Fused ring kif18a inhibitor compounds, pharmaceutical compositions, and methods of making and using the same

Abstract

The present application provides a compound shown in formula (I), a pharmaceutical composition and a preparation method and application thereof. The compound has good KIF18A inhibition effect, can be combined with microtubules to form a complex to regulate KIF18A protein, is used for treating KIF18A-mediated diseases and / or diseases, such as tumor diseases, and is used for preparing medicines for such diseases or diseases.

Description

[0001] This invention claims the following:

[0002] Priority is claimed for the earlier application filed with the China National Intellectual Property Administration on September 30, 2022, with patent application number 202211217025.8 and title "Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions and their preparation methods and applications".

[0003] Priority is claimed for the earlier application filed with the China National Intellectual Property Administration on November 10, 2022, with patent application number 202211406766.0 and title "Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions and their preparation methods and applications".

[0004] Priority is claimed for the earlier application filed with the China National Intellectual Property Administration on February 16, 2023, with patent application number 202310125351.4 and title "Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions and their preparation methods and applications".

[0005] Priority is claimed for the earlier application filed with the China National Intellectual Property Administration on September 20, 2023, with patent application number 202311219723.6 and title "Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions and their preparation methods and applications".

[0006] The entire contents of the prior application are incorporated herein by reference. Technical Field

[0007] This invention belongs to the pharmaceutical field, specifically relating to a fused-ring KIF18A inhibitor compound, a pharmaceutical composition, its preparation method, and its application. Background Technology

[0008] Cancer is one of the most serious diseases affecting human health, with mortality and morbidity rates often ranking among the highest of all diseases. Although the quality of life for some patients has been greatly improved with the continuous development and progress of medical technology and drug research, there are still many unmet clinical needs in the search for effective treatments or cures for different cancers, and new targets will provide new possibilities for future cancer drug development.

[0009] Cancer cells exhibit unregulated cell proliferation due to damage or loss of one or more genes that regulate the cell cycle. Various kinases and kinesins have been identified as playing key roles in the regulation and progression of cell cycle and mitosis in both normally dividing cells and cancer cells.

[0010] Kinesin molecules are kinetic proteins that use intracellular microtubules as their orbital pathways; also known as molecular motors, they convert ATP energy into mechanical energy and are closely related to eukaryotic cell division, mitosis, meiosis, tissue and organ growth and development, neuronal development, and signal transduction. Kinesin members share a relatively conserved motor domain. Based on the location of the motor domain in the molecule, the kinesin family is broadly divided into three categories: N-type kinesins, where the amino (-NH2) terminal region of the polypeptide chain contains a motor domain; M-type kinesins, where the middle region contains a motor domain; and C-type kinesins, where the carboxyl (-COOH) terminal region contains a motor domain.

[0011] Chromosomal instability is a hallmark of cancer, caused by errors in chromosome segregation during mitosis. Targeting chromosomal instability is an emerging therapeutic strategy in drug development. KIF18A, a member of the N-type Kinesin-8 kinesin family, has been shown to play a role in maintaining the integrity of the bipolar spindle and promoting the survival of chromosomally unstable cancer cells. Mitosis is an effective intervention point, and many antimitotic drugs are used clinically to treat human cancers. The most widely used microtubule inhibitors both stabilize microtubules and prevent microtubule assembly. Currently, antimitotic drugs have limitations due to their narrow therapeutic window, and these issues necessitate the development of new targets to address them.

[0012] Although tubulin inhibitors are widely used as standard treatments for various types of human cancer, these drugs can cause collateral damage to normal cells, including myelosuppression and neurotoxicity. Since KIF18A may not be essential in normal diploid somatic cells (KIF18A knockout mice are viable but have reproductive defects, suggesting that KIF18A is not a necessary gene for normal somatic cell division), targeting KIF18A could potentially significantly reduce its toxicity, thus improving the therapeutic safety window of tubulin-targeting drugs in clinical practice.

[0013] KIF18A protein is highly expressed in various tumors, including colorectal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, and ovarian cancer. KIF18A plays a crucial role in the occurrence, development, and metastasis of breast cancer, and its high expression predicts poor prognosis. KIF18A is essential for the proliferation of chromosomally unstable cells derived from triple-negative breast cancer or colorectal cancer, but is not required in diploid cells. Knockout of the KIF18A gene leads to infertility in male mice, but female mice are unaffected. 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 in breast cancer, with a risk factor of 3.2. Furthermore, inhibiting KIF18A expression not only affects its key function 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 apoptosis.

