Novel heterocyclic compound and application thereof

Abstract

The invention provides an RET selective inhibitor and a preparation method and application thereof. The present invention provides compounds of formula (II) and pharmaceutically acceptable salts, solvates, polymorphs or isomers thereof, pharmaceutical compositions comprising these compounds, and the use of such compounds in the treatment of RET-related diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a class of compounds, as well as pharmaceutically acceptable salts, hydrates, solvates or prodrugs of said compounds, methods for their preparation, and their use as therapeutic agents, particularly as RET inhibitors, and in medicaments for the treatment and / or prevention of hyperproliferative diseases such as cancer, and / or virus-induced infectious diseases, and / or cardiovascular diseases and / or diseases caused by fungi. Background Technology

[0002] The REarranged during Transfection (RET) gene is located on chromosome 10 and encodes the RET protein, a proto-oncogene tyrosine protein kinase receptor located on the cell membrane. Abnormalities in the RET gene can lead to abnormal RET dimerization or promote ligand-independent activation, which is a carcinogenic driver of various solid tumors. Point mutations in the RET gene, such as V804M, M918T, C620R / S, C634R / W / Y, can promote abnormal RET dimerization or enhance its ATP-binding ability, which are commonly seen in patients with multiple endocrine tumors (MEN2A, MEN2B) and 50% of patients with medullary thyroid carcinoma (MTC) (Ann Oncol. 2016; 27(7): 1286-1291). Chromosomal rearrangements of the RET gene can lead to the fusion of the RET gene kinase domain with different genes such as kif5b, ccdc6, and ncoa4, resulting in high RET expression or ligand-independent dimerization activation, which becomes a driving factor for malignant tumor proliferation (Transl Lung Cancer Res. 2015; 4(2): 156-164). RET gene fusions are present in 5%-40% of papillary thyroid carcinomas (PTCs) and 1-2% of non-small cell lung cancer (NSCLC) patients (Oncologist. 2013; 18(7): 865-875); as well as various solid tumors, including prostate cancer and breast cancer (Clin Cancer Res, 2017. 23(8): p. 1988-1997; Front Oncol, 2023. 13: p. 1090757). NSCLC patients with RET fusion-positive NSCLC tend to be younger (≤60 years old), have little or no history of smoking, have later disease stages, and have a higher risk of brain metastasis (Nat Rev ClinOncol, 2018.15(3): p.151-167; Journal of Clinical Oncology, 2012.30(35): p.4352-4359).

[0003] In 2012, RET fusion positivity was first identified as a driving factor for NSCLC (Nat Med, 2012.18(3): p.375-7.). At that time, only a few multi-kinase inhibitors (MKIs) with RET inhibitory activity were available, including Cabozantinib and Vandetanib. Preliminary data showed that MKIs had limited clinical benefits for RET fusion-positive NSCLC. Therefore, the development of highly selective RET inhibitors has attracted the research interest of many pharmaceutical companies. The development of specific RET inhibitors has become an emerging treatment approach to improve the therapeutic effect of RET-driven cancers.

[0004] Recently, two selective RET inhibitors, selpercatinib and pralsetinib, have demonstrated rapid and durable clinical efficacy in advanced RET-altered solid tumors with fewer treatment-related adverse events. Pralsetinib is currently approved in China for locally advanced or metastatic RET fusion-positive NSCLC following platinum-based chemotherapy.

[0005] Despite the encouraging efficacy of highly selective RET tyrosine kinase inhibitors (RET-TKIs), drug resistance remains a significant and unavoidable problem. In 2020, the first report of RET fusion-positive NSCLC patients developing RET G810 solvent front mutations after selpercatinib treatment led to resistance and disease progression (J ThoracOncol. 2020 April; 15(4): 541-549). In 2021, RET fusion-positive NSCLC patients were also reported to develop RET L730 roof mutations after pralsetinib treatment, resulting in pralsetinib resistance (Precision Oncology (2021) 5: 48). It is anticipated that as more patients receive RET-selective inhibitors, the increasing number of resistant patients will become a major issue. Therefore, there is a significant clinical need to develop next-generation RET inhibitors targeting RET resistance mutations to effectively overcome the resistance problem in these patients. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a novel compound, and its pharmaceutically acceptable salts, solvates, polymorphs, or isomers as agents capable of inhibiting proteins such as RET mutations, reducing or inhibiting RET mutations in cells, and applicable in drugs for the treatment and / or prevention of diseases mediated by proteins such as RET, particularly hyperproliferative diseases or induced infectious diseases and / or cardiovascular diseases and / or diseases caused by fungi.

[0007] In one aspect, the present invention provides compounds of formula (II), or pharmaceutically acceptable salts, solvates, polymorphs, or isomers thereof:

[0008]

[0009] in,

[0010] Ring B is or The * key is connected to L1.

[0011] L1 is -L2-L4-, -L2-L5-, -L2-L6-, -L2-L3-L4-, -L2-L3-L5-, or -L2-L3-L6-,

[0012] L2 is a 5-6 membered heteroaryl ring.

[0013] L3 is a 4-8 membered heterocyclic ring.

[0014] L4 is -CH2-.

[0015] L5 is -CH2-NR-,

[0016] L6 is -(CO)-NR-,

[0017] L is or

[0018] X is N or CH.

[0019] Y is either N or CH.

[0020] R1 is H or methyl.

[0021] R2 is methyl, or

[0022] R1 and R2 are connected together to form a 5-6 member heterocyclic ring.

[0023] Ring A is a 6-10 aryl group or a 5-12 heteroaryl group, wherein the aryl and heteroaryl groups may optionally be converted by halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted,

[0024] R 10 It is a 5-6 member heteroaryl or -OC 1-6 Alkyl groups, wherein the heteroaryl group may optionally be replaced by halogen, -CN, -OH, -NH2, -OC 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), or C 1-6 Alkyl substitution,

[0025] R can be independently H or C. 1-6 alkyl,

[0026] n is 0 or 1.

[0027] In some embodiments, ring A is a 5-6 membered heteroaryl group, which may optionally be replaced by a halogen, -CN, -OH, -NH2, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

[0028] In another aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof.

[0029]

[0030] in,

[0031] L is or

[0032] X is N or CH.

[0033] Y is either N or CH.

[0034] R1 is H or methyl.

[0035] R2 is methyl, or

[0036] R1 and R2 are connected together to form a 5-6 member heterocyclic ring.

[0037] Ring A is a 6-10 aryl group or a 5-12 heteroaryl group, wherein the aryl and heteroaryl groups may optionally be converted by halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

[0038] In some embodiments, ring A is a 5-6 membered heteroaryl group, which may optionally be replaced by a halogen, -CN, -OH, -NH2, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

[0039] In some embodiments, the present invention provides the following compounds, or pharmaceutically acceptable salts, solvates, polymorphs, or isomers thereof:

[0040]

[0041]

[0042] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, and optionally comprising a pharmaceutically acceptable carrier.

[0043] In another aspect, the present invention provides a method for treating RET-related diseases, the method comprising administering to a subject an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph or isomer thereof, or a combination thereof; in some embodiments, the RET-related diseases are lung cancer, papillary thyroid carcinoma, medullary thyroid carcinoma, differentiated thyroid carcinoma, recurrent thyroid carcinoma, refractory differentiated thyroid carcinoma, type 2A or 2B multiple endocrine tumors (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer, colorectal cancer, papillary renal cell carcinoma, gastrointestinal mucosal gangliocytoma, and cervical cancer, preferably RET fusion-type lung cancer or medullary thyroid carcinoma, more preferably small cell lung cancer, non-small cell lung cancer, bronchioloalveolar carcinoma, or lung adenocarcinoma.

[0044] In some embodiments of the present invention, the object of the present invention is a mammal including humans, preferably humans.

[0045] 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 RET-related diseases; in some embodiments, the RET-related diseases are lung cancer, thyroid cancer, medullary thyroid carcinoma, type II multiple endocrine tumors, pheochromocytoma, parathyroid hyperplasia, breast cancer, colorectal cancer, papillary renal cell carcinoma, gastrointestinal mucosal gangliocytoma, chronic myeloid leukemia, salivary gland cancer, ovarian cancer, cervical cancer, or prostate cancer, preferably RET fusion lung cancer or medullary thyroid carcinoma, more preferably small cell lung cancer, non-small cell lung cancer, or bronchioloalveolar lung cancer or lung adenocarcinoma. Invention Details

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] Some chemical terms

[0051] 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.

[0052] 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.

[0053] 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-.

[0054] 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.

[0055] 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.

[0056] The terms “halogen,” “halogenated,” or “halogenated” refer to bromine, chlorine, fluorine, or iodine.

[0057] As used herein, the terms “aromatic,” “aromatic ring,” “aromatic,” “aromatic,” and “aromatic cyclic” refer to a planar ring or ring portion of one or more rings having a delocalized electronic conjugated system containing 4n+2 electrons, where n is an integer. An aromatic ring 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 compound includes all carbocyclic rings (such as benzene rings) and rings containing one or more heteroatoms (such as pyridine).

[0058] The term "heteroatom" or "heteroatom" as used herein, alone or as part of other components, 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.

[0059] The term “dense” or “dense ring” as used alone or in combination in this article refers to a ring structure in which two or more rings share one or more bonds.

[0060] The term “spiral” or “spiroring” as used alone or in combination in this article refers to a ring structure in which two or more rings share one or more atoms.

[0061] The term "alkyl" as used hereby, either alone or as part of other components (e.g., monoalkylamino), 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.

[0062] The term "alkenyl" as used alone or in combination herein 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 may 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, for example, "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.

[0063] The term "alkynyl" as used alone or in combination herein 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 as defined herein is accompanied by a numerical range, such as "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.

[0064] The term "cycloalkyl" as used herein, alone or as part of other components, refers to a stable, monovalent, non-aromatic monocyclic or polycyclic hydrocarbon group containing only carbon and hydrogen atoms. It may include fused ring, spirocyclic, 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, 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.

[0065] The terms “heterocyclic group,” “heterocyclic alkyl group,” and “heterocycle” as used alone or as part of other components herein 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 stated, 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. Heterocyclic groups 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 can contain one or more aromatic rings or heteroaromatic rings, as long as the atoms connected to the rest of the molecule are from non-aromatic rings. For the purposes of this application, the heterocyclic group is preferably a stable 4-11 valent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, and more preferably a stable 4-8 valent 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, imidazolinyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinoneyl, pyranyl, pyrazolyl, pyrrolidinyl, quinazinyl, quininecycloyl, tetrahydrofuranyl, tetrahydropyranyl, etc.

[0066] The term "aryl" refers 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.

[0067] The term "heteroaryl" refers 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.

[0068] 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.

[0069] Intermediate compounds and polymorphs of the compounds of this invention are also within the scope of this invention.

[0070] Unless otherwise specified, the olefin double bonds contained in the compounds of this invention include E and Z isomers.

[0071] 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.

[0072] The term “pharmaceutically acceptable salt” as used in this article includes both salts with added acid salts and salts with added alkali salts.

[0073] "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. "Pharmaceutically 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 inorganic bases 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.

[0074] Organic bases that form salts include, but are not limited to, primary amines, secondary amines, tertiary amines, and cyclic amines, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purines, piperazine, piperidine, choline, and caffeine. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0075] 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.

[0076] 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 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] "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.

[0081] 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.

[0082] The term "treatment" as used in this article refers to the treatment of relevant diseases or conditions in mammals, particularly humans, including...

[0083] (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.

[0084] (ii) To suppress a disease or symptom, that is, to control its development;

[0085] (iii) To alleviate the disease or symptom, that is, to make the disease or symptom subside;

[0086] (iv) Relieve symptoms caused by disease or illness.

[0087] 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.

[0088] 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 the 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.

[0089] 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 a preferred embodiment, the compounds and compositions discussed herein are administered orally.

[0090] Preparation of the compounds of the present invention

[0091] The following reaction route illustrates a method for preparing the compounds of the present invention.

[0092] 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.

[0093] 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

[0094] The following non-limiting embodiments are merely illustrative and do not limit the invention in any way.

[0095] 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.

[0096] 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.

[0097] 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.

[0098] NMR data ( 1H 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 shape 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).

[0099] Example 1: (S)-1-(5-cyano-2-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-N-(1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-4-methoxypiperidine-4-carboxamide

[0100]

[0101] Step 1: Synthesis of Compound II

[0102] Under nitrogen protection, compound I (500 mg) and SEM-C1 (512 mg) were added to 10 mL of DMF, and NaH (226 mg) was added at 0 °C. The mixture was stirred at 25 °C for 2 hours. Ice water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (780 mg). ESI-MS m / z: 308.2 [M+H] + .

[0103] Step 2: Synthesis of Compound III

[0104] Under nitrogen protection, compound II (400 mg) was added to 6 mL of THF, cooled to -78 °C, and 2 M LDA (1 mL) was slowly added dropwise, followed by stirring for 30 minutes. Iodine (495 mg) was added, and the mixture was stirred for 30 minutes at -78 °C. The reaction was quenched by adding saturated ammonium chloride aqueous solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (416 mg). ESI-MS m / z: 434.2 [M+H] + .

[0105] Step 3: Synthesis of Compound V

[0106] Under nitrogen protection, compound III (430 mg), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-pyrazole (250 mg), Pd(dppf)Cl2 (55 mg), and K2CO3 (0.52 g) were added to 10 mL of dioxane and 2 mL of water, heated to 80 °C, and stirred for 2 hours. Water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (251 mg). ESI-MS m / z: 388.2 [M+H] + .

[0107] Step 4: Synthesis of Compound VII

[0108] Compound V (387 mg), (S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-4-methoxypiperidine-4-carboxamide (390 mg), and DIEA (387 mg) were added to 10 mL of n-butanol, heated to 160 °C, and stirred for 4 hours. The mixture was then cooled to room temperature, concentrated under reduced pressure, and the residue was separated by column chromatography to obtain the target compound (496 mg). ESI-MS m / z: 485.2 [M+H] + .

[0109] Step 5: Synthesis of Example 1

[0110] Compound VII (69.8 mg) was added to TFA (10 mL) and DCM (10 mL) and stirred at 25 °C for 2 hours. The solvent was removed by vortexing, and then methanol (10 mL) and K₂CO₃ (1.2 g) were added, and the mixture was stirred at 25 °C for 4 hours. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by column chromatography to obtain the target compound (40 mg). ESI-MS m / z: 569.3 [M+H] + .

[0111] Example 2: (S)-4-(4-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-5-oxo-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-2-(1-methyl-1H-pyrazin-4-yl)-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0112]

[0113] The synthesis method is the same as in Example 1. ESI-MS m / z: 581.3 [M+H] + .

[0114] Example 3: (S)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)pyridin-3-yl)-2-(1-methyl-1H-pyrazin-4-yl)-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0115]

[0116] Step 1: Synthesis of Compound III

[0117] Under nitrogen protection, compound I (390 mg), compound II (260 mg), Pd(dppf)Cl2 (50 mg), and potassium carbonate (417 mg) were added to 5 mL of dioxane and 2 mL of water, and heated to 80 °C with stirring for 2 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (277 mg). ESI-MS m / z: 448.2 [M+H] + .

[0118] Step 2: The synthesis method is the same as in Example 1. ESI-MS m / z: 635.3 [M+H] + .

[0119] Step 3: The synthesis method is the same as in Example 1. ESI-MS m / z: 505.3 [M+H] + .

[0120] Example 4: (S)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)(methyl)amino)pyridin-3-yl)-2-(1-methyl-1H-pyrazin-4-yl)-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0121]

[0122] The synthesis method is as described in Example 1. ESI-MS m / z: 581.3 [M+H] + .

[0123] Example 5: (S)-1-(6-cyano-2-(1-methyl-1H-pyrazol-4-yl)-3H-imidazo[4,5-b]pyridin-7-yl)-N-(1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-4-methoxypiperidine-4-carboxamide

[0124]

[0125] Step 1: Synthesis of Compound III

[0126] Compound I (600 mg) and 1-methyl-1H-pyrazole-4-carboxaldehyde (242 mg) were added to 10 mL of EtOH, followed by the addition of 1 M Na₂S₂O₄ (1.47 mg). The mixture was heated to 80 °C and stirred for 12 hours. After cooling to room temperature, ice water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extracts were washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (429 mg). ESI-MS m / z: 360.2 [M+H] + .

[0127] Step 2: Synthesis of Compound IV

[0128] Under nitrogen protection, compound III (360 mg) and SEM-Cl (249 mg) were added to 10 mL of DMF, and NaH (166 mg) was added at 0 °C. The mixture was stirred at 25 °C for 2 hours. Ice water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (441 mg). ESI-MS m / z: 490.2 [M+H] + .

[0129] Step 3: Synthesis of Compound V

[0130] Under nitrogen protection, compound IV (490 mg), Pd(PPh3)4 (100 mg), and Zn(CN)2 (232 mg) were added to 10 mL of NMP and heated to 130 °C with stirring for 12 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (163 mg). ESI-MS m / z: 389.2 [M+H] + .

[0131] Step 4: Synthesis of Compound VII

[0132] Compound V (390 mg), (S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-4-methoxypiperidine-4-carboxamide (400 mg), and DIEA (387 mg) were added to 10 mL of n-butanol and heated to 160 °C with stirring for 4 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure, and the residue was separated by column chromatography to obtain the target compound (528 mg). ESI-MS m / z: 700.4 [M+H]+ .

[0133] Step 5: Synthesis of Example 5

[0134] The synthesis method is the same as in Example 1. ESI-MS m / z: 570.4 [M+H] + .

[0135] Example 6: (S)-7-(4-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-5-oxo-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-2-(1-methyl-1H-pyrazin-4-yl)-3H-imidazo[4,5-b]pyridin-6-nitrile

[0136]

[0137] The synthesis method is the same as in Example 5. ESI-MS m / z: 582.3 [M+H] + .

[0138] Example 7: (S)-7-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)pyridin-3-yl)-2-(1-methyl-1H-pyrazin-4-yl)-3H-imidazo[4,5-b]pyridin-6-nitrile

[0139]

[0140] The synthesis method is the same as in Example 5. ESI-MS m / z: 582.3 [M+H] + .

[0141] Example 8: (S)-7-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)(methyl)amino)pyridin-3-yl)-2-(1-methyl-1H-pyrazin-4-yl)-3H-imidazo[4,5-b]pyridin-6-nitrile

[0142]

[0143] The synthesis method is the same as in Example 5. ESI-MS m / z: 582.3 [M+H] + .

[0144] Example 9: 7-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-2-(1-methyl-1H-pyrazol-4-yl)-3H-imidazo[4,5-b]pyridin-6-nitrile

[0145]

[0146] Step 1: Synthesis of Compound III

[0147] Under nitrogen protection, compound I (390 mg), compound II (452 ​​mg), Pd(dppf)Cl2 (50 mg), and potassium carbonate (417 mg) were added to 5 mL of dioxane and 2 mL of water, and heated to 80 °C with stirring for 2 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by column chromatography to obtain the target compound (324 mg). ESI-MS m / z: 649.2 [M+H] + .

[0148] Step 2: Synthesis of Example 9

[0149] The synthesis method is the same as in Example 1. ESI-MS m / z: 519.3 [M+H] + .

[0150] Example 10: 7-(5-(6-(6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyrazin-2-yl)-2-(1-methyl-1H-pyrazol-4-yl)-3H-imidazo[4,5-b]pyridin-6-nitrile

[0151]

[0152] The synthesis method is the same as in Example 9. ESI-MS m / z: 520.3 [M+H] + .

[0153] Example 11: 5-Chloro-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine

[0154]

[0155] Step 1: Synthesis of Compound III

[0156] Under nitrogen protection, compound I (165 mg), compound II (452 ​​mg), Pd(dppf)Cl2 (50 mg), and potassium carbonate (417 mg) were added to 5 mL of dioxane and 2 mL of water, and heated to 80 °C with stirring for 2 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to obtain compound III (206 mg). ESI-MS m / z: 443.2 [M+H] + .

[0157] Step 2: Synthesis of Example 11

[0158] The synthesis method is as described in Example 1. ESI-MS m / z: 504.3 [M+H] + .

[0159] Example 12: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-5-methyl-N-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine

[0160]

[0161] The synthesis method is the same as in Example 11. ESI-MS m / z: 484.3 [M+H] + .

[0162] Example 13: 5-Fluoro-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine

[0163]

[0164] The synthesis method is the same as in Example 11. ESI-MS m / z: 488.3 [M+H] + .

[0165] Example 14: 5-Chloro-N-(1-(difluoromethyl)-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-2-amine

[0166]

[0167] The synthesis method is the same as in Example 11. ESI-MS m / z: 540.3 [M+H]+ .

[0168] Example 15: 2-(4-((5-chloro-4-(6-(((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-ylpyrimidin-2-yl)amino)-1H-pyrazol-1-yl)ethane-1-ol

[0169]

[0170] The synthesis method is the same as in Example 11. ESI-MS m / z: 534.4 [M+H] + .

[0171] Example 16: 5-Chloro-N-(1-cyclopropyl-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidine-2-amine

[0172]

[0173] The synthesis method is the same as in Example 11. ESI-MS m / z: 530.4 [M+H] + .

[0174] Example 17: 5-Chloro-N-(1-isopropyl-1H-pyrazol-4-yl)--4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-2-amine

[0175]

[0176] The synthesis method is the same as in Example 11. ESI-MS m / z: 532.4 [M+H] + .

[0177] Example 18: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidine-5-nitrile

[0178]

[0179] Step 1: Synthesis of Compound III

[0180] Compound I (217 mg), compound II (97 mg), and DIEA (387 mg) were added to 5 mL of methanol and heated to 65 °C with stirring for 2 hours. After cooling to room temperature, 15 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to obtain compound III (206 mg). ESI-MS m / z: 235.2 [M+H] + .

[0181] Step 2: Synthesis of Example 18

[0182] The synthesis method is the same as in Example 1. ESI-MS m / z: 495.3 [M+H] + .

[0183] Example 19: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)-5-(trifluoromethyl)pyrimidin-2-amine

[0184]

[0185] The synthesis method is the same as in Example 11. ESI-MS m / z: 538.4 [M+H] + .

[0186] Example 20: 5-chloro-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-(oxecyclobutane-3-yl)-1H-pyrazol-4-yl)pyrimidin-2-amine

[0187]

[0188] The synthesis method is as described in Example 11. ESI-MS m / z: 546.4 [M+H] + .

[0189] Example 21: 5-Chloro-N-(1-(2-(dimethylamino)ethyl)-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl)pyrimidine-2-amine

[0190]

[0191] The synthesis method is the same as in Example 11. ESI-MS m / z: 561.4 [M+H] +.

[0192] Example 22: 5-Chloro-N-(1-(2,2-difluoroethyl)-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-2-amine

[0193]

[0194] The synthesis method is the same as in Example 11. ESI-MS m / z: 554.4 [M+H] + .

[0195] Example 23: 2-((1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-5-nitrile

[0196]

[0197] The synthesis method is the same as in Example 17. ESI-MS m / z: 525.4 [M+H] + .

[0198] Example 24: 3-(4-((5-chloro-4-(6-(((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-ylpyrimidin-2-yl)amino)-1H-pyrazol-1-yl)propane-1,2-diol

[0199]

[0200] The synthesis method is the same as in Example 11. ESI-MS m / z: 564.4 [M+H] + .

[0201] Example 25: N-(5-chloro-4-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-2-yl)isothiazolyl-4-amine

[0202]

[0203] The synthesis method is the same as in Example 11. ESI-MS m / z: 507.4 [M+H] + .

[0204] Example 26: 4-(5-(6-(6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyrazin-2-yl)-2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidine-5-nitrile

[0205]

[0206] The synthesis method is the same as in Example 17. ESI-MS m / z: 496.4 [M+H] + .

[0207] Example 27: 4-(6-((6-methoxypyridin-3-yl)methyl)amino)pyridin-3-yl-2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-5-nitrile

[0208]

[0209] The synthesis method is the same as in Example 17. ESI-MS m / z: 414.4 [M+H] + .

[0210] Example 28: 2-Chloro-N-(1-(5-cyano-2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)pyridin-2-yl)piperidin-4-yl)-6-fluorobenzamide

[0211]

[0212] The synthesis method is the same as in Example 17. ESI-MS m / z: 532.4 [M+H] + .

[0213] Example 29: N-((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)methyl)-4-methoxy-1-(2-(((1-methyl-1H-pyrazin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

[0214]

[0215] Step 1: Synthesis of Compound III

[0216] Under nitrogen protection, compound I (318 mg), compound II (372 mg), Pd2(dba)3 (50 mg), Xant-Phos (42 mg), and potassium carbonate (490 mg) were added to 5 mL of dioxane and heated to 100 °C with stirring for 6 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to obtain compound III (492 mg). ESI-MS m / z: 615.2 [M+H] + .

[0217] Step 2: Synthesis of Compound V

[0218] Compound III (610 mg), compound IV (120 mg), and DIEA (500 mg) were added to 5 mL of isopropanol and heated to 80 °C with stirring for 1 hour. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to give compound V (430 mg). ESI-MS m / z: 676.4 [M+H] + .

[0219] Step 3: Synthesis of Example 29

[0220] The synthesis method is the same as in Example 1. ESI-MS m / z: 546.4 [M+H] + .

[0221] Example 30: 4-(6-(((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)methyl)amino)pyridin-3-acyl)-2-((1-methyl-1H-pyrazin-4-yl)amino)pyrimidine-5-nitrile)

[0222]

[0223] The synthesis method is the same as in Example 17. ESI-MS m / z: 468.4 [M+H] + .

[0224] Example 31: 4-(6-(((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)methyl)(methyl)amino)pyridin-3-yl)-2-((1-methyl-1H-pyrazin-4-yl)amino)pyrimidin-5-nitrile)

[0225]

[0226] The synthesis method is as described in Example 17. ESI-MS m / z: 482.4 [M+H] + .

[0227] Example 32: 2-(isothiazo-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0228]

[0229] The synthesis method is as described in Example 1. ESI-MS m / z: 521.4 [M+H] + .

[0230] Example 33: 2-(isothiazolyl-4-ylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]pyrimidin-5-nitrile

[0231]

[0232] The synthesis method is the same as in Example 17. ESI-MS m / z: 498.4 [M+H] + .

[0233] Example 34: 2-(1-((R)-2,3-dihydroxypropyl)-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0234]

[0235] The synthesis method is the same as in Example 17. ESI-MS m / z: 578.4 [M+H] + .

[0236] Example 35: 2-(1-((S)-2,3-dihydroxypropyl)-1H-pyrazol-4-yl)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0237]

[0238] The synthesis method is the same as in Example 17. ESI-MS m / z: 578.4 [M+H] + .

[0239] Example 36: 2-(4-((4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)amino)-1H-pyrazol-1-yl)ethane-1-ol

[0240]

[0241] Step 1: Synthesis of Compound III

[0242] Under nitrogen protection, compound I (318 mg), compound II (422 mg), K3(PO4)4 (700 mg), and Pd(dppf)Cl2 (50 mg) were added to 5 mL of dioxane and 3 mL of water, heated to 80 °C, and stirred for 2 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to obtain compound III (360 mg). ESI-MS m / z: 578.2 [M+H] + .

[0243] Step 2: Synthesis of Compound V

[0244] Compound III (577 mg), compound IV (128 mg), Pd2(dba)3 (50 mg), Xant-Phos (42 mg), and potassium carbonate (490 mg) were added to 5 mL of dioxane and heated to 100 °C with stirring for 12 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to give compound V (406 mg). ESI-MS m / z: 669.4 [M+H] + .

[0245] Step 3: Synthesis of Example 36

[0246] The synthesis method is the same as in Example 1. ESI-MS m / z: 539.4 [M+H] + .

[0247] Example 37: (S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-4-methoxy-1-(2-((1-methyl-1H-pyrazin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

[0248]

[0249] The synthesis method is the same as in Example 28. ESI-MS m / z: 560.4 [M+H] + .

[0250] Example 38: (S)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)pyridin-3-yl)-2-(((1-methyl-1H-pyrazin-4-yl)amino)pyrimidine-5-nitrile)

[0251]

[0252] The synthesis method is the same as in Example 3. ESI-MS m / z: 482.4 [M+H] + .

[0253] Example 39: (R)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)pyridin-3-yl)-2-(((1-methyl-1H-pyrazin-4-yl)amino)pyrimidine-5-nitrile)

[0254]

[0255] The synthesis method is the same as in Example 3. ESI-MS m / z: 482.4 [M+H] + .

[0256] Example 40: (S)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)(methyl)amino)pyridin-3-yl)-2-((1-methyl-1H-pyrazin-4-yl)amino)pyrimidin-5-nitrile)

[0257]

[0258] The synthesis method is the same as in Example 3. ESI-MS m / z: 496.4 [M+H] + .

[0259] Example 41: 4-(5-(6-(6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyrazin-2-yl)-N-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0260]

[0261] The synthesis method is the same as in Example 36. ESI-MS m / z: 510.4 [M+H]+ .

[0262] Example 42: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0263]

[0264] The synthesis method is the same as in Example 36. ESI-MS m / z: 509.4 [M+H] + .

[0265] Example 43: 4-(6-(6-((5-methoxypyridin-2-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl)-N-(1-methyl-1H-pyrazol-4-yl)-1H-pyrazino[3,4-d]pyrimidin-6-amine

[0266]

[0267] The synthesis method is as described in Example 36. ESI-MS m / z: 510.4 [M+H] + .

[0268] Example 44: 5-chloro-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0269]

[0270] The synthesis method is as described in Example 36. ESI-MS m / z: 543.4 [M+H] + .

[0271] Example 45: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-6-methyl-N-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0272]

[0273] The synthesis method is the same as in Example 36. ESI-MS m / z: 523.4 [M+H] + .

[0274] Example 46: 5-Fluoro-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-N-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0275]

[0276] The synthesis method is the same as in Example 36. ESI-MS m / z: 527.4 [M+H] + .

[0277] Example 47: 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl]-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-nitrile

[0278]

[0279] The synthesis method is the same as in Example 36. ESI-MS m / z: 534.4 [M+H] + .

[0280] Example 48: (S)-4-(4-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-5-oxo-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-2-((1-methyl-1H-pyrazin-4-yl)amino)pyrimidin-5-nitrile

[0281]

[0282] The synthesis method is the same as in Example 5. ESI-MS m / z: 558.4 [M+H] + .

[0283] Example 49: ((S)-4-(6-((1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)amino)pyridin-3-yl)-N-(1-methyl-1H-pyrazin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine

[0284]

[0285] The synthesis method is the same as in Example 36. ESI-MS m / z: 496.4 [M+H] + .

[0286] Example 50: (S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-1-methoxy-4-(2-((1-methyl-1H-pyrazin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)cyclohexane-1-carboxamide

[0287]

[0288]

[0289] Step 1: Synthesis of Compound III

[0290] Under nitrogen protection, compound I (318 mg), compound II (296 mg), K₂CO₃ (700 mg), and Pd(dppf)Cl₂ (50 mg) were added to 5 mL of dioxane and 3 mL of water, and heated to 80 °C with stirring for 2 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extract was washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to obtain compound III (280 mg). ESI-MS m / z: 452.2 [M+H] + .

[0291] Step 2: Synthesis of Compound IV

[0292] Compound III (452 ​​mg), 1-methyl-1H-pyrazole-4-amine (100 mg), Pd2(dba)3 (50 mg), Xant-Phos (42 mg), and potassium carbonate (490 mg) were added to 5 mL of dioxane and heated to 100 °C with stirring for 12 hours. After cooling to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The extracts were washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was separated by column chromatography to give compound IV (305 mg). ESI-MS m / z: 513.4 [M+H] + .

[0293] Step 3: Synthesis of Compound V

[0294] Compound IV (300 mg) and Pd / C (10 mg) were added to 5 mL of ethanol and heated to 50 °C for 12 hours. After cooling to room temperature, the mixture was filtered to give a white solid (V) (220 mg). ESI-MS m / z: 515.4 [M+H] + .

[0295] Step 4: Synthesis of Compound VI

[0296] Compound V (514 mg) and sodium hydroxide (200 mg) were added to 5 mL of methanol and 5 mL of water, and the mixture was heated to 60 °C and stirred for 4 hours. After cooling to room temperature, the methanol was removed by concentration under reduced pressure. The pH was adjusted to 4 with dilute hydrochloric acid, and the mixture was extracted with ethyl acetate (50 mL × 3). The extracts were washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain compound VI (450 mg). ESI-MS m / z: 501.4 [M+H] + .

[0297] Step 5: Synthesis of Compound VII

[0298] The synthesis method is the same as in Example 1. ESI-MS m / z: 689.4 [M+H] + .

[0299] Step 6: Synthesis of Example 50

[0300] The synthesis method is the same as in Example 1. ESI-MS m / z: 559.4 [M+H] + .

[0301] Example 51: (S)-4-(5-cyano-2-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-N-(1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-1-methoxycyclohexane-1-carboxamide

[0302]

[0303] The synthesis method is as described in Example 50. ESI-MS m / z: 545.4 [M+H] + .

[0304] Example 52: (S)-N-(1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-1-methoxy-4-(2-((1-methyl-1H-pyrazin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)cyclohexane-1-carboxamide

[0305]

[0306] The synthesis method is the same as in Example 50. ESI-MS m / z: 559.4 [M+H] + .

[0307] Example 53: (1s,4R)-4-(5-cyano-2-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-N-((S)-1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-1-methoxycyclohexane-1-carboxamide

[0308]

[0309] The synthesis method is the same as in Example 50. ESI-MS m / z: 568.4 [M+H] + .

[0310] Example 54: (1r,4S)-4-(5-cyano-2-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-4-yl)-N-((S)-1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-1-methoxycyclohexane-1-carboxamide

[0311]

[0312] The synthesis method is the same as in Example 50. ESI-MS m / z: 568.4 [M+H] + .

[0313] Example 55: (S)-1-(5-cyano-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-(1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-4-methoxypiperidine-4-carboxamide

[0314]

[0315] The synthesis method is the same as in Example 5. ESI-MS m / z: 585.4 [M+H] + .

[0316] Example 56: 4-(4-(6-(6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)-2-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrolo[2,3-b]pyridin-5-nitrile

[0317]

[0318] The synthesis method is the same as in Example 3. ESI-MS m / z: 558.4 [M+H] + .

[0319] Example 57: (S)-4-(5-cyano-2-((1-methyl-1H-pyrazol-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-(1-(6-(4-fluoro-1H-pyrazin-1-yl)pyridin-3-yl)ethyl)-1-methoxycyclohexane-1-carboxamide

[0320]

[0321] The synthesis method is the same as in Example 50. ESI-MS m / z: 584.4 [M+H] + .

[0322] Biological testing

[0323] 1. Experiment on the anti-tumor proliferation activity of tumor cells:

[0324] Compound in BaF3-KIF5B-RET G810R Cell proliferation activity assay

[0325] The KIF5B-RET(K15;R12) fusion gene was constructed into plasmid pCDHL. A site-directed mutagenesis method was used to introduce a G mutation at position 810 of the RET protein, changing it to R. This plasmid was then co-transfected with pSPAX2 and pMD2G (4:2:1) plasmids into 293T cells. Lentiviral particles were packaged, and the lentiviral suspension was collected 48 hours post-transfection. BaF3 cells were then infected with puromycin (purchased from Solarbio) at a final concentration of 1 μg / ml for selection. One week later, IL-3-independent BaF3 cells were selected for monoclonal selection to construct BaF3-KIF5B-RET. G810R Cell lines.

[0326] BaF3-KIF5B-RET G810R Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, purchased from Biological Industries, BI) and 1% penicillin / streptomycin antibiotics (P / S, purchased from Life Technology) at 37°C and 5% CO2. The day before compound detection, BaF3-KIF5B-RET... G810RCells 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. The compound and cells were incubated together in a cell culture incubator for 72 hours (3 days). Then, 40 μL of Cell-Titer Glo (G7570, purchased from Promega) reagent was added for a second incubation of 5–10 minutes. Fluorescence values ​​were read on Envision, and the IC50 of the compound's inhibitory effect on cell proliferation was calculated using GraphPad Prism software. 50 Value. Tests have shown that the compounds of this invention exhibit excellent cellular activity.

[0327] 2. In vitro enzyme activity assay method:

[0328] Compounds on RET G810R In vitro enzyme activity assay

[0329] The RET (658-1114aa) gene was cloned and ligated into the pFASTBac plasmid. Site-directed mutagenesis was performed using primers RET G810R F: GGAGTACGCCAAATACcggTCCCTGCGGGGCTT and RET G810R R: AAGCCCCGCAGGGAccgGTATTTGGCGTACTCC to introduce a G-to-R mutation at position 810 of the RET protein. Expression was then performed in insect Sf9 cells. Cells were collected and resuspended in Lysis Buffer (50mM Tris-HCl, pH 7.5, 150mM NaCl, 0.1mM EDTA, 0.5% Triton X-100, 20% Glycerol, 1mM DTT, 1% protease inhibitor), followed by sonication lysis. The lysate was passed through a GSTrap HP (GE Healthcare) affinity column and washed with wash buffer (50mM Tris-HCl, pH 7.5, 150mM NaCl, 0.1mM EDTA, 0.5% Triton X-100, 20% Glycerol, 1mM DTT, 1% protease inhibitor). GST-RET was washed with EDTA, 20% Glycerol, and 1 mM DTT to remove contaminating proteins, and then eluted with Eluent Buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1 mM EDTA, 20% Glycerol, 20 mM Glutathione, and 1 mM DTT). G810R The protein was concentrated, aliquoted, flash-frozen in liquid nitrogen, and stored at -80°C.

[0330] The compound in this patent affects RET G810R 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 MgCl2, 1 mM DTT, 0.001% Tween 20, 100 μg / ml BSA, 20 nM SEB) and mixed thoroughly. 2.5 μL was then added to a 384-well plate (OptiPlate-384, purchased from PerkinElmer), followed by 5 μL of GST-RET. G810R (Final concentration: 0.01 nM) After centrifugation and remixing, 2.5 μL of ATP (final concentration: 10 μM) and TK Peptide Substrate mixture (final concentration: 1 μM, purchased from Cisbio) was added to initiate 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, fluorescence values ​​were read on an Envision (purchased from PerkinElmer) microscope (excitation at 320 nm, detection of emission at 665 nm and 620 nm, the ratio of which is the enzyme activity signal). RET was measured at 7 concentrations for each compound. G810R The enzymatic activity signal was obtained, and the IC50 of the compound was calculated using GraphPad Prism software. 50 Value. Testing showed that the compounds of this invention possess excellent enzymatic activity.

Claims

1. A compound of formula (II), or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof: in, Ring B is or The * key is connected to L1. L1 is -L2-L4-, -L2-L5-, -L2-L6-, -L2-L3-L4-, -L2-L3-L5-, or -L2-L3-L6, L2 is a 5-6 membered heteroaryl ring. L3 is a 4-8 membered heterocyclic ring. L4 is -CH2-. L5 is -CH2-NR-, L6 is -(CO)-NR-, L is or X is either N or CH. Y is either N or CH. R1 is H or methyl. R2 is methyl, or R1 and R2 are connected together to form a 5-6 member heterocyclic ring. Ring A is a 6-10 aryl group or a 5-12 heteroaryl group, wherein the aryl and heteroaryl groups may optionally be converted by halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted, and the alkyl, alkenyl, alkynyl, or heterocyclic group may optionally be replaced by halogen, -CN, -OH, -NH2, -CF3, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted, R 10 It is a 5-6 member heteroaryl or -OC 1-6 Alkyl groups, wherein the heteroaryl group may optionally be replaced by halogen, -CN, -OH, -NH2, -OC 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), or C 1-6 Alkyl substitution, R can be independently H or C. 1-6 alkyl, n is 0 or 1.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, wherein ring A is a 5-6 membered heteroaryl group, said heteroaryl group optionally coated with a halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, having the structure shown in formula (I): in, L is or X is either N or CH. Y is either N or CH. R1 is H or methyl. R2 is methyl, or R1 and R2 are connected together to form a 5-6 member heterocyclic ring. Ring A is a 6-10 aryl group or a 5-12 heteroaryl group, wherein the aryl and heteroaryl groups may optionally be converted by halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted with a halogen, -CN, -OH, -NH2, -CF3, or -OC group. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

4. The compound according to claim 3, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, wherein ring A is a 5-6 membered heteroaryl group, said heteroaryl group optionally coated with a halogen, -CN, -OH, -NH2, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group is substituted, and the alkyl, alkenyl, alkynyl, or heterocyclic group may optionally be replaced by halogen, -CN, -OH, -NH2, -CF3, or -OC. 1-6 Alkyl, -NH(C) 1-6 alkyl), -N(C) 1-6 Alkyl)(C 1-6 Alkyl), C 1-6 Alkyl, 3-8 membered cycloalkyl, or 3-8 membered heterocyclic substituted.

5. The following compounds, or their pharmaceutically acceptable salts, solvates, polymorphs, or isomers:

6. A pharmaceutical composition comprising a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, and optionally comprising a pharmaceutically acceptable carrier.

7. Use of the compound of any one of claims 1-5, or a pharmaceutically acceptable salt, solvate, polymorph, or isomer thereof, or the composition of claim 6, in the preparation of a medicament for treating RET-related diseases.

8. The use according to claim 7, wherein the RET-related disease is lung cancer, thyroid cancer, medullary thyroid carcinoma, type II multiple endocrine tumor, pheochromocytoma, parathyroid hyperplasia, breast cancer, colorectal cancer, papillary renal cell carcinoma, gastrointestinal mucosal gangliocytoma, chronic myeloid leukemia, salivary gland cancer, ovarian cancer, cervical cancer, or prostate cancer.

9. The use according to claim 7, wherein the RET-related disease is small cell lung cancer, non-small cell lung cancer, or bronchiolar lung cancer or lung adenocarcinoma.

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