Novel heterocyclic compound

Abstract

The present invention relates to a novel heterocyclic compound which is a compound having FGFR protein activity. Specifically, the invention relates to a compound shown as a formula (II), or pharmaceutically acceptable salt, solvate, polymorphic substance or isomer thereof, and application of the compound in preparation of medicines for treating diseases related to FGFR (Factor Growth Factor Receptor).

Description

Technical Field

[0001] This invention relates to compounds that inhibit the activity of FGFR proteins, as well as methods for preparing these compounds and the use of pharmaceutical compositions thereof. Background Technology

[0002] Fibroblast growth factor receptors (FGFRs) are transmembrane polypeptide tyrosine kinases, of which five (FGFR1–FGFR5) have been identified. FGFR2 is primarily distributed in tissues originating from the endoderm, such as the stomach, liver, pancreas, esophagus, and bile ducts in the digestive system. Previous studies have shown that FGFR2 is associated with various tumors. FGFR2 gene amplification has been reported in triple-negative breast cancer and gastric cancer with poor prognosis. FGFR2 gene mutations have been found in endometrial cancer, lung cancer, gastric cancer, and urothelial carcinoma. FGFR2 fusion proteins are present in lung adenocarcinoma, squamous cell carcinoma, thyroid cancer, prostate cancer, and cholangiocarcinoma. In colorectal cancer, FGFR2 can upregulate programmed cell death ligand 1 (PD-L1) expression through the JAK / STAT3 pathway, regulating tumor immune evasion, and FGFR2 expression levels are closely related to prognosis. Studies have found FGFR2 gene fusions in 10-20% of patients with intrahepatic cholangiocarcinoma. FGFR3 abnormalities, including point mutations, fusions, amplifications, and overexpression, may be associated with various tumors, and are found in approximately 15%-20% of advanced urothelial carcinomas, ~15% of uterine carcinosarcomas, ~5% of endometrial cancers, and other solid tumors. With the deepening understanding of tumorigenesis, the FGFR (fibroblast growth factor receptor) family has received widespread attention due to its aberrant expression in various tumors. In particular, FGFR2 and FGFR3 have been found to exhibit significant mutations in many types of cancer, and these mutations are closely related to tumor growth, spread, and resistance to traditional treatments. Therefore, developing drugs that can effectively inhibit FGFR2 and FGFR3 has become an urgent clinical need. While first-generation pan-FGFR inhibitors have demonstrated some clinical efficacy, the side effects (hyperphosphatemia, tissue mineralization) caused by their inhibition of FGFR1 severely limit their safety window, thus affecting the clinical application of these drugs. This invention aims to develop a new generation of oral, highly active, and highly selective small-molecule FGFR2 / 3 inhibitors. These inhibitors will significantly reduce FGFR1 inhibition while maintaining high activity against FGFR2 and FGFR3, and are expected to address various drug resistance issues. Summary of the Invention

[0003] This invention provides an FGFR inhibitor, which is a compound represented by general formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof. This invention also provides a series of compounds represented by general formula (I) and their pharmaceutically acceptable salts, solvates, polymorphs, or isomers, pharmaceutical compositions comprising these compounds, and methods of treating FGFR-related diseases with such compounds.

[0004] In one aspect, the present invention provides compounds of formula (II) or pharmaceutically acceptable salts, solvates, polymorphs or isomers thereof:

[0005]

[0006] in,

[0007] R is Or -(CO)-NH-R 10 ,

[0008] R 10 C 1-6 Alkyl group, -(CH2) 0-2 -(3-8 membered cycloalkyl), or -(CH2) 0-2 -(3-8 membered heterocyclic group), wherein the alkyl, cycloalkyl, and heterocyclic group are optionally converted by halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl, halogenated C 1-6 Alkyl, Halogenated C 2-6 alkenyl or halogenated C 2-6 Alkyne substitution,

[0009] The C-ring and D-ring are each independently a 6-10 member aromatic ring or a 5-12 member heteroaromatic ring.

[0010] Ring A and ring B are fused.

[0011] Ring A is a 5-6 membered heteroaromatic ring or a 5-6 membered heterocyclic ring.

[0012] Ring B can be a benzene ring, a 5-6 membered heteroaromatic ring, a 5-6 membered carbon ring, or a 5-6 membered heterocycle, and ring B may be optionally halogenated, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, C 1-6 Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclic, or 5-6 membered heteroaryl substitution,

[0013] R2 and R3 are each independently halogenated, -CN, -OH, -NH2, or C. 1-6 alkyl,

[0014] R4, R5, R6, and R7 are each independently H, halogen, -CN, -OH, -NH2, and -OC, respectively. 1-6 Alkyl, or C 1-6 alkyl,

[0015] R8 and R9 are each independently H, -OH, -NH2, or C. 1-6 Alkyl, or

[0016] R7 and R8 can be optionally connected together to form a 6-10 element ring.

[0017] R1 is or

[0018] p is 0, 1, or 2.

[0019] q can be 0, 1, or 2.

[0020] In some embodiments, R4, R5, R6, and R7 are each independently H, a halogen, -CN, -OH, -NH2, or C. 1-6 alkyl.

[0021] In some implementations, R 10 It is -CH2-CF3.

[0022] In another aspect, the present invention provides compounds of formula (I) or pharmaceutically acceptable salts, solvates, polymorphs or isomers thereof:

[0023]

[0024] in,

[0025] The C-ring and D-ring are each independently a 6-10 member aromatic ring or a 5-12 member heteroaromatic ring.

[0026] Ring A and ring B are fused.

[0027] Ring A is a 5-6 membered heteroaromatic ring or a 5-6 membered heterocyclic ring.

[0028] Ring B can be a benzene ring, a 5-6 membered heteroaromatic ring, a 5-6 membered carbon ring, or a 5-6 membered heterocyclic ring.

[0029] R2 and R3 are each independently halogenated, -CN, -OH, -NH2, or C. 1-6 alkyl,

[0030] R4, R5, R6, and R7 are each independently H, halogen, -CN, -OH, -NH2, or C. 1-6 alkyl,

[0031] R8 and R9 are each independently H, -OH, -NH2, or C.1-6 Alkyl, or

[0032] R7 and R8 can be optionally connected together to form a 6-10 element ring.

[0033] R1 is or

[0034] p is 0, 1, or 2.

[0035] q can be 0, 1, or 2.

[0036] In some embodiments, the C ring is a 5-6 membered heteroaromatic ring, preferably a pyrimidine ring.

[0037] In some implementations, the D ring is a benzene ring.

[0038] In some embodiments, ring A is a 5-6 membered heteroaromatic ring, and ring B is a benzene ring or a pyridine ring.

[0039] In some implementations, ring A is Ring B is a pyridine ring.

[0040] In some implementations, R4, R5, R6, and R7 are H.

[0041] In some implementations, R8 and R9 are H.

[0042] In some implementations, p is 0.

[0043] In some implementations, q is 1.

[0044] In some implementations, R1 is

[0045] In some embodiments, the present invention provides the following compounds

[0046]

[0047]

[0048]

[0049] Or its pharmaceutically acceptable salts, solvates, polymorphs or isomers.

[0050] Another aspect of the present invention relates to pharmaceutical compositions comprising the compounds of the present invention or pharmaceutically acceptable salts, solvates, polymorphs or isomers thereof, and pharmaceutically acceptable carriers.

[0051] In another aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, or a combination thereof, in the preparation of a medicament for treating diseases associated with FGFR; preferably, the FGFR-associated disease is a tumor; in some embodiments, the tumor has mutations in FGFR2 N549, V561, V565, N550, N540, V555, E566, K660, V550. In some embodiments, the FGFR-associated cancer is cholangiocarcinoma, urothelial carcinoma, lung cancer, bladder cancer, cervical cancer, endometrial cancer, breast cancer, thyroid cancer, colorectal cancer, gastric cancer, liver cancer, ovarian cancer, colorectal cancer, pancreatic cancer, gallbladder cancer, leukemia, multiple myeloma, Hodgkin's lymphoma, or melanoma.

[0052] The present invention also provides a use for treating diseases related to abnormal expression, mutation, or abnormal expression and activity of the FGFR receptor or its corresponding ligand.

[0053] The present invention also relates to a method for treating tumors resistant to FGFR inhibitors, the method comprising administering to a subject an effective amount of the compound of the present invention or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, or a combination thereof; in some embodiments, the FGFR-related diseases are cholangiocarcinoma, urothelial carcinoma, lung cancer, bladder cancer, cervical cancer, endometrial cancer, breast cancer, thyroid cancer, colorectal cancer, gastric cancer, liver cancer, ovarian cancer, colorectal cancer, pancreatic cancer, gallbladder cancer, leukemia, multiple myeloma, Hodgkin lymphoma, melanoma, etc., preferably liver cancer and cholangiocarcinoma.

[0054] In some embodiments of the present invention, the object of the present invention is a mammal, including humans. Invention Details

[0055] Exemplary embodiments utilizing the principles of the invention are set forth in the following detailed description of the invention. The features and advantages of the invention can be better understood by referring to the following summary of the invention.

[0056] It should be understood that the scope of protection of each aspect of the present invention is determined by the claims, and the methods and structures within the scope of these claims, as well as their equivalents, are all within the scope of these claims.

[0057] Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent applications, and publications cited in this document are incorporated herein in their entirety through reference.

[0058] It should be understood that the above summary and the following detailed description are exemplary and explanatory, and not intended to limit any subject matter of the invention. Unless otherwise specified, the singular form includes the plural form. Unless otherwise specified, the use of "or" or "or" means "and / or". Furthermore, the use of the term "comprising" and other forms such as "including," "containing," and "containing" are not limiting.

[0059] Some chemical terms

[0060] The terms “optional,” “optional,” or “optionally” mean that the event or condition described below may or may not occur, including both the occurrence and non-occurrence of the event or condition. For example, “optionally substituted alkyl” means “unsubstituted alkyl” or “substituted alkyl.” Furthermore, the optionally substituted group can be unsubstituted (e.g., -CH2CH3), fully substituted (e.g., -CF2CF3), monosubstituted (e.g., -CH2CH2F), or any level between monosubstituted and fully substituted (e.g., -CH2CHF2, -CF2CH3, -CFHCHF2, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution mode that is spatially impossible and / or cannot be synthesized is introduced.

[0061] Unless otherwise stated, conventional methods within the scope of the art, such as mass spectrometry, nuclear magnetic resonance, high-performance liquid chromatography, infrared and ultraviolet / visible spectroscopy, and pharmacological methods, are employed. Unless specifically defined herein, the terminology, experimental procedures, and techniques used herein in analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and patient treatment. For example, reactions and purifications can be carried out using the manufacturer's instructions for use of reagent kits, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein are generally carried out according to conventional methods well known in the art, based on descriptions in several summary and more specific documents cited and discussed herein. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural moieties and compounds.

[0062] When a substituent is described using a conventional chemical formula written from left to right, it also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0063] As used in this article, the terms "group" and "chemical group" refer to a specific part or functional group of a molecule. Chemical groups are often considered as chemical entities that are embedded in or attached to a molecule.

[0064] Some chemical groups named herein may be indicated by abbreviations to represent the total number of carbon atoms. For example, C1-C6 alkyl describes an alkyl group having a total of 1 to 6 carbon atoms, as defined below. The total number of carbon atoms indicated by the abbreviations does not include carbon atoms on possible substituents.

[0065] The terms “halogen,” “halogenated,” or “halogenated” refer to bromine, chlorine, fluorine, or iodine.

[0066] The compounds of the present invention may contain one or more (e.g., one, two, three, or four) isotopic substitutions. For example, in said compounds, H may be any isotopic form, including 1 H, 2 H (D or deuterium) and 3 H (T or tritium); C can be any isotopic form, including 12 C 13 C and 14 C and O can be any isotopic form, including 16 O and 18 O etc.

[0067] The terms "aromatic," "aromatic ring," "aromatic," "aromatic," and "aromatic ring" refer to a planar ring or ring portion having a delocalized electronic conjugated system containing 4n+2 electrons, where n is an integer. Aromatic rings can be formed from 5, 6, 7, 8, 9, or more than 9 atoms. Aromatic compounds can be optionally substituted and can be monocyclic or fused polycyclic. The term aromatic compounds includes all carbocyclic rings (such as benzene rings) and rings containing one or more heteroatoms (such as pyridine).

[0068] The term "heteroatom" or "hetero" refers to an atom other than carbon and hydrogen. Heteroatoms are independently selected from, but not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, selenium, and tin. In embodiments where two or more heteroatoms are present, the two or more heteroatoms may be identical to each other, or some or all of the two or more heteroatoms may be different from each other.

[0069] The term "dense" or "dense ring" refers to a ring structure in which two or more rings share one or more bonds.

[0070] The term "spiral" or "spiral ring" refers to a ring structure in which two or more rings share one or more atoms.

[0071] The term "alkyl" refers to a monovalent saturated hydrocarbon with optional substituted straight or optional substituted branched chains having 1-12 carbon atoms, preferably 1-8 carbon atoms, more preferably 1-6 carbon atoms, and connected to other parts of the molecule by single bonds, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, n-octyl, n-nonyl, n-decyl, etc.

[0072] The term "alkenyl" refers to a monovalent hydrocarbon group of optional substituted straight or optional substituted branched form, having one or more C=C double bonds and having 2 to about 10 carbon atoms, more preferably 2 to about 6 carbon atoms. The double bonds in these groups can be in cis or trans conformations and should be understood to include both isomers. Examples include, but are not limited to, vinyl (CH=CH2), 1-propenyl (CH2CH=CH2), isopropenyl (C(CH3)=CH2), butenyl, and 1,3-butadienyl. When alkenyl as defined herein appears in numerical ranges, e.g., "C2-C6 alkenyl" or "C..." 2-6 "Alkenyl" refers to an alkenyl group that can be composed of 2, 3, 4, 5, or 6 carbon atoms. In this article, alkenyl also includes cases where no numerical range is specified.

[0073] The term "alkynyl" refers to an optionally substituted straight-chain or branched monovalent hydrocarbon group having one or more C≡C triple bonds and having 2 to about 10 carbon atoms, more preferably 2 to about 6 carbon atoms. Examples include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1,3-butadiynyl. When alkynyl is used in this definition and a numerical range is included, for example, "C2-C6 alkynyl" or "C 2-6 "Alynyl" refers to an alkynyl group that can be composed of 2, 3, 4, 5, or 6 carbon atoms. In this article, alkynyl also includes cases where no numerical range is specified.

[0074] The terms "aryl" and "aromatic ring" refer to a fully carbon monocyclic or fused ring having a fully conjugated π-electron system, having 6-14 carbon atoms, preferably 6-12 carbon atoms, and most preferably 6 carbon atoms. The aryl group can be unsubstituted or substituted with one or more substituents, examples of which include, but are not limited to, alkyl, alkyloxy, aryl, aralkyl, amino, halogen, hydroxyl, sulfonyl, sulfinyl, phosphoryl, and heterocyclic groups. Non-limiting examples of unsubstituted aryl groups include, but are not limited to, phenyl, naphthyl, and anthraceneyl.

[0075] The terms "heteroaryl" and "heteroary ring" refer to a monocyclic or fused ring with 5-12 ring atoms, having 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, of which 1, 2, 3, or 4 are selected from N, O, and S, and the remaining ring atoms are C, and possessing a fully conjugated π-electron system. Heteroaryl groups can be unsubstituted or substituted, and the substituents include, but are not limited to, alkyl, alkyloxy, aryl, aralkyl, amino, halogen, hydroxyl, cyano, nitro, carbonyl, and heterocyclic groups. Non-limiting examples of unsubstituted heteroaryl groups include, but are not limited to, pyrrole, furanyl, thiophene, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, and triazineyl.

[0076] The term "cycloalkyl" as used alone or as part of other components herein refers to a stable, monovalent, non-aromatic monocyclic or polycyclic hydrocarbon group containing only carbon and hydrogen atoms. It may include fused ring, spiro ring, or bridged ring systems, containing 3-15 cyclic carbon atoms, preferably 3-10 cyclic carbon atoms, more preferably 3-8 cyclic carbon atoms, and may be saturated or unsaturated, and is linked to other parts of the molecule by single bonds. " Non-limiting examples of "cycloalkyl" include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0077] The term "carbon ring" refers to a structure consisting of a covalently closed ring of carbon, which can be saturated or partially unsaturated. A carbon ring can be formed by 3, 4, 5, 6, 7, 8, 9, or more atoms. The distinction between a carbon ring and a heterocycle lies in the fact that the ring skeleton of a heterocycle contains at least one atom different from carbon. The term "carbon ring" as used herein can refer to a monocyclic or polycyclic ring, including spirocyclic, fused, and bridged rings. Carbon rings can be arbitrarily substituted. Preferably, a "carbon ring" as used herein contains about 5 to about 20, 5 to 10, 5-8, or 5-6 skeletal ring atoms.

[0078] The terms "heterocyclic alkyl," "heterocyclic group," and "heterocycle" refer to a stable 3-18 member monovalent non-aromatic ring comprising 2-12 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, the heterocyclic group can be a monocyclic, bicyclic, tricyclic, or tetracyclic system, which may contain fused rings, spirocyclic, or bridged ring systems. The nitrogen, carbon, or sulfur on the heterocyclic group may be selectively oxidized, the nitrogen atom may be selectively quaternized, and the heterocyclic group may be partially or completely saturated. The heterocyclic group can be connected to the rest of the molecule via a single bond through a carbon atom or heteroatom on the ring. Heterocyclic groups containing fused rings may contain one or more aromatic or heteroaromatic rings, provided that the atoms connected to the rest of the molecule are atoms from non-aromatic rings. For the purposes of this application, the heterocyclic group is preferably a stable 4-11 valent monovalent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur; more preferably, it is a stable 4-8 valent monovalent non-aromatic monocyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocyclic groups include azirheptanyl, azirheptanyl, decahydroisoquinolinyl, dihydrofuranyl, dihydroindolyl, dioxopentyl, 1,1-dioxo-thiomorpholinyl, imidazoalkyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinoneyl, pyranyl, pyrazolyl, pyrrolidinyl, quinazinyl, quininecycloyl, tetrahydrofuranyl, tetrahydropyranyl, etc.

[0079] The term "polymorph" or "polymorphism" as used in this invention refers to the fact that the compounds of this invention have multiple crystal lattice forms. Some compounds of this invention may have more than one crystal form, and this invention covers all polymorphs or mixtures thereof.

[0080] Intermediate compounds and polymorphs of the compounds of this invention are also within the scope of this invention.

[0081] Unless otherwise specified, the olefin double bonds contained in the compounds of this invention include E and Z isomers.

[0082] It should be understood that the compounds of the present invention may contain asymmetric centers. These asymmetric centers may independently be R or S configurations. Some compounds of the present invention may also exhibit cis-trans isomerism, which will be apparent to those skilled in the art. It should be understood that the compounds of the present invention include their individual geometric isomers and stereoisomers, as well as mixtures thereof, including racemic mixtures. These isomers can be isolated from mixtures thereof by implementing or modifying known methods, such as chromatography and recrystallization techniques, or they can be prepared separately from suitable isomers of their intermediates.

