CN114057749B - Preparation method and application of irreversible alkyne heterocyclic compound FGFR inhibitor - Google Patents

Abstract

The invention discloses a preparation method and application of an irreversible alkyne heterocyclic compound FGFR inhibitor, which are compounds shown in a general formula (I) and pharmaceutically acceptable salts thereof, a pharmaceutical composition containing the compounds and/or pharmaceutically acceptable salts thereof, and application of the compounds or pharmaceutically acceptable salts in medicaments for treating or preventing FGFR kinase-related diseases, especially tumors, and the preparation method of the pharmaceutical composition containing alkyne heterocyclic template compounds and the pharmaceutically acceptable salts thereof.

Description

Preparation method and application of irreversible alkyne heterocyclic compound FGFR inhibitor

Technical field:

the present invention relates to irreversible alkyne heterocyclic compound FGFR inhibitors; a pharmaceutical composition comprising said acetylenic heterocyclic compound or a pharmaceutically acceptable salt thereof; a process for the preparation of the acetylenic heterocyclic compound or a pharmaceutically acceptable salt thereof; use of said acetylenic heterocyclic compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said acetylenic heterocyclic compound or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment and/or prevention of FGFR-related disorders, in particular tumors.

The background technology is as follows:

fibroblast growth factor receptor (Fibroblast Growth Factor Receptor, FGFR) is a class of Receptor Tyrosine Kinases (RTKs), and the FGFR family mainly includes four subtypes FGFR1-, FGFR2, FGFR3, and FGFR 4. FGFR1 is a transmembrane protein belonging to the receptor tyrosine kinase and consists of three major components: namely, extracellular, transmembrane and cellular segments, the extracellular segment being the binding region for the ligand fibroblast growth factor (Fibroblast growth factor, FGFs). FGFs are also a polygene family, and there are 19 members, FGF1 also called acidic fibroblast growth factor (aFGF), FGF2 also called basic fibroblast growth-related document factor (bFGF), which have biological activities that stimulate the growth of fibroblasts, vascular endothelial cells, smooth muscle cells and nerve cells.

However, when FGFR is mutated or overexpressed, excessive activation of FGFR signaling pathways is caused, and normal cell canceration is further induced. Wherein, excessive activation of RAS-RAF-MAPK stimulates cell proliferation and differentiation; excessive activation of PI3K-AKT leads to inhibition of apoptosis; SATA is closely related to promoting tumor invasion and metastasis, enhancing tumor immune escape ability; the PLC gamma signal path is an important path for tumor cell metastasis regulation. According to a study published in Clinical CancerResearch in 2015, next Generation Sequencing (NGS) against 4853 types of solid tumors showed that FGFR aberrations (absences) were found and activated abnormally in approximately 7.1% of cancers, most of which were gene amplification (66%), followed by mutation (26%) and rearrangement (8%). FGFR aberration is present in almost all malignant tumors detected, and abnormal activation of FGFR is also found in tumors such as urinary 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, which occur at a high incidence.

There are some non-FGFR specific drugs on the market such as lenretini of Sunitinib, eisai and nintedanib of Boehringer Ingelheimr of pfizer. Whereas FDA approved FGFR inhibitors are only Balversa (Erdafitinib) and Pemazyre (pemigatinib). Small molecule inhibitors of FGFR1/2/3 that enter the clinic are: informatinib (BGJ 398) and AZD4547, fisogatinib (BLU-554), roblitinib (FGF 401), H3B6527, lucitanib (E-3810), futibatinib (TAS-120), RPN1371, ICP-192, derazantinib, 3D185, BPI-17509, HMPL-453.

Although the development of FGFR inhibitors attracts numerous corporate layouts at home and abroad, although 2 FGFR inhibitors have been marketed, new compounds are still required to be developed due to the prospects they show in the treatment of various malignant tumors. Through continuous efforts, the invention designs irreversible inhibitors which have independent intellectual property and show excellent activity on FGFR-1-4 protein kinase.