[0014] Therefore, the development of KIF18A protein inhibitors may be a new breakthrough in cancer drugs. Summary of the Invention

[0015] To address the aforementioned technical problems, the present invention provides a compound of formula (I), its racemic mixture, stereoisomer, tautomer, isotope label, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound:

[0016]

[0017] Wherein, A is selected from unsubstituted or arbitrarily substituted by one, two or more Rs. a Replacement Or a fused ring group 1; said fused ring group 1 comprises two, three, or four C groups independently selected from saturated or partially unsaturated C groups. 3-14 Carbon ring, C 6-14 Aromatic rings, 5-14 quinone heterocyclic rings, and 3-14 quinone heterocyclic rings; each R a They may be identical or different, independently selected from H, OH, halogen, cyano, NH2, NO2, unsubstituted or optionally substituted by one, two or more R groups. a1 The following groups are substituted: C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl; or, two R atoms attached to the same carbon atom in a ring. a Together with the carbon atoms it is attached to, they form saturated or partially unsaturated C atoms. 3-14 Carbon rings; each R a1 They may be the same or different, and are independently selected from H, OH, halogens, cyano groups, NH2, NO2, and C.1-12 Alkyl, C 1-12 Alkoxy, C 3-12 cycloalkyl;

[0018] X1, X2, and X3 may be the same or different, and are independently selected from N or CR0; R0 is selected from H, halogen, cyano, and C. 1-12 Alkyl, Halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkoxy;

[0019] B is selected from unsubstituted or arbitrarily assigned to one, two or more R. b The following groups are substituted: C 1-12 Alkyl, Halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkoxy, C 3-12 cycloalkyl, C 3-12 Cycloalkyloxy, C 3-12 Cycloalkylthio groups, 3-14 membered heterocyclic groups; each R b They may be the same or different, and are independently selected from halogens, cyano groups, and C. 1-12 Alkyl, Halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkoxy;

[0020] Ring G is selected from unsubstituted or arbitrarily replaced by one, two or more R. g The substituted fused ring group 2, wherein the fused ring group 2 is formed by the fusion of ring G1 and ring G2; ring G1 is selected from C 6-14 Aromatic rings, 5-14 heterocyclic aromatic rings, 3-14 heterocyclic rings; ring G2 is selected from C 3-14 Carbon ring, C 6-14 Aromatic rings, 5-14 quinone heteroaromatic rings, 3-14 quinone heterocyclic rings; M and E are preferably connected to ring G1; each R g They may be the same or different, and are independently selected from halogens, cyano groups, and C. 1-12 Alkyl, Halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkoxy;

[0021] E is selected from unsubstituted or optionally by one, two or more R. e The following groups are substituted: -NH-S(=O)2-R e1 -S(=O)2-NH-R e2 -S(=O)(=NH)-R e3 -N(R) e4 (R) e5 ), 3-14 membered heterocyclic groups; each R e They may be the same or different, and are independently selected from OH, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkoxy, halogenated C 1-12 Alkyl, Halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, -N(R) e6 (R) e7 );

[0022] R e1 R e2 R e3 R e4 R e5 R e6 R e7 Whether the two are the same or different, they are selected independently from H and C. 1-12 Alkyl, C 1-12 Alkoxy, hydroxy C 1-12 Alkyl, Halogenated C 1-12 Alkyl, Halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups, C 1-12 Alkoxy-C 1-12 alkyl;

[0023] M is selected from unsubstituted or optionally by one, two or more Rs. m The following groups are substituted: C 3-12 cycloalkyl, C 3-12 Cycloalkenyl, 3-14 membered heterocyclic groups; each R m They may be the same or different, and are independently selected from halogens, cyano groups, and C. 1-12 Alkyl, Halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, cyano C 1-12 Alkyl group.

[0024] According to some implementation schemes, A is selected from unsubstituted or optionally by one, two or more R. a Replacement Or a fused ring group 1; said fused ring group 1 comprises two, three, or four C groups independently selected from saturated or partially unsaturated C groups. 3-8 Carbon ring, C 6-10 Aromatic rings, 5-10 quinary heterocyclic rings, and 3-8 quinary heterocyclic rings; each R a They may be the same or different, and are independently selected from H, OH, halogens, cyano groups, NH2, NO2, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 cycloalkyl, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, cyano C 1-6 Alkyl, cyano C 1-6 Alkoxy, C 3-8 cycloalkyl-C 1-6 alkoxy group; or, two R groups attached to the same carbon atom in a ring. a Together with the carbon atoms it is attached to, they form saturated or partially unsaturated C atoms. 3-8 Carbon rings;

[0025] According to some implementation schemes, A is selected from...

[0026] Wherein, T is selected from CH2, CH, NH, NR a Or O;

[0027] Z is selected from CH2, CH, NH, NR a Or O;

[0028] X is selected from N or CH;

[0029] n is selected from 0, 1, 2, 3, 4 or 5;

[0030] p and q are independently selected from 0, 1, 2, and 3, and p and q are not both 0 at the same time;

[0031] r and s are independently selected from 0, 1, 2, and 3, and r and s are not both 0 at the same time.

[0032] According to some implementation schemes, X1 and X2 may be the same or different, and are independently selected from N or CR0.

[0033] According to some implementation schemes, X1 and X2 may be the same or different, and are independently selected from N or CH.

[0034] According to some implementation schemes, X2 and X3 are not both N.

[0035] According to some implementation schemes, 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, halogens, and C. 1-6 alkyl;

[0036] According to some implementation schemes, 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 and C. 1-6 alkyl;

[0037] According to some implementation schemes, B is selected from C. 1-6 Alkyl, C 3-8 cycloalkyl, C 3-8 Cycloalkyloxy groups, 3-8 membered heterocyclic groups, and halogenated 3-8 membered heterocyclic groups.

[0038] According to some implementation schemes, B is selected from a halogenated 6-membered N-containing heterocyclic group; for example, a halogenated piperidinyl group.

[0039] According to some implementation schemes, B is selected from...

[0040] According to some implementation schemes, A is selected from unsubstituted or optionally by one, two or more R. a Replacement Or a fused ring group 1; the fused ring group 1 is formed by the fusion of two, three, four or more rings selected from benzene ring, pyridine ring, imidazole ring, piperazine ring, dihydropyridine ring, tetrahydropyridine ring, dihydropyrrole ring, tetrahydropyrrole ring, dihydropyran ring, cyclobutene ring, cyclopentene ring, cyclohexene ring, and cycloheptene ring.

[0041] According to some implementation schemes, A is selected from unsubstituted or optionally by one, two or more R. a The following groups are substituted:

[0042]

[0043] According to some implementation schemes, each R a The same or different, independently selected from H, OH, F, Cl, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclopropylmethoxy; or, two R atoms attached to the same carbon atom of a ring. a Together with the carbon atoms it is attached to, they form cyclopropane rings, cyclobutane rings, cyclopentane rings, cyclohexane rings, and cycloheptane rings.

[0044] According to some implementation schemes, A is selected from...

[0045] According to some implementation schemes, ring G is selected from unsubstituted or optionally replaced by one, two or more Rs. g The substituted fused ring group 2, wherein the fused ring group 2 is formed by the fusion of ring G1 and ring G2; ring G1 is selected from C 6-10 Aromatic rings, 5-10 degree heterocyclic aromatic rings, 5-10 degree heterocyclic rings; ring G2 is selected from C 3-10 Carbon ring, C 6-10 Aromatic rings, 5-10 degree heterocyclic aromatic rings, 3-10 degree heterocyclic rings;

[0046] According to some implementation schemes, 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, and thiophene ring; preferably, it is a benzene ring or a pyridine ring.

[0047] According to some implementation schemes, ring G2 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, and methylimidazolium ring.

[0048] According to some implementation schemes, ring G is selected from...

[0049] According to some implementation schemes, E is selected from unsubstituted or optionally replaced by one or two R. e The following groups are substituted: -NH-S(=O)2-R e1 -S(=O)2-NH-R e2 -S(=O)(=NH)-R e3 ;R e1 R e2 R e3 Whether the two are the same or different, they are selected independently from H and C. 1-6 Alkyl, hydroxyl C 1-6 Alkyl, Halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, cyano C 1-6 Alkyl, cyano C 1-6 Alkoxy, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 1-6 Alkoxy-C 1-6 Alkyl; each R e They may be the same or different, and are independently selected from OH, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6Alkoxy, cyano C 1-6 Alkyl, cyano C 1-6 Alkyl group.

[0050] According to some implementation schemes, E is selected from...

[0051] According to some implementation schemes, M is selected from unsubstituted or optionally replaced by one, two or more R. m Replacement C 3-8 cycloalkyl, C 3-8 Cycloalkenyl or nitrogen-containing 3-8 membered heterocyclic groups; each R m They may be the same or different, and are independently selected from H, halogen, cyano, and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, C 1-6 Alkoxy, cyano C 1-6 alkoxy group; or, two R groups attached to the same carbon atom in a ring. m Together with the carbon atoms it is attached to, they form saturated or partially unsaturated C atoms. 3-8 Carbon rings;

[0052] According to some implementation schemes, M is selected from unsubstituted or optionally replaced by one, two or more R. m Replacement

[0053] According to some embodiments, the compound shown in formula (I) is selected from the following structures:

[0054]

[0055] Among them, A, M, E, ring G, ring G1, and ring G2 independently have the definitions described in this paper. It represents a carbon-carbon single bond or a carbon-carbon double bond.

[0056] According to some embodiments, the compound represented by formula (I) has the following structure:

[0057]

[0058] Among them, B, M, E, ring G, X, Z, T, X1, X2, X3, R a n, p, q, r, and s each have their own independent definitions as described in this paper.

[0059] According to some embodiments, the compound represented by formula (I) has the following structure:

[0060]

[0061] Among them, B, ring G, X, Z, T, X1, X2, X3, Ra n, r, s, p, and q each have their own definitions as described in this paper.

[0062] According to some embodiments, the compound represented by formula (I) has the following structure:

[0063]

[0064] Among them, A, ring G1, and ring G2 have the definitions described herein independently of each other.

[0065] According to some embodiments, the compound represented by formula (I) has the following structure:

[0066]

[0067] Here, A and ring G2 are independently defined as described in this paper.

[0068] According to some embodiments, the compound represented by formula (I) has the following structure:

[0069]

[0070] Among them, B, ring G2, X, Z, T, X1, X2, X3, R a n, r, s, p, and q each have their own definitions as described in this paper.

[0071] According to some embodiments, the compound represented by formula (I) has the following structure:

[0072]

[0073] Among them, rings G2, X1, X2, and X3 have the definitions described herein independently of each other.

[0074] According to some embodiments, the compound shown in formula (I) is selected from the following structures:

[0075]

[0076]

[0077]

[0078]

[0079] The present invention also provides a method for preparing the compound shown in formula (I), comprising the following steps:

[0080]

[0081] (1) Compound a reacts with compound A-NH2 to give compound b;

[0082] (2) Compound b reacts with compound MH to give compound c;

[0083] (3) Compound c reacts with compound EH to give the compound shown in formula (I);

[0084] Among them, 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.

[0085] The present invention also provides a pharmaceutical composition comprising at least one of the following: a compound of formula (I), a racemic mixture, a stereoisomer, a tautomer, an isotope label, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof, in a therapeutically effective amount.

[0086] According to embodiments of the present invention, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients.

[0087] According to embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0088] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the following: a compound of formula (I), its racemic mixture, stereoisomer, tautomer, isotope label, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound.

[0089] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of the above-described pharmaceutical composition.

[0090] The cancers mentioned include colorectal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, and ovarian cancer.

[0091] In some implementations, the patient includes mammals, preferably humans.

[0092] The present invention also provides at least one of the following: a compound of formula (I) for treating tumor diseases, a racemic mixture, a stereoisomer, a tautomer, an isotope label, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof, or a pharmaceutical composition thereof.

[0093] The present invention also provides the use of at least one of the compounds of formula (I), racemates, stereoisomers, tautomers, isotope labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof in the preparation of pharmaceuticals.

[0094] According to embodiments of the present invention, the use may be in the preparation of medicaments for treating KIF18A-mediated conditions and / or diseases, such as in the preparation of KIF18A inhibