TW202415648A - Polycyclic kif18a inhibitor, and pharmaceutical composition, preparation method and use thereof - Google Patents

Abstract

The present disclosure provides a compound shown in general formula (I), pharmaceutical composition, preparation method and use thereof. The compound has a good KIF18A inhibitory effect, and can regulate the KIF18A protein alone or by forming binding complexes with microtubules, for the treatment of KIF18A mediated diseases and/or conditions, such as tumor diseases, and the preparation of drugs for such diseases or conditions.

Description

Fused-ring KIF18A inhibitor compounds, drug compositions, and preparation methods and uses thereof

The present invention claims: The priority of the prior application filed with the National Intellectual Property Administration of China on September 30, 2022, with patent application number 202211217025.8, entitled "Fused-ring KIF18A inhibitor compounds, drug compositions, and preparation methods and applications thereof"; The priority of the prior application filed with the National Intellectual Property Administration of China on November 10, 2022, with patent application number 202211406766.0, entitled "Fused-ring KIF18A inhibitor compounds, drug compositions, and preparation methods and applications thereof"; Priority to the prior application filed with the National Intellectual Property Administration of China on February 16, 2023, with patent application number 202310125351.4, entitled "Fused-ring KIF18A inhibitor compounds, drug compositions, and preparation methods and applications thereof"; Priority to the prior application filed with the National Intellectual Property Administration of China on September 20, 2023, with patent application number 202311219723.6, entitled "Fused-ring KIF18A inhibitor compounds, drug compositions, and preparation methods and applications thereof"; The full text of the prior application is incorporated into the present invention by reference.

The present invention belongs to the field of medicine, and specifically relates to a fused-ring KIF18A inhibitor compound, a drug composition, and a preparation method and use thereof.

Cancer is one of the most serious diseases affecting human health, and its mortality and morbidity rates are often among the highest among all diseases. Although the quality of life of some patients has been greatly improved with the continuous development and progress of medical technology and drug research and development, there are still more unmet clinical needs in the search for effective treatments or cures for different cancers, and more new targets will provide new possibilities for future cancer drug research and development.

Cancer cells display unregulated cell proliferation due to damage or loss of one or more genes that regulate the cell cycle. Various kinases and drivers have been identified as playing key roles in the regulation and progression of the cell cycle and mitosis in both normally dividing cells and cancer cells.

Kinesin molecules are motor proteins that use intracellular microtubules as tracks, also known as molecular motors, which can convert ATP energy into mechanical energy. They are closely related to mitosis and meiosis, the growth and development of tissues and organs, and the development and signal transduction of neurons in eukaryotic cells. Kinesin members share a relatively conserved motor domain. According to the location of the motor domain in the molecule, the kinesin family is roughly divided into three categories: N-type kinesin, that is, the amino (-NH ₂ ) terminal region of the protein polypeptide chain has a motor domain; M-type kinesin, that is, the middle region has a motor domain; C-type kinesin, that is, the carboxyl (-COOH) terminal region has a motor domain.

Chromosomal instability is a hallmark of cancer and results from errors in chromosome segregation during mitosis. Targeting chromosomal instability is an emerging therapeutic strategy in drug development. KIF18A is a member of the N-type Kinesin-8 family of kinesins and has been shown to play a role in maintaining the integrity of bipolar spindles and promoting the viability of chromosomally unstable cancer cells. Mitosis is an effective intervention point and many anti-mitotic drugs are used in the clinical treatment of human cancers. The most widely used tubulin inhibitors can both stabilize microtubules and prevent their assembly. Current anti-mitotic drugs have limitations with a narrow therapeutic window, and these issues require the development of new targets to address them.

Although tubulin inhibitors are widely used as standard treatments for a variety of human cancer types, these drugs have collateral damage to normal cells, including bone marrow suppression and neurotoxicity. Since KIF18A may not be essential in normal diploid somatic cells (KIF18A knockout mice are viable but have reproductive defects, indicating that KIF18A is not an essential gene for normal somatic cell division), targeting KIF18A may significantly reduce its toxicity, which is beneficial to improve the therapeutic safety window of tubulin-targeted drugs in clinical practice.

KIF18A protein is highly expressed in a variety of tumors, including colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, and ovarian cancer. KIF18A plays a key role in the occurrence, development, and metastasis of breast cancer, and its high expression predicts poor prognosis in patients. KIF18A is required for the proliferation of chromosomally unstable cells derived from triple-negative breast cancer or colorectal cancer, but KIF18A is not required in diploid cells. Knockout of the KIF18A gene can cause infertility in male mice, but female mice are not affected. KIF18A mRNA expression is significantly associated with higher tumor grade and larger tumors in breast cancer patients, and KIF18A is an independent predictor of lymph node metastasis of breast cancer with a risk coefficient of 3.2. In addition, inhibiting KIF18A expression not only affects the key function of KIF18A in cell mitosis, but also reduces cancer cell migration by stabilizing leading-edge microtubules, ultimately leading to inactivation of the PI3K-AKT signaling pathway and inducing cell apoptosis.

Therefore, the development of KIF18A protein inhibitors may be a new breakthrough in cancer drugs.

In order to improve the above technical problems, the present invention provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: (I) wherein A is selected from unsubstituted or optionally substituted with one, two or more _Ra or a fused cyclic group 1; the fused cyclic group 1 comprises two, three or four rings independently selected from saturated or partially unsaturated C _3-14 carbon rings, C _6-14 aromatic rings, 5-14 membered heteroaromatic rings, 3-14 membered heterocyclic rings; each _{Ra is} the same or different and independently selected from H, OH, halogen, nitrile, NH ₂ , NO ₂ , the following groups which are unsubstituted or optionally substituted by one, two or more _Ra1 : C _1-12 alkyl, C _1-12 alkoxy, C _3-12 cycloalkyl; or, two _Ras connected to the same ring carbon atom together with the carbon atom to which they are connected form a saturated or partially unsaturated C _3-14 carbon ring; each R _a1 is the same or different and is independently selected from H, OH, halogen, nitrile, _NH2 , _NO2 , _C1-12 alkyl, _C1-12 alkoxy, _C3-12 cycloalkyl; _X1 , _X2 , _X3 are the same or different and are independently selected from N or _CR0 ; _R0 is selected from H, halogen, nitrile, _C1-12 alkyl, halogenated _C1-12 alkyl, nitrile C1-12 alkyl, _C1-12 alkoxy, halogenated _C1-12 alkoxy, nitrile _C1-12 alkoxy; B is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Rb : _C1-12 alkyl, halogenated _C1-12 alkyl, nitrile _{C1-12 alkyl} , _C1-12 alkoxy, halogenated C1-12 alkoxy, nitrile _C1-12 alkoxy R _is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, _{nitrile C 1-12 alkyl, C 1-12 alkoxy, halogenated C 1-12 alkoxy , nitrile C 1-12} alkoxy; Ring G is selected from a fused cyclic group 2 which _is unsubstituted or optionally substituted with one, two or more R _g , and the fused cyclic group 2 is formed by condensing Ring _{G 1 and} Ring G ₂ ; Ring G ₁ is selected from a C _6-14 aromatic ring, a 5-14 heteroaromatic ring, and a 3-14 heterocyclic ring; Ring G _{R 2} is selected from C _3-14 carbocyclic ring, C _6-14 aromatic ring, 5-14 member heteroaromatic ring, 3-14 member heterocyclic ring; M and E are preferably connected to ring G ₁ ; each R _g is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, nitrile C _1-12 alkyl, C _1-12 alkoxy, halogenated C _1-12 alkoxy, nitrile C _1-12 alkoxy; E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : -NH-S(=O) _{2 -Re1} , -S(=O) ₂ -NH- _Re2 , -S(=O)(=NH) _-Re3 , -N( _Re4 )( _Re5 ), a 3-14 membered heterocyclic group; each _{Re is} the same or different and is independently selected from OH, halogen, nitrile, C _1-12 alkyl, C _1-12 alkoxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkoxy, nitrile C _1-12 alkyl, nitrile C _1-12 alkoxy, -N( _Re6 )( _Re7 ); _Re1 , _Re2 , _Re3 , _Re4 , _Re5 , _Re6 , _Re7 are the same or different and are independently selected from H, C _1-12 alkyl, C _1-12 alkoxy, hydroxyl C _1-12 alkyl, halogenated C _1-12 alkyl, halogenated C _1-12 alkoxy, nitrile C _1-12 alkyl, nitrile C _1-12 alkoxy, C C _3-12 cycloalkyl, 3-14 membered heterocyclic group, C _1-12 alkoxy-C _1-12 alkyl; M is selected from the following groups which are unsubstituted or optionally substituted by one, two or more R _m : C _3-12 cycloalkyl, C _3-12 cycloalkenyl, 3-14 membered heterocyclic group; each R _m is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, nitrile C _1-12 alkyl, C _1-12 alkoxy, nitrile C _1-12 alkoxy.

According to some embodiments, A is selected from unsubstituted or optionally substituted with one, two or more _Ra or a fused cyclic group 1; the fused cyclic group 1 comprises two, three or four rings independently selected from saturated or partially unsaturated C _3-8 carbon rings, C _6-10 aromatic rings, 5-10 membered heteroaromatic rings, and 3-8 membered heterocyclic rings; each _{Ra is} the same or different and independently selected from H, OH, halogen, nitrile, NH ₂ , NO ₂ , C _1-6 alkyl, C _1-6 alkoxy, C _3-8 cycloalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, nitrile C _1-6 alkyl, nitrile C _1-6 alkoxy, and C _3-8 cycloalkyl-C _1-6 alkoxy; or, two R a connected to the same ring carbon atom are selected from: _a together with the carbon atom to which it is connected form a saturated or partially unsaturated C _3-8 carbon ring;

According to some implementations, A is selected from , , , wherein T is selected from CH ₂ , CH, NH, NR _a or O; Z is selected from CH ₂ , CH, NH, NR _a or O; X is selected from N or CH; n is selected from 0, 1, 2, 3, 4 or 5; p and q are independently selected from 0, 1, 2, 3, and p and q are not simultaneously 0; r and s are independently selected from 0, 1, 2, 3, and r and s are not simultaneously 0.

According to some embodiments, X ₁ and X ₂ are the same or different and are independently selected from N or CR ₀ .

According to some embodiments, X ₁ and X ₂ are the same or different and are independently selected from N or CH.

According to some implementation schemes, X ₂ and X ₃ are not N at the same time.

According to some embodiments, when _X1 and _X2 are N, _X3 is _CR0 ; when _X1 is N or _CR0 and _X2 is _CR0 , _X3 is N or _CR0 ; when _X1 is N or _CR0 and _X2 is N, _X3 is _CR0 ; _R0 is selected from H, halogen, _C1-6 alkyl;

According to some embodiments, when _X1 and _X2 are N, _X3 is _CR0 ; when _X1 is N or CH and _X2 is CH, _X3 is N or _CR0 ; when _X1 is CH and _X2 is N, _X3 is _CR0 ; when _X1 is N and _X2 is _CR0 , _X3 is N; _R0 is selected from H, _C1-6 alkyl;

According to some embodiments, B is selected from C _1-6 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-8 membered heterocyclic group, halogenated 3-8 membered heterocyclic group.

According to some embodiments, B is selected from a halogenated 6-membered N-containing heterocyclic group; for example, a halogenated piperidinyl group.

According to some implementations, B is selected from .

According to some embodiments, A is selected from unsubstituted or optionally substituted with one, two or more _Ra , , , , Or a fused ring group 1; the fused ring group 1 is formed by condensing two, three, four or more rings selected from a benzene ring, a pyridine ring, an imidazole ring, a piperazine ring, a dihydropyridine ring, a tetrahydropyridine ring, a dihydropyrrole ring, a tetrahydropyrrole ring, a dihydropyran ring, a cyclobutene ring, a cyclopentene ring, a cyclohexene ring and a cycloheptene ring.

According to some embodiments, A is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Ra : , , , , , , , , , .

According to some embodiments, each Ra _is the same or different and is independently selected from H, OH, F, Cl, nitrile, methyl, ethyl, isopropyl, methoxy, ethoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclopropylmethoxy; or, two _Ra connected to the same ring carbon atom together with the carbon atom to which they are connected form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, or a cycloheptane ring.

According to some implementations, A is selected from , , , , , , , , , , , , , , .

According to some embodiments, ring G is selected from a fused cyclic group 2 which is unsubstituted or optionally substituted by one, two or more _Rg , and the fused cyclic group 2 is formed by condensing ring _G1 and ring _G2 ; ring _G1 is selected from _C6-10 aromatic ring, 5-10 member heteroaromatic ring, 5-10 member heterocyclic ring; ring _G2 is selected from _C3-10 carbon ring, _C6-10 aromatic ring, 5-10 member heteroaromatic ring, 3-10 member heterocyclic ring;

According to some embodiments, ring _G1 is selected from cyclopentene ring, cyclohexene ring, dihydrofuran ring, dihydropyran ring, imidazole ring, triazole ring, benzene ring, pyridine ring, dihydropyridine ring, pyrrole ring, pyrazole ring, furan ring, thiophene ring; preferably, it is a benzene ring or a pyridine ring.

According to some embodiments, ring _G2 is selected from a cyclopentene ring, a cyclohexene ring, a dihydrofuran ring, a dihydropyran ring, an imidazole ring, a triazole ring, a benzene ring, a pyridine ring, a pyrrole ring, a pyrazole ring, a furan ring, a thiophene ring, and a methylimidazole ring.

According to some implementation schemes, Ring G is selected from , , , , , , , , , , , , , .

According to some embodiments, E is selected from the following groups which are unsubstituted or optionally substituted with one or two _Re : -NH-S(=O) ₂ - _Re1 , -S(=O) ₂ -NH- _Re2 , -S(=O)(=NH) _-Re3 ; _Re1 , _Re2 , and _Re3 are the same or different and are independently selected from H, _C1-6 alkyl, hydroxyl _C1-6 alkyl, halogenated _C1-6 alkyl, halogenated _C1-6 alkoxy, nitrile _C1-6 alkyl, nitrile _C1-6 alkoxy, _C3-8 cycloalkyl, 3-8 membered heterocyclic group, and _C1-6 alkoxy- _C1-6 alkyl; each _{Re is} the same or different and is independently selected from OH, halogen, nitrile, _C1-6 alkyl, _C1-6 alkoxy, halogenated C1-6 C _1-6 alkyl, halogenated C _1-6 alkoxy, nitrile C _1-6 alkyl, nitrile C _1-6 alkoxy.

According to some implementations, E is selected from .

According to some embodiments, M is selected from _C3-8 cycloalkyl, _C3-8 cycloalkenyl or nitrogen-containing 3-8 membered heterocyclic group which is unsubstituted or optionally substituted by one, two or more _Rm ; each _Rm is the same or different and is independently selected from H, halogen, nitrile, _C1-6 alkyl, halogenated _C1-6 alkyl _, nitrile C1-6 alkyl, _C1-6 alkoxy, nitrile _C1-6 alkoxy; or, two _Rm attached to the same ring carbon atom together with the carbon atom to which they are attached form a saturated or partially unsaturated _C3-8 carbon ring;

According to some embodiments, M is selected from unsubstituted or optionally substituted with one, two or more _R , or .

According to some embodiments, the compound represented by formula (I) is selected from the following structures: (IA-1) (IA-2) (IA-3) (IA-4) wherein A, M, E, Ring G, Ring _G1 , and Ring _G2 independently have the definitions described herein, Indicates a carbon-carbon single bond or a carbon-carbon double bond.

According to some embodiments, the compound shown in formula (I) has the following structure: (IB-1) (IB-2) (IB-3) (IB-4) wherein B, M, E, ring G, X, Z, T, X ₁ , X ₂ , X ₃ , _Ra , n, p, q, r, and s independently have the meanings described herein.

According to some embodiments, the compound shown in formula (I) has the following structure: (IC-1) (IC-2) (IC-3) wherein B, ring G, X, Z, T, X ₁ , X ₂ , X ₃ , _Ra , n, r, s, p, and q independently have the meanings described herein.

According to some embodiments, the compound shown in formula (I) has the following structure: , (IIA-1) (IIA-2) wherein A, Ring G ₁ , and Ring G ₂ independently have the definitions described herein.

According to some embodiments, the compound shown in formula (I) has the following structure: , (IIB-1) (IIB-2) wherein A and ring _G2 independently have the definitions described herein.

According to some embodiments, the compound shown in formula (I) has the following structure: , (IIC-1) (IIC-2) (IIC-3) wherein B, Ring _G2 , X, Z, T, _X1 , _X2 , _X3 , _Ra , n, r, s, p, and q independently have the definitions described herein.

According to some embodiments, the compound shown in formula (I) has the following structure: (III) wherein Ring G ₂ , X ₁ , X ₂ , and X ₃ independently have the definitions described herein.

According to some embodiments, the compound represented by formula (I) is selected from the following structures: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

The present invention also provides a method for preparing the compound represented by formula (I), comprising the following steps: (1) Compound a reacts with compound A- _NH2 to obtain compound b; (2) Compound b reacts with compound MH to obtain compound c; (3) Compound c reacts with compound EH to obtain a compound represented by formula (I); wherein A, E, M and ring G independently have the definitions described above; L is selected from halogens, such as Cl, Br, I; Q is selected from halogens, such as F, Cl, Br.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.

According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

According to the embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.

The present invention also provides a method for treating tumor diseases, comprising administering to a patient an effective preventive or therapeutic amount of the above-mentioned pharmaceutical composition.

The tumor diseases include intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer and ovarian cancer.

In some embodiments, the patient comprises a mammal, preferably a human.

The present invention also provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or at least one of its prodrug compounds, or a pharmaceutical composition thereof for treating tumor diseases.

The present invention also provides the use of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof in the preparation of a drug.

According to an embodiment of the present invention, the use may be for preparing a medicament for treating KIF18A-mediated disorders and/or diseases, such as for preparing a KIF18A inhibitor drug.

According to an embodiment of the present invention, the disease is, for example, cancer, including intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.

Beneficial Effects

The compound of the present invention has a good KIF18A inhibitory effect. The compound can regulate KIF18A protein alone or form a binding complex with microtubules to treat KIF18A-mediated disorders and/or diseases, such as tumor diseases, and to prepare drugs for such disorders or diseases.

The present invention creatively obtains a class of novel structural compounds through structural optimization, which not only have good KIF18A inhibitory effect and OVCAR-3 in vitro cell activity, but also have significantly improved physicochemical properties (solubility, permeability), and significantly improved OVCAR-3 in vivo efficacy.

Definitions and Explanations

Unless otherwise stated, the definitions of groups and terms described in the present invention specification and the scope of the patent application, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined and combined with each other. The group definitions and compound structures after such combination and combination should be understood to be within the scope described in the present invention specification and/or the scope of the patent application.

Unless otherwise stated, the numerical ranges described in this specification and patent application are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-14" is equivalent to describing each integer value in the numerical range "1-14", namely, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14.

The term "optionally" (or "optionally", "optionally") in the general formula definition of the present invention means substituted by zero, one or more substituents, for example, "optionally substituted by one, two or more R" means that it may not be substituted by R (unsubstituted) or may be selectively substituted by one, two or more R.

"More" means three or more.

The term "C _1-12 alkyl" is understood to mean straight chain and branched alkyl groups having 1 to 12 carbon atoms, "C _1-8 alkyl" means straight chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and "C _1-6 alkyl" means straight chain and branched alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl or the like or isomers thereof.

The term "C _3-12 cycloalkyl" should be understood to mean a saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) alkyl or tricyclic alkane having 3 to 12 carbon atoms, preferably "C _3-10 cycloalkyl", more preferably "C _3-8 cycloalkyl". The term "C _3-12 cycloalkyl" should be understood to mean a saturated monovalent monocyclic, bicyclic (such as bridged ring, spiro ring) alkyl or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C The _3-12- cycloalkyl group may be a monocyclic alkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic alkyl group such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2 .2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonanyl, 2,6-diazaspiro[3,4]octanyl, or a tricyclic alkyl group such as adamantyl.

The term "C _3-12 cycloalkenyl" should be understood to mean a monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) or tricyclic alkene containing a carbon-carbon double bond, which has 3 to 12 carbon atoms, preferably a "C _3-10 cycloalkenyl", and more preferably a "C _3-8 cycloalkenyl", which may have 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C _3-12 cycloalkenyl group may be a monocyclic alkyl group such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl or cyclodecenyl, or a bicyclic alkyl group such as spiro[2.5]oct-5-enyl, spiro[3.5]non-6-enyl, spiro[4.5]dec-7-enyl.

The term "C _6-14 aryl" should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 14 carbon atoms, which may be a single aromatic ring or a fused polyaromatic ring, preferably a "C _6-10 aryl". The term " _C6-14 aryl" is to be understood as preferably meaning a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" _C6-14 aryl"), in particular a ring having 6 carbon atoms (" _C6 aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms (" _C9 aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms (" _C10 aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms (" _C13 aryl"), such as fluorenyl, or a ring having 14 carbon atoms (" _C14 aryl"), such as anthracenyl. When the C _6-20 aryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution position, for example, it may be ortho-, para- or meta-substituted.

The term "5-14 membered heteroaryl" is to be understood as including monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5-10 membered heteroaryl". The term "5-14 membered heteroaryl" is to be understood as including monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, in each case, may additionally be benzo-fused. "Heteroaryl" also refers to a group in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclic rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3- , 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, -acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-oxadiazole, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthroline, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl , 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]-pyranyl 2-, 3-, 4-, 5-, 7-, or 8-5H-pyrido[2,3-d]-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]oxazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9-imidazo[2,3-d]oxazinyl, -furano[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4- , 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-14 membered heteroaryl group is linked to other groups to form the compound of the present invention, the carbon atoms on the 5-14 membered heteroaryl ring may be linked to the other groups, or the hetero atoms on the 5-14 membered heteroaryl ring may be linked to the other groups. When the 5-14 membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution position, for example, a hydrogen atom connected to a carbon atom on the heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on the heteroaryl ring may be substituted.

Unless otherwise defined, the term "3- to 14-membered heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6- or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring (e.g., fused ring, bridged ring, spiro ring) or a 10-, 11-, 12-, 13- or 14-membered tricyclic ring system, and contains at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O) ₂ -. Preferably, the heterocyclic group can be selected from "3-10 membered heterocyclic groups". The term "3-10 membered heterocyclic group" means a saturated or unsaturated non-aromatic ring or ring system, and contains at least one heteroatom selected from O, S and N. The heterocyclic group can be connected to the rest of the molecule through any of the carbon atoms or the nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. In particular, the heterocyclic group may include, but is not limited to: a 4-membered ring, such as azacyclobutane, oxacyclobutane; a 5-membered ring, such as a tetrahydrofuranyl, a dioxacyclopentane, a pyrrolidinyl, an imidazolidinyl, a pyrazolidinyl, a pyrrolinyl; or a 6-membered ring, such as a tetrahydropyranyl, a piperidinyl, a morpholinyl, a dithianyl, a thiomorpholinyl, a piperazinyl or a trithianyl; or a 7-membered ring, such as a diazacycloheptanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, for example but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring. The heterocyclic group may be partially unsaturated, i.e. it may contain one or more double bonds, for example but not limited to dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, for example but not limited to dihydroisoquinolinyl. When the 3-14 membered heterocyclic group is connected with other groups to form the compound of the present invention, the carbon atom on the 3-14 membered heterocyclic group may be connected with the other groups, or the heterocyclic atom on the 3-14 membered heterocyclic group may be connected with the other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl may be connected with the other groups. Or when the 3-14 membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and the carbon atom at the para position thereof may be connected with the other groups.

The term "spiro" refers to a ring system in which two rings share one ring atom.

The term "fused ring" refers to a ring system in which two rings share two ring atoms.

The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

The term "halogen" refers to fluorine, chlorine, bromine and iodine.

"Halogenation" refers to substitution with one or more halogens.