Disclosure of Invention

Wherein:

X ₁ ，X ₂ ，X ₃ can be independently selected from N, CR ¹ ；

Ring B is a benzene ring or a 5-6 membered heteroaromatic ring, wherein the benzene ring and the heteroaromatic ring are optionally substituted with one or more G ¹ Substituted;

R ¹ independently selected from H, D, cyano, halogen, C _1-6 Alkyl, COOH, CONH2, NHCOH, CONHR ² 、OR ² or-NHR ² ；；

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

u is independently selected from-C _0-4 Alkyl-, -CR ⁷ R ⁸ -、-C _1-2 Alkyl (R) ⁷ )(OH)-、-C(O)-、-CR ⁷ R ⁸ O-、-OCR ⁷ R ⁸ -、-SCR ⁷ R ⁸ -、-CR ⁷ R ⁸ S-、-NR ⁷ -、-NR ⁷ C(O)-、-C(O)NR ⁷ -、-NR ⁷ C(O)NR ⁸ -、-CF ₂ -、-O-、-S-、-S(O) _m -、-NR ⁷ S(O) ₂ -、-S(O) ₂ NR ⁷ -；

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 spiroheterocyclyl, aryl or heteroaryl, wherein said cycloalkyl, heterocycloalkyl, spiroheterocyclyl, fused ring, fused heterocyclyl, spiroheterocyclyl, aryl or heteroaryl is optionally substituted with one or more G ² Substituted;

z is independently selected from cyano, -NR ⁹ CN、Bond a is a double bond or a triple bond;

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

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

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

R ⁹ independently selected from H, D, C _1-6 Alkyl, C _3-6 Cycloalkyl or 3-6 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted with 1 or more G ⁵ Substituted;

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

R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ 、R ¹⁰ 、R ¹¹ 、R ¹² 、R ¹³ 、R ¹⁴ and R is ¹⁵ Each independently selected from hydrogen, D, cyano, halogen, C _1-6 Alkyl, C _3-8 Cycloalkyl or 3-8 membered monocyclic heterocyclyl, monocyclic heteroaryl or phenyl; and m is 1 or 2.

The compounds according to the general structural formula (I) of the present invention include one of the compounds numbered 1-8, but are not limited to the following:

the present invention provides methods of the above-described novel FGFR inhibitors or isomers, hydrates, solvates, polymorphs, pharmaceutically acceptable salts thereof.

The compounds of the invention are useful for the treatment or prophylaxis of FGFR-associated tumors, such as non-small cell lung cancer, esophageal cancer, melanoma rhabdomyosarcoma, cell 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 invention provides a method of treating or preventing a FGFR mediated disease (e.g., neoplastic), comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the 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 invention relates to a compound of formula (I) or a prodrug, stable isotope derivative, polymorph, solvate, pharmaceutically acceptable salt, isomer, or mixture thereof for use as a medicament or in medicine for treating or preventing FGFR mediated diseases, such as tumors 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, diaphragmatic cancer, bladder cancer, pancreatic cancer, lung cancer, prostate cancer.

The invention further relates to a pharmaceutical composition comprising the compound of the invention or a prodrug thereof, a stable isotope derivative, a pharmaceutically acceptable salt isomer and a mixture thereof, and a pharmaceutically acceptable carrier, diluent and excipient.

Another aspect of the invention relates to the use of a compound of formula (I) or a prodrug stable isotope derivative thereof, a pharmaceutically acceptable salt, an isomer, a mixture thereof, or a pharmaceutical composition thereof in the manufacture of a medicament, wherein the medicament is for the treatment or prevention of FGFR mediated diseases such as tumors and inflammatory diseases.

According to the present invention, the drug may be 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 used in the specification and claims.

The expression "C" as used herein has the following meaning _x-y "means a range of carbon number wherein x and y are integers, e.g. C _3-8 Cycloalkyl means cycloalkyl having 3 to 8 carbon atoms, i.e. cycloalkyl having 3, 4, 5, 6, 7 or 8 carbon atoms. It is also to be understood that "C _3-8 "also includes any subrange therein, e.g. C _3-7 、C _3-6 、C _4-7 、C _4-6 、C _5-6 Etc.

"alkyl" refers to a straight or branched hydrocarbon 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 hydrocarbon group containing at least one carbon-carbon double 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 alkenyl groups include vinyl, 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 hydrocarbon group containing at least one carbon-carbon triple bond and typically from 2 to 20 carbon atoms, for example from 2 to 8 carbon atoms, from 2 to 6 carbon atoms, or from 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 3 to 14 carbon ring atoms. Cycloalkyl groups may be monocyclic, typically containing 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 decalin, which cycloalkyl groups may be substituted or unsubstituted.

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

"spiroheterocyclyl" refers to a 5 to 20 membered, monocyclic, polycyclic heterocyclic group sharing one atom (referred to as the spiro atom) between the rings, wherein one or more of the ring atoms is selected from nitrogen, oxygen or a heteroatom of S (O) (where m is an integer from 0 to 2) and the remaining ring atoms are carbon. These may contain one or more double bonds, but the electronic system in which none of the rings has complete conjugation is preferably 6 to 14 membered, more preferably 7 to 10 membered. The spirocycloalkyl group is classified into a single spiroheterocyclyl group, a double spiroheterocyclyl group or a multiple spiroheterocyclyl group according to the number of common spiro atoms between rings, with single spirocycloalkyl groups and double spirocycloalkyl groups being preferred. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro-cyclic group. Non-limiting examples of spirocycloalkyl groups include:

"fused heterocyclyl" refers to a 5 to 20 membered, polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system, one or more of which may contain one or more double bonds, but none of which has complete conjugationPi electron systems in which one or more ring atoms are selected from nitrogen, oxygen or S (O) _m (wherein m is an integer from 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The number of constituent rings may be classified into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclyl groups include:

"aryl" or "aryl" refers to an aromatic ring or fused polycyclic group containing from 6 to 14 carbon atoms, preferably 6 to 10 members, such as phenyl and naphthyl, most preferably phenyl, which may be fused to heteroaryl, heterocyclyl or cycloalkyl rings, wherein the ring attached to the parent structure is an aryl ring, 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, and at least one aromatic ring. Unless otherwise indicated, heteroaryl groups may be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include fused or bridged ring systems, so long as the point of attachment to the rest of the molecule is an aromatic ring atom, the nitrogen, carbon, and sulfur atoms of the heteroaromatic ring may be selectively oxidized, and the nitrogen atom may be selectively quaternized. For the purposes of the present invention, heteroaryl groups are preferably stable 4-11 membered monoaromatic rings which contain 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably stable 5-8 membered monoaromatic rings which contain 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heteroaryl groups include acridinyl, azepinyl, benzimidazolyl, benzindolyl, benzodioxinyl, benzodioxanyl, benzofuranonyl, benzofuranyl, benzonaphtofuranyl, benzopyronyl, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazole, furyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quininyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, and the like. In this application, heteroaryl is preferably a 5-8 membered heteroaryl group comprising 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably pyridinyl, pyrimidinyl, thiazolyl. The heteroaryl group may be substituted or unsubstituted.

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

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

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

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

The term "spiro" or "spiro" 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 instances where the event or circumstance occurs or does not occur, e.g., an "optionally alkyl-substituted heterocyclic group" 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, in the group are independently substituted with a corresponding number of substituents. It goes without saying thatThe person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort, the substituents being in their possible chemical positions. For example, a carbon atom having a free amine or hydroxyl group bonded to an unsaturated (e.g., olefinic) bond may be unstable. The substituents include, but are not limited to, hydroxy, amino, halogen, cyano, C _1-6 Alkyl, C _1-6 Alkoxy, C _2-6 Alkenyl, C _2-6 Alkynyl, C _3-8 Cycloalkyl groups, and the like.

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

"isomer" refers to a compound having the same molecular formula but differing in the nature or sequence of their atoms bonded or the spatial arrangement of their atoms, and is referred to as an "isomer" and an isomer differing in the spatial arrangement of its atoms is referred to as a "stereoisomer". 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 in the "R" or "S" configuration. Optical isomers include enantiomers and diastereomers, and methods for preparing and separating optical isomers are known in the art.

The compounds of the invention may also exist as geometric isomers. 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 Z or E configuration, and substituents around cycloalkyl or heterocycle are designated as 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 limited to any one tautomeric or stereoisomeric form used in the naming or chemical formulae of the compounds.

"isotopes" are all isotopes of atoms that are present in compounds of the 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, each 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 F ³⁶ Cl. Isotopically-labeled 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 an appropriate isotopically-labeled reagent in place of a non-isotopically-labeled reagent. Such compounds have a variety of potential uses, for example as standards and reagents in assaying biological activity. In the case of stable isotopes, such compounds have the potential to advantageously alter biological, pharmacological or pharmacokinetic properties.

By "prodrug" is meant that the compounds of the invention may be administered in the form of a prodrug. Prodrugs refer to derivatives of the biologically active compounds of the present invention which are converted under physiological conditions in vivo, e.g., by oxidation, reduction, hydrolysis, etc. (each of which is performed with or without the aid of an enzyme). Examples of prodrugs are the following compounds: wherein the amine groups in the compounds of the invention are acylated, alkylated or phosphorylated, such as eicosanoylamino, propylamino, pivaloyloxymethylamino, or wherein the hydroxyl groups are acylated, alkylated, phosphorylated or converted to borates, such as acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaryloxy, propylaminooxy, or wherein the carboxyl groups are esterified or amidated, or wherein the sulfhydryl groups form disulfide bridges with carrier molecules, such as peptides, that selectively deliver the drug to the target and/or cytosol of the cell, these compounds may be prepared from the compounds of the invention according to well known methods.

"pharmaceutically acceptable salts" or "pharmaceutically acceptable" refer to those prepared 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 encompasses their corresponding pharmaceutically acceptable salts. Thus, the compounds according to the invention containing acidic groups may be present in salt form and may 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 salts or salts with amines or organic amines, such as primary, secondary, tertiary, cyclic amines, etc., for example, ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purine, piperazine, piperidine, choline, and caffeine, and particularly preferred organic bases are salts of isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. The compounds of the invention containing basic groups may be present in salt form and may 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 includes, in addition to the salt forms mentioned, also internal salts or betaines. The individual 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, references in the present application to "a compound", "a compound of the invention" or "a compound of the invention" include all such compound forms, e.g., prodrugs, stable isotope derivatives, pharmaceutically acceptable salts, isomers, meso, racemates, enantiomers, diastereomers, and mixtures thereof.

Herein, the term "tumor" includes benign tumors and malignant tumors (e.g., cancers).

As used herein, the term "cancer" includes various malignant tumors that FGFR kinase references take place in, including but not limited to non-small cell lung cancer, esophageal cancer, melanoma, striated muscle grenade, cell cancer, 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 cancer), more particularly liver cancer, gastric cancer, and bladder cancer.

The term "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein refers 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 of a disease or any other desired alteration of a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to provide clinically significant relief from a disorder. Effective amounts suitable in any individual case can be determined using techniques such as a dose escalation test.

The term "polymorph" or "polymorphic form" as used herein means that a compound of the present invention has a plurality of crystalline forms, some compounds of the present invention may have more than one crystalline form, and the present invention encompasses all polymorphic forms or mixtures thereof.

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

Crystallization often yields solvates of the compounds of the present invention, and the term "solvate" as used herein refers to a complex composed of one or more molecules of the compounds 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 is also possible. 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 simply accidentally retain water or a mixture of water with some other solvent, the compounds of the invention may be reacted in one solvent or precipitated or crystallized in one solvent. Solvates of the compounds of the present invention are also included within the scope of the present invention.

The term "acceptable" in relation to a formulation, composition or ingredient as used herein means that there is no sustained detrimental effect on the overall health of the subject being treated.

The term "pharmaceutically acceptable" as used herein refers to a material (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 material can be administered to an individual without causing an adverse biological reaction or interacting in an adverse manner with any of the components contained in the composition.

"pharmaceutically acceptable carrier" includes, but is not limited to, adjuvants, carriers, excipients, adjuvants, deodorants, diluents, preservatives, dyes/colorants, flavor enhancers, surfactants and wetting agents, dispersing agents, suspending agents, stabilizer isotonic agents, solvents, or emulsifiers that have been approved by the relevant government administration for use in humans and domestic animals.

The terms "subject," "patient," "subject," or "individual" as used herein refer to an individual having 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: human, 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, guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment of the related 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, including

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

(ii) Inhibiting the disease or disorder, i.e., controlling its progression;

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

(iv) Relieving symptoms caused by diseases or symptoms.

The terms "disease" and "disorder" as used herein may be used interchangeably or differently and, because some specific diseases or disorders have not yet been known to cause a disease (and therefore the cause of the disease is not yet known), they cannot be considered as a disease but rather can be considered as an unwanted condition or syndrome, more or less specific symptoms of which have been confirmed by clinical researchers.

The terms "administering," "administering," and the like as used herein refer to methods that enable delivery of a compound or composition to a desired site for biological action. Including, but not limited to, oral routes, duodenal routes, 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, however, should not be regarded as limiting the scope of the invention in any way. The compounds described herein may also be synthesized by synthetic techniques known to those skilled in the art, or by a combination of methods known in the art and methods described herein. The product obtained in each step is obtained using separation techniques known in the art including, but not limited to, extraction, filtration, distillation, crystallization, chromatographic separation, and the like. The starting materials and chemical reagents required for the synthesis can be synthesized conventionally according to the literature (reaxys) or purchased.

Unless otherwise indicated, temperatures are degrees celsius. Reagents were purchased from commercial suppliers such as chemlocks Inc, astatech Inc or michelin and these reagents were used directly without further purification unless otherwise indicated.

Unless otherwise indicated, the following reactions were carried out at room temperature, in anhydrous solvents, under positive pressure of nitrogen or gas, or using dry tubes; glassware drying and/or heat drying.

Column chromatography purification uses 200-300 mesh silica gel from the Qingdao marine chemical plant unless otherwise indicated; preparation of thin layer chromatography A thin layer chromatography silica gel prefabricated plate (HSGF 254) manufactured by Kagaku chemical industry research institute of tobacco, inc.; MS was determined using a Therno LCD Fleet type (ESI) liquid chromatograph-mass spectrometer.

Nuclear magnetic data [ ] ¹ H NMR) using Bruker Avance-400MHz or Varian Oxford-400Hz nuclear magnetic instruments with CDCl as solvent for the nuclear magnetic data ₃ 、CD ₃ OD、D ₂ O、DMSO-d ₆ Etc., based on tetramethylsilane (0.000 ppm) or on residual solvent (CDCl) _3: 7.26ppm；CD ₃ OD:3.31ppm；D ₂ O:4.79ppm；DMSO-d ₆ 2.50 ppm) when peak shape diversity is indicated, the following abbreviations represent the different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (doublet), dt (doublet). If the coupling constant is given, it is in Hertz (Hz).

Detailed Description

The invention is further illustrated by the following specific examples.

Example 1: preparation of (S) -1- (1-acryloylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-difluorophenylethynyl) -4-amino-7-hydroxy-1H-pyrrolo [2,3-d ] pyridazine (Compound 1)

Step 1: synthesis of Compound 1b

The compound 3-cyano-1H-pyrrole-2-methyl is added into a reaction bottleEthyl acetate 1a (1.64 g,10.0 mmol), hydrazine hydrate 5mL, ethanol 50mL, and the mixture was warmed to reflux overnight with stirring. Cooled to room temperature and the solvent was evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 1b (0.64 g, yield 43%) as a white solid. LC/MS (ESI) m/z=151.1 [ M+H ]] ⁺ .

Step 2: synthesis of Compound 1c

In a reaction flask, 10mL of Compound 1b (0.6 g,4.0 mmol) and 10mL of N, N-dimethylformamide were added in portions NBS (1.07 g,6.0 mmol), and the mixture was reacted at 50℃for 4 hours under stirring. Cooled to room temperature, the reaction mixture was poured into 50mL of water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated under reduced pressure. The residue was purified by column chromatography to give compound 1c (0.63 g, yield 69%) as a white solid. LC/MS (ESI) m/z=229.0 [ M+H ]] ⁺ .

Step 3: synthesis of Compound 1d

Into a reaction flask were charged compound 1c (0.46 g,2.0 mmol), 3, 5-dimethoxyphenylacetylene (0.48 g,3.0 mmol), ditriphenylphospholpalladium dichloride (140 mg,0.2 mmol), cuprous iodide (38 mg,0.2 mmol), triethylamine (1.01 g,10.0 mmol) and N, N-dimethylformamide 15mL. The reaction was carried out overnight at 90℃with stirring by 3 nitrogen substitutions. Cooled to room temperature, the reaction 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 under reduced pressure. The residue was purified by column chromatography to give compound 1d (0.46 g, yield 74%) as a yellow solid. LC/MS (ESI) m/z=311.1 [ M+H ]] ⁺ .

Step 4: synthesis of Compound 1e

To the reaction flask were added (R) -1-tert-butoxycarbonyl-3-hydroxypiperidine (241 mg,1.2 mmol), triphenylphosphine (315 mg,1.2 mmol) and THF 10mL, followed by DIAD (243 mg,1.2 mmol). The yellow solution was stirred for 5-10 minutes, then intermediate 1d (310 mg,1.0 mmol) was added and the reaction was stirred at room temperature for 12 hours. The solvent was distilled off under reduced pressure to give a brown oil, and the residue was purified by column chromatography to give compound 1e (345 mg, yield 70%) as a yellow solid. LC/MS (ESI) m/z=494.2 [ M+H ]] ⁺ .

Step 5: synthesis of Compound 1f

To the reaction flask was added intermediate 1e (298 mg,0.6 mmol), ethyl acetate 1mL,4N HCl in 1, 4-dioxane, 1mL. The mixture was stirred at room temperature for 2 hours, the reaction mixture 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 evaporated to dryness under reduced pressure. Compound 1f (227 mg, 96% yield) was obtained and used directly in the next step, LC/MS (ESI): m/z=394.2 [ M+H)] ⁺ .

Step 6: synthesis of Compound 1

To the reaction flask was added compound 1f (197mg, 0.5 mmol), triethylamine (76 mg,0.75 mmol), 2mL of dichloromethane, and after cooling in an ice-water bath, a solution of acryloyl chloride (78 mg,0.75 mmol) in 0.5mL of dichloromethane was slowly added dropwise. Stirring was continued for 4 hours after the addition was completed. The reaction mixture was quenched with methanol and evaporated to dryness under reduced pressure. The residue was purified by column chromatography to give compound 1 (95 mg, yield 44%) as an off-white solid. ¹ H NMR(400MHz,DMSO)δ:11.58(s,1H),8.06-8.11(d,1H),7.14-7.10(s,1H),6.69-6.55(m,1H),6.22-6.15(m,1H),6.18-5.97(m,1H),5.99-5.68(m,1H),5.60(br,2H),4,14-4.13(m,1H),3.90(s,6H),3.54-3.99(m,3H),2.47-2.39(m,2H)；LC/MS(ESI):m/z＝470.0[M+H] ⁺ .

Example 2: preparation of (S) -1- (1-but-2-ynylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-difluorophenylethynyl) -4-amino-7-hydroxy-1H-pyrrolo [2,3-d ] pyridazine (Compound 2)

Using a method similar to example 1 (intermediate was changed to but-2-ynoyl chloride), compound 2 (112 mg, 52% yield) was obtained as a yellow solid. LC/MS (ESI) m/z=482.0 [ M+H ]] ⁺ .

Example 3: preparation of (S) -1- (1-acryloylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-dichlorophenylethynyl) -4-amino-7-hydroxy-1H-pyrrolo [2,3-d ] pyridazine (Compound 3)

Using a method similar to example 1 (intermediate changed to 3, 5-dimethoxy-2, 6-dichlorophenylacetylene), compound 3 (95 mg, 41% yield) was obtained as an off-white solid. LC/MS (ESI) m/z=503.0 [ M+H ]] ⁺ .

Example 4: preparation of (S) -1- (1-but-2-ynylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-dichlorophenylethynyl) -4-amino-7-hydroxy-1H-pyrrolo [2,3-d ] pyridazine (Compound 4)

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Using a method similar to example 2 (intermediate changed to 3, 5-dimethoxy-2, 6-dichlorophenylacetylene), compound 4 (134 mg, 57% yield) was obtained as a yellow solid. LC/MS (ESI) m/z=514.0 [ M+H ]] ⁺ .

Example 5: preparation of (S) -1- (1-acryloylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-difluorophenylethynyl) -4-amino-7-hydroxy-1H-pyrrolo [2,3-d ] pyridazine (Compound 5)

Compound 5 (73 mg, 34% yield) was obtained as a yellow solid in a similar manner to example 1. ¹ LC/MS(ESI):m/z＝471.0[M+H] ⁺ .

Example 6: preparation of (S) -1- (1-but-2-ynylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-difluorophenylethynyl) -4-amino-7-hydroxy-1H-pyrazolo [2,3-d ] pyridazine (Compound 6)

Using a method similar to example 2 (intermediate was changed to but-2-ynoyl chloride), compound 6 (129 mg, 58% yield) was obtained as a yellow solid. LC/MS (ES)I):m/z＝483.0[M+H] ⁺ .

Example 7: preparation of (S) -1- (1-acryloylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-difluorophenylethynyl) -4-amino-7-hydroxy-1H-pyrazolo [2,3-d ] pyridazine (Compound 7)

Using a method similar to example 1 (intermediate changed to 3, 5-dimethoxy-2, 6-dichlorophenylacetylene), compound 7 (88 mg, 38% yield) was obtained as a yellow solid. LC/MS (ESI) m/z=503.0 [ M+H ]] ⁺ .

Example 8: preparation of (S) -1- (1-but-2-ynylpyrrolidin-3-yl) -3- (3, 5-dimethoxy-2, 6-dichlorophenylethynyl) -4-amino-7-hydroxy-1H-pyrazolo [2,3-d ] pyridazine (Compound 8)

Using a method similar to example 2 (intermediate was changed to but-2-ynoyl chloride), compound 8 (134 mg, 57% yield) was obtained as a yellow solid. LC/MS (ESI) m/z=515.0 [ M+H ]] ⁺ .

Example 9: inhibition assay for in vitro Activity of kinases FGFR1, FGFR2, FGFR3 and FGFR4

FGFR1, FGFR2, FGFR3 and FGFR4 protein kinase activity was determined using Caliper mobility shift detection technology (Caliper mobility shift assay). Compounds were dissolved in DMSO and diluted in kinase buffer, and 5. Mu.L of 5-fold final concentration of compound (10% DMS 0) was added to 384-well plates. After adding 10. Mu.L of 2.5-fold enzyme (FGFR 1, FGFR2, FGFR3 and FGFR4 respectively) solution, incubation was performed for 10 minutes at room temperature, and then 10. Mu.L of 2.5-fold substrate (FAM-labeled peptide and ATP) solution was added. After incubation at 28℃for 30-60 min, the reaction was stopped by adding 25. Mu.L of stop solution (pH 7.5100mM HEPES,0.015%Brij-35,0.2%Coating Reagent#3,50mM EDTA). Conversion data was read on Caliper EZ Reader II (Caliper Life Sciences). Conversion into inhibition data (% inhibition= (max—sample conversion)Conversion)/(max-min) 100). Where max refers to the conversion of the DMSO control and min refers to the conversion of the no enzyme activity control. Curves were plotted with compound concentration and inhibition as the abscissa and ordinate, fitted using XLFit exceladd-in version4.3.1 software and IC calculated ₅₀ . The assay results are shown in the following table, which shows the activity data of compounds 1-8 for kinases FGFR1, FGFR2, FGFR3 and FGFR 4. Active utilization IC ₅₀ Characterization, wherein "A" represents IC ₅₀ Less than or equal to 10nM; "B" means 10<IC ₅₀ Less than or equal to 100nM; "C" means 100<IC ₅₀ Less than or equal to 500nM; "D" means 500<IC ₅₀ ≤2000nM。

Table 1 inhibitory Activity against FGFR1, FGFR2, FGFR3 and FGFR4

Example 21: human hepatoma cell Hep3B survival assay

The human liver cancer Hep3B cell line is derived from ATCC. Cells were cultured in DMEM broth, additionally supplemented with fetal bovine serum (10% FBS), penicillin-streptomycin (100,000U/L). The cells were maintained in medium at 37 ℃, 95% humidity and 5% carbon dioxide. In the experiment, hep3B cells were plated in 96-well plates at a density of 3000 cells per well, the cell suspension volume was 100PL per well, and cells were cultured overnight to allow cell attachment. The next day, each compound was diluted with DMSO in a three-fold gradient, 1PL compound in DMSO was added to the cell culture medium, and three parallel sub-wells were set for each concentration of each compound with IM DMSO as a control. The cells were then placed in a 37℃incubator, after 72 hours of compound treatment, 50. Mu.L CellTiter-Glo (Promega, madisonWI) was added to the cell culture medium, and the Relative Luminescence Units (RLU) of each well were determined and the cell viability and compound activity (IC) were calculated ₅₀ ) Wherein "A" represents IC ₅₀ Less than or equal to 10nM; "B" means 10<IC ₅₀ Less than or equal to 100nM; "C" means 100<IC ₅₀ Less than or equal to 500nM; "D" means 500<IC ₅₀ Less than or equal to 2000nM. The results of the inhibitory activity of the compounds of the examples on Hep3B cells are shown in table 2 below:

TABLE 2 inhibitory Activity on Hep3B cells

Sample numbering	IC 50 (nM)
		1	A
3	A
		5	A
7	A

Claims (5)

1. An irreversible alkyne heterocyclic compound FGFR inhibitor selected from the compounds shown below:

or a pharmaceutically acceptable salt thereof, and mixtures thereof.

2. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of FGFR mediated diseases.

4. The use of claim 3, 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.

5. The use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier in the manufacture of a medicament for the treatment or prevention of a FGFR mediated related disease.