomethylpiperidine), compound 16(156mg, yield 42%) was obtained as a pale yellow colorAnd (3) a solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.07(s,1H),7.97(s,1H),6.78(d,2H),6.45(t,1H),6.25(dd,1H),6.08(dd,1H),5.76(s,1H),5.40(dd,1H),3.81(s,6H),3.58-3.23(m,4H),2.96-2.71(m,2H),2.09-1.53(m,5H)；LC/MS(ESI):m/z＝450.2[M+H] ⁺ .

Example 17: preparation of 2- (1-acryloylpyrrolidine-3-methylamino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 17)

Using a method similar to example 10 (intermediate was changed to 1-tert-butoxycarbonyl-3-aminomethylpyrrolidine), compound 17(162mg, yield 45%) was obtained as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.13(s,1H),8.07(s,1H),7.97(s,1H),6.78(d,2H),6.45(t,1H),6.23(dd,1H),6.11(dd,1H),5.80(s,1H),5.42(dd,1H),3.81(s,6H),3.46-3.19(m,4H),3.12-2.85(m,2H),1.89-1.51(m,3H)；LC/MS(ESI):m/z＝436.2[M+H] ⁺ .

Example 18: preparation of 2- (1-acryloyl azetidine-3-methylamino) -5-carbamoyl-6- (3, 5-dimethoxyphenylethynyl) pyrazine (Compound 18)

Using a method similar to example 10 (intermediate was changed to 1-tert-butoxycarbonyl-3-aminomethylazetidine), compound 18(97mg, yield 21%) was obtained as a yellow solid. ¹ H NMR(400MHz,DMSO-d ₆ )δ:8.12(s,1H),8.06(s,1H),7.99(s,1H),6.76(d,2H),6.45(t,1H),6.18(dd,1H),6.03(dd,1H),5.86(s,1H),5.33(dd,1H),3.81(s,6H),3.87-3.32(m,5H),2.23-1.86(m,2H)；LC/MS(ESI):m/z＝422.2[M+H] ⁺ .

Example 19: in vitro activity inhibition of kinases FGFR1, FGFR2, FGFR3 and FGFR4

Using a Caliper mobility shift assay technique) FGFR1, FGFR2, FGFR3 and FGFR4 protein kinase activities were determined. Compounds were dissolved in DMSO and diluted with kinase buffer, and 5 μ L of compound (10% DMS0) at 5-fold final reaction concentration was added to the 384-well plate. After adding 10. mu.L of a 2.5-fold enzyme solution (FGFR 1, FGFR2, FGFR3 and FGFR4, respectively), the mixture was incubated at room temperature for 10 minutes, and then 10. mu.L of a 2.5-fold substrate (FAM-labeledpeptide and dATP) solution was added. After incubation at 28 ℃ for 30-60 minutes, 25. mu.L of stop buffer (pH 7.5100mM HEPES, 0.015% Brij-35, 0.2% Coating Reagent #3,50mM EDTA) was added to stop the reaction. Conversion data were read on a Caliper EZ Reader II (Caliper life Sciences). The conversion was converted to inhibition data (% inhibition ═ max-sample conversion)/(max-min) × 100). Wherein max refers to the conversion rate of a DMSO control, and min refers to the conversion rate of an enzyme-free control. Taking the concentration and the inhibition rate of the compound as horizontal and vertical coordinates, drawing a curve, fitting the curve by using XLFit excel add-in version4.3.1 software and calculating IC ₅₀ . The results of the assay are shown in the following table showing activity data of compounds 1-18 on the kinases FGFR1, FGFR2, FGFR3 and FGFR 4. Active utilization of IC ₅₀ Characterization, wherein "A" represents IC ₅₀ Less than or equal to 10 nM; "B" means 10<IC ₅₀ Less than or equal to 100 nM; "C" means 100<IC ₅₀ Less than or equal to 500 nM; "D" means 500<IC ₅₀ ≤2000nM。

Claims (7)

1. A compound having the general formula I or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, polymorph or isomer thereof,

wherein:

ring B is a phenyl ring or a 5-6 membered heteroaromatic ring wherein the above phenyl and heteroaromatic rings are optionally substituted by one or more G ¹ Substituted;

R ¹ ，R ² independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -OR ⁴ 、-NR ⁴ R ⁵ 、-C(O)NR ⁴ R ⁵ Wherein said alkyl, cycloalkyl OR heterocycloalkyl is optionally substituted by cyano, halogen, -OR ⁶ 、-NR ⁶ R ⁷ 、C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocycloalkyl;

x is independently selected from-C _0-4 Alkyl-, -CR ⁸ R ⁹ -、-C _1-2 Alkyl (R) ⁸ )(OH)-、-C(O)-、-CR ⁸ R ⁹ O-、-OCR ⁸ R ⁹ -、-SCR ⁸ R ⁹ -、-CR ⁸ R ⁹ S-、-NR ⁸ -、-NR ⁸ C(O)-、-C(O)NR ⁸ -、-NR ⁸ C(O)NR ⁹ -、-CF ₂ -、-O-、-S-、-S(O) _m -、-NR ⁸ S(O) ₂ -、-S(O) ₂ NR ⁸ -；

Y is absent or C is selected _3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclyl, 5-12 membered spiro cyclic, aromatic or heteroaromatic, wherein said cycloalkyl, heterocycloalkyl, spiro cyclic, fused heterocyclyl, spiro heterocyclic, aromatic or heteroaromatic is optionally substituted with one or more G ² Substituted;

z is independently selected from cyano, -NR ¹⁰ CN、

Bond a is a double or triple bond;

when a is a double bond, R ^a 、R ^b And R ^c Each independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl. Wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ³ Substituted;

R ^a and R ^b Or R ^b And R ^c Optionally together with the carbon atom to which they are attached to form an optionally heteroatom-containing radical3-6 membered ring of (a);

when bond a is a triple bond, R ^a And R ^c Is absent, R ^b Independently selected from H, cyano, halogen, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl by one or more G ⁴ Substituted;

R ¹⁰ is independently selected from H, C _1-6 Alkyl radical, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl, wherein said alkyl, cycloalkyl and heterocyclyl are optionally substituted by 1 or more G ⁵ Substituted;

G ¹ 、G ² 、G ³ 、G ⁴ and G ⁵ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-to 8-membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹¹ 、-OC(O)NR ¹¹ R ¹² 、-C(O)OR ¹¹ 、-C(O)NR ¹¹ R ¹² 、-C(O)R ¹¹ 、-NR ¹¹ R ¹² 、-NR ¹¹ C(O)R ¹² 、-NR ¹¹ C(O)NR ¹² R ¹³ 、-S(O) _m R ¹¹ or-NR ¹¹ S(O) _m R ¹² Wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by 1 or more cyano, halogen, C _1-6 Alkyl radical, C _2-6 Alkenyl radical, C _2-6 Alkynyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, C _6-10 Aryl, 5-10 membered heteroaryl, -OR ¹⁴ 、-OC(O)NR ¹⁴ R ¹⁵ 、-C(O)OR ¹⁴ 、-C(O)NR ¹⁴ R ¹⁵ 、-C(O)R ¹⁴ 、-NR ¹⁴ R ¹⁵ 、-NR ¹⁴ C(O)R ¹⁵ 、-NR ¹⁴ C(O)NR ¹⁵ R ¹⁶ 、-S(O) _m R ¹⁴ or-NR ¹⁴ S(O) _m R ¹⁵ Substituted with the substituent(s);

R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ 、R ⁹ 、R ¹¹ 、R ¹² 、R ¹³ 、R ¹⁴ 、R ¹⁵ and R ¹⁶ Each independently selected from cyano, halogen, C _1-6 Alkyl radical, C _3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl; and m is 1 or 2.

2. A compound according to claim 1 or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, polymorph or isomer thereof and a mixture form thereof.

3. Selected from the following compounds

Or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, isomer, and mixtures and forms thereof.

4. A pharmaceutical composition comprising a compound of claims 1-3 or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of the claims or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, solvate, polymorph or isomer thereof in the manufacture of a medicament for the treatment of an FGFR-mediated disease.

6. The use of claim 5, wherein the FGFR-mediated disease is one or more of non-small cell lung cancer, esophageal cancer, melanoma, gastric cancer, multiple myeloma, liver cancer, cholangiocarcinoma, prostate cancer, skin cancer, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, breast cancer, colon cancer, glioma, and rhabdomyosarcoma.

7. A compound according to any one of claims 5 or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, and a pharmaceutically acceptable carrier for use as a medicament.