When Ring G is selected from When there are substituents such as , it has three connection sites. Generally speaking, the three connection sites can be connected to A, M, and E in the general structure respectively. For example, the connection position 1 is connected to A, the connection position 2 is connected to M, and the connection position 3 is connected to E. When other substituents are selected for ring G and ring _G1 , the same explanation as above shall apply.

It is known to those skilled in the art that when A is selected from unsubstituted or optionally substituted with one, two or more R _a When X 1 is CH, _Ra can be located at any position on the ring where X ₁ is located. For example, when X ₁ is CH, _Ra can replace the H on X ₁ to form CR _a .

Those skilled in the art will appreciate that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form an acid addition salt; if these compounds have an acidic center, they may form an alkali addition salt; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amine group), they may also form an internal salt.

The compounds of the invention may exist in the form of solvates (e.g. hydrates), wherein the compounds of the invention contain polar solvents as structural elements of the crystal lattice of the compounds, in particular water, methanol or ethanol, for example. The polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric amount.

According to their molecular structure, the compounds of the present invention may be chiral, and therefore various mirror image isomers may exist. Therefore, these compounds may exist in the form of racemates or optically active forms. The compounds of the present invention cover isomers or mixtures and racemates thereof in which each chiral carbon is in R or S configuration. The compounds of the present invention or their intermediates can be separated into mirror image isomer compounds by chemical or physical methods known to those of ordinary skill in the art to which the present invention belongs, or used in this form for synthesis. In the case of racemic amines, non-mirror image isomers are prepared from the mixture by reaction with an optically active resolution agent. Examples of suitable resolution agents are optically active acids, such as tartaric acid in its R and S forms, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, apple acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids. Column chromatography can also be advantageously performed for resolution of the image with the aid of optically active resolution agents (e.g. dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, e.g. hexane/isopropanol/acetonitrile.

The corresponding stable isomers can be separated according to known methods, for example, by extraction, filtration or column analysis.

The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

The term "therapeutically effective amount" refers to the amount of an active compound or drug that elicits the biological or medicinal response that a researcher, veterinarian, physician or other clinician is seeking in a tissue, system, animal, individual or human, and includes one or more of the following: (1) Preventing disease: e.g., preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed the pathology or symptoms of the disease. (2) Inhibiting disease: e.g., inhibiting the disease, disorder or condition (i.e., preventing further development of the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition. (3) Relieving disease: e.g., relieving the disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition.

Specific implementation methods

The following will further explain the technical solution of the present invention in combination with specific embodiments. It should be understood that the following embodiments are only for illustrative purposes to illustrate and explain the present invention, and should not be interpreted as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope of protection intended by the present invention.

Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Embodiment 1

5-{6-azaspiro[2.5]octan-6-yl} -N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (Compound 1)

The first step is the synthesis of 2-bromo-4-fluoro-5-methoxybenzaldehyde (compound 1-2):

To a solution of 4-fluoro-3-methoxybenzaldehyde (10 g, 64.876 mmol, 1 eq) in water (20 mL) was added potassium bromide (38.60 g, 324.380 mmol, 5 eq) at room temperature, followed by bromine (9.98 mL, 194.628 mmol, 3 eq). Stirring was continued at room temperature for 15 hours. The precipitated solid was collected by filtration and washed with water (3×5 mL). Drying under vacuum gave solid 2-bromo-4-fluoro-5-methoxybenzaldehyde (15 g, 99.22%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.13 (d, J = 0.8 Hz, 1H), 7.83 (dd, J = 10.4, 1.6 Hz, 1H), 7.57-7.54 (dd, J = 10.4, 1.6 Hz, 1H), 3.92 (s, 3H).

Step 2 Synthesis of 3-(2-bromo-4-fluoro-5-methoxyphenyl)propionic acid (Compound 1-4):

Under nitrogen protection, add Mach's acid (compound 1-3) (4947.77 mg, 34.329 mmol, 1 eq) to a solution of 2-bromo-4-fluoro-5-methoxybenzaldehyde (8 g, 34.329 mmol, 1 eq) in formic acid (10 mL) at room temperature, and then add triethylamine (14.32 mL, 102.987 mmol, 3 eq) dropwise. The reaction solution was heated to 100°C and stirred for 10 hours. After the reaction was completed, quench with water at room temperature, extract with ethyl acetate (3×200 mL), combine the organic phases, wash with saturated sodium chloride solution (1×300 mL), and dry over anhydrous sodium sulfate. Filter, and concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether:ethyl acetate (1:1) to give 3-(2-bromo-4-fluoro-5-methoxyphenyl)propanoic acid (2.8 g, 29.44%).

MS(ESI, m/z ): 275.05 [MH] ^- , RT(min):0.886.

Step 3 Synthesis of 4-bromo-6-fluoro-7-methoxy-2,3-dihydroindan-1-one (Compound 1-5):

Under nitrogen protection, trifluoromethanesulfonic acid (3.19 mL, 36.090 mmol, 5 eq) was added dropwise to a solution of 3-(2-bromo-4-fluoro-5-methoxyphenyl)propanoic acid (2.0 g, 7.218 mmol, 1 eq) in 1,2-dichloroethane (12 mL) at room temperature. The reaction solution was heated to 80°C and microwaved for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with dichloromethane (3×60 mL). The organic phases were combined, backwashed with saturated brine (1×60 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 4-bromo-6-fluoro-7-methoxy-2,3-dihydroindan-1-one (1.5 g, 80.22%).

MS (ESI, m/z ): 259.15 [M+H] ⁺ , RT (min): 1.030.

¹ H NMR (400 MHz, CDCl ₃ - d )δ 7.51(d, J = 11.2 Hz, 1H), 4.09(s, 3H), 2.99-2.96(m, 2H), 2.77-2.74(m, 2H).

Step 4 Synthesis of 7-bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (Compound 1-6):

Under nitrogen protection, triethylsilane (3.74 mL, 23.160 mmol, 3 eq) was added to a solution of 4-bromo-6-fluoro-7-methoxy-2,3-dihydroindan-1-one (2 g, 7.720 mmol, 1 eq) in trifluoroacetic acid (8.80 mL) at 0°C. The reaction mixture was then heated and stirred at 50°C for 16 hours, and the reaction system was monitored by thin layer column chromatography. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3×60 mL). The organic phases were combined, backwashed with saturated brine (1×60 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 7-bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (1.5 g, 79.28%).

¹ H NMR (400 MHz, deuterated chloroform) δ 7.06 (d, J = 10.8 Hz, 1H), 3.91 (s, 3H), 3.03-2.99 (m, 2H), 2.90-2.83 (m, 2H), 2.14-2.07 (m, 2H).

Step 5 Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-ol (Compound 1-7):

7-Bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (800 mg, 3.264 mmol, 1 eq) and hydrogen bromide in acetic acid (8.00 mL) were heated and stirred at 100°C for 2 hours. The reaction solution was cooled to room temperature and the reaction mixture was diluted with ethyl acetate (50 mL). The reaction mixture was washed with distilled water (1×30 mL), saturated sodium bicarbonate (1×30 mL), and saturated brine (1×30 mL) in sequence. The organic phase was dried over anhydrous sodium sulfate, and the resulting mixture was filtered. The filtrate was concentrated under reduced pressure and the crude product was directly used in the next step without further purification.

MS(ESI, m/z ): 231.05 [MH] ^- , RT(min):1.041

Step 6 Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yl trifluoromethanesulfonate (Compound 1-9):

Under nitrogen protection, pyridine (1.54 g, 19.474 mmol, 1.50 eq) was added to a solution of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-ol (3 g, 12.983 mmol, 1 eq) in dichloromethane (30.00 mL) at 0°C and the mixture was stirred for 10 minutes. Then trifluoromethanesulfonic anhydride (Compound 1-8) (3.29 mL, 19.474 mmol, 1.50 eq) was added and stirred at room temperature overnight. The reaction solution was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3×80 mL). The organic phases were combined, backwashed with saturated brine (1×100 mL), and dried over anhydrous sodium sulfate. The obtained mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yl trifluoromethanesulfonate (2 g, 42.42%).

¹ H NMR (400 MHz, deuterated chloroform) δ 7.23 (d, J = 9.2 Hz, 1H), 3.18-3.14 (m, 2H), 2.99-2.95 (m, 2H), 2.23-2.16 (m, 2H).

Step 7 Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid methyl ester (Compound 1-10):

Under nitrogen protection, sodium acetate (74.19 mg, 0.331 mmol, 0.1 eq), bis(diphenylphosphinobutane) (140.94 mg, 0.331 mmol, 0.1 eq) and triethylamine (1148.39 µl, 8.263 mmol, 2.5 eq) were added to a solution of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yl trifluoromethanesulfonate (1.2 g, 3.305 mmol, 1 eq) in dimethylsulfoxide/methanol (7.5 mL, 3/2, v/v) at room temperature. The reaction solution was heated to 80°C and carbon monoxide was introduced for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid methyl ester (400 mg, 44.32%).

¹ H NMR (400 MHz, deuterated chloroform) δ 7.14 (d, J = 10.0 Hz, 1H), 3.91 (s, 3H), 3.25 (t, J = 7.6 Hz, 2H), 2.90 (t, J = 7.6 Hz, 2H), 2.16-2.08 (m, 2H).

Step 8 Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid (Compound 1-11):

Lithium hydroxide (131.54 mg, 5.493 mmol, 3 eq) was added to a solution of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid methyl ester (500 mg, 1.831 mmol, 1 eq) in tetrahydrofuran/water (4 mL, 3/1, v/v) at room temperature. The reaction mixture was stirred at room temperature for 16 hours, and the reaction mixture was diluted with water at room temperature and acidified to pH 5-6 with 1 mol hydrochloric acid solution. The reaction mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (1×40 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product/the resulting mixture was not further purified and was directly used in the next step.

MS(ESI, m/z ): 257.10 [MH] ^- , RT(min):1.053

Step 9 Synthesis of 7-bromo-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (Compound 1-13):

Under nitrogen protection, tetramethylchloroformamidine hexafluorophosphate (2166.02 mg, 7.720 mmol, 4 eq) and N-methylimidazole (1584.61 mg, 19.300 mmol, 10 eq) were added to a dichloromethane solution (5 mL) of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid (500 mg, 1.930 mmol, 1 eq) and 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-amine (Compound 1-12) (660.76 mg, 2.895 mmol, 1.5 eq) at room temperature. Stir at room temperature for 1 hour. The reaction mixture was diluted with water at room temperature and extracted with dichloromethane (3×30 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 7-bromo- N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (160 mg, 17.67%).

MS(ESI, m/z ): 469.25 [M+H] ⁺ , RT(min):1.455.

Step 10 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-2,3-dihydro-1H-indene-4-carboxamide (Compound 1-15):

Under nitrogen protection, sodium tert-butoxide (196.59 mg, 2.046 mmol, 6 eq) was added to a solution of 7-bromo- N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (160 mg, 0.341 mmol, 1 eq) and 6-azaspiro[2.5]octane hydrochloride (Compound 1-14) (251.69 mg, 1.705 mmol, 5 eq) in N,N -dimethylacetamide (10 mL) at room temperature. The reaction mixture was heated to 140°C and heated for 3 days. The reaction mixture was cooled to room temperature and quenched with water. The reaction mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (1×30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-2,3-dihydro-1H-indene-4-carboxamide (48 mg, 25.12%).

MS(ESI, m/z ): 560.60 [M+H]+, RT(min):1.494.

¹ H NMR (400 MHz, deuterated chloroform) δ 12.69(s, 1H), 7.46(s, 1H), 7.27(s, 1H), 3.99(d, J = 7.2 Hz, 4H), 3.47(t, J = 7.6 Hz, 2H), 3.03(t, J = 5.4 Hz, 4H), 2.93(t, J = 7.6 Hz, 2H), 2.37(s, 3H), 2.10(q, J = 7.6 Hz, 2H), 1.99(s, 4H), 1.55(s, 4H), 0.38(s, 4H).

Step 11 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl} -N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro-1H-indene-4-carboxamide (Compound 1):

Under nitrogen protection, dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphane (Ephos, 8.59 mg, 0.016 mmol , 0.2 eq), methanesulfonic acid {dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl- [1,1'-biphenyl]-2-yl)phosphine}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (Ephos-Pd-G4, 7.37 mg, 0.008 mmol, 0.1 eq), cesium carbonate (78.48 mg, 0.240 mmol, 3 eq) and 2-hydroxyethanesulfonamide (Compound 1-16) (12.06 mg, 0.096 mmol, 1.2 eq). The reaction solution was heated to 100°C for 1 hour. The reaction solution was cooled to room temperature, and the reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: XBridge Prep OBD C18 Column, 30*150 mm, 5μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL/min; elution gradient: 48% B to 80% B, 80% B in 8 min; detection wavelength: UV 220 nm; retention time (min): 7.88). Compound 1 (21.93 mg, 44.81%) was obtained.

MS(ES, m/z ): 605.45 [M+H] ⁺ , RT(min): 1.894.

¹ H NMR: (400 MHz, DMSO- d ₆ ) δ 12.92(s, 1H), 9.39(s, 1H), 7.38(s, 1H), 7.27(s, 1H), 4.99(s, 1H), 3.90(s, 4H), 3.77(t, J = 6.5 Hz, 2H), 3.31(s, 2H), 3.22(t, J = 7.5 Hz, 2H), 2.94(s, 4H), 2.85(t, J = 7.5 Hz, 2H), 2.31(s, 3H), 2.05-1.86(m, 6H), 1.65(s, 4H), 0.36(s, 4H).

Embodiment 2

5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro-1H-indene-4-carboxamide (Compound 9)

The first step is the synthesis of 7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (Compound 9-2):

Under nitrogen protection, N-methylimidazole (475.38 mg, 5.790 mmol, 10 eq) was added to a solution of 6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-amine (197.37 mg, 0.868 mmol, 1.5 eq), N ,N,N',N' -tetramethylchloroformamidine hexafluorophosphate (649.81 mg, 2.316 mmol, 4 eq) and 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid (150 mg, 0.579 mmol, 1.00 eq) in dichloromethane (7.5 mL) at room temperature. The reaction mixture was heated to 60°C and stirred for 1 hour. The reaction mixture was cooled to room temperature and extracted with dichloromethane (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to obtain 7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (180 mg, 66.38%).

MS:(ESI, m/z ): 468.30 [M+H] ⁺ , RT(min):1.397.

Step 2 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (Compound 9-3):

Under nitrogen protection, sodium tert-butoxide (82.08 mg, 0.855 mmol, 5 eq) was added to a solution of 7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-fluoro-2,3-dihydro-1H-indene-4-carboxamide (80 mg, 0.171 mmol, 1 eq) and 6-azaspiro[2.5]octane hydrochloride (126.11 mg, 0.855 mmol, 5 eq) in N,N -dimethylacetamide (4 mL) at room temperature. The reaction mixture was heated to 140°C and stirred for 48 hours. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (3 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative column chromatography with petroleum ether/ethyl acetate (1:1) to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (50 mg, 52.31%).

MS:(ESI, m/z ): 559.45 [M+H] ⁺ , RT(min):1.590.

Step 3 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro-1H-indene-4-carboxamide (Compound 9):

Under nitrogen protection, dicyclohexyl(3-isopropoxy- 2 ',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphane (8.6 mg, 0.016 mmol, 0.2 eq) was added to a solution of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (45 mg, 0.080 mmol, 1 eq) and 2-hydroxyethanesulfonamide (20.13 mg, 0.160 mmol, 2 eq) in 1,4-dioxane (4.5 mL) at room temperature. eq), methanesulfonic acid {bicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphine} (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) (7.39 mg, 0.008 mmol, 0.1 eq) and cesium carbonate (78.62 mg, 0.240 mmol, 3 eq). The reaction solution was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 5-{6-azaspiro[2.5]octan-6-yl}- N -[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (8.8 mg, 17.98%).

MS:(ESI, m/z ): 604.40 [M+H] ⁺ , RT(min): 1.861.

¹ H NMR: (400 MHz, DMSO -d ₆ )δ 12.16(s, 1H), 9.31(s,1H), 7.46(s, 1H), 7.22(s, 1H), 6.53(s, 1H), 4.96(s, 1H), 3.77(t, J = 6.5 Hz, 2H), 3.71(t, J = 5.8 Hz, 4H), 3.33(s, 1H), 3.29(s, 1H), 3.21(t, J = 7.5 Hz, 2H), 2.94(t, J = 5.3 Hz, 4H), 2.85(t, J = 7.5 Hz, 2H), 2.26(s, 3H), 2.05-1.88(m, 6H), 1.63(s, 4H), 0.34(s, 4H).

Embodiment 3

5-(6-Azaspiro[2.5]octan-6-yl) -N- [6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 8)

The first step is the synthesis of 5-(6-azaspiro[2.5]octane-6-yl)-7-bromo-2,3-dihydro- 1H -indene-4-carboxylic acid (Compound 8-1):

Under nitrogen protection, sodium tert-butoxide (77.50 mg, 0.805 mmol, 5 eq) was added to a solution of 7-bromo-5-fluoro-2,3-dihydro- 1H -indene-4-carboxylic acid (50 mg, 0.161 mmol, 1 eq) in N -methylpyrrolidone (1 mL) at room temperature. After stirring for 2 minutes, 6-azaspiro[2.5]octane hydrochloride (89.66 mg, 0.805 mmol, 5 eq) was added at room temperature. The reaction system was heated to 160°C and reacted for 4 hours. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (3:1) to obtain 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo-2,3-dihydro- 1H -indene-4-carboxylic acid (30 mg, 53.11%).

MS:(ESI,m/z):377.00[M+H] ⁺ , RT(min):0.827

Step 2 Synthesis of 5-(6-azaspiro[2.5]octane-6-yl)-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (Compound 8-3):

Under nitrogen protection, 6-(4,4-difluoropiperidin-1-yl)pyridin - 2-amine (9.13 mg, 0.044 mmol, 1.5 eq), tetramethylchlorouronium hexafluorophosphate (64.09 mg, 0.228 mmol, 4 eq), and N-methylimidazole (46.88 mg, 0.570 mmol, 10 eq) were added to a solution of 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (20 mg, 0.057 mmol, 1 eq) in dichloromethane (1 mL) at room temperature. The reaction mixture was heated to 80°C for 1 hour. The reaction mixture was cooled to room temperature and extracted with dichloromethane (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (3:1) to obtain 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (30 mg, 96.32%).

MS:(ESI, m/z):365.90[M+H] ⁺ , RT(min):1.140

Step 3 Synthesis of 5-(6-azaspiro[2.5]octan-6-yl) -N- [6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 8):

Under nitrogen protection, dicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl- [ 1,1'-biphenyl]-2-yl)phosphine (4.90 mg, 0.009 mmol , 0.2 eq), methanesulfonic acid {bicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphine} (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) (4.21 mg, 0.005 mmol, 0.1 eq), cesium carbonate (44.80 mg, 0.138 mmol, 3 eq) and 2-hydroxyethanesulfonamide (8.60 mg, 0.069 mmol, 1.5 eq). The reaction solution was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture is filtered, the filtrate is compressed and concentrated. The crude product was purified by HPLC under the following conditions (column chromatography column Kinete 5 μm EVO C18, 30 mm * 150 mm, mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile, flow rate: 60 mL/min, elution gradient: 40% B to 75% B in 8 min; 254 nm; Rt: 6.9 min) to obtain 5-(6-azaspiro[2.5]octan-6-yl) -N- [6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (2.79 mg, 10.29%).

MS:(ESI, m/z):590.05[M+H] ⁺ , RT(min):1.874

¹ H-NMR: (400 MHz, DMSO- d ₆ )δ 12.30(s, 1H), 9.34(s, 1H), 7.61 – 7.54(m, 2H), 7.22(s, 1H), 6.70 – 6.63(m, 1H), 4.99(s, 1H), 3.77(t, J = 6.5 Hz, 2H), 3.72(t, J = 5.7 Hz, 4H), 3.30(s, 2H), 3.22(t, J = 7.5 Hz, 2H), 3.03 – 2.89(m, 4H), 2.83(t, J = 7.5 Hz, 2H), 2.09 – 1.90(m, 6H), 1.64(s, 4H), 0.35(s, 4H).

Embodiment 4

5-{6-azaspiro[2.5]octan-6-yl}- N -[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 13)

The first step is the synthesis of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (Compound 13-2):

Under nitrogen protection, to a solution of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-2,3-dihydro- 1H -indene-4-carboxylic acid (25 mg, 0.071 mmol, 1 eq) and 6-(4,4-difluoropiperidin-1-yl)pyrazin-2-amine (18.35 mg, 0.085 mmol, 1.2 eq) in dichloromethane (5 mL) were added N , N , N , N -tetramethylchloroformamidine hexafluorophosphate (80.11 mg, 0.284 mmol, 4 eq) and N-methylimidazole (58.60 mg, 0.710 mmol, 10 eq) at room temperature. The reaction solution was heated to 80°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 5 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, and petroleum ether/ethyl acetate (3:1) was used to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (25 mg, 39.19%).

MS:(ESI, m/z):548.05 [M+H] ⁺ , RT(min):1.255

Step 2 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 13):

Under nitrogen protection, dicyclohexyl (3-isopropoxy- 2 ',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine (2.45 mg, 0.005 mmol, 0.1 eq) was added to a solution of 5-{6-azaspiro[2.5] octan -6-yl}-7-bromo-N-[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (25 mg, 0.046 mmol, 1 eq) and 2-hydroxyethanesulfonamide (11.6 mg, 0.092 mmol, 2 eq) in 1,4-dioxane (5 mL) at room temperature. eq) and methanesulfonic acid {biscyclohexyl [3-isopropoxy-2', 4', 6'-triisopropyl-(1,1'-biphenyl)-2-yl] phosphine} (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) (8.40 mg, 0.009 mmol, 0.2 eq) and cesium carbonate (44.72 mg, 0.138 mmol, 3 eq), the reaction solution was heated to 100 ° C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 5 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (2 × 5 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC under the following conditions: column specification: Ultimate 5 μM XB-C18; mobile phase A: water (10 mmol/L sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution gradient: 35% B to 60% B in 22 min; detection wavelength: 254nm/220nm; retention time (min): 7.55, to obtain 5-{6-azaspiro[2.5]octan-6-yl}- N -[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (2.23 mg, 8.17%).

MS:(ESI, m/z):591.20 [M+H] ⁺ , RT(min):1.754

¹ H NMR (400 MHz, DMSO- d ₆ )δ 12.99(s, 1H), 8.76(s, 1H), 8.16(s, 1H), 7.24(s, 1H), 5.00(s, 1H), 3.77(d, J = 6.5 Hz, 6H), 3.24(m, 4H), 2.95(d, J = 5.3 Hz, 4H), 2.80(t, J = 7.6 Hz, 2H), 2.05(m, 4H), 1.95(m, 2H), 1.64(s, 4H), 0.36(s, 4H).

Embodiment 5

5-(6-Azaspiro[2.5]octan-6-yl) -N- [2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 27)

The first step is the synthesis of 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo- N- [2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-2,3-dihydro- 1H -indene-4-carboxamide (Compound 27-2):

Under nitrogen protection, N, N, N', N'-tetramethylchloroformamidine hexafluorophosphate (80.11 mg, 0.284 mmol, 4 eq) and 1 - methylimidazole (58.60 mg, 0.710 mmol, 10 eq) were added to a solution of 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (25 mg, 0.071 mmol, 1 eq) and 2- (4,4-difluoropiperidin- 1 -yl)-3-fluoropyridin- 4 -amine (18.15 mg, 0.078 mmol, 1.1 eq) in dichloromethane (2 mL) at room temperature. The temperature was raised to 60°C and stirred for 1 hour. The reaction mixture was diluted with water (10 mL). Extract with dichloromethane (3 × 10 mL). Combine the organic phases, backwash with saturated brine (1 × 10 mL), and dry over anhydrous sodium sulfate. After filtering the resulting mixture, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain 5-(6-azaspiro[2.5]octan-6-yl)-7-bromo- N- [2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-2,3-dihydro- 1H -indene-4-carboxamide (35 mg, 85.25%).

