Kinesin KIF18a inhibitor and method for using same

Abstract

A compound represented by formula (I) serving as a KIF18A inhibitor, a pharmaceutical composition containing the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, and a use of the compound represented by formula (I) and the pharmaceutical composition in the treatment of cell proliferative diseases, for example, cancers.

Description

Kinesin KIF18A inhibitors and methods of use thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to the following five Chinese invention patent applications, the entire contents of which are hereby incorporated by reference in their entirety:

[0003] Patent application No. 202410362057.X filed with the State Intellectual Property Office of China on March 27, 2024,

[0004] Patent application No. 202410799138.6 filed with the State Intellectual Property Office of China on June 19, 2024,

[0005] Patent application No. 202510032448.X filed with the State Intellectual Property Office of China on January 8, 2025, and

[0006] Patent application No. 202510338448.2 submitted to the State Intellectual Property Office of China on March 20, 2025. Technical Field

[0007] The present disclosure belongs to the field of medicinal chemistry, and particularly relates to a compound as a KIF18A inhibitor, a pharmaceutical composition containing the compound, and a method for treating and preventing cell proliferative diseases, such as cancer, using the compound and the pharmaceutical composition. Background Art

[0008] KIF18A is a mitotic motor protein that regulates chromosome arrangement during cell mitosis, avoids errors in the mitotic process, and ensures the normal progress of mitosis.

[0009] The vast majority of human cells are diploid, and there are multiple cell cycle checkpoints to ensure that the genome remains stable during continuous cell division. The inactivation of some key proteins related to cell cycle checkpoints can lead to abnormal mitosis and the doubling of the entire genome, thereby converting a natural diploid cell into a chromosomally unstable tetraploid cell. Whole-genome doubling (WGD) events (at least one WGD has occurred) are widely present in clinical tumor samples and are an important feature of chromosomally unstable tumors. The results showed that relative to diploid cells, KIF18A expression levels were significantly increased in WGD cells. + KIF18A knockout in all 10 cell lines revealed that the expression of KIF18A in the WT cells was significantly increased compared to that in the WGD cells. - Cell line, WGD + Cell viability was significantly reduced, and live cell imaging showed that WGD occurred after KIF18A loss.+ Compared with WGD- cells, breast cancer cells showed increased spindle length and chromosome hyperoscillation, which triggered abnormal events such as chromosome shedding and mitosis prolongation. The above data revealed that the loss of KIF18A would cause WGD + Abnormal mitosis of tumor cells inhibits WGD + Therefore, KIF18A is an attractive therapeutic target and its inhibition may help to specifically target cells with WGD. + characteristic tumors without affecting the normal diploid cells of human tissues. Summary of the Invention

[0010] The present disclosure provides a compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0011] wherein Ring A is a 5-6 membered heteroaryl group, and the 5-6 membered heteroaryl group contains at least 3 heteroatoms selected from N, O or S;

[0012] Ring B is a 4-6 membered heterocyclic group, and the 4-6 membered heterocyclic group contains at least one heteroatom selected from N, O or S;

[0013] Ring C is a 6-8 membered heterocyclic group, and the 6-8 membered heterocyclic group contains at least one heteroatom selected from N, O or S;

[0014] X 1 、X 2 、Y 1 and Y 2 Each independently selected from N or CR x ; the R x Selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 alkoxy;

[0015] R 1 and R 2 are each independently selected from halogen, C 1-6 Alkyl or C 3-6 Cycloalkyl, and the C 1-6 Alkyl or C 3-6 The cycloalkyl group is optionally substituted with one or more radicals selected from OH, halogen, -NH2 or C 1-4 substituted by an alkoxy substituent;

[0016] R 3 Halogen, OH, C 1-6 Alkyl, C 3-6 Cycloalkyl or

[0017] R3a and R 3b are each independently selected from H, halogen or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more radicals selected from halogen, hydroxy, amino, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 3-6 substituted by a cycloalkyl group, a 3-8 membered heterocyclyl group, a phenyl group or a 5-6 membered heteroaryl group;

[0018] R 4 Selected from -NHSO2-C 1-4 Alkyl, and the C 1-4 The alkyl group is optionally substituted with OH or halogen;

[0019] m, n and p are each independently selected from 0, 1, 2 or 3.

[0020] In the compound represented by formula (I) of the present disclosure, its stereoisomers or pharmaceutically acceptable salts thereof, R 3 for means two R on the same ring atom 3 Groups together form

[0021] In some embodiments, the ring A is a 5-membered heteroaryl group and contains at least 3 heteroatoms selected from N, O, or S. In some embodiments, the ring A is a 5-membered heteroaryl group and contains 3 heteroatoms selected from N, or contains 2 heteroatoms selected from N and 1 heteroatom selected from O or S, or contains 4 heteroatoms selected from N. In some embodiments, the ring A is a 5-membered heteroaryl group and contains 3 heteroatoms selected from N. In some embodiments, the ring A is selected from In some embodiments, Ring A is In some embodiments, Ring A is In some embodiments, the ring A is selected from

[0022] In some embodiments, the R 1 Selected from C 1-6 Alkyl, and the C 1-6 Alkyl is optionally substituted with one or more substituents selected from OH or halogen; and / or m is selected from 0 or 1. In some embodiments, the R 1 Selected from methyl, CHF2 or m is selected from 0 or 1. In some embodiments, the R 1 It is a methyl group.

[0023] In some embodiments, the structural unit Selected from In some embodiments, the structural unit for In some embodiments, the structural unit for In some embodiments, the structural unit Selected from In some embodiments, the structural unit Selected from

[0024] In some embodiments, the ring B is a 6-membered heterocyclyl containing one heteroatom selected from N. In some embodiments, the ring B is

[0025] In some embodiments, the R 2 is halogen; and / or n is selected from 0, 1 or 2, or n is 2. In some embodiments, the R 2 is selected from F, Cl or Br; n is selected from 0, 1 or 2.

[0026] In some embodiments, the structural unit Selected from

[0027] In some embodiments, the ring C is a 6-, 7-, or 8-membered heterocyclyl containing one heteroatom selected from N. In some embodiments, the ring C is selected from

[0028] In some embodiments, the R 3 for and / or R 3a and R 3b are each independently selected from H or halogen; and / or p is selected from 0, 1 or 2; or p is selected from 0 or 1. In some embodiments, the R 3 for p is selected from 0, 1 or 2.

[0029] In some embodiments, the structural unit Selected from

[0030] In some embodiments, the X 1 、X 2 Each independently selected from CR x ; and / or said R xIn some embodiments, X 1 、X 2 are each independently selected from CH or N; or, X 1 、X 2 In some embodiments, the X 1 、X 2 are each independently selected from CH, -CF or N; or, X 1 CH, X 2 In some embodiments, Y 1 、Y 2 are each independently selected from CH or N; or, Y 1 N, Y 2 CH; or, Y 1 N, Y 2 is N.

[0031] In some embodiments, R 4 Selected from -NHSO2-C 1-4 Alkyl, and the C 1-4 Alkyl is optionally substituted with one or more OH. 4 Selected from -NHSO2-CH2CH2OH, -NHSO2-CH3.

[0032] In some embodiments, the compound represented by formula (I) of the present disclosure, its stereoisomer or pharmaceutically acceptable salt thereof is selected from the compound represented by formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe), formula (IIf) or formula (IIg), its stereoisomer or pharmaceutically acceptable salt thereof:

[0033] Among them, ring A, ring B, ring C, R 1 、R 2 、R 3 、R 4 、X 1 、X 2 , m, p and n are as defined above in formula (I).

[0034] In some embodiments, the compound of formula (I) of the present disclosure, its stereoisomers, or pharmaceutically acceptable salts thereof are selected from the compound of formula (III), its stereoisomers, or pharmaceutically acceptable salts thereof:

[0035] Among them, the structural unit Selected from

[0036] Structural unit Selected from

[0037] X 2 selected from CH or -CF;

[0038] R 4 Selected from -NHSO2-CH2CH2OH or -NHSO2-CH3.

[0039] In some embodiments, the structural unit Selected from Alternatively, the structural unit Selected from

[0040] In some embodiments, the structural unit Selected from And the structural unit Selected from Alternatively, the structural unit Selected from

[0041] In some embodiments, the structural unit for And the structural unit Selected from

[0042] In some embodiments, the compounds of the present disclosure, their stereoisomers, or pharmaceutically acceptable salts thereof are selected from the following compounds, their stereoisomers, or pharmaceutically acceptable salts thereof:

[0043] On the other hand, the present disclosure also relates to a pharmaceutical composition comprising a compound represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the compound of the present disclosure, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0044] In some embodiments, the content of the compound, its stereoisomer, or pharmaceutically acceptable salt thereof in the pharmaceutical composition is selected from 0.1 mg to 1000 mg. In some embodiments, the content of the compound, its stereoisomer, or pharmaceutically acceptable salt thereof in the pharmaceutical composition is 1% to 95%.

[0045] In some embodiments, in the pharmaceutical composition, the pharmaceutically acceptable carrier includes one or more of a filler, a disintegrant, a binder, a glidant, and a lubricant.

[0046] On the other hand, the present disclosure also relates to the use of a compound represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for treating or preventing a disease mediated by KIF18A.

[0047] On the other hand, the present disclosure also relates to a compound represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for treating or preventing a disease mediated by KIF18A.

[0048] In another aspect, the present disclosure also relates to a method for treating or preventing a disease mediated by KIF18A in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0049] In another aspect, the present disclosure also relates to the use of a compound represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in treating or preventing diseases mediated by KIF18A.

[0050] In some embodiments, the disease mediated by KIF18A is a tumor. In some embodiments, the tumor is a chromosomally unstable tumor. In some embodiments, the tumor is selected from breast cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, head and neck cancer, gastric cancer, esophageal cancer, or skin cancer. In some embodiments, the tumor is selected from breast cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, or head and neck cancer. In some embodiments, the lung cancer is selected from squamous cell lung cancer, adenocarcinoma of the lung, large cell lung cancer, or small cell lung cancer.

[0051] definition

[0052] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered ambiguous or unclear without a specific definition, but should be understood according to its ordinary meaning in the art.

[0053] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0054] The term "pharmaceutically acceptable salt" refers to a derivative obtained by reacting a compound of the present invention with a relatively non-toxic acid or base. These salts can be prepared during the synthesis, separation, and purification of the compound, or can be prepared by reacting the purified free form of the compound with a suitable acid or base. When the compound contains a relatively acidic functional group (e.g., -COOH, -OH, -SO3H, etc.), it reacts with an appropriate inorganic or organic cation (base) to obtain a base addition salt, including salts formed with alkali metals or alkaline earth metals, ammonium salts formed with amines or their derivatives, salts formed with amino acids, etc. When the compound contains a relatively basic functional group (e.g., -NH2, etc.), it reacts with an appropriate inorganic or organic anion (acid) to obtain an acid addition salt, including salts formed with an inorganic acid or organic acid (e.g., carboxylic acid, etc.).

[0055] The term "pharmaceutically acceptable carrier" refers to a substance generally accepted in the art for delivering a biologically active agent to an animal, particularly a mammal, and includes, for example, adjuvants, excipients, or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, and dispersants, depending on the mode of administration and the nature of the dosage form. Pharmaceutically acceptable carriers are formulated within the purview of those skilled in the art based on a number of factors, including, but not limited to, the type and nature of the active agent being formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the intended therapeutic indication. Pharmaceutically acceptable carriers can include both aqueous and non-aqueous media, as well as materials required for the formulation of a variety of solid and semisolid dosage forms. In addition to the active agent, such carriers include a variety of different ingredients and additives, and the inclusion of such additional ingredients in a formulation for various reasons (e.g., stabilization of the active agent, binders, etc.) is well known to those skilled in the art.

[0056] The term "prophylactically or therapeutically effective amount" refers to a sufficient amount of a compound of the present disclosure, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof to treat a disorder at a reasonable effect / risk ratio applicable to any medical treatment and / or prevention. However, it should be recognized that the total daily dose of the compound or pharmaceutically acceptable salt thereof and the composition represented by any one of Formula (I), Formula (IIa) to Formula (IIg) and Formula (III) disclosed herein must be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific therapeutically effective dose level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the patient's age, weight, general health, sex, and diet; the administration time, route of administration, and excretion rate of the specific compound employed; the duration of treatment; drugs used in combination with or concurrently with the specific compound employed; and similar factors well known in the medical field.

[0057] The “isomers” described in the present disclosure include geometric isomers and stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic mixtures and other mixtures thereof, all of which fall within the scope of the present disclosure. The term “enantiomer” refers to stereoisomers that are mirror images of each other. The term “tautomer” refers to a type of functional group isomer that has different hydrogen attachment points due to one or more double bond displacements, for example, a ketone and its enol form are keto-enol tautomers. The term “diastereomer” refers to a stereoisomer in which a molecule has two or more chiral centers and is not a mirror image between the molecules. The term “cis-trans isomer” refers to different spatial configurations in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely. The term “atropisomer” refers to stereoisomers that can be separated because single bond rotation is hindered or rotates very slowly. Stereoisomers of the disclosed compounds can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. For example, one enantiomer of a compound disclosed herein can be prepared by asymmetric catalysis or chiral auxiliary derivatization; or by chiral resolution techniques to obtain a single stereoisomer from a mixture; or directly from chiral starting materials. Separation of optically pure compounds disclosed herein is typically accomplished using preparative chromatography, employing chiral columns, to achieve the separation of chiral compounds.

[0058] The absolute stereoconfiguration of a compound can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction can be used, or the chiral structure of the starting materials and the reaction mechanism of asymmetric synthesis can be used to confirm the absolute configuration of the compound. Alternatively, after resolution, the stereoconfiguration can be determined by comparison with products with a known absolute configuration. Compounds labeled "absolute configuration unknown / undetermined" herein are typically resolved from racemic compounds into individual isomers by chiral preparative SFC, followed by characterization and testing.

[0059] The term "isotopic derivative" refers to a compound in which at least one atom is replaced by an atom having the same atomic number but a different atomic mass. Examples of isotopes that can be introduced into the compounds of the present disclosure include stable and radioactive isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine, and iodine, for example, 2 H (deuterium, D), 3 H (tritium, T), 11 C. 13 C. 14 C. 15 N. 17 O. 18 O. 32 p、 33 p、 33 S. 34 S. 35 S. 36 S. 18 F. 36 Cl, 82 Br, 123 I. 124 I. 125 I. 129 I and 131 I, etc., preferably deuterium. Compared to non-deuterated drugs, deuterated drugs have one or more advantages such as reduced toxic side effects, increased drug stability, enhanced therapeutic efficacy, and extended drug biological half-life. All isotopic composition changes of the compounds of the present disclosure, whether radioactive or not, are included in the scope of the present disclosure. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom, wherein the deuterium replacement can be partial or complete, and partial deuterium replacement means that at least one hydrogen is replaced by at least one deuterium.

[0060] The term "nitrogen oxide" or "N-oxide" refers to a derivative formed by further oxidation of the nitrogen atom in a nitrogen-containing group. Common N-oxides include N-oxides of tertiary amines or nitrogen atoms in nitrogen-containing heterocycles. Synthesis methods of N-oxides are well known to those skilled in the art and include oxidation of heterocycles and tertiary amines using peroxyacids such as peracetic acid and m-chloroperbenzoic acid, hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxiranes such as dimethyldioxirane.

[0061] The term "prodrug" refers to certain derivatives of the disclosed compounds that have little or no pharmacological activity themselves, which have cleavable groups and decompose into the disclosed compounds through solvent decomposition or under physiological conditions. The types of prodrugs include, but are not limited to, amides, esters, anhydrides, salts, and the like. The "ester" refers to a derivative formed with a suitable alcohol when the disclosed compound contains an acidic group (such as a carboxylic acid); or a derivative formed with a suitable acid (including an organic acid or an inorganic acid) when the disclosed compound contains a hydroxyl group. Methods for preparing prodrugs are well known to those skilled in the art.

[0062] The compounds of the present disclosure can exist in unsolvated as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are also encompassed within the scope of the present disclosure.

[0063] The compounds of the present disclosure include stereoisomers, isotopic derivatives, solvates, N-oxides, prodrugs or pharmaceutically acceptable salt forms thereof.

[0064] The compounds of the present disclosure and their salts may exist as isotopic derivatives, and the compounds of the present disclosure include various isotopic derivatives and mixtures thereof.

[0065] The compounds of the present disclosure and their salts may exist in the form of solvates, such as hydrates, and the compounds of the present disclosure include various solvates and mixtures thereof.

[0066] The compounds of the present disclosure and their salts may exist in the form of "N-oxides", and the compounds of the present disclosure include various N-oxides and mixtures thereof.

[0067] The compounds described in the present disclosure, and their pharmaceutically acceptable salts, isotopic derivatives, solvates, and N-oxides have the same or similar biological activities and are all included within the scope of the present disclosure.

[0068] The term "optionally substituted" as used herein refers to two situations in which one or more hydrogen atoms of the substituted group may be "substituted" or "unsubstituted" by one or more substituents.

[0069] When the substituent appears in the structure Indicates that the atom is a bonding atom, for example Indicates that the C atom on the pyrimidine ring is a bonding atom. A dash "-" in a substituent structure indicates the point of attachment for the substituent, for example, -CH3 is attached through a C atom. Indicates the absolute configuration of a stereocenter, i.e., R or S configuration. It represents cis or trans configuration. Double real bonds or double imaginary bonds both represent cis configuration, and one real and one imaginary bond represent trans configuration.

[0070] When a substituent's bond can cross-link to a ring, it means that the substituent can be bonded to any atom on the ring. The substituent R can be substituted at any position on the benzene ring.

[0071] When the structural unit When m is 0, the ring A is substituted with H by default; when m is 1, 2 or 3, R 1 is a substituent other than H. When a substituent is listed without indicating the atom via which the substituent is attached to a given group or a given formula, then the substituent may be attached via any bondable atom thereof.

[0072] When any variable (such as R d ) appears more than once in a compound's composition or structure, its definition is independent in each instance. For example, Indicates that the cyclopentyl group is surrounded by 3 R d is replaced, and each R d There are independent options.

[0073] Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine or iodine atom.

[0074] Unless otherwise specified, the term "alkyl" refers to a group derived from a branched or straight chain saturated aliphatic alkane having the specified number of carbon atoms by removing one hydrogen. For example, "C 1-10 "Alkyl" refers to C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 Alkyl, "C 1-6 Alkyl", "C 1-4 Alkyl", "C 1-3 "alkyl"; specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, sec-butyl, 2-methylbutyl, 1,1-dimethylbutyl, etc.

[0075] Unless otherwise specified, the term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by a halogen atom. 1-6 Alkyl, more preferably halogenated C 1-4 Examples of haloalkyl groups include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, and the like. Alkyl groups are as defined above.

[0076] Unless otherwise specified, the term "alkoxy" refers to an alkyl group as defined herein attached to another group through an oxygen atom, i.e., "alkyl-O-". 1-6 Alkoxy" (structure is C 1-6 Alkyl-O-), "C 1-4 "alkoxy", specific examples include but are not limited to methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, etc.; preferably, the "alkoxy" described in the present disclosure is preferably C 1-4 Alkoxy, more preferably C 1-3 Alkoxy.