[0083] The term “pharmaceutically acceptable salt” as used in this article includes both salts with added acid salts and salts with added alkali salts.

[0084] "Pharmaceutically acceptable salts" refer to salts that retain the biological potency and properties of the free base of a compound, are not biologically or otherwise undesirable, and are formed with inorganic acids, such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or organic acids, such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, decanoic acid, hexanoic acid, carbonic acid, cinnamic acid, citric acid, etc. "Pharmaceutical acceptable base salts" refer to salts that retain the biological potency and properties of the free acid of a compound, and are not biologically or otherwise undesirable. These salts are prepared by reacting a free acid with an inorganic or organic base. Salts formed by reacting with an inorganic base include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, etc. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and manganese salts.

[0085] Organic bases that form salts include, but are not limited to, primary amines, secondary amines, tertiary amines, cyclic amines, etc., such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purine, piperazine, piperidine, choline, and caffeine. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0086] Crystallization often produces solvates of the compounds of this invention. As used herein, the term "solvate" refers to a combination of one or more molecules of the compounds of this invention and one or more solvent molecules.

[0087] The solvent can be water, in which case the solvate is a hydrate. Alternatively, it can be an organic solvent. Therefore, the compounds of this invention can exist as hydrates, including monohydrates, dihydrates, hemihydrates, trihydrates, tetrahydrates, etc., and the corresponding solvated forms. The compounds of this invention can be true solvates, but in other cases, they may simply retain water or a mixture of water and some other solvents by chance. The compounds of this invention can react in a solvent or precipitate or crystallize in a solvent. The solvates of the compounds of this invention are also included within the scope of this invention.

[0088] As used herein, the term "pharmaceutical composition" refers to a formulation containing the compounds of the present invention and a medium generally accepted in the art for delivering biologically active compounds to mammals, such as humans. This medium includes all pharmaceutically acceptable carriers.

[0089] As used in this article, the term "acceptable" in relation to formulations, compositions, or ingredients means that it does not have a lasting harmful effect on the overall health of the treated subject.

[0090] As used herein, the term "pharmaceutically acceptable" means a substance (such as a 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., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0091] "Pharmaceutically acceptable carriers" include, but are not limited to, adjuvants, carriers, excipients, auxiliaries, deodorants, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants and wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that have been approved by the relevant government administrative departments for use in humans and domesticated animals.

[0092] As used herein, the terms “subject,” “patient,” “object,” or “individual” refer to an individual suffering from a disease, disorder, or symptom, including both mammals and non-mammals. Examples of mammals 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, and pigs; domesticated animals such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0093] The term "treatment" as used in this article refers to the treatment of relevant diseases or conditions in mammals, particularly humans, including...

[0094] (i) To prevent mammals, especially those previously exposed to a disease or condition but not yet diagnosed with it, from developing the corresponding disease or condition.

[0095] (ii) To suppress a disease or symptom, that is, to control its development;

[0096] (iii) To alleviate the disease or symptom, that is, to make the disease or symptom subside;

[0097] (iv) Relieve symptoms caused by disease or illness.

[0098] The terms “disease” and “symptom” used in this article may be used interchangeably or have different meanings, because some specific diseases or symptoms do not yet have known causative factors (so the cause of the disease is still unclear), so they cannot be recognized as diseases but can only be regarded as unwanted conditions or syndromes. These syndromes have more or less some specific symptoms that have been confirmed by clinical researchers.

[0099] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to an amount of at least one drug or compound that, when taken, is sufficient to alleviate, to some extent, one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief 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 containing the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0100] As used herein, the terms “administration,” “application,” “dosage,” etc., refer to methods that deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral administration, duodenal administration, parenteral administration (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), local administration, and rectal administration. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

[0101] Preparation of the compounds of the present invention

[0102] The following reaction route illustrates a method for preparing the compounds of the present invention.

[0103] It should be understood that, in the following description, the combination of substituents and / or variables of the molecular formula is permitted only in the case of forming a stable compound.

[0104] Those skilled in the art will also understand that, in the processes described below, the functional groups of the intermediate compounds may need to be protected by suitable protecting groups. These functional groups include hydroxyl, amino, mercapto, and carboxyl groups. Suitable hydroxyl protecting groups include trialkylsilyl or diarylalkylsilyl (e.g., tert-butylmethylsilyl, tert-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, etc. Suitable amino, amidine, and guanidine protecting groups include tert-butyloxycarbonyl, benzyloxycarbonyl, etc. Suitable mercapto protecting groups include -C(O)-R″ (R″ represents alkyl, aryl, or arylalkyl), p-methoxybenzyl, triphenylmethyl, etc. Suitable carboxyl protecting groups include alkyl, aryl, or arylalkyl esters. Protecting groups can be added or removed using standard techniques known to those skilled in the art. Example

[0105] The following non-limiting embodiments are merely illustrative and do not limit the invention in any way.

[0106] Unless otherwise specified, temperatures are in Celsius. Reagents were purchased from commercial suppliers such as Sinopharm Chemical Reagents Beijing Co., Ltd., Alfa Aesar, or Beijing Bailingwei Technology Co., Ltd., and these reagents are ready for use without further purification, unless otherwise specified.

[0107] Unless otherwise specified, the following reactions are carried out at room temperature, in anhydrous solvents, under positive pressure of nitrogen or argon, or using a drying tube; the reaction flask is fitted with a rubber diaphragm to allow for the addition of substrates and reagents via syringe; glassware is dried by drying and / or heating.

[0108] Unless otherwise specified, column chromatography purification used 200-300 mesh silica gel from Qingdao Ocean Chemical Plant; preparative thin-layer chromatography used thin-layer chromatography silica gel pre-plates (HSGF254) produced by Yantai Chemical Industry Research Institute; MS determination was performed using a ThermoLCQ Fleet (ESI) liquid chromatography-mass spectrometry system; optical rotation determination was performed using an SGW-3 automatic polarimeter from Shanghai Shenguang Instrument Co., Ltd.

[0109] NMR data ( 1 H NMR was performed using a Varian instrument at 400 MHz. Solvents used for NMR data included CDCl3, CD3OD, D2O, and DMSO-d6, with tetramethylsilane (0.00 ppm) or residual solvents as the reference (CDCl3: 7.26 ppm; CD3OD: 3.31 ppm; D2O: 4.79 ppm; d6-DMSO: 2.50 ppm). When indicating peak diversity, the following abbreviations are used to represent different peak shapes: s (singleton), d (doublet), t (triplet), q (quartet), m (multiplex), br (broad peak), dd (double doublet), dt (double triplet). If coupling constants are given, they are expressed in Hertz (Hz).

[0110] Example 1: 6-(4-acrylamidophenyl)-7-(4-((4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0111]

[0112] Step 1: 6-amino-7-(4-(4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-nitrile (1c)

[0113] At room temperature, compounds 1a (1.75 g) and 1b (2.15 g) were dissolved in methanol (30 mL), and acetone nitrile (1.0 g) and 1,8-diazabicyclo[5.4.0]undec-7-ene (5.0 g) were added. The mixture was heated to 70 °C and stirred for 1 hour. After cooling to room temperature, 20 mL of ethanol was added, resulting in a large amount of precipitate. The precipitate was filtered and dried under vacuum to give compound 1c (2.9 g).

[0114] Step 2: 6-Iodo-7-(4-(4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-nitrile (1d)

[0115] At room temperature, compound 1c (398 mg) and diiodomethane (2.67 g) were dissolved in chloroform (10 mL), and isoamyl nitrite (1 mL) was slowly added, followed by stirring for 12 hours. The reaction solution was concentrated under reduced pressure and then poured into a saturated sodium bicarbonate aqueous solution (50 mL). The solution was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 30:1 (V:V)) to give compound 1d (225 mg).

[0116] Step 3: 6-Iodo-7-(4-(4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide (1e)

[0117] At 0°C, 30% hydrogen peroxide (1 mL) was slowly added dropwise to a DMSO (10 mL) solution of compound 1d (225 mg) and potassium carbonate (300 mg), and the mixture was reacted at room temperature for 2 hours. The reaction solution was poured into water (50 mL), extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1 (V:V)) to give compound 1e (133 mg).

[0118] Step 4: 6-(4-acrylamidophenyl)-7-(4-((4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2] ′ [1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide (compound 1)

[0119] Under nitrogen protection, compound 1e (53 mg), compound 1f (30 mg), and potassium carbonate (28 mg) were dissolved in a mixed solvent (10 mL) of dioxane and water (4:1). Tetra(triphenylphosphine)palladium (11 mg) was added, and the mixture was heated to 90 °C and stirred for 3 hours. After cooling to room temperature, saturated sodium chloride aqueous solution (100 mL) was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1 (V:V)) to obtain compound 1 (44 mg).

[0120] MSm / z [LC-MS]: 547.17 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.36 (s, 1H), 8.42-8.44 (m, 2H), 7.75 (d, J=8.8Hz, 2H), 7.44 ( d, J=8.8Hz, 2H), 7.44 (d, J=8.8Hz, 2H), 7.33 (dd, J=8.8Hz, 4.8Hz, 1H), 7.19 (d, J=8.8H z, 2H), 7.13 (d, J=5.2Hz, 1H), 7.03 (dd, J=8.0Hz, 1.2Hz, 1H), 6.44 (dd, J=17.2Hz, 10.0 Hz, 1H), 6.26 (dd, J=16.8Hz, 2.0Hz, 1H), 5.77 (dd, J=10.0Hz, 2.0Hz, 1H), 2.37 (s, 3H).

[0121] Example 2: 6-(4-(2-fluoroacrylamido)phenyl)-7-(4-(4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0122]

[0123] Under nitrogen protection, compound 1e (53 mg), compound 2a (29 mg), and potassium carbonate (28 mg) were dissolved in a mixed solvent (10 mL) of dioxane and water (4:1). Tetra(triphenylphosphine)palladium (11 mg) was added, and the mixture was heated to 90 °C and stirred for 2 hours. After cooling to room temperature, saturated sodium chloride aqueous solution (100 mL) was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1 (V:V)) to obtain compound 1 (32 mg).

[0124] MSm / z [LC-MS]: 565.16 [M+H] + .

[0125] Example 3: 6-(4-(2-fluoroacrylamido)phenyl)-7-(4-(4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0126]

[0127] Under nitrogen protection, compound 1e (53 mg), compound 3a (29 mg), and potassium carbonate (28 mg) were dissolved in a mixed solvent (10 mL) of dioxane and water (4:1). Tetra(triphenylphosphine)palladium (11 mg) was added, and the mixture was heated to 90 °C and stirred for 2 hours. After cooling to room temperature, saturated sodium chloride aqueous solution (100 mL) was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane: methanol = 20:1 (V:V)) to obtain compound 1 (31 mg).

[0128] MSm / z [LC-MS]: 561.18 [M+H] + .

[0129] Example 4: 6-(4-acrylamidophenyl)-7-(3-fluoro-4-((4-methylpyrimidin-2-yl)oxy)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0130]

[0131] The synthesis method of Example 1 was used to obtain compound 4 (24 mg) using 4a as the starting material.

[0132] MSm / z [LC-MS]: 565.16 [M+H] + . 1 H NMR (400MHz, CDCl3), 8.42 (dd, J=4.8Hz, 1.6Hz, 1H), 8.34 (d, J=5.2Hz, 1H), 7.68 (d, J=8. 0Hz, 2H), 7.33-7.35 (m, 3H), 7.23-7.27 (m, 1H), 7.20 (dd, J=8.8Hz, 1.2Hz, 1H), 7.13-7.17 (m, 2H), 7.09 (dd, J=8.0Hz, 4.8Hz, 1H), 6.93 (d, J=5.2Hz, 1H), 6.48 (d, J=16.8Hz, 1H), 6. 27 (dd, J=16.8Hz, 10.0Hz, 1H), 5.84 (d, J=10.0Hz, 1H), 5.31-5.39 (brS, 2H), 2.48 (s, 3H).

[0133] Example 5: 7-(3-fluoro-4-((4-methylpyrimidin-2-yl)oxy)phenyl)-6-(4-(2-fluoroacrylamido)phenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0134]

[0135] The synthesis method of Example 2 was used to obtain compound 5 (14 mg) using 4 d as the starting material.

[0136] MSm / z [LC-MS]: 583.15 [M+H] + . 1 H NMR (400MHz, DMSO), 10.34 1 H NMR (400MHz, CDCl3), (s, 1H), 8.44-8.48 (m, 2H), 8.21 (dd, J=4.8Hz, 1.6Hz, 1H), 7.67 (d, J=8.4Hz, 2H), 7.32- 7.39 (m, 6H), 7.16-7.20 (m, 3H), 5.70 (dd, J=47.6Hz, 3.6Hz, 1H), 5.42 (dd, J=16.0Hz, 3.6Hz, 1H), 2.39 (s, 3H).

[0137] Example 6: 7-(3-fluoro-4-((4-methylpyrimidin-2-yl)oxy)phenyl)-6-(4-methacrylamidophenyl)pyrrolo[2′,1′:2,3]thiazo[5,4-b]pyridine-8-carboxamide

[0138]

[0139] Compound 6 (12 mg) was obtained by using 4 d as the starting material in the synthesis method of Example 3.

[0140] MS m / z [LC-MS]: 579.17 [M+H] + .

[0141] The following compounds were synthesized using a method similar to that used in Examples 1-6:

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150] Example 31: 8-(4-acrylamidophenyl)-7-(3-methoxy-4-((2,2,2-trifluoroethyl)carbamoyl)phenyl)pyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridine-6-carboxamide

[0151]

[0152] Step 1: Methyl 4-(8-amino-6-cyanopyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridin-7-yl)-2-methoxybenzoate (31c)

[0153] Compounds 31a (1.75 g) and 31b (1.94 g) were dissolved in methanol (50 mL) at room temperature, followed by the addition of acetone nitrile (1.5 g) and 1,8-diazabicyclo[5.4.0]undec-7-ene (5.0 g). The reaction mixture was heated to 70 °C and stirred for 2 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 25:1 (V:V)) to give compound 31c (0.9 g).

[0154] Step 2: Methyl 4-(6-cyano-8-iodopyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridin-7-yl)-2-methoxybenzoate (31d)

[0155] At room temperature, compound 31c (378 mg) and diiodomethane (2.67 g) were dissolved in acetonitrile (20 mL), and isoamyl nitrite (2 mL) was added. The mixture was then heated to 50 °C and stirred for 1 hour. The reaction solution was concentrated under reduced pressure and then poured into a saturated sodium bicarbonate aqueous solution (50 mL). The solution was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol = 30:1 (V:V)) to give compound 31d (99 mg).

[0156] Step 3: 4-(3-carbamoyl-1-iodobenzo[d]pyrrolo[2,1-b]thiazolyl-2-yl)benzoic acid (31e)

[0157] At 0°C, 30% hydrogen peroxide (1 mL) was added dropwise to a DMSO (10 mL) solution of compound 31d (99 mg) and potassium carbonate (150 mg), and the mixture was reacted at room temperature for 5 hours. The reaction solution was poured into water (50 mL), the pH was adjusted to 4 with dilute hydrochloric acid (1 M), and then extracted with n-butanol. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 31e (53 mg).

[0158] Step 4: 8-Iodo-7-(3-methoxy-4-(2,2,2-trifluoroethyl)carbamoyl)phenyl)pyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridine-6-carboxamide (31f)

[0159] At room temperature, compound 31e (53 mg) and 2,2,2-trifluoroethylamine (20 mg) were dissolved in DMF (50 mL), HATU (80 mg) and DIEA (100 mg) were added, and the mixture was stirred at room temperature for 12 hours. A saturated aqueous sodium chloride solution (50 mL) was added, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol = 20:1 (V:V)) to give compound 31f (24 mg).

[0160] Step 5: 8-(4-acrylamidophenyl)-7-(3-methoxy-4-((2,2,2-trifluoroethyl)carbamoyl)phenyl)pyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridine-6-carboxamide (compound 31)

[0161] Under nitrogen protection, compound 31f (24 mg), compound 1f (15 mg), and potassium carbonate (20 mg) were dissolved in a mixed solvent (5 mL) of dioxane and water (4:1). Tetra(triphenylphosphine)palladium (6 mg) was added, and the mixture was heated to 90 °C and stirred for 5 hours. After cooling to room temperature, saturated sodium chloride aqueous solution (100 mL) was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by thin-layer chromatography (dichloromethane:methanol = 15:1 (V:V)) to give compound 31 (13 mg).

[0162] MSm / z [LC-MS]: 594.15 [M+H] + . 1H NMR (400MHz, DMSO), 10.19 (s, 1H), 8.66 (t, J=5.6Hz, 1H), 8.44 (dd, J=7.6Hz, 1.6Hz, 1H), 8.22 (dd, J=4.8Hz, 1.6 Hz, 1H), 6.82-7.74 (brs, 1H), 7.68 (d, J=8.0Hz, 1H), 7.57 (d, J=8.8Hz, 2H), 7.37 (dd, J=8.0Hz, 4.8Hz, 1H), 7.32 (d, J=8.8Hz, 2H), 7.06 (d, J=1.2Hz, 1H), 6.98 (dd, J=8.0Hz, 1.2Hz, 1H), 6.41 (dd, J=16.8Hz, 10.4Hz, 1H), 6.24( dd, J=16.8Hz, 1.6Hz, 1H), 5.74 (dd, J=10.4Hz, 1.6Hz, 1H), 5.10-5.62 (brs, 1H), 4.02-4.11 (m, 2H), 3.73 (s, 3H).

[0163] Example 32: 8-(4-(2-fluoroacrylamido)phenyl)-7-(3-methoxy-4-((2,2,2-trifluoroethyl)carbamoyl)phenyl)pyrrolo[2′,1′:2,3]thiazo[4,5-b]pyridine-6-carboxamide

[0164]

[0165] Under nitrogen protection, compound 31f (57 mg), compound 2a (29 mg), and potassium carbonate (28 mg) were dissolved in a mixed solvent (10 mL) of dioxane and water (4:1). Tetra(triphenylphosphine)palladium (11 mg) was added, and the mixture was heated to 90 °C and stirred for 2 hours. After cooling to room temperature, saturated sodium chloride aqueous solution (100 mL) was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol = 20:1 (V:V)) to give compound 32 (22 mg).

[0166] MSm / z [LC-MS]: 612.15 [M+H] + . 1H NMR (400MHz, DMSO), 10.31 (s, 1H), 8.66 (t, J=6.4Hz, 1H), 8.44 (dd, J=8.4Hz, 1.6Hz, 1H), 8.22 (dd, J=4.8Hz, 1.6Hz, 1H), 7.68 (d, J=8.8Hz, 1H), 7.62 (d, J=8.4Hz, 2H), 7. 34-7.39 (m, 3H), 7.07 (d, J = 1.2Hz, 1H), 6.98 (dd, J = 8.4Hz, 1.6Hz, 1H), 5.69 (dd, J = 4 8.0Hz, 3.6Hz, 1H), 5.42 (dd, J=15.6Hz, 4.0Hz, 1H), 4.02-4.11 (m, 2H), 3.73 (s, 3H).

[0167] The following compounds were synthesized using a method similar to that used in Examples 31-32:

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174] Bioactivity experiment

[0175] 1. Compounds on FGFR2 WT In vitro enzyme activity assay

[0176] The compound in this patent affects FGFR2. WT Enzymatic activity inhibits IC 50 Value determination was performed using homogeneous time-resolved fluorescence (HTRF). The compound was serially diluted 5-fold (total of 7 concentrations) starting at 0.2 mM with 100% DMSO. 2 μL of each concentration was added to 48 μL of reaction buffer (50 mM HEPES pH 7.5, 0.1 mM Na3VO4, 5 mM MnCl2, 1 mM DTT, 0.001% Tween 20, and 0.01% BSA) and mixed thoroughly. 2.5 μL of the diluted compound was added to a 384-well plate (OptiPlate-384, purchased from PerkinElmer), followed by 5 μL of GST-FGFR2. WT(399-821aa, final concentration 0.3nM), centrifuged and mixed, then pre-incubated at 23°C for 1 hour. 2.5 μL of ATP (final concentration 1 μM) and TK Peptide Substrate mixture (final concentration 1 μM, purchased from Cisbio) was added to initiate the reaction, for a total reaction volume of 10 μL. The 384-well plate was incubated at 23°C for 1 hour, then 5 μL of TK Antibody (purchased from Cisbio) and 5 μL of Streptavidi N-XL665 (purchased from Cisbio) were added to stop the reaction. After a second incubation of 1 hour, fluorescence values ​​were read on an Envision (purchased from PerkinElmer) microscope (320 nm excitation, detection of emission at 665 nm and 620 nm, the ratio of which represents the enzyme activity signal). FGFR2 was measured at 7 concentrations for each compound. WT The enzymatic activity signal was obtained, and the IC50 of the compound was calculated using GraphPad Prism software. 50 value.

[0177] 2. Compounds on FGFR2 N549K In vitro enzyme activity assay

[0178] The compound in this patent affects FGFR2. N549K Enzymatic activity inhibits IC 50 Value determination was performed using homogeneous time-resolved fluorescence (HTRF). The compound was serially diluted 5-fold (total of 7 concentrations) starting at 0.2 mM with 100% DMSO. 2 μL of each concentration was added to 48 μL of reaction buffer (50 mM HEPES pH 7.5, 0.1 mM Na3VO4, 5 mM MnCl2, 1 mM DTT, 0.001% Tween 20, and 0.01% BSA) and mixed thoroughly. 2.5 μL of the diluted compound was added to a 384-well plate (OptiPlate-384, purchased from PerkinElmer), followed by 5 μL of GST-FGFR2. N549K(399-821aa, final concentration 0.5nM), centrifuged and mixed, pre-incubated at 23°C for 1 hour, then 2.5 μL of ATP (final concentration 30nM) and TK Peptide Substrate mixture (final concentration 1 μM, purchased from Cisbio) was added to start the reaction, with a total reaction volume of 10 μL. The 384-well plate was incubated at 23°C for 2 hours, then 5 μL of TK Antibody (purchased from Cisbio) and 5 μL of Streptavidi N-XL665 (purchased from Cisbio) were added to stop the reaction. After a second incubation of 1 hour, fluorescence values ​​were read on an Envision (purchased from PerkinElmer) (320nm excitation, detection of emission at 665nm and 620nm, the ratio of which is the enzyme activity signal). FGFR2 was measured at 7 concentrations for each compound. N549K The enzymatic activity signal was obtained, and the IC50 of the compound was calculated using GraphPadPrism software. 50 value.

[0179] 3. Compounds on FGFR3 WT In vitro enzyme activity assay

[0180] The compound in this patent affects FGFR3. WT The IC50 value of enzyme activity inhibition was determined using homogeneous time-resolved fluorescence (HTRF). The compound was serially diluted 5-fold with 100% DMSO starting at 0.2 mM (total of 7 concentrations). 2 μL of each concentration was added to 48 μL of reaction buffer (50 mM HEPES pH 7.5, 0.1 mM Na3VO4, 5 mM MnCl2, 1 mM DTT, 0.001% Tween 20, and 0.01% BSA) and mixed thoroughly. 2.5 μL was added to a 384-well plate (OptiPlate-384, purchased from PerkinElmer), followed by 5 μL of GST-FGFR3. WT(436-806aa, final concentration 1.0nM), centrifuged and mixed, the 384-well plate was pre-incubated at 23°C for 1 hour, then 2.5 μL of ATP (final concentration 15 μM) and TK Peptide Substrate mixture (final concentration 1 μM, purchased from Cisbio) was added to start the reaction, with a total reaction volume of 10 μL. The 384-well plate was incubated at 23°C for 2 hours, then 5 μL of TK Antibody (purchased from Cisbio) and 5 μL of Streptavidin-XL665 (purchased from Cisbio) were added to stop the reaction. After incubation for another 1 hour in an incubator, fluorescence values ​​were read on an Envision (purchased from PerkinElmer) instrument (excitation at 320 nm, detection of emission at 665 nm and 620 nm, the ratio of which is the enzyme activity signal). The enzymatic activity signal of FGFR3w for each compound was measured at 7 concentrations, and the IC50 value of the compound was calculated using GraphPadPrism software.

[0181] 4. Compounds on FGFR1 WT In vitro enzyme activity assay

[0182] The compound in this patent affects FGFR1. WTThe IC50 value of enzyme activity inhibition was determined using homogeneous time-resolved fluorescence (HTRF). The compound was serially diluted 5-fold with 100% DMSO starting at 0.2 mM (totaling 7 concentrations). 2 μL of each concentration was added to 48 μL of reaction buffer (50 mM HEPES pH 7.5, 0.1 mM Na3VO4, 5 mM MnCl2, 1 mM DTT, 0.001% Tween 20, and 0.01% BSA) and mixed thoroughly. Add 2.5 μL of the diluted compound to a 384-well plate (OptiPlate-384, purchased from PerkinElmer), then add 5 μL of GST-FGFR1WT (398-822aa, final concentration 1.0 nM), centrifuge to mix, and incubate at 23°C for 1 hour. Then add 2.5 μL of a mixture of ATP (final concentration 2 μM) and TK Peptide Substrate (final concentration 1 μM, purchased from Cisbio) to start the reaction, for a total reaction volume of 10 μL. Incubate the 384-well plate at 23°C for 2 hours, then add 5 μL of TK Antibody (purchased from Cisbio) and 5 μL of Streptavidin-XL665 (purchased from Cisbio) to stop the reaction. After a second 1-hour incubation in an incubator, fluorescence values ​​were read on an Envision (PerkinElmer) microscope (excitation at 320 nm, detection of emission at 665 nm and 620 nm, the ratio of which represents the enzyme activity signal). FGFR1 was measured at seven concentrations for each compound. WT The enzymatic activity signal was obtained, and the IC50 of the compound was calculated using GraphPadPrism software. 50 value.

[0183] 5. Compounds on FGFR2 N549K -PHGDH / 3T3 cell proliferation activity assay

[0184] FGFR2 fusion cell line constructed using lentiviral infection N549K -PHGDH / 3T3 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS, purchased from Gibco) and 1% penicillin / streptomycin antibiotics (P / S, purchased from LifeTechnology) at 37°C and 5% CO2. The day before compound detection, FGFR2... N549K-PHGDH / 3T3 cells were seeded at a concentration of 1000 cells / 195 μL / well in 96-well plates (#3917, purchased from Corning). After 24 hours, the compound was serially diluted 3-fold with 100% DMSO, starting at 10 mM (totaling 10 concentrations). 2 μL of each concentration was then added to 48 μL of DMEM medium for further dilution. 5 μL of each diluted compound was added to the seeded cell suspension, and the compound and cells were incubated together in a cell culture incubator for 72 hours (3 days). After aspirating the culture medium, 25 μL of Cell-Titer Glo (G7570, purchased from Promega) reagent was added, and the cells were incubated again for 5-10 minutes. The cells were then cultured in CLARIO star... Plus Fluorescence values ​​were read on a multi-functional microplate reader, and the IC50 of the compound on cell proliferation was calculated using GraphPad Prism software. 50 value.

[0185] Table 1. Protein activity and cell inhibitory activity data of some examples

[0186]

[0187]

[0188] The result "A" indicates IC. 50 <1nM; "B" indicates IC 50 <10nM; "C" indicates IC 50 >20 nM; "-" indicates untested

Claims

1. A compound of formula (II) or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof: in, R is Or -(CO)-NH-R 10 , R 10 C 1-6 Alkyl group, -(CH2) 0-2 -(3-8 membered cycloalkyl), or -(CH2) 0-2 -(3-8 membered heterocyclic group), wherein the alkyl, cycloalkyl, and heterocyclic group are optionally converted by halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl, halogenated C 1-6 Alkyl, Halogenated C 2-6 alkenyl or halogenated C 2-6 Alkyne substitution, The C-ring and D-ring are each independently a 6-10 member aromatic ring or a 5-12 member heteroaromatic ring. Ring A and ring B are fused. Ring A is a 5-6 membered heteroaromatic ring or a 5-6 membered heterocyclic ring. Ring B can be a benzene ring, a 5-6 membered heteroaromatic ring, a 5-6 membered carbon ring, or a 5-6 membered heterocycle, and ring B may be optionally halogenated, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, C 1-6 Alkyl, 3-8 membered cycloalkyl, 3-8 membered heterocyclic, or 5-6 membered heteroaryl substitution, R2 and R3 are each independently halogenated, -CN, -OH, -NH2, or C. 1-6 alkyl, R4, R5, R6, and R7 are each independently H, halogen, -CN, -OH, -NH2, and -OC. 1-6 Alkyl, or C 1-6 alkyl, R8 and R9 are each independently H, -OH, -NH2, or C. 1-6 Alkyl, or R7 and R8 can be optionally connected together to form a 6-10 element ring. R1 is or p is 0, 1, or 2. q can be 0, 1, or 2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, wherein R4, R5, R6, and R7 are each independently H, a halogen, -CN, -OH, -NH2, or C. 1-6 alkyl.

3. The compound according to claim 2, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, wherein R 10 It is -CH2-CF3.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, having the structure shown in formula (I): in, The C-ring and D-ring are each independently a 6-10 member aromatic ring or a 5-12 member heteroaromatic ring. Ring A and ring B are fused. Ring A is a 5-6 membered heteroaromatic ring or a 5-6 membered heterocyclic ring. Ring B can be a benzene ring, a 5-6 membered heteroaromatic ring, a 5-6 membered carbon ring, or a 5-6 membered heterocyclic ring. R2 and R3 are each independently halogenated, -CN, -OH, -NH2, or C. 1-6 alkyl, R4, R5, R6, and R7 are each independently H, halogen, -CN, -OH, -NH2, or C. 1-6 alkyl, R8 and R9 are each independently H, -OH, -NH2, or C. 1-6 Alkyl, or R7 and R8 can be optionally connected together to form a 6-10 element ring. R1 is or p is 0, 1, or 2. q can be 0, 1, or 2.

5. The compound according to claim 4 or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, wherein the C ring is a 5-6 membered heteroaromatic ring and the D ring is a benzene ring.

6. The compound according to claim 4 or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, wherein ring A is a 5-6 membered heteroaromatic ring and ring B is a benzene ring or a pyridine ring.

7. The compound according to claim 4 or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, wherein R4, R5, R6 and R7 are H, and R8 and R9 are H.

8. The compound according to claim 4 or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, wherein p is 0 and q is 1.

9. The compound according to claim 4, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, wherein R1 is...

10. The following compounds or their pharmaceutically acceptable salts, solvates, polymorphs or isomers 11. A pharmaceutical composition comprising the compound according to any one of claims 1-10, or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, and a pharmaceutically acceptable carrier.

12. Use of the compound of any one of claims 1-10 or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, or the pharmaceutical composition of claim 11 in the preparation of a medicament for treating diseases associated with FGFR.

13. The use according to claim 12, wherein the disease associated with FGFR is cholangiocarcinoma, urothelial carcinoma, lung cancer, bladder cancer, cervical cancer, endometrial cancer, breast cancer, thyroid cancer, intestinal cancer, gastric cancer, liver cancer, ovarian cancer, colorectal cancer, pancreatic cancer, gallbladder cancer, leukemia, multiple myeloma, Hodgkin's lymphoma, or melanoma.

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