MS:(ESI, m/z ):563.15 [M+H] ⁺ , RT(min):1.184

Step 2 Synthesis of 5-(6-azaspiro[2.5]octan-6-yl) -N- [2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 27):

Under nitrogen protection, cesium carbonate (52.04 mg, 0.159 mmol , 3 eq), dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine (5.69 mg, 0.053 mmol, 1 eq) and 2-hydroxyethanesulfonamide (13.33 mg, 0.106 mmol, 2 eq) were added to a solution of 5-(6-azaspiro[2.5] octan -6-yl)-7-bromo-N-[2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-2,3-dihydro-1H-indene-4-carboxamide (30 mg, 0.053 mmol, 1 eq) and 2-hydroxyethanesulfonamide (13.33 mg, 0.106 mmol, 2 eq) in 1,4-dioxane (2 mL) at room temperature. 0.011 mmol, 0.2 eq) and methanesulfonic acid {bicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphine} (2'-methylamino-1,1'-biphenyl-2-yl) palladium (II) (4.89 mg, 0.005 mmol, 0.1 eq). After the addition was complete, the system was stirred at 100°C for 1 hour. After the reaction was completed, water (10 mL) was added to quench the reaction and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by HPLC under the following conditions: column specifications: XBridge BEH Shield RP18 5 µm, 30 mm *150 mm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL/min; elution gradient: 50% B to 75% B in 8 min; 254 nm; Rt: 6.9 min. 5-(6-azaspiro[2.5]octan-6-yl) -N- [2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (17.1 mg, 52.75%) was obtained.

MS:(ESI, m/z ):608.45 [M+H] ⁺ , RT(min):1.736

¹ H NMR (400 MHz, DMSO- d ₆ )δ 11.50(s, 1H), 9.33(s, 1H), 8.00 – 7.85(m, 2H), 7.18(s, 1H), 4.99(s, 1H), 3.78(t, J = 6.5 Hz, 2H), 3.54(t, J = 5.7 Hz, 4H), 3.30(t, J = 6.4 Hz, 2H), 3.10(t, J = 7.5 Hz, 2H), 2.94(t, J = 5.2 Hz, 4H), 2.85(t, J = 7.5 Hz, 2H), 2.14 – 2.01(m, 4H), 2.01 – 1.91(m, 2H), 1.46(s, 4H), 0.32(s, 4H).

Embodiment 6

5-{6-azaspiro[2.5]octan-6-yl}- N -[6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 28)

The first step is the synthesis of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (Compound 28-2):

Under nitrogen protection, 6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin - 2-amine (23.77 mg, 0.103 mmol, 1.2 eq), tetramethylchloroformamidine hexafluorophosphate (96.13 mg, 0.344 mmol, 4 eq) and N-methylimidazole (70.33 mg, 0.860 mmol, 10 eq) were added to a solution of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (30 mg, 0.086 mmol, 1 eq) in dichloromethane (1 mL) at room temperature, and the mixture was stirred at 80°C for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated brine (1 × 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (8:1) to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (50 mg, 91.17%).

MS:(ESI, m/z ): 562.65[M+H] ⁺ , RT(min):1.806

Step 2 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethylsulfonylamino)-2,3-dihydro- 1H -indene-4-carboxamide (Compound 28):

Under nitrogen protection, to a solution of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-2,3-dihydro- 1H -indene-4-carboxamide (50 mg, 0.089 mmol, 1 eq) in 1,4-dioxane (2 mL) was added methanesulfonic acid {bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphane}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (8.15 mg, 0.009 mmol, 0.1 eq), dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphane (9.49 mg, 0.018 mmol, 0.2 eq), cesium carbonate (86.74 mg, 0.267 mmol, 3 eq) and 2-hydroxyethane-1-sulfonamide (16.66 mg, 0.134 mmol, 1.5 eq). The reaction mixture was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (2 × 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC under the following conditions: chromatographic column specifications: YMC Triart C18 EXRs 5 µm, 30 mm * 150 mm; mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL/min; elution gradient: 60% B to 85% B in 8 min; detection wavelength: 220 nm; retention time (min): 7.8. Obtained 5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro- 1H -indene-4-carboxamide (10.08 mg, 18.69%)

MS:(ESI, m/z ): 608.45 [M+H] ⁺ , RT(min):1.863

¹ H NMR: (400 MHz, DMSO -d ₆ )δ 12.43(s, 1H), 9.34(s, 1H), 7.78 – 7.68(m, 1H), 7.62 – 7.52(m, 1H), 7.23(s, 1H), 4.97(s, 1H), 3.77(s, 2H), 3.64 – 3.48(m, 4H), 3.30(s, 2H), 3.25 – 3.18(m, 2H), 3.01 – 2.89(m, 4H), 2.88 – 2.79(m, 2H), 2.18 – 2.03(m, 4H), 2.02 – 1.91(m, 2H), 1.63(s, 4H), 0.35(s, 4H).

Embodiment 7

7-{6-azaspiro[2.5]octan-6-yl} -N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethylsulfonamido)imidazo[1,2-a]pyridine-8-carboxamide (Compound 29)

The first step is the synthesis of 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylic acid methyl ester (compound 29-3):

Under nitrogen protection, a solution of 2-bromo-1,1-dimethoxyethane (3.2 g, 19.001 mmol, 2.1 eq) in hydrobromic acid (20 mL) was stirred at room temperature for 30 minutes, and then ethanol (15 mL) was added. Then sodium bicarbonate (5.3 g, 63.336 mmol, 7 eq) and methyl 2-amino-4,6-dichloropyridine-3-carboxylate (2 g, 9.048 mmol, 1 eq) were added to the above system. The reaction solution was heated to 60°C and stirred for 1 hour. The reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3×80 mL). The organic phases were combined, backwashed with saturated brine (2×80 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (3:1) to obtain 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylic acid methyl ester (1.5 g, 67.65%).

MS:(ESI, m/z ): 244.08[M+H] ⁺ , RT(min):0.886

Step 2 Synthesis of 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylic acid (Compound 29-4):

Lithium hydroxide (199.37 mg, 8.325 mmol, 3 eq) was added to a mixed solution of methyl 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylate (680 mg, 2.775 mmol, 1 eq) in water (5 mL) and tetrahydrofuran (5 mL) at room temperature, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was acidified to pH 6 with 1N hydrochloric acid solution. The resulting mixture was concentrated to give crude 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylic acid (1.2 g).

MS:(ESI, m/z ): 230.90 [M+H] ⁺ , RT(min):0.498

Step 3 Synthesis of 5,7-dichloro- N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]imidazo[1,2-a]pyridine-8-carboxamide (Compound 29-5):

Under nitrogen protection, 6-(4,4-difluoropiperidin-1-yl)-4-methylpyrimidin-2-amine (944.33 mg, 4.156 mmol, 1.2 eq), N , N , N ', N' -tetramethylchloroformamidine hexafluorophosphate (3886.32 mg, 13.852 mmol, 4 eq) and N -methylimidazole (2843.13 mg, 34.630 mmol, 10 eq) were added to a solution of 5,7-dichloroimidazo[1,2-a]pyridine-8-carboxylic acid (800 mg, 3.463 mmol, 1 eq) in dichloromethane (4 mL) at room temperature. The mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate (3×50 mL). The organic phases were combined, backwashed with saturated brine (1×50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to obtain 5,7-dichloro- N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]imidazo[1,2-a]pyridine-8-carboxamide (350 mg, 22.91%).

MS:(ESI, m/z ): 441.25 [M+H] ⁺ , RT(min):1.198

Step 4 Synthesis of ethyl 2-[(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]aminoformyl}imidazo[1,2-a]pyridin-5-yl)sulfonyl]acetate (Compound 29-7):

Under nitrogen protection, ethyl 2- aminosulfonylacetate (170.49 mg, 1.020 mmol, 1.5 eq), palladium (II) methanesulfonate (62.45 mg, 0.068 mmol, 0.1 eq) and dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine (2'-methylamino-1,1'-biphenyl-2-yl) were added to a solution of 5,7-dichloro-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]imidazo[1,2-a]pyridine-8-carboxamide (300 mg, 0.680 mmol, 1 eq) in 1,4-dioxane (5 mL) at room temperature. eq), dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphine (72.72 mg, 0.136 mmol, 0.2 eq) and cesium carbonate (664.54 mg, 2.040 mmol, 3 eq). The reaction solution was heated to 100°C for 1 hour. The reaction solution was cooled to room temperature, and the reaction mixture was extracted with ethyl acetate (3×40 mL). The organic phases were combined, backwashed with saturated brine (1×50 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to obtain ethyl 2-[(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]aminoformyl}imidazo[1,2-a]pyridin-5-yl)sulfamoyl]acetate (150 mg, 38.57%).

MS:(ESI, m/z ): 572.45 [M+H] ⁺ , RT(min):1.094

Step 5 Synthesis of ethyl 2-[(7-{6-azaspiro[2.5]octan-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]aminoformyl}imidazo[1,2-a]pyridin-5-yl]sulfonyl]acetate (Compound 29-8):

Under nitrogen protection, 6-azaspiro[2.5]octane (34.99 mg, 0.315 mmol, 1.5 eq) and triethylamine (63.69 mg, 0.630 mmol, 3 eq) were added to a solution of ethyl 2-[(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]aminoformyl}imidazo[1,2-a]pyridin-5-yl)sulfonamide]acetate (120 mg, 0.210 mmol, 1 eq) in tert-butanol (1 mL) at room temperature. The reaction mixture was heated to 100°C and stirred for 2 days. The reaction mixture was cooled to room temperature and the reaction mixture was extracted with ethyl acetate (3×20 mL). The organic phases were combined, backwashed with saturated brine (2×20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (1:1) to obtain ethyl 2-[(7-{6-azaspiro[2.5]octan-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]aminoformyl}imidazo[1,2-a]pyridin-5-yl]sulfonylamine]acetate (60 mg, 44.22%).

MS:(ESI, m/z ): 647.45 [M+H] ⁺ , RT(min):1.240

Step 6 Synthesis of 7-{6-azaspiro[2.5]octan-6-yl} -N- [2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethylsulfonylamino)imidazo[1,2-a]pyridine-8-carboxamide (Compound 29):

Under nitrogen protection, lithium aluminum hydroxide (5.28 mg, 0.140 mmol, 1.5 eq) was added to a solution of ethyl 2-[(7-{6-azaspiro[2.5]octan-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl]sulfamoyl]acetate (60 mg, 0.093 mmol, 1 eq) in tetrahydrofuran (3.00 mL) at 0°C and stirred for 30 minutes at room temperature. The reaction solution was quenched with sodium sulfate decahydrate at 0°C. Filter and concentrate under reduced pressure. The crude product was purified by HPLC under the following conditions (chromatographic column specifications: XBridge BEH Shield RP18 5 μm, 30 mm *150 mm; mobile phase A: water (10mmol/L sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60ml/min; elution gradient: 40% B to 60% B in 10 min; detection wavelength: 254nm/220nm; retention time (min): 8.33). 7-{6-azaspiro[2.5]octan-6-yl}- N -[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethylsulfonamido)imidazo[1,2-a]pyridine-8-carboxamide (1.8 mg, 3.21%) was obtained.

MS:(ES, m/z ): 605.45 [M+H] ⁺ , RT(min): 1.559.

¹ H NMR: (400 MHz, DMSO- d ₆ ) δ 12.31(s, 1H), 11.77(s, 1H), 7.83(s, 1H), 7.48-7.28(m, 2H), 7.11(s, 1H), 4.15(s, 2H), 3.98(s, 4H), 3.70-3.35(m, 4H), 3.32-3.01(m, 4H), 2.55(s, 2H), 2.38(s, 3H), 1.98(s, 4H), 0.43(s, 4H).

Embodiment 8

N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-(2-hydroxyethanesulfonamido)-7-{6-azaspiro[2.5]octan-6-yl}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (Compound 30)

The first step is the synthesis of methyl 4,6-dichloro-2-[[( E )-(hydroxyamino)methylene]amino]pyridine-3-carboxylate (Compound 30-2):

Under nitrogen protection, add N , N -dimethylformamide dimethyl acetal (2.18 mL, 16.286 mmol, 1.20 eq) to a solution of 2-amino-4,6-dichloropyridine-3-carboxylic acid methyl ester (3 g, 13.572 mmol, 1 eq) in isopropanol (60.00 mL) at room temperature, heat to 70°C and stir for 3 hours, cool to room temperature, add hydroxylamine hydrochloride (1.13 g, 16.286 mmol, 1.2 eq) to the reaction solution, stir for 3 hours at room temperature. Filter, collect the filter cake and wash with isopropanol (3 × 50 mL). The product was methyl 4,6-dichloro-2-[[( E )-(hydroxyamino)methylene]amino]pyridine-3-carboxylate (1.5 g, 41.85%).

MS:(ESI, m/z ): 264.10 [M+H] ⁺ , RT(miN):0.949.

Step 2 Synthesis of 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (Compound 30-3):

Under nitrogen protection, add trifluoroacetic anhydride (1.79 g, 8.521 mmol, 1.5 eq) in acetonitrile (5 mL) to a solution of methyl 4,6-dichloro-2-[[( E )-(hydroxyamino)methylene]amino]pyridine-3-carboxylate (1.5 g, 5.681 mmol, 1 eq) in acetonitrile (10 mL) at 0°C. Heat to room temperature and stir for 4 hours. Concentrate the reaction solution under reduced pressure. Neutralize to pH = 7 with saturated sodium bicarbonate aqueous solution. Extract with dichloromethane (3 × 20 mL). Combine the organic phases, backwash with saturated brine (1 × 20 mL), and dry over anhydrous sodium sulfate. Filter the resulting mixture, and concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (1:2) to obtain 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (1.1 g, 77.13%).

MS:(ESI, m/z ): 246.20 [M+H] ⁺ , RT(min):0.828.

Step 3 Synthesis of 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (Compound 30-5):

Under nitrogen protection, add triethylamine (740.29 mg, 7.317 mmol, 3 eq) to a solution of 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (600 mg, 2.439 mmol, 1 eq) in tert-butanol (6 mL) at room temperature. Stir the mixture for 2 minutes, then add 3,4-dimethoxybenzylamine (16.31 mg, 0.097 mmol, 1.2 eq) dropwise at room temperature. Heat to 60°C and stir for 2 hours. Cool the reaction mixture to room temperature. Dilute with water (20 mL) and extract with ethyl acetate (3 × 20 mL). Combine the organic phases, backwash with saturated brine (1 × 20 mL), and dry over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (1:3) to obtain 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (700 mg, 68.56%).

MS:(ESI, m/z ): 377.35 [M+H] ⁺ , RT(min):1.062.

Step 4 Synthesis of 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid (Compound 30-6):

Under nitrogen protection, lithium hydroxide (114.41 mg, 4.776 mmol, 3 eq) was added in batches to a solution of 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid methyl ester (600 mg, 1.592 mmol, 1 eq) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL) at room temperature. The temperature was raised to 60°C and stirred for 1 hour. The reaction mixture was acidified to pH 4 with 1N hydrochloric acid solution. A white solid precipitated, which was filtered and the filter cake was collected to obtain 1 g of crude product.

MS:(ESI, m/z ): 363.60 [M+H] ⁺ , RT(min):1.000.

Step 5 Synthesis of 7-chloro- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (Compound 30-7):

Under nitrogen protection, N, N, N', N'-tetramethylchloroformamidine hexafluorophosphate (1.5 g, 5.512 mmol, 4 eq) and N-methylimidazole (1.1 g, 13.780 mmol, 10 eq) were added in portions to a solution of 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino } -[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid (500 mg, 1.378 mmol, 1 eq) and 6-(4,4-difluoropiperidin- 1 -yl ) -4-methylpyridin-2-amine (375.87 mg, 1.654 mmol , 1.2 eq) in N , N -dimethylformamide (10 mL) at room temperature. The temperature was raised to 60°C and the reaction was stirred for 1 hour. The reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with dichloromethane (3 × 20 mL). The organic phases were combined, backwashed with saturated brine (1 × 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (3:1) to obtain 7-chloro- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (500 mg, 60.25%).

MS:(ESI, m/z ): 572.50 [M+H] ⁺ , RT(min):1.370.

Step 6 Synthesis of 7-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (Compound 30-8):

Under nitrogen protection, 6-azaspiro[2.5]octane hydrochloride (309.75 mg, 2.098 mmol, 2 eq) was added in portions to a solution of 7-chloro- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (600 mg, 1.049 mmol, 1 eq) and triethylamine (318.43 mg, 3.147 mmol, 3 eq) in tert-butanol (10 mL) at room temperature. The temperature was raised to 120°C and stirred for 2 hours. The reaction solution was cooled to room temperature and diluted with water (10 mL). Extract with dichloromethane (3 × 10 mL). Combine the organic phases, backwash with saturated brine (1 × 10 mL), and dry over anhydrous sodium sulfate. After filtering the resulting mixture, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with dichloromethane/methanol (10:1) to obtain compound 30-8 (445 mg, 62.32%).

MS:(ESI, m/z ): 647.60 [M+H] ⁺ , RT(min):1.172.

Step 7 Synthesis of 5-amino-7-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (Compound 30-9):

Under nitrogen protection, a solution of compound 30-8 (410 mg, 0.634 mmol, 1 eq) in trifluoroacetic acid (5 mL, 67.315 mmol, 106.18 eq) was stirred at 60°C for 1 hour. The reaction solution was cooled to room temperature, and the residue was concentrated under reduced pressure. Alkalize to pH = 8 with saturated sodium bicarbonate aqueous solution. Extract with ethyl acetate (3 × 20 mL). Combine the organic phases, backwash with saturated brine (1 × 20 mL), and dry over anhydrous sodium sulfate. After filtering the obtained mixture, the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column chromatography under the following conditions: column specifications (C18 column), mobile phase, water and acetonitrile, 0% to 80% gradient in 10 minutes, UV254 nano detector. Compound 30-9 (310 mg, 88.63%) was obtained.

MS:(ESI, m/z ): 497.30[M+H] ⁺ , RT(min):1.410.

Step 8 Synthesis of ethyl 2-[(7-{6-azaspiro[2.5]octan-6-yl}-8-{[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]aminoformyl}-[1,2,4]triazolo[1,5-a]pyridin-5-yl)sulfonyl]acetate (Compound 30-11):

Under nitrogen protection, ethyl 2-(chlorosulfonyl)acetate (75.16 mg, 0.402 mmol, 2 eq) was added dropwise to a solution of compound 30-9 (100 mg, 0.201 mmol, 1 eq) and triethylamine (61.14 mg, 0.603 mmol, 3 eq) in dichloromethane (3 mL) at 0°C. The mixture was heated to room temperature and stirred for 2 hours. Diluted with water (10 mL) and extracted with dichloromethane (3 × 10 mL). The organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column chromatography under the following conditions: C18 column, mobile phase, water and acetonitrile, 10% to 70% gradient in 10 minutes, UV254 nano detector to obtain compound 30-11 (80 mg, 61.43%).

MS:(ESI, m/z ): 647.30[M+H] ⁺ , RT(min):1.100.

Step 9 Synthesis of N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-(2-hydroxyethylsulfonylamino)-7-{6-azaspiro[2.5]octan-6-yl}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (Compound 30):

Under nitrogen protection, at 0°C, a solution of lithium aluminum tetrahydrofuran (6.16 mg, 0.162 mmol, 1.5 eq) was added dropwise to a solution of compound 30-11 (70 mg, 0.108 mmol, 1 eq) in tetrahydrofuran (3 mL). After the addition, the system was stirred for 1 hour at 0°C. The reaction mixture was quenched with ice water at 0°C. The reaction mixture was extracted with ethyl acetate (3 × 10 mL), the organic phases were combined, backwashed with saturated brine (1 × 10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by HPLC under the following conditions: column specifications: SuNfire C18 5 μm, 30 mm * 150 mm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 60 mL/min; elution gradient: 35% B to 68% B in 8 min; 254nm/220nm; Rt: 7.42 min. Compound 30 (23.73 mg, 35.42%) was obtained.

MS:(ESI, m/z ): 605.20[M+H] ⁺ , RT(min):1.720.

¹ H NMR (400 MHz, DMSO- d ₆ )δ 14.04(s, 1H), 11.77(s, 1H), 8.96(s, 1H), 7.51(s, 1H), 6.87(s, 1H), 6.59(s, 1H), 4.96(s, 1H), 3.83(t, J = 6.6 Hz, 2H), 3.72(t, J = 6.1 Hz, 4H), 3.21(t, J = 6.7 Hz, 2H), 3.10(s, 4H), 2.27(s, 3H), 1.97(t, J = 15.6 Hz, 4H), 1.72(s, 4H), 0.39(s, 4H).

Embodiment 9

5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-3-methyl-1,3-benzodiazole-4-carboxamide (Compound 31)

The first step is the synthesis of methyl 4-bromo-2,6-difluoro-3-nitrobenzoate (compound 31-2):

Under nitrogen protection, nitric acid (1.25 mL) was added to a sulfuric acid solution (17 mL) of methyl 4-bromo-2,6-difluorobenzoate (5 g, 20 mmol, 1 eq) at room temperature, and the reaction solution was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature, extracted with dichloromethane (3 × 100 mL), the organic phases were combined, backwashed with saturated brine (2 × 120 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product of methyl 4-bromo-2,6-difluoro-3-nitrobenzoate (6.01 g) was obtained.

¹ H NMR: (400 MHz, DMSO- d ₆ )δ 8.11 (dd, J = 9.5, 2.1 Hz, 1H), 3.93 (s, 3H).

Step 2 Synthesis of 4-bromo-6-fluoro-2-methylamino-3-nitrobenzoic acid methyl ester (Compound 31-3):

Under nitrogen protection, methylamine (472 mg, 15 mmol, 1.5 eq) was added to a methanol solution (30 mL) of methyl 4-bromo-2,6-difluoro-3-nitrobenzoate (3 g, 10 mmol, 1 eq) at room temperature, and the reaction solution was stirred at 60°C for 16 hours. The reaction solution was cooled to room temperature, quenched with water, and extracted with ethyl acetate (3 × 80 mL). The organic phases were combined and backwashed with saturated brine (2 × 80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain methyl 4-bromo-6-fluoro-2-methylamino-3-nitrobenzoate (2.21 g, 71%).

MS:(ESI, m/z ):306.85 [M+H] ⁺ , RT(min):1.180

Step 3 Synthesis of 6-{6-azaspiro[2.5]octane-6-yl}-4-bromo-2-methylamino-3-nitrobenzoic acid methyl ester (Compound 31-4):

Under nitrogen protection, 6-azaspiro[2.5]octane hydrochloride (217 mg, 2 mmol, 1.2 eq) and triethylamine (824 mg, 8 mmol, 5 eq) were added to a solution of methyl 4-bromo-6-fluoro-2-methylamino-3-nitrobenzoate (500 mg, 1.6 mmol, 1 eq) in tert-butanol (15 mL) at room temperature. The reaction mixture was heated to 100°C and stirred for 16 hours. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined and backwashed with saturated brine (2 × 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain methyl 6-{6-azaspiro[2.5]octan-6-yl}-4-bromo-2-methylamino-3-nitrobenzoate (460 mg, 71%).

MS:(ESI, m/z ): 398.35 [M+H] ⁺ ,RT(min):1.380

Step 4 Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid methyl ester (Compound 31-6):

Under nitrogen protection, trimethyl orthoformate (3.5 g, 33 mmol, 30 eq), zinc powder (433 mg, 6.6 mmol, 6 eq), lithium bromide (115 mg, 1.3 mmol, 1.2 eq) and formic acid (508 mg, 11 mmol, 10 eq) were added to a solution of methyl 6-{6-azaspiro[2.5]octan-6-yl}-4-bromo-2-methylamino-3-nitrobenzoate (440 mg, 1.1 mmol, 1 eq) in ethanol (3 mL) at room temperature. After the addition, the reaction was stirred at room temperature for 1 hour. The mixture was quenched with water and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined and backwashed with saturated brine (2 × 30 mL). The mixture was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid methyl ester (190 mg, 45.5%).

MS:(ESI, m/z ):378.30 [M+H] ⁺ ,RT(min):1.193

Step 5 Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid (Compound 31-7):

Under nitrogen protection, lithium hydroxide (31.7 g, 1.3 mmol, 5 eq), distilled water (2 mL) and methanol (2 mL) were added to a tetrahydrofuran solution (2 mL) of 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid methyl ester (100 mg, 0.3 mmol, 1 eq) at room temperature. The reaction mixture was heated to 60°C and stirred for 72 hours. The reaction solution was extracted with ethyl acetate (2 × 10 mL), the aqueous phase was collected, the pH was adjusted to 6 with 1 M dilute hydrochloric acid, extracted with ethyl acetate (2 × 10 mL), the organic phases were combined and backwashed with saturated brine (2 × 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid (101 mg, 105%).

MS:(ESI, m/z ): 364.30 [M+H] ⁺ ,RT(min):0.763

Step 6 Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-3-methyl-1,3-benzodiazole-4-carboxamide (Compound 31-8):

Under nitrogen protection, 6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-amine (112 mg, 0.5 mmol, 3 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (185 mg, 0.7 mmol, 4 eq) and N -methylimidazole (135 mg, 1.7 mmol, 10 eq) were added to a dichloromethane solution (4 mL) of 5- {6- azaspiro [2.5]octan-6-yl}-7-bromo-3-methyl-1,3-benzodiazole - 4 -carboxylic acid (60 mg, 0.16 mmol, 1 eq) at room temperature. The reaction solution was heated to 60°C and stirred for 1 hour. The reaction solution was cooled to room temperature, extracted with ethyl acetate (3 × 10 mL), the organic phases were combined, backwashed with saturated brine (2 × 10 mL), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo- N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-3-methyl-1,3-benzodiazole-4-carboxamide (72 mg, 76%).

MS:(ESI, m/z ): 573.30 [M+H] ⁺ , RT(min):1.392

Step 7 Synthesis of 5-{6-azaspiro[2.5]octan-6-yl} -N- [6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonylamino)-3-methyl-1,3-benzodiazole-4-carboxamide (Compound 31):

Under nitrogen protection, 2-hydroxyethanesulfonamide (26 mg, 0.2 mmol, 1.2 eq), methanesulfonic acid {bicyclohexyl[3 - isopropoxy-2',4',6'-triisopropyl-(1,1'-biphenyl)-2-yl]phosphane}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (16.02 mg, 0.17 mmol, 1 eq) and 5-{6-azaspiro[2.5]octan-6-yl}-7-bromo-N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-3-methyl-1,3-benzotriazole-4-carboxamide (100 mg, 0.17 mmol, 1 eq) were added to a solution of 1,4-dioxane (1 mL) at room temperature. 0.017 mmol, 0.1 eq), dicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphane (18.65 mg, 0.035 mmol, 0.2 eq), cesium carbonate (170.44 mg, 0.522 mmol, 3 eq), the reaction solution was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined and backwashed with saturated brine (2 × 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography to obtain 5-{6-azaspiro[2.5]octan-6-yl}- N -[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-3-methyl-1,3-benzodiazole-4-carboxamide (7.08 mg, 6.3%).

MS:(ESI, m/z ): 618.70 [M+H] ⁺ ,RT(min):1.903

¹ H NMR: (400 MHz, DMSO -d ₆ )δ 10.73(s, 1H), 9.15(s, 1H), 8.18(s, 1H), 7.42(s, 1H), 7.15(s, 1H), 6.57(s, 1H), 5.05(s, 1H), 3.83(d, J = 6.6 Hz, 2H), 3.80(d, J = 3.9 Hz, 3H), 3.69(t, J = 5.5 Hz, 4H), 3.45(t, J = 6.6 Hz, 2H), 2.96(t, J = 5.2 Hz, 4H), 2.29(s, 3H), 2.05 – 1.85(m, 4H), 1.45(s, 4H), 0.29(s, 4H).

Biological Evaluation

Test example 1

Test Name: Imaging-based Nuclear Count Analysis (NCA) in OVCAR-3 cells

Day 0 Compound dilution and handling

a) The final tested concentrations of AM-5308 are: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5 nM.

b) The final test concentrations of the test compounds were: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5 nM.

c) The cells were cultured in a 37°C, 5% CO ₂ incubator for 4 days.

d) DMSO concentration is 0.1%.

Day 1: Inoculate cells into 384-well cell culture plates

a) When the confluence of cells reaches 80%-90%, process the cells.

b) Resuspend the cells in medium, then count and dilute the cells at the desired density.

c) Add 30 µL/well of the cell suspension containing the appropriate cells to a 384-well plate: 600 cells/well.

Day 4 Testing

a) Add 30 µL of 8% fixative (final concentration 4%) and incubate the plate at room temperature for 30 minutes.

b) Centrifuge plate, 1000 RPM, 30 s.

c) Wash twice with 60 µl/well PBS.

d) After fixation, cells were permeabilized and stained in 60 µL of wash buffer (1% BSA, 0.2% Triton X-100, 1X PBS) containing 2 µg/mL Hoechst 33342 DNA stain.

e) Seal the plate and incubate at room temperature in the dark for 1 hour.

f) Wash 3 times with PBS.

g) Add 50 µL PBS/well and scan the plate using HCS.

h) Data acquisition and testing.

Data Analysis

Inhibition rate (%) = 100-(compound well reading - low reading control well reading)/(high reading control well reading - low reading control well reading)* 100

High reading control wells: cells plus 30 nL DMSO; low reading control wells: 10 μM AM-5308.

GraphPad Prism 8 software was used to calculate IC ₅₀ (nM) and plot the compound effect-dose curves.

Table 1 Biological activity data of the compounds of the present invention Compound No. NCA(OVCAR-3) IC ₅₀₍ nM) 1 65.25 8 40.12 9 45.54 13 24.27 27 64.8 28 56.89

Test example 2

Test Name: ADP-Glo™ Kinase Assay

Steps:

1) Prepare 1× reaction buffer.

2) Use Echo 655 to transfer 100 nL of the diluted compound stock solution to each well of the reaction plate. The final DMSO concentration is 1%.

3) Seal the reaction plate with sealing film and centrifuge at 1000 g for 1 minute.

4) Prepare 2× enzyme solution using 1× reaction buffer.

5) Add 5 μL of 2× enzyme solution to each well of the reaction plate. Seal the plate with a sealing film and centrifuge at 1000g for 1 minute, then place at room temperature for 15 minutes.

6) Prepare 2× ATP solution using 1× reaction buffer.

7) Add 5 μL of 2× ATP solution to the reaction plate and centrifuge at 1000g for 1 minute to start the reaction.

8) Incubate at room temperature for 60 minutes.

9) Add 10 μL ADP Glo reagent. Centrifuge at 1000 g for 1 min and incubate at room temperature for 60 min.

10) Add 20 μL of kinase assay reagent. Centrifuge at 1000 g for 1 minute and incubate at room temperature for 60 minutes.

11) Centrifuge at 1000 g for 1 min.

12) Read the luminescence signal on Envision 2104.

Data Analysis:

The inhibition percentage is calculated as follows:

%inhibition = 100-(Signal _cmpd -Signal _{Ave_PC} )/(Signal _{Ave_VC} -Signal _{Ave_PC} )× 100

Signal _cmpd : Average value of the test compound on the plate.

Signal _{Ave_PC} : Average value of positive controls on the reaction plate.

Signal _{Ave_VC} : The average value of negative controls on the reaction plate.

Calculate _IC50 and dose-effect curve of the proposed compound:

The IC ₅₀ values of the compounds were obtained using GraphPad 8.0 using a nonlinear fitting formula.

3) Quality Control

Z factor ＞ 0.5; S/B ＞ 2.

Table 2 Enzyme activity data of the compounds of the present invention Compound No. ADP-Glo IC50(nM) 9 24.58

Test Example 3 Effect of the Compounds of the Invention on Tumor Regression

To demonstrate the effect of KIF18A inhibitors on tumor regression, OVCAR-3 cells (ATCC) were selected for the experiment. 0.1 mL of approximately 5 × 10 ⁶ OVCAR-3 cells were injected subcutaneously into the right flank of female nude mice. According to the tumor volume (average tumor volume was 150 mm ³ ), the animals were randomly divided into 6 groups (solvent group, compound 9 was divided into three dose groups of 10, 20 or 30 mg/kg, and AMG650 was divided into two dose groups of 15 or 30 mg/kg), with 10 animals in each group. Starting from the 28th day after tumor inoculation, the drug was orally administered once a day. The tumor volume was measured using an electronic vernier caliper (Chengdu Sanhe Measuring Instrument Co., Ltd.). The tumor volume and animal weight were measured twice a week (28 days at the beginning of the study and 46 days at the end of the study), and the tumor growth inhibition rate of the compound was calculated.

The formula for calculating tumor volume is: 1/2×a×b ² , where a and b are the measured length and width of the tumor, respectively.

The calculation formula for tumor inhibition rate %TGI is: 1-(TV _Tn -TV _T0 )/(TV _Cn -TV _C0 )× 100%, TVC is the average tumor volume of the negative control group, and TVT is the average tumor volume of the treatment group.

GraphPad Prism software (V9.5.0) was used to plot the data, and the tumor volume and body weight data were expressed as the mean plus or minus the standard error of the mean. As shown in Figure 1, the TGI of the compound 9 dose groups of 10, 20, and 30 mg/kg were 47.35%, 106.60%, and 115.58%, respectively; the TGI of AMG650 15 and 30 mg/kg were 72.58% and 113.15%, respectively. All dose groups of compound 9 had no significant effect on the weight changes of animals in the solvent group, and no obvious toxicity was observed.

The above results show that compound 9 of the present invention can induce tumor regression in female nude mouse xenograft tumor models caused by human plasma ovarian cancer OVCAR-3 cells (TP53 mutation, CCNE1 amplification). Compound 9 at doses of 20 and 30 mg/kg can significantly inhibit tumor growth in the subcutaneous transplant tumor model of OVCAR-3 mice, and the inhibitory effect of compound 9 at a dose of 20 mg/kg on subcutaneous transplant tumors in mice is equivalent to that of AMG650 30 mg/kg.

As shown in Table 3, the tumor-to-blood ratio (ratio of tumor tissue exposure to plasma exposure) of compound 9 is better than that of AMG650, indicating that a higher proportion of the drug enters the tumor tissue to exert its effect. At the same time, compared with AMG650, the lower absolute exposure of compound 9 can reduce the potential cumulative toxicity of the drug.

Table 3. Exposure levels in mouse plasma and tumor tissue

The above is an exemplary description of the implementation of the technical solution of the present invention. It should be understood that the protection scope of the present invention is not limited to the above implementation. Any modification, equal replacement, improvement, etc. made by a person with ordinary knowledge in the technical field to which the present invention belongs within the spirit and principles of the present invention shall be included in the protection scope of the patent scope of the present invention.

without

FIG1 is a line graph showing the effects of compound 9 of the present invention on tumor volume and body weight of mice bearing an OVCAR-3 mouse xenograft tumor model.

Claims (10)

A compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: ( I ) wherein A is selected from unsubstituted or optionally substituted with one, two or more _Ra or a fused cyclic group 1; the fused cyclic group 1 comprises two, three or four rings independently selected from saturated or partially unsaturated C _3-14 carbon rings, C _6-14 aromatic rings, 5-14 membered heteroaromatic rings, 3-14 membered heterocyclic rings; each _{Ra is} the same or different and independently selected from H, OH, halogen, nitrile, NH ₂ , NO ₂ , the following groups which are unsubstituted or optionally substituted by one, two or more _Ra1 : C _1-12 alkyl, C _1-12 alkoxy, C _3-12 cycloalkyl; or, two _Ras connected to the same ring carbon atom together with the carbon atom to which they are connected form a saturated or partially unsaturated C _3-14 carbon ring; each R _a1 is the same or different and is independently selected from H, OH, halogen, nitrile, _NH2 , _NO2 , _C1-12 alkyl, _C1-12 alkoxy, _C3-12 cycloalkyl; _X1 , _X2 , _X3 are the same or different and are independently selected from N or _CR0 ; _R0 is selected from H, halogen, nitrile, _C1-12 alkyl, halogenated _C1-12 alkyl, nitrile C1-12 alkyl, _C1-12 alkoxy, halogenated _C1-12 alkoxy, nitrile _C1-12 alkoxy; B is selected from the following groups which are unsubstituted or optionally substituted with one, two or more _Rb : _C1-12 alkyl, halogenated _C1-12 alkyl, nitrile _{C1-12 alkyl} , _C1-12 alkoxy, halogenated C1-12 alkoxy, nitrile _C1-12 alkoxy R _is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, _{nitrile C 1-12 alkyl, C 1-12 alkoxy, halogenated C 1-12 alkoxy , nitrile C 1-12} alkoxy; Ring G is selected from a fused cyclic group 2 which _is unsubstituted or optionally substituted with one, two or more R _g , and the fused cyclic group 2 is formed by condensing Ring _{G 1 and} Ring G ₂ ; Ring G ₁ is selected from a C _6-14 aromatic ring, a 5-14 heteroaromatic ring, and a 3-14 heterocyclic ring; Ring G _{R 2} is selected from C _3-14 carbocyclic ring, C _6-14 aromatic ring, 5-14 member heteroaromatic ring, 3-14 member heterocyclic ring; M and E are preferably connected to ring G ₁ ; each R _g is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, nitrile C _1-12 alkyl, C _1-12 alkoxy, halogenated C _1-12 alkoxy, nitrile C _1-12 alkoxy; E is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Re : -NH-S(=O) _{2 -Re1} , -S(=O) ₂ -NH- _Re2 , -S(=O)(=NH) _-Re3 , -N( _Re4 )( _Re5 ), a 3-14 membered heterocyclic group; each _{Re is} the same or different and is independently selected from OH, halogen, nitrile, C _1-12 alkyl, C _1-12 alkoxy, halogenated C _1-12 alkyl, halogenated C _1-12 alkoxy, nitrile C _1-12 alkyl, nitrile C _1-12 alkoxy, -N( _Re6 )( _Re7 ); _Re1 , _Re2 , _Re3 , _Re4 , _Re5 , _Re6 , _Re7 are the same or different and are independently selected from H, C _1-12 alkyl, C _1-12 alkoxy, hydroxyl C _1-12 alkyl, halogenated C _1-12 alkyl, halogenated C _1-12 alkoxy, nitrile C _1-12 alkyl, nitrile C _1-12 alkoxy, C C _3-12 cycloalkyl, 3-14 membered heterocyclic group, C _1-12 alkoxy-C _1-12 alkyl; M is selected from the following groups which are unsubstituted or optionally substituted by one, two or more R _m : C _3-12 cycloalkyl, C _3-12 cycloalkenyl, 3-14 membered heterocyclic group; each R _m is the same or different and is independently selected from halogen, nitrile, C _1-12 alkyl, halogenated C _1-12 alkyl, nitrile C _1-12 alkyl, C _1-12 alkoxy, nitrile C _1-12 alkoxy. The compound as claimed in claim 1, wherein A is selected from unsubstituted or optionally substituted by one, two or more _Ra or a fused cyclic group 1; the fused cyclic group 1 comprises two, three or four rings independently selected from saturated or partially unsaturated C _3-8 carbon rings, C _6-10 aromatic rings, 5-10 membered heteroaromatic rings, and 3-8 membered heterocyclic rings; each _{Ra is} the same or different and independently selected from H, OH, halogen, nitrile, NH ₂ , NO ₂ , C _1-6 alkyl, C _1-6 alkoxy, C _3-8 cycloalkyl, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, nitrile C _1-6 alkyl, nitrile C _1-6 alkoxy, and C _3-8 cycloalkyl-C _1-6 alkoxy; or, two R a connected to the same ring carbon atom are selected from: _a together with the carbon atom to which it is connected form a saturated or partially unsaturated C _3-8 carbon ring; preferably, A is selected from , , , ; wherein T is selected from CH ₂ , CH, NH, NR _a or O; Z is selected from CH ₂ , CH, NH, NR _a or O; X is selected from N or CH; n is selected from 0, 1, 2, 3, 4 or 5; p and q are independently selected from 0, 1, 2, 3, and p and q are not 0 at the same time; r and s are independently selected from 0, 1, 2, 3, and r and s are not 0 at the same time; preferably, X ₁ and X ₂ are the same or different and are independently selected from N or CR ₀ ; preferably, X ₁ and X ₂ are the same or different and are independently selected from N or CH; preferably, X ₂ and X ₃ are not N at the same time; preferably, when X ₁ and X ₂ are N, X ₃ is CR ₀ ; when X ₁ is N or CR ₀ and X ₂ is CR ₀ , X ₃ is N or CR ₀ ; when X ₁ is N or CR ₀ , X ₂ is N, X ₃ is CR ₀ ; R ₀ is selected from H, halogen, C _1-6 alkyl; preferably, when X ₁ and X ₂ are N, X ₃ is CR ₀ ; when X ₁ is N or CH, X ₂ is CH, X ₃ is N or CR ₀ ; when X ₁ is CH, X ₂ is N, X ₃ is CR ₀ ; when X ₁ is N, X ₂ is CR ₀ , X ₃ is N; R ₀ is selected from H, C _1-6 alkyl; preferably, B is selected from C _1-6 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-8 member heterocyclic group, halogenated 3-8 member heterocyclic group; Preferably, B is selected from a halogenated 6-membered N-containing heterocyclic group; for example, a halogenated piperidinyl group; Preferably, B is selected from Preferably, A is selected from unsubstituted or optionally substituted with one, two or more _Ra , , , , or a fused ring group 1; the fused ring group 1 is formed by condensing two, three, four or more rings selected from a benzene ring, a pyridine ring, an imidazole ring, a piperazine ring, a dihydropyridine ring, a tetrahydropyridine ring, a dihydropyrrole ring, a tetrahydropyrrole ring, a dihydropyran ring, a cyclobutene ring, a cyclopentene ring, a cyclohexene ring, and a cycloheptene ring; preferably, A is selected from the following groups which are unsubstituted or optionally substituted by one, two or more _Ra : , , , , , , , , , ; Preferably, each _{Ra is} the same or different and is independently selected from H, OH, F, Cl, nitrile, methyl, ethyl, isopropyl, methoxy, ethoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclopropylmethoxy; or, two _Ras connected to the same ring carbon atom together with the carbon atom to which they are connected form a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, or a cycloheptane ring; Preferably, A is selected from , , , , , , , , , , , , , , . The compound as claimed in claim 1 or 2, wherein ring G is selected from a fused cyclic group 2 which is unsubstituted or optionally substituted by one, two or more _Rg , and the fused cyclic group 2 is formed by condensing ring _G1 and ring _G2 ; ring _G1 is selected from _C6-10 aromatic ring, 5-10 member heteroaromatic ring, 5-10 member heterocyclic ring; ring _G2 is selected from _C3-10 carbon ring, _C6-10 aromatic ring, 5-10 member heteroaromatic ring, 3-10 member heterocyclic ring; preferably, ring G ₁ is selected from cyclopentene ring, cyclohexene ring, dihydrofuran ring, dihydropyran ring, imidazole ring, triazole ring, benzene ring, pyridine ring, dihydropyridine ring, pyrrole ring, pyrazole ring, furan ring, thiophene ring; preferably a benzene ring or a pyridine ring; preferably, ring G ₂ is selected from cyclopentene ring, cyclohexene ring, dihydrofuran ring, dihydropyran ring, imidazole ring, triazole ring, benzene ring, pyridine ring, pyrrole ring, pyrazole ring, furan ring, thiophene ring, methylimidazole ring; preferably, ring G is selected from , , , , , , , , , , , , , . The compound as described in any one of claims 1 to 3, wherein E is selected from the following groups which are unsubstituted or optionally substituted by one or two _Re : -NH-S(=O) ₂ - _Re1 , -S(=O) ₂ -NH- _Re2 , -S(=O)(=NH) _-Re3 ; _Re1 , _Re2 , and _Re3 are the same or different and are independently selected from H, _C1-6 alkyl, hydroxyl _C1-6 alkyl, halogenated _C1-6 alkyl, halogenated _C1-6 alkoxy, nitrile _C1-6 alkyl, nitrile _C1-6 alkoxy, _C3-8 cycloalkyl, 3-8 membered heterocyclic group, and _C1-6 alkoxy- _C1-6 alkyl; each _{Re is} the same or different and is independently selected from OH, halogen, nitrile, _C1-6 alkyl, _C1-6 alkoxy, halogenated C1-6 _{Preferably ,} E _is selected _from . The compound as described in any one of claims 1 to 4, wherein M is selected from C _3-8 cycloalkyl, C _3-8 cycloalkenyl or nitrogen-containing 3-8 membered heterocyclic group which is unsubstituted or optionally substituted by one, two or more R _m ; each R _m is the same or different and is independently selected from H, halogen, nitrile, C _1-6 alkyl, halogenated C _1-6 alkyl, nitrile C _1-6 alkyl, C _1-6 alkoxy, nitrile C _1-6 alkoxy; or, two R _m connected to the same ring carbon atom together with the carbon atom to which they are connected form a saturated or partially unsaturated C _{3-8 carbocyclic ring; preferably, M is selected from C 3-8 cycloalkyl, C 3-8} cycloalkenyl or nitrogen-containing 3-8 membered heterocyclic group which is unsubstituted or optionally substituted by one, two or more R _m , or . The compound according to any one of claims 1 to 5, wherein the compound has the following structure: (IA-1) (IA-2) (IA-3) (IA-4) wherein A, M, E, Ring G, Ring G ₁ , and Ring G ₂ independently have the definitions as described in any one of claims 1 to 5, represents a carbon-carbon single bond or a carbon-carbon double bond; Preferably, the compound represented by formula (I) has the following structure: (IB-1) (IB-2) (IB-3) (IB-4) wherein B, M, E, ring G, X, Z, T, X ₁ , X ₂ , X ₃ , _Ra , n, p, q, r, s are independently defined as in any one of claims 1 to 5; preferably, the compound represented by formula (I) has the following structure: (IC-1) (IC-2) (IC-3) wherein B, ring G, X, Z, T, X ₁ , X ₂ , X ₃ , _Ra , n, r, s, p, q are independently defined as in any one of claims 1 to 5; preferably, the compound represented by formula (I) has the following structure: , (IIA-1) (IIA-2) wherein A, ring G ₁ and ring G ₂ independently have the definitions as described in any one of claims 1 to 5; preferably, the compound represented by formula (I) has the following structure: , (IIB-1) (IIB-2) wherein A and ring _G2 independently have the definitions as described in any one of claims 1 to 5; preferably, the compound represented by formula (I) has the following structure: , (IIC-1) (IIC-2) (IIC-3) wherein B, ring G ₂ , X, Z, T, X ₁ , X ₂ , X ₃ , _Ra , n, r, s, p, q are independently defined as in any one of claims 1 to 5; preferably, the compound represented by formula (I) has the following structure: (III) wherein ring G ₂ , X ₁ , X ₂ , and X ₃ independently have the meanings as defined in any one of claims 1 to 5. The compound as described in any one of claims 1 to 6, wherein the compound is selected from the following structures: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , . A method for preparing the compound as described in any one of claims 1 to 7, comprising the following steps: (1) Compound a reacts with compound A- _NH2 to obtain compound b; (2) Compound b reacts with compound MH to obtain compound c; (3) Compound c reacts with compound EH to obtain a compound represented by formula (I); wherein A, E, M and ring G independently have the definitions as described in any one of claim 1 to 7; L is selected from halogens, such as Cl, Br, I; Q is selected from halogens, such as F, Cl, Br. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound as described in any one of claims 1 to 7, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof. A compound as described in any one of claims 1 to 7, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, for use in the preparation of a drug; Preferably, the use may be for the preparation of a drug for treating KIF18A-mediated conditions and/or diseases, such as for the preparation of a KIF18A inhibitor drug; Preferably, the disease is, for example, cancer, including intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.