[0077] Unless otherwise specified, the term "haloalkoxy" refers to a group in which one or more hydrogen atoms in an alkoxy group are replaced by halogen. Preferably, the "haloalkoxy" described in the present disclosure is preferably a "haloC 1-6 Alkoxy", "halogenated C 1-4 Alkoxy". Specific examples of the present disclosure include: fluoromethoxy (including monofluoromethoxy, difluoromethoxy, trifluoromethoxy), -OCH2CF3, -OCHFCH3, etc. Alkoxy is as defined above.

[0078] Unless otherwise specified, the term "cycloalkyl" refers to a saturated cyclic alkyl derived from a cycloalkane by removing a hydrogen atom, including a monocyclic or polycyclic saturated hydrocarbon group; the polycyclic saturated hydrocarbon group refers to a polycyclic group formed by two or more cyclic alkyl structures connected by spiro, bridge, condensed, etc. The carbon atoms in the cycloalkyl group can be further oxidized to form C(O). Unless otherwise specified, the "monocyclic cycloalkyl" described herein can be understood as a monocyclic cycloalkyl group. When it is polycyclic, it will be specifically specified as a spiro, condensed or bridged ring group. The cycloalkyl group includes "C 3-8 Cycloalkyl", "C 3-6 Cycloalkyl", "C 3-5 Preferably, the cycloalkyl group is a monocyclic, saturated structure; specific examples include but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0079] Unless otherwise specified, the term "heterocyclyl" refers to a saturated cyclic group derived from the replacement of one or more ring carbon atoms in a cycloalkyl group by a heteroatom and / or a heteroatom group. The heteroatom and / or heteroatom group is generally selected from N, O, S, NO, SO, S(O)2, P(O), and NR (R is H or other substituents defined herein), wherein the carbon atoms in the heterocyclic ring are optionally oxidized, i.e., to form -C(O); preferably, the heteroatom is independently selected from 1-3 N and / or O. The heterocyclyl includes a monoheterocyclyl, a fused heterocyclyl, a spiroheterocyclyl and a bridged heterocyclyl; the heterocyclyl includes "3-8 membered heterocyclyl", "3-6 membered heterocyclyl", "3-5 membered heterocyclyl", "4-6 membered heterocyclyl", "5-6 membered heterocyclyl". Specific examples include, but are not limited to, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, wait.

[0080] Unless otherwise specified, the term "fused heterocyclic group" refers to a saturated non-aromatic cyclic group formed by two or more cyclic structures sharing two adjacent atoms, containing at least one ring atom as a heteroatom; the heteroatom is generally selected from N, O, and S; the ring carbon atoms and heteroatoms in the fused heterocyclic ring can be further oxidized to form a cyclic group containing C(O), NO, SO, or S(O)2 groups, which is also included in the definition of the heterocyclic group described in the present disclosure. The condensation mode disclosed herein can be a 5-6-membered heterocyclyl and a 5-6-membered heterocyclyl, a 5-6-membered heterocyclyl and a 5-6-membered cycloalkyl, a benzo 5-6-membered heterocyclyl, a benzo 5-6-membered saturated heterocyclyl, a 5-6-membered heteroaryl and a 5-6-membered heterocyclyl, a 5-6-membered heteroaryl and a 5-6-membered saturated heterocyclyl, a benzo 5-6-membered heterocyclyl and a 5-6-membered heterocyclyl, a 5-6-membered heteroaryl and a 5-6-membered heterocyclyl, a 5-6-membered heteroaryl and a 5-6-membered heterocyclyl, a benzo 5-6-membered cycloalkyl and a 5-6-membered heterocyclyl, and a 5-6-membered heteroaryl and a 5-6-membered cycloalkyl and a 5-6-membered heterocyclyl.

[0081] Unless otherwise specified, the term "spiroheterocyclyl" refers to a cyclic structure derived from the replacement of at least one carbon atom in a "spiroheterocyclyl" by a heteroatom / heteroatom group, wherein the heteroatom / heteroatom group is selected from N, O, S, P, NO, SO, S(O)2, P(O) and NR, and R is H or any substituent group that may exist, including but not limited to a cyclic structure formed by a heterocyclic spiroheterocycle and a heterocyclic spiroheteroalkane. The spiroheterocycle preferably contains 1-2 heteroatoms selected from NR and / or O, more preferably 1 NR and 0-1 NR or O heteroatoms. The spiroheterocycle is preferably a "nitrogen-containing spiroheterocycle", which refers to a spiroheterocycle in which at least one ring atom is NR. The spiroheterocycle includes a 7-11-membered spiroheterocycle, a 7-9-membered spiroheterocycle, a 7-11-membered nitrogen-containing spiroheterocycle, and a 7-9-membered nitrogen-containing spiroheterocycle. Specific examples include but are not limited to: wait.

[0082] Unless otherwise specified, the term "bridged heterocyclic group" refers to a cyclic structure derived from a "bridged heterocyclic group" in which at least one carbon atom is replaced by a heteroatom / heteroatom group, wherein the heteroatom / heteroatom group is selected from N, O, S, P, NO, SO, S(O)2, P(O) and NR, and R is H or any substituent group. The bridged heterocyclic ring is preferably a "nitrogen-containing bridged heterocyclic ring", which refers to a heterocyclic ring in which at least one ring atom is NR. The bridged heterocyclic ring includes "6-10 membered bridged heterocyclic rings", "6-8 membered bridged heterocyclic rings", etc. Specific examples include but are not limited to: wait.

[0083] Unless otherwise specified, the term "aryl" refers to an unsaturated, usually aromatic hydrocarbon group which may be a single ring or multiple rings fused together. Preferably, the term "aryl" refers to a carbocyclic ring system wherein the ring atoms are carbon atoms. 5-10 Aryl or C 6-10 Aryl, more preferably C 5-8 Aryl or C 6-8 Aryl, most preferably monocyclic C 5-6 Aryl or C6 aryl; examples of aryl include, but are not limited to, phenyl and naphthyl.

[0084] Unless otherwise specified, the term "condensed ring aromatic group" refers to a condensed ring group formed by two or more aromatic rings sharing two adjacent carbon atoms, including naphthyl, anthracenyl and phenanthrenyl.

[0085] The "heteroaryl" group described in the present disclosure refers to a monocyclic group with aromatic properties in which at least one ring atom is a heteroatom and / or a heteroatom group, wherein the heteroatom and / or heteroatom group is generally selected from N, O, S, P, NO, SO, S(O)2, P(O) and NR, R is H or any substituent group, wherein the carbon atoms in the heterocyclic ring are optionally oxidized, i.e., to form -C(O); preferably, the heteroatoms are independently selected from 1-3 N and / or O. The heteroaryl group includes "5-6 membered heteroaryl"; specific examples include, but are not limited to, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, and pyrimidinyl.

[0086] The "5-6 membered heteroaryl" described in the present disclosure preferably contains at least 3 heteroatoms selected from N, O, and S. Specific examples include but are not limited to

[0087] Combinations of substituents and / or variables described herein are permissible only if these combinations result in stable compounds or useful synthetic intermediates. A stable compound or structure is one that is sufficiently robust to survive chemical reactions, to be isolated to a useful degree of purity, and to be formulated into an efficacious therapeutic agent. DETAILED DESCRIPTION

[0088] In the embodiments of the present disclosure, the naming of the title compound is converted from the compound structure with the help of Chemdraw. If there is an inconsistency between the compound name and the compound structure, it can be determined by integrating relevant information and reaction routes; if it cannot be confirmed by other means, the given compound structure shall prevail. The preparation method of some compounds in this disclosure quotes the preparation method of the aforementioned similar compounds. Those skilled in the art should know that when using or referring to the preparation method cited, the feed ratio of the reactants, the reaction solvent, the reaction temperature, etc. can be appropriately adjusted according to the different reactants.

[0089] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

[0090] Experimental instruments:

[0091] The structures of the compounds disclosed herein are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS), or ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker Neo 400M or Bruker Ascend 400 NMR instrument, using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), deuterated chloroform (CDCl3), and heavy water (D2O) as the internal standard, with tetramethylsilane (TMS).

[0092] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1260-6125B single quadrupole mass spectrometer with a Welch Biomate column (C18, 2.7 μm, 4.6 × 50 mm) or a Waters H-Class SQD2 with a Welch Ultimate column (XB-C18, 1.8 μm, 2.1 × 50 mm) mass spectrometer (electrospray ionization as the ion source).

[0093] Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters UPLC H-class SQD mass spectrometer (electrospray ionization as the ion source).

[0094] HPLC analysis was performed using Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high performance liquid chromatography.

[0095] Preparative HPLC used a Waters 2555-2489 (10 μm, ODS 250 cm×5 cm) or a GILSON Trilution LC, and a Welch XB-C18 column (5 μm, 21.2×150 mm).

[0096] Thin layer chromatography silica gel plates use GF254 silica gel plates from Yantai Jiangyou Silica Gel Development Co., Ltd. or GF254 silica gel plates from Rushan Shangbang New Materials Co., Ltd. The specifications used for TLC are 0.15mm-0.20mm, and the preparative type is 20×20cm. Column chromatography generally uses 200-300 mesh silica gel from Chenghua as a carrier.

[0097] The starting materials in the examples of this disclosure are known and commercially available, or can be synthesized using or according to methods known in the art. Unless otherwise specified, all reactions of this disclosure were carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, using dry solvents, and reaction temperatures in degrees Celsius or °C.

[0098] The abbreviations and corresponding names used in the examples are as follows:

[0099] Example 1 N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0100] Step A: To a solution of 5-bromo-6-fluoropyridin-2-amine (20 g, 0.1 mol) in 1,4-dioxane (500 mL) were added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (7.7 g, 0.01 mol), diboronic acid pinacol ester (38 g, 0.15 mol) and potassium acetate (29 g, 0.3 mol) in sequence at room temperature. After nitrogen replacement, the mixture was stirred at 110 ° C for 16 h. After the reaction, the reaction mixture was cooled to room temperature and extracted with ethyl acetate (500 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain 6-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (16 g). MS (ESI) M / Z: 239.2 [M+H] + .

[0101] Step B: To a solution of 6-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (3 g, 12.6 mmol) in 1,4-dioxane / water (volume ratio 5 / 1, 50 mL) were added 4-bromo-1-methyl-1H-1,2,3-triazole (3 g, 18.8 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.84 g, 2.52 mmol) and potassium carbonate (5.2 g, 37.8 mmol) in sequence at room temperature. After nitrogen substitution, the mixture was reacted at 100 ° C for 3 h. After the reaction, the reaction mixture was cooled to room temperature, quenched with ice water, extracted with ethyl acetate (100 mL × 3), and the organic phases were combined, washed with saturated brine (100 mL × 3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (5% methanol in dichloromethane) to obtain 6-fluoro-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (1.3 g). MS (ESI) M / Z: 194.1 [M+H] + .

[0102] Step C: To a solution of 6-fluoro-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (1.3 g, 6.7 mmol) in dimethyl sulfoxide (15 mL) were added 4,4-difluoropiperidine (2.43 g, 20.1 mmol) and potassium carbonate (2.8 g, 20.1 mmol) in sequence. After nitrogen substitution, the mixture was microwaved at 160°C for 16 h. After cooling to room temperature, the reaction solution was directly purified by C18 reverse phase column (acetonitrile / water = 40 / 60) to obtain 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (0.6 g). MS (ESI) M / Z: 295.2 [M+H] + .

[0103] Step D: To a solution of 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (0.12 g, 0.34 mmol) in tetrahydrofuran (2 mL) was added thionyl chloride (0.12 g, 1.02 mmol) dropwise at room temperature. A catalytic amount of N,N-dimethylformamide (3 mg, 0.034 mmol) was then added to the reaction system, and the reaction was allowed to react at 70°C for 2 h. The reaction system was then concentrated to dryness to obtain a residue. A solution of 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (0.1 g, 0.34 mmol) in tetrahydrofuran (2 mL) was then added to the residue obtained in the previous step. Potassium tert-butoxide (1.7 mL, 1.7 mmol, 1 M solution in tetrahydrofuran) was then added to the reaction system, and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice water and extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (5% methanol in dichloromethane) to obtain the target compound, N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (68 mg). MS (ESI) M / Z: 634.2 [M+H] + .

[0104] Step E: To a solution of N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (68 mg, 0.1 mmol) in N,N-dimethylformamide (2 mL) were added 2-hydroxyethane-1-sulfonamide (25 mg, 0.2 mmol), cuprous iodide (19 mg, 0.1 mmol), sarcosine (9 mg, 0.1 mmol) and potassium phosphate (85 mg, 0.4 mmol) in sequence at room temperature. After nitrogen substitution, the reaction system was placed at 120 ° C for 16 hours. The reaction system was cooled to room temperature, and the reaction solution was directly purified by high performance liquid chromatography to obtain the target product, N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (18.2 mg). MS (ESI) M / Z: 631.3 [M+H] + . 1 H NMR (400MHz, DMSO) δ13.29(s,1H),8.53(s,1H),8.26(d,J=8.3Hz,1H),8.06(dd,J=14.8,8.5Hz,2H),7.26(d,J=1.9Hz,1H),7.12(dd,J=8.6,2. 0Hz,1H),4.13(s,3H),3.76(t,J=6.5Hz,2H),3.36(s,2H),3.18–3.14( m,4H),3.00(s,4H),2.21–2.12(m,4H),1.97–1.49(m,4H),0.40(s,4H).

[0105] Example 2 N-(6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0106] Step A: To a solution of methyl 6-bromo-5-fluoropicolinate (2 g, 8.6 mmol) and 4H-1,2,4-triazole (0.59 g, 8.6 mmol) in N,N-dimethylformamide (30 mL) was added potassium carbonate (3.56 g, 25.8 mmol) at room temperature, followed by stirring at 80°C for 2 h. After completion of the reaction, the reaction mixture was cooled to room temperature, quenched with ice water, and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain the target compound, methyl 6-bromo-5-(4H-1,2,4-triazol-4-yl)picolinate (2 g). MS (ESI) M / Z: 283.0 [M+H] + .

[0107] Step B: To a solution of methyl 6-bromo-5-(4H-1,2,4-triazol-4-yl)picolinate (2 g, 7.1 mmol) in N,N-dimethylformamide (30 mL) at room temperature were added 4,4-difluoropiperidine (1 g, 8.5 mmol) and potassium carbonate (2.9 g, 21.3 mmol). The mixture was then stirred in an oil bath at 100°C for 4 h. After completion of the reaction, the reaction mixture was cooled to room temperature, quenched with ice water (50 mL), and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain the target compound, methyl 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)picolinate (2 g). MS (ESI) M / Z: 324.0 [M+H] + .

[0108] Step C: To a solution of methyl 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)picolinate (2 g, 6.2 mmol) in tetrahydrofuran / methanol / water (4 / 2 / 1) (20 mL) was added lithium hydroxide (0.74 g, 31 mmol) at room temperature. The mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was rotary evaporated to remove the solvent to obtain a residue. The pH was adjusted to 5-6 with dilute hydrochloric acid (1 M). A solid precipitated and the filter cake was filtered and washed with methyl tert-butyl ether. The filter cake was then lyophilized to obtain the target compound, 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)picolinic acid (1.5 g). MS (ESI) M / Z: 310.0 [M+H] + .

[0109] Step D: To a solution of 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)picolinic acid (1.5 g, 4.8 mmol) in N,N-dimethylformamide (30 mL) were added diisopropylethylamine (0.89 g, 6.9 mmol) and diphenylphosphoryl azide (1.9 g, 6.9 mmol) at room temperature. The mixture was moved into an oil bath and stirred at 90°C for 5 mins. Water (0.43 g, 24 mmol) was then added and the reaction was continued to stir at this temperature for 2 h. The reaction mixture was cooled to room temperature and quenched with ice water (50 mL). The mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the target compound 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-amine (1.2 g). MS (ESI) M / Z: 281.0 [M+H] + .

[0110] Step E:

[0111] At room temperature, thionyl chloride (0.64 g, 5.4 mmol) was added dropwise to a solution of 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (0.64 g, 1.8 mmol) in tetrahydrofuran (10 mL), and then a catalytic amount of N,N-dimethylformamide (13 mg, 0.18 mmol) was added to the reaction system. The reaction system was placed at 70 ° C. for 2 hours. The reaction system was then directly concentrated to obtain a residue, and then a solution of 6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-amine (0.5 g, 1.8 mmol) in tetrahydrofuran (5 mL) was added to the residue obtained in the previous step, and tert-butanol was added. Potassium (9 mL, 9 mmol, 1 M tetrahydrofuran solution) was added to the reaction system and stirred at room temperature for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the target compound, N-(6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.5 g). MS (ESI) M / Z: 620.2 [M+H] + .

[0112] Step F: To a solution of N-(6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.5 g, 0.81 mmol) in N,N-dimethylformamide (5 mL) were added 2-hydroxyethane-1-sulfonamide (202 mg, 1.62 mmol), cuprous iodide (153 mg, 0.81 mmol), sarcosine (72 mg, 0.81 mmol) and potassium phosphate (686 mg, 3.24 mmol) in sequence at room temperature. After nitrogen substitution, the reaction system was placed at 120°C for 16 hours. The reaction system was cooled to room temperature, and the reaction solution was directly purified by high performance liquid chromatography to obtain the target product, N-(6-(4,4-difluoropiperidin-1-yl)-5-(4H-1,2,4-triazol-4-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (107.52 mg). MS (ESI) M / Z: 617.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ13.39(s,1H),9.02(s,1H),8.29(s,1H),8.09(d,J=8.6 Hz,1H),7.99(d,J=8.4Hz,1H),7.83(d,J=8.4Hz,1H),7.29(d,J=1.8Hz,1H),7.1 4(dd,J=8.6,1.9Hz,1H),3.77(t,J=6.5Hz,2H),3.36(t,J=6.5Hz,2H),3.10–3.0 4(m,4H),3.03-2.99(m,4H),2.02–1.93(m,4H),1.91–1.32(m,4H),0.40(s,4H).

[0113] Example 3 2-(3-Azabicyclo[3.2.1]oct-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)benzamide

[0114] Step A: Methyl 4-bromo-2-fluorobenzoate (5 g, 21.46 mmol), 3-azabicyclo[3.2.1]octane hydrochloride (4.75 g, 32.19 mmol), and N,N-diisopropylethylamine (13.87 g, 107.30 mmol) were dissolved in dimethyl sulfoxide (100 mL). The reaction mixture was heated to 120°C and stirred for 12 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate / water (200 mL / 100 mL). The mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by flash silica gel column chromatography (ethyl acetate / petroleum ether = 5%) to give methyl 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromobenzoate (6.4 g) as a yellow solid. MS (ESI) M / Z: 324.0 [M+H] + .

[0115] Step B: To a mixture of methyl 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromobenzoate (6.4 g, 19.74 mmol) in tetrahydrofuran (80 mL) and water (20 mL) was added sodium hydroxide (4.74 g, 118.44 mmol), and the reaction mixture was heated to 80°C and stirred for 36 hours. The reaction mixture was then cooled to room temperature, concentrated under reduced pressure, and the pH was adjusted to 5-6 with hydrochloric acid (2 M). A white solid precipitated, which was filtered and dried to obtain 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromobenzoic acid (5.2 g) as a white solid. MS (ESI) M / Z: 310.0 [M+H] + .

[0116] Step C: Under nitrogen, to a mixed solution of 6-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (4 g, 16.80 mmol), 3-bromo-1-methyl-1H-1,2,4-triazole (3.27 g, 20.16 mmol) and potassium carbonate (6.97 g, 50.40 mmol) in dioxane (60 mL) and water (15 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (1.37 g, 1.68 mmol). The reaction mixture was purged with nitrogen three times and then reacted at 100°C for 4 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate / water (200 mL / 100 mL). The mixed solution was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by flash silica gel column chromatography (methanol / dichloromethane = 5%) to give 6-fluoro-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-amine (800 mg). MS (ESI) M / Z: 194.1 [M+H]+ .

[0117] Step D: A solution of 6-fluoro-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-amine (400 mg, 2.07 mmol), 4,4-difluoropiperidine (0.75 g, 6.21 mmol), and N,N-diisopropylethylamine (1.34 g, 10.35 mmol) in dimethyl sulfoxide (10 mL) was heated in a microwave oven at 160°C for 5 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate / water (100 mL / 50 mL). The mixture was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by flash silica gel column chromatography (methanol / dichloromethane = 5%) to afford 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-amine (290 mg) as a yellow solid. MS (ESI) M / Z: 295.1 [M+H] + .

[0118] Step E: 2-(3-Azabicyclo[3.2.1]octan-3-yl)-4-bromobenzoic acid (0.30 g, 0.97 mmol) was dissolved in N,N-dimethylformamide (10 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.50 g, 1.32 mmol) was added. The reaction solution was stirred for 10 minutes, and then 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-amine (260 mg, 0.88 mmol) and N,N-diisopropylethylamine (0.23 g, 1.76 mmol) were added. The reaction solution was heated and stirred at 50°C in a microwave for 4 hours. The reaction mixture was then cooled to room temperature and diluted with ethyl acetate / water (100 ml / 50 ml). The mixed solution was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by flash silica gel column chromatography (ethyl acetate / petroleum ether = 50%) to obtain 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromo-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)benzamide (370 mg, yield = 71.41%). MS (ESI) M / Z: 586.2 [M+H] + .

[0119] Step F: To a solution of 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromo-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)benzamide (60 mg, 0.10 mmol) and 2-hydroxyethane-1-sulfonamide (37.55 mg, 0.30 mmol) in dimethyl sulfoxide (3 mL) were added cuprous iodide (38.09 mg, 0.20 mmol), potassium phosphate (42.45 mg, 0.20 mmol) and (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (28.45 mg, 0.20 mmol) in sequence. The reaction mixture was then heated to 150°C under nitrogen protection in a microwave for 1 hour. The reaction mixture was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (50 mL), and the organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified and separated by high-performance liquid chromatography (formic acid) to obtain 2-(3-azabicyclo[3.2.1]octan-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)benzamide (19.95 mg). MS (ESI) M / Z: 631.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.85 (s, 1H), 8.53 (s, 1H), 8.11 (d, J = 8.4Hz, 1H), 7.94 (d,J=8.4Hz,1H),7.71(d,J=8.5Hz,1H),7.14(d,J=2.1Hz,1H),6.98(dd,J=8.4, 2.0Hz,1H),3.92(s,3H),3.75(t,J=6.6Hz,2H),3.29(t,J=6.6Hz,6H),3.05-2.9 5(m,2H),2.83-2.79(m,2H),2.25(s,2H),2.08-1.99(m,6H),1.65-1.46(m,4H).

[0120] Example 4 N-(6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0121] Step A: To a solution of 2,6-dibromo-3-nitropyridine (10 g, 35.7 mmol) and 4,4-difluoropiperidine (4.3 g, 35.7 mmol) in tetrahydrofuran (100 mL) was added potassium carbonate (14.5 g, 107.1 mmol) at room temperature, followed by stirring at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched with ice water and extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain the target compound, 6-bromo-2-(4,4-difluoropiperidin-1-yl)-3-nitropyridine (6 g). MS (ESI) M / Z: 322.0 [M+H] + .

[0122] Step B: Under ice bath, to a solution of 6-bromo-2-(4,4-difluoropiperidin-1-yl)-3-nitropyridine (6 g, 18.6 mmol) in N,N-dimethylformamide (60 mL) was added bipyridine (0.3 g, 1.9 mmol). After complete dissolution, B2(OH)4 (8.3 g, 93 mmol) was added to the system in small amounts and batches. During this period, the reaction solution changed from light yellow to dark brown and then back to light yellow within 10 minutes, and the reaction was completed. After the reaction mixture was cooled to room temperature, it was quenched with ice water (100 mL) and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain the target compound 6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridin-3-amine (2.9 g). MS (ESI) M / Z: 292.0 [M+H] + .

[0123] Step C: To a solution of 6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridin-3-amine (2.9 g, 1 mmol) in acetonitrile (40 mL) was added tert-butyl nitrite (1.5 g, 1.5 mmol) under ice-cooling. The mixture was stirred for 10 minutes, and then trimethylsilyl azide (1.7 g, 1.5 mmol) was added. The mixture was stirred for 1 hour. The reaction mixture was cooled to room temperature, quenched with ice water (100 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with saturated brine (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain the target compound, 3-azido-6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridine (1.8 g). MS (ESI) M / Z: 318.0 [M+H]+ .

[0124] Step D: To a solution of 3-azido-6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridine (1.8 g, 5.7 mmol) in tetrahydrofuran (20 mL) at room temperature were added 2-methylbut-3-yn-2-ol (0.5 g, 6 mmol), copper acetate (0.1 g, 0.6 mmol), sodium ascorbyl palmitate (0.1 g, 0.6 mmol) and water (0.5 g, 28.5 mmol) in sequence. After nitrogen replacement, the reaction was stirred at room temperature for 16 hours. The reaction mixture was quenched with ice water (50 mL) and extracted with dichloromethane / methanol = 10 / 1 (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the target compound 2-(1-(6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridin-3-yl)-1H-1,2,3-triazol-4-yl)propan-2-ol (1.2 g). MS (ESI) M / Z: 402.0 [M+H] + .

[0125] Step E: To a solution of 2-(1-(6-bromo-2-(4,4-difluoropiperidin-1-yl)pyridin-3-yl)-1H-1,2,3-triazol-4-yl)propan-2-ol (1.2 g, 3 mmol) in 1,4-dioxane (20 mL) were added tris(dibenzylideneacetone)dipalladium (0.24 g, 0.3 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (0.22 g, 0.45 mmol), cesium carbonate (2.9 g, 9 mmol) and tert-butyl carbamate (0.7 g, 6 mmol) at room temperature. After nitrogen substitution, the reaction was continued with stirring at 100°C for 2 hours. After the reaction, the reaction mixture was cooled to room temperature, quenched with ice water (50 mL), and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 3), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the target compound (tert-butyl 6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)carbamate (1 g). MS (ESI) M / Z: 439.2 [M+H] + .

[0126] Step F: To a solution of tert-butyl (6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)carbamate (1 g, 2.3 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (5 mL) at room temperature. The mixture was stirred at room temperature for 1 hour. After the reaction, the solvent was removed by vortexing to obtain the target compound, 2-(1-(6-amino-2-(4,4-difluoropiperidin-1-yl)pyridin-3-yl)-1H-1,2,3-triazol-4-yl)propan-2-ol, as a trifluoroacetate salt (0.8 g). MS (ESI) M / Z: 339.0 [M+H] + .

[0127] Step G: To a solution of 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (0.86 g, 2.4 mmol) in tetrahydrofuran (10 mL) was added dropwise thionyl chloride (0.86 g, 7.2 mmol) at room temperature. A catalytic amount of N,N-dimethylformamide (15 mg, 0.2 mmol) was then added to the reaction system, and the reaction system was placed at 70°C for 2 hours. The reaction system was then concentrated directly to obtain a residue, and a solution of trifluoroacetate of 2-(1-(6-amino-2-(4,4-difluoropiperidin-1-yl)pyridin-3-yl)-1H-1,2,3-triazol-4-yl)propan-2-ol (0.8 g, 2.4 mmol) in tetrahydrofuran (10 mL) was added to the residue obtained in the previous step. Potassium tert-butoxide (12 mL, 12 mmol, 1 M tetrahydrofuran solution) was then added to the reaction system, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the target compound, N-(6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.8 g). MS (ESI) M / Z: 678.2 [M+H] + .

[0128] Step H: To a solution of N-(6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.2 g, 0.3 mmol) in N,N-dimethylformamide (5 mL) were added 2-hydroxyethane-1-sulfonamide (75 mg, 0.6 mmol), cuprous iodide (57 mg, 0.3 mmol), sarcosine (27 mg, 0.3 mmol) and potassium phosphate (254 mg, 1.2 mmol) in sequence at room temperature. After nitrogen substitution, the reaction system was placed at 120 ° C for 16 hours. The reaction system was cooled to room temperature, and the reaction solution was directly purified by high performance liquid chromatography to obtain the target product, N-(6-(4,4-difluoropiperidin-1-yl)-5-(4-(2-hydroxypropan-2-yl)-1H-1,2,3-triazol-1-yl)pyridin-2-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (83.95 mg). MS (ESI) M / Z: 675.2 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ13.44(s,1H),8.30(s,1H),8.10(d,J=8.6Hz,1H),8. 00(d,J=8.4Hz,1H),7.88(d,J=8.4Hz,1H),7.29(d,J=1.9Hz,1H),7.15(dd,J=8 .7,2.0Hz,1H),5.27(s,1H),3.77(t,J=6.5Hz,2H),3.36(t,J=6.5Hz,2H),3.03 -2.97(m,8H),2.01–1.94(m,4H),1.90–1.61(m,4H),1.54(s,6H),0.40(s,4H).

[0129] The following target compounds were prepared by referring to the synthesis methods of the above examples:

[0130] Example 17 2-(3-Azabicyclo[3.2.1]octan-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-(methylsulfonamido)benzamide

[0131] Step A: To a solution of 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (1 g, 3.1 mmol) in acetonitrile (20 mL) were added N-methylimidazole (0.75 g, 9.3 mmol), tetramethylchlorouronium hexafluorophosphate (1.74 g, 6.2 mmol) and 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-(methylsulfonamido)benzoic acid (0.9 g, 3.1 mmol) in sequence at room temperature, and the reaction system was placed at 70°C for 6 h. The reaction system was cooled to room temperature, the reaction solution was directly concentrated, and the residue was purified by silica gel column chromatography to obtain the target compound 2-(3-azabicyclo[3.2.1]octan-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-4-(methylsulfonamido)benzamide (645 mg). MS (ESI) M / Z: 601.3 [M+H] + .

[0132] 1 H NMR (400MHz, DMSO-d6) δ10.95(s,1H),10.15(s,1H),8.57(s,1H),8.35(d,J=8.4Hz, 1H),8.15(d,J=8.4Hz,1H),7.81(d,J=8.4Hz,1H),7.23(s,1H),7.08(dd,J=8.5,1.7 Hz,1H),4.18(s,3H),3.18-3.15(m,4H),3.14(s,3H),3.08-3.06(m,2H),2.89-2.86 (m,2H),2.33-2.29(m,2H),2.24–2.14(m,4H),2.05-2.01(m,2H),1.66–1.53(m,4H).

[0133] Example 23 2-(3-Azabicyclo[3.2.1]octan-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluoro-4-(methylsulfonamido)benzamide

[0134] Step A: To a solution of methyl 4-bromo-2,5-difluorobenzoate (2 g, 8 mmol) and 3-azabicyclo[3.2.1]octane hydrochloride (1.3 g, 8.8 mmol) in N-methylpyrrolidone (30 mL) was added potassium carbonate (3.3 g, 24 mmol) at room temperature. The mixture was then stirred in a sealed container at 120°C for 4 h. The reaction mixture was cooled to room temperature, quenched with ice water, and extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using 10% ethyl acetate in petroleum ether to yield the target compound, methyl 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromo-5-fluorobenzoate (1.6 g). MS (ESI) M / Z: 342.0 [M+H] + .

[0135] Step B: To a solution of methyl 2-(3-azabicyclo[3.2.1]octan-3-yl)-4-bromo-5-fluorobenzoate (0.6 g, 1.75 mmol) in N,N-dimethylformamide (10 mL) were added methylsulfonamide (0.5 g, 5.25 mmol), CuI (0.33 g, 1.75 mmol), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (124 mg, 0.87 mmol) and potassium phosphate (1.5 g, 7 mmol) in an ice bath. After nitrogen substitution, the reaction system was stirred at 120°C for 6 h. The reaction system was cooled to room temperature, quenched with ice water, and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using 5% methanol and dichloromethane as the eluent to obtain the target compound, methyl 2-(-3-azabicyclo[3.2.1]octan-3-yl)-5-fluoro-4-(methylsulfonamido)benzoate (0.4 g). MS (ESI) M / Z: 357.2 [M+H] + .

[0136] Step C: To a solution of methyl 2-((1R,5S)-3-azabicyclo[3.2.1]octan-3-yl)-5-fluoro-4-(methylsulfonamido)benzoate (0.4 g, 1.1 mmol) in tetrahydrofuran / methanol / water (10 mL) was added sodium hydroxide (0.22 g, 5.5 mmol) at room temperature, followed by stirring at 70°C for 2 h. The reaction mixture was cooled to room temperature and the pH was adjusted to a weakly acidic state with hydrochloric acid (1 M). A white solid precipitated, which was filtered and the filter cake was lyophilized to yield the target compound, 2-(3-azabicyclo[3.2.1]octan-3-yl)-5-fluoro-4-(methylsulfonamido)benzoic acid (0.25 g). MS (ESI) M / Z: 343.2 [M+H] + .

[0137] Step D: To a solution of 2-(3-azabicyclo[3.2.1]octan-3-yl)-5-fluoro-4-(methylsulfonamido)benzoic acid (0.15 g, 0.44 mmol) in acetonitrile (10 mL) were added N-methylimidazole (108 mg, 1.32 mmol), tetramethylchlorouronium hexafluorophosphate (246 mg, 0.88 mmol) and 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (130 mg, 0.44 mmol) in sequence at room temperature, and the reaction system was placed at 70 ° C for 6 h. The reaction system was cooled to room temperature, the reaction solution was directly concentrated, and the residue was purified by silica gel column chromatography to obtain the target compound 2-(-3-azabicyclo[3.2.1]octan-3-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluoro-4-(methylsulfonamido)benzamide (65 mg). MS (ESI) M / Z: 619.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.34(s,1H),10.31(s,1H),8.53(s,1H),8.29(d,J=8. 4Hz,1H),8.09(d,J=8.4Hz,1H),7.62(d,J=11.5Hz,1H),7.43(d,J=7.3Hz,1H),4 .13(s,3H),3.13-3.10(m,4H),3.05(s,3H),2.96-2.93(m,2H),2.87-2.84(m,2 H),2.26-2.23(m,2H),2.19–2.08(m,4H),2.01-1.97(m,2H),1.66–1.53(m,4H).

[0138] Example 28 2-(4-(difluoromethylene)piperidin-1-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluoro-4-(methylsulfonamido)benzamide

[0139] Step A: To a solution of 4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)-5-fluorobenzoic acid (0.2 g, 0.56 mmol) in acetonitrile (10 mL) were added N-methylimidazole (138 mg, 1.68 mmol), tetramethylchlorouronium hexafluorophosphate (314 mg, 1.12 mmol) and 6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-amine (165 mg, 0.56 mmol) in sequence at room temperature, and the reaction system was placed at 70 ° C for 4 h. The reaction system was cooled to room temperature, and the reaction solution was directly concentrated and purified by silica gel column chromatography (10% methanol / dichloromethane) to obtain the target compound, 4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluorobenzamide (0.28 g). MS (ESI) M / Z: 626.2 [M+H] + .

[0140] Step B: To a solution of 4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluorobenzamide (80 mg, 0.13 mmol) in dimethyl sulfoxide (2 mL) were added methylsulfonamide (37 mg, 0.39 mmol), cuprous iodide (25 mg, 0.13 mmol), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (10 mg, 0.07 mmol) and potassium phosphate (110 mg, 0.52 mmol) in sequence at room temperature. After nitrogen substitution, the reaction system was placed under microwave at 100 ° C for 4 h. The reaction system was cooled to room temperature, and the reaction solution was directly purified by preparative HPLC to obtain the target product, 2-(4-(difluoromethylene)piperidin-1-yl)-N-(6-(4,4-difluoropiperidin-1-yl)-5-(1-methyl-1H-1,2,3-triazol-4-yl)pyridin-2-yl)-5-fluoro-4-(methylsulfonamido)benzamide (17.13 mg). MS (ESI) M / Z: 641.2 [M+H].

[0141] 1H NMR (400MHz, DMSO-d6) δ13.37(s,1H),10.44(s,1H),8.52(s,1H),8.30(d,J=8.0Hz,1H),8.03(d,J=7.6Hz,1H),7.8 4(d,J=11.6Hz,1H),7.44(d,J=6.2Hz,1H),4.13(s,3H),3.08-3.01(m,11H),2.59-2.56(m,4H),2.13-2.10(m,4H).

[0142] Biological test evaluation:

[0143] Test Example 1: Evaluation of the inhibitory effect of the compounds disclosed herein on human KIF18A actin activity

[0144] This experiment used the ADP-Glo ​​method to test the inhibitory effect of the disclosed compounds on human KIF18A actin activity, and obtained the half-maximal concentration IC of the compounds for inhibiting KIF18A actin activity. 50 .

[0145] Positive reference molecule: AMG-650, whose structure is as follows:

[0146] The abbreviations used in the test examples and their corresponding names are as follows:

[0147] 1. Experimental Materials

[0148] 1) MgCl2, KCl, EGTA, DTT, BSA, Tween-20 and PIPES (pH 6.9) buffer were purchased from Sigma to prepare assay buffer.

[0149] 2) KIF18A was purchased from CP

[0150] 3) MT (tubulin) was purchased from Cytoskeleton

[0151] 4) Paclitaxel was purchased from MCE

[0152] 5) Preparation of assay buffer:

[0153] 50 mM PIPES, pH 6.9

[0154] 50 mM KCl

[0155] 0.5 mM EGTA

[0156] 5mM MgCl2

[0157] 1mM DTT

[0158] 0.01% BSA

[0159] 0.005% Tween-20

[0160] 5μM Paclitaxel

[0161] 6) ADP-Glo ​​reagent, purchased from Promega.

[0162] 2. Experimental Methods

[0163] a) Use Echo650 to transfer the test compound or DMSO to a 384-well assay microplate. The test compound is serially diluted with DMSO from the highest test concentration to 3-fold, and DMSO is added to ensure that the final DMSO concentration in each well is consistent (1%).

[0164] b) Add 2.5 μL of KIF18A diluted in assay buffer (10 nM KIF18A) and incubate at room temperature for 30 minutes;

[0165] c) Add 2.5 μL of substrate mixture diluted in assay buffer, consisting of 180 μM ATP (from ADP-Glo ​​reagent) and 0.1 mg / mL MT, and incubate at room temperature for 60 minutes.

[0166] d) Add 5 μL of ADP-Glo ​​R1, centrifuge briefly, and incubate at room temperature for 2 hours;

[0167] e) Add 10 μL of ADP-Glo ​​R2, centrifuge briefly, and incubate at room temperature for 1 hour;

[0168] f) Chemiluminescent signals were detected using Envision microplate reader (PerkinElmer).

[0169] g) GraphPad Prism 6 software was used for data analysis to obtain the IC values ​​of the compounds. 50 .

[0170] 3. Experimental Results

[0171] The results of the inhibition of the human KIF18A protein activity by the compounds disclosed herein are shown in Table 1.

[0172] Table 1 Inhibition of KIF18A protein activity

[0173] Conclusion: The compounds disclosed herein have a strong inhibitory effect on the activity of human KIF18A protein.

[0174] Test Example 2: Evaluation of the inhibitory effect of the compounds disclosed herein on the proliferation of ovarian cancer cells NCIOVCAR3

[0175] This study used the CellTiter-Glo (CTG) method to test the inhibitory effect of compounds on NCIOVCAR3 cell proliferation and to obtain the half-maximal concentration IC of the compounds that inhibited cell growth. 50 .

[0176] 1. Experimental Materials

[0177] a) 1640 culture medium, fetal bovine serum (FBS), and Penicillin-Streptomycin were purchased from GIBCO

[0178] b) CellTiter-Glo reagent, purchased from Promega.

[0179] 2. Experimental Methods

[0180] a) Day 0: NCIOVCAR3 cells were seeded into 384-well plates at a density of 500 cells per well, with 50 μL per well.

[0181] b) Day 1: Add the test compound to the culture plate using TECAN at a final DMSO concentration of 0.5%, add DMSO to the blank control wells at a final concentration of 0.5%, and incubate the culture plate in a cell culture incubator for 120 hours (37°C, 5% CO2).

[0182] c) Day 6: Add 50 μL of Cell Titer-Glo reagent to each well, shake at 500 rpm for 2 minutes, and incubate at room temperature in the dark for 10 minutes to stabilize the luminescence signal.

[0183] d) Chemiluminescent signals were detected using Envision microplate reader (PerkinElmer).

[0184] e) GraphPad Prism 6 software was used for data analysis to obtain the IC values ​​of the compounds. 50 .

[0185] 3. Experimental Results

[0186] The results of the inhibition of the proliferation of ovarian cancer cells NCIOVCAR3 by the compounds disclosed herein are shown in Table 2.

[0187] Table 2 Results of inhibition of NCIOVCAR3 cell proliferation

[0188] Conclusion: The compounds disclosed herein have a strong inhibitory effect on the proliferation of ovarian cancer cell line NCIOVCAR3.

[0189] Test Example 3: Evaluation of the inhibitory effect of the disclosed compounds on the proliferation of chromosomally unstable tumor cells

[0190] The inhibitory activity of the disclosed compounds on the proliferation of chromosomally unstable tumors such as OVCAR8, HT29, MDAMB157, BT549, NCIH23, and Calu6 was tested using methods well known to those skilled in the art, such as the CTG method.

[0191] 3.1 Experimental Materials

[0192] 3.2 Experimental steps

[0193] 1) Cell Culture: All cell culture procedures were performed according to the manufacturer's instructions. Specifically, using HT29 cells as an example, HT29 cells were cultured in RPMI1640 medium supplemented with 10% FBS and passaged every 3-4 days. Passaging: Thoroughly disperse the cells, collect the cell suspension, and centrifuge at 800 rpm for 3 minutes. Discard the supernatant and resuspend the cells in fresh medium. Subculture was performed at a ratio of approximately 1:4.

[0194] 2) HT29 cells were harvested, resuspended in culture medium, counted, and seeded into 96-well white clear-bottom plates, with 800 cells per well in 100 μL of culture medium.

[0195] 3) Using an Echo 650, transfer the disclosed compounds to the 96-well plate at a top concentration of 2 μM, with 3-fold dilutions for a total of 9 concentration points. The total DMSO content was 0.5%. The plate was incubated in a 37°C, 5% CO2 incubator for 7 days.

[0196] 4) After incubation, add 50 μL of CTG to each well, shake for 10 minutes, and then read the chemiluminescence value using Envision.

[0197] GI 50 Calculation: The reading of the well containing only DMSO before the addition of the test compound on the day of addition of the test compound is the low control, and the reading of the well containing only DMSO on the day of the end of drug treatment is the high control. The inhibition rate of the compound = 100% × ((RLU high control -RLU low control )–(RLU compound –RLU low control )) / (RLU high control –RLU low control ); see Table 3 for experimental results.

[0198] Table 3 Inhibitory activity of the compounds disclosed herein on the proliferation of chromosomally unstable tumors

[0199] The results showed that compared with AMG-650, the compound of Example 23 had a stronger inhibitory effect on the proliferation of chromosomally unstable tumors.

[0200] Test Example 4: In vivo pharmacodynamic study of the compounds of the present disclosure in an OVCAR3 transplant tumor model

[0201] 4.1 Experimental animals and breeding environment

[0202] Species: Mouse

[0203] Strain: BALB / c Nude mice

[0204] Arrival age: 6-8 weeks

[0205] Gender: Female

[0206] Weight: 19-24 grams

[0207] Supplier: Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd.

[0208] Animal Certificate: SCXK(Zhejiang)2024-0001 20240603Ab220619000250

[0209] 4.2 Experimental methods and steps

[0210] 1) Cell culture

[0211] Human ovarian cancer OVCAR3 cells (Ovarian Carcinoma, ATCC, Cat No. HTB-161, Lot No. 4470175) were cultured as monolayers in RPMI high-glucose 1640 medium supplemented with 20% heat-inactivated fetal bovine serum, 10 μg / mL insulin, 100 U / mL penicillin, and 100 U / mL streptomycin at 37°C and 5% CO2. Cells were passaged two to three times weekly. When cells reached the exponential growth phase, they were harvested, counted, and plated.

[0212] 2) Tumor cell inoculation and grouping

[0213] will contain 10×10 6 100 μL PBS of 100 OVCAR3 cells and 100 μL Matrigel (1:1) were mixed in equal volumes and inoculated subcutaneously on the back of the right forelimb of each mouse. On the 31st day after inoculation, the average tumor volume reached 132 mm 3 At 4 pm, the mice were randomly divided into groups and drug administration began. Each group consisted of 6 mice, and the drug was orally administered once a day for a total of 28 days. The specific groupings are shown in Table 4.

[0214] Table 4. Experimental animal groups

[0215] Note: The vehicle group consisted of 5% DMSO + 10% Solutol + 85% Saline.

[0216] 3) Tumor measurement and experimental indicators

[0217] Tumor diameter was measured twice a week using a vernier caliper. Tumor volume was calculated using the formula: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.

[0218] The tumor inhibition efficacy of the compound was evaluated using the tumor inhibition rate (TGI) (%). TGI (%) was calculated as follows: TGI (%) = [1 - (mean tumor volume of a treatment group at the end of dosing - mean tumor volume of the treatment group at the start of dosing) / (mean tumor volume of the solvent control group at the end of treatment - mean tumor volume of the solvent control group at the start of treatment)] × 100%.

[0219] The tumor inhibition effects of each experimental group are shown in Table 5.

[0220] Table 5 Inhibition results of OVCAR3 subcutaneous xenograft tumor proliferation

[0221] The experimental results showed that the compound of Example 23 had a better in vivo efficacy against the OVCAR3 transplanted tumor model than the positive reference AMG-650.

[0222] Test Example 5:

[0223] Activity tests were performed using Compound 1 and Compound 5 having the following structures disclosed in CN116554151A as control compounds.

[0224] This study used the ADP-Glo ​​method to test the inhibitory effect of the compound on human KIF18A actin activity and obtained the half-maximal concentration IC of the compound to inhibit KIF18A actin activity. 50 .

[0225] 1. Experimental Materials

[0226] 1) MgCl2, KCl, EGTA, DTT, BSA, Tween-20 and PIPES (pH 6.9) buffer were purchased from Sigma to prepare assay buffer.

[0227] 2) KIF18A was purchased from CP

[0228] 3) MT (tubulin) was purchased from Cytoskeleton, Lot#031

[0229] 4) Paclitaxel was purchased from MCE

[0230] 5) ADP-Glo ​​reagent, purchased from Promega.

[0231] 6) Preparation of assay buffer:

[0232] 50 mM PIPES, pH 6.9

[0233] 50 mM KCl

[0234] 0.5 mM EGTA

[0235] 5mM MgCl2

[0236] 1mM DTT

[0237] 0.01% BSA

[0238] 0.005% Tween-20

[0239] 5μM Paclitaxel

[0240] 2. Experimental Methods

[0241] a) Use Echo650 to transfer the test compound or DMSO into a 384-well microplate. The test compound is transferred to the highest well of the microplate with DMSO.

[0242] b) Perform a 3-fold serial dilution of the test concentration and add DMSO to ensure that the final DMSO concentration in each well is consistent (1%);

[0243] c) Add 2.5 μL of KIF18A diluted in assay buffer (10 nM KIF18A) and incubate at room temperature for 15 minutes;

[0244] d) Add 2.5 μL of substrate mixture diluted in assay buffer, consisting of 180 μM ATP (from ADP-Glo ​​reagent) and 0.1 mg / mL MT, and incubate at room temperature for 60 minutes.

[0245] e) Add 5 μL of ADP-Glo ​​R1, centrifuge briefly, and incubate at room temperature for 2 hours;

[0246] f) Add 10 μL of ADP-Glo ​​R2, centrifuge briefly, and incubate at room temperature for 1 hour;

[0247] g) Chemiluminescent signals were detected using Envision microplate reader (PerkinElmer).

[0248] h) GraphPad Prism 6 software was used for data analysis to obtain the IC values ​​of the compounds. 50 .

[0249] 3. Experimental Results

[0250] The results of the inhibition of the compounds on the activity of human KIF18A protein are shown in Table 6.

[0251] Table 6 Inhibition of KIF18A protein activity

[0252] Conclusion: As can be seen from Table 6, our molecule has stronger inhibitory activity against KIF18A protein.

[0253] Test Example 6: Pharmacokinetics of the compound in mice

[0254] Mice were used as test animals to study the pharmacokinetic behavior of the compound in plasma at a dose of 1 mg / kg intravenous injection and 10 mg / kg oral administration.

[0255] 1. Experimental Plan

[0256] 1.1 Investigational Drugs:

[0257] The compound of Example 23 of the present disclosure and compound 1 and compound 7 of CN116554151A.

[0258] 1.2 Experimental animals

[0259] Twelve male CD-1 mice (3 mice / group) were obtained from Shanghai Jihui Experimental Animal Breeding Co., Ltd.

[0260] 1.3 Administration:

[0261] Twelve male CD-1 mice were used; the mice in the IV administration group were fed freely, and the mice in the PO administration group were fasted. The IV administration dose was 1 mg / kg, and the administration volume was 5 mL / kg; the PO administration dose was 10 mg / kg, and the administration volume was 10 mL / kg.

[0262] 1.4 Experimental Equipment

[0263] Centrifuge (5810R) was purchased from Eppendorf, pipettes were purchased from Eppendorf, and vortexer was purchased from Scientific Industries.

[0264] 1.5 Sample collection

[0265] After administration to mice, 0.1 mL of blood was collected from the saphenous vein at 0.0833 (IV), 0.25, 0.5, 1, 2, 4, 8 and 24 hours, placed in EDTA-K2 tubes, centrifuged at 4600 rpm for 5 minutes at 4°C, and the plasma was separated and stored at -80°C.

[0266] 1.6 Sample processing

[0267] 1) Take 50 μL of plasma sample, add 200 μL of methanol:acetonitrile (v:v=1:1), precipitate, mix, and centrifuge at 2773×g for 15 minutes.

[0268] 2) Take 50 μL of the treated supernatant solution, dissolve it in diluent (methanol:water, v:v=1:1, containing 0.1% formic acid), and analyze the concentration of the test compound by LC / MS / MS.

[0269] 1.7 Liquid phase analysis

[0270] Liquid phase: Shimadzu LC-30AD or LC-40D XS

[0271] Mass spectrometer: AB Sciex 5500

[0272] Chromatographic column: Phenomenex Kinetex C18 (2.6μm, 5mm*3.0mm) or ACE C4 (50mm*2.1mm) or Waters HSS T3 (2.5μm, 50mm*2.1mm)

[0273] Mobile phase: A: 0.1% formic acid in 5 mM ammonium acetate, B: 0.05% or 0.1% formic acid in acetonitrile

[0274] Flow rate: 0.6 mL / min

[0275] Gradient: Gradient elution 0-3 minutes.

[0276] 2. Experimental Results and Analysis

[0277] The main pharmacokinetic parameters were calculated using WinNonlin software. The pharmacokinetic parameters of intravenous injection and oral administration of the drug in mice are shown in Tables 7 and 8.

[0278] Table 7 Pharmacokinetic parameters of compounds injected intravenously into mice

[0279] Table 8 Pharmacokinetic parameters of compounds administered orally to mice

[0280] The experimental results show that the compound T in Example 23 of the present disclosure 1 / 2 The AUC and AUC were significantly better than those of compound 1 and compound 7 of CN116554151A, and showed good pharmacokinetic properties in mice.

Claims

1. A compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: in, Ring A is a 5-6 membered heteroaryl group, and the 5-6 membered heteroaryl group contains at least 3 heteroatoms selected from N, O or S; Ring B is a 4-6 membered heterocyclic group, and the 4-6 membered heterocyclic group contains at least one heteroatom selected from N, O or S; Ring C is a 6-8 membered heterocyclic group, and the 6-8 membered heterocyclic group contains at least one heteroatom selected from N, O or S; X 1 、X 2 、Y 1 and Y 2 Each independently selected from N or CR x ; the R x Selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 alkoxy; R 1 and R 2 are each independently selected from halogen, C 1-6 Alkyl or C 3-6 Cycloalkyl, and the C 1-6 Alkyl or C 3-6 The cycloalkyl group is optionally substituted with one or more radicals selected from OH, halogen, -NH2 or C 1-4 substituted by an alkoxy substituent; R 3 Halogen, OH, C 1-6 Alkyl, C 3-6 Cycloalkyl or R 3a and R 3b are each independently selected from H, halogen or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with one or more radicals selected from halogen, hydroxy, amino, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 3-6 substituted by a cycloalkyl group, a 3-8 membered heterocyclyl group, a phenyl group or a 5-6 membered heteroaryl group; R 4 Selected from -NHSO2-C 1-4 Alkyl, and the C 1-4 The alkyl group is optionally substituted with OH or halogen; m, n and p are each independently selected from 0, 1, 2 or 3.

2. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein The ring A is a 5-membered heteroaryl group and contains at least 3 heteroatoms selected from N, O or S; or, the ring A is a 5-membered heteroaryl group and contains 3 heteroatoms selected from N, or contains 2 heteroatoms selected from N and 1 heteroatom selected from O or S, or contains 4 heteroatoms selected from N; or, the ring A is selected from Alternatively, the ring A is selected from 3. The compound according to claim 1 or 2, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The R 1 Selected from C 1- 6 alkyl, and the C 1-6 The alkyl group is optionally substituted with one or more substituents selected from OH or halogen, and / or m is selected from 0 or 1; or, the R 1 Selected from methyl, CHF2 or and m is selected from 0 or 1; or, said R 1 is methyl, and m is selected from 0 or 1.

4. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The structural unit Selected from Alternatively, the structural unit Selected from Alternatively, the structural unit Selected from 5. The compound according to any one of claims 1 to 4, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The ring B is a 6-membered heterocyclic group containing one heteroatom selected from N; or, the ring B is 6. The compound according to any one of claims 1 to 5, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The R 2 is halogen, and / or n is selected from 0, 1 or 2; or, said R 2 is selected from F, Cl or Br, and n is selected from 0, 1 or 2.

7. The compound according to any one of claims 1 to 6, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The structural unit Selected from 8. The compound according to any one of claims 1 to 7, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The ring C is a 6-, 7- or 8-membered heterocyclic group containing a heteroatom selected from N; or, the ring C is selected from 9. The compound according to any one of claims 1 to 8, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The R 3 for R 3a and R 3b are each independently selected from H or halogen, and p is selected from 0, 1 or 2; or, said R 3 for p is selected from 0, 1 or 2.

10. The compound according to any one of claims 1 to 9, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The structural unit Selected from 11. The compound according to any one of claims 1 to 10, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The X 1 、X 2 Each independently selected from CR x , and the R x is selected from H or halogen; or, said X 1 、X 2 are each independently selected from CH, -CF or N; or, X 1 CH, X 2 It is CH or -CF.

12. The compound according to any one of claims 1 to 11, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The Y 1 、Y 2 are each independently selected from CH or N; or, Y 1 N, Y 2 CH; or, Y 1 N, Y 2 is N.

13. The compound according to any one of claims 1 to 12, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The R 4 Selected from -NHSO2-C 1-4 Alkyl, and the C 1-4 The alkyl group is optionally substituted with one or more OH groups; or, the R 4 Selected from -NHSO2-CH2CH2OH or -NHSO2-CH3.

14. The compound according to any one of claims 1 to 13, its stereoisomer or a pharmaceutically acceptable salt thereof, which is selected from: in, Ring A, Ring B, Ring C, R 1 、R 2 、R 3 、R 4 、X 1 、X 2 , m, p and n are as defined in any one of claims 1-13.

15. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, which is selected from: in, Structural unit Selected from Alternatively, the structural unit Selected from Alternatively, the structural unit Selected from Structural unit Selected from X 2 selected from CH or -CF; R 4 Selected from -NHSO2-CH2CH2OH or -NHSO2-CH3.

16. The compound according to any one of claims 1, 5-7 and 11-15, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from And the structural unit Selected from structural unit Selected from or, Structural unit for And the structural unit Selected from 17. A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, selected from:

18. A pharmaceutical composition comprising the compound according to any one of claims 1 to 17, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

19. Use of the compound according to any one of claims 1 to 17, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 18 in the preparation of a medicament for treating a disease mediated by KIF18A.

20. A method for treating or preventing a disease mediated by KIF18A, comprising administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1 to 17, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 18.

21. The compound according to any one of claims 1 to 17, its stereoisomer or pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 18 for use in treating or preventing a disease mediated by KIF18A.

22. The use according to claim 19, the method according to claim 20, or the compound, stereoisomer thereof, or pharmaceutically acceptable salt or pharmaceutical composition according to claim 21, wherein the disease mediated by KIF18A is a tumor; or the disease mediated by KIF18A is a tumor with chromosomal instability.

23. The use or method according to claim 22, or the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, or pharmaceutical composition, wherein the tumor is selected from breast cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, head and neck cancer, gastric cancer, esophageal cancer, or skin cancer; preferably, the lung cancer is selected from squamous cell lung carcinoma, lung adenocarcinoma, large cell lung cancer, or small cell lung cancer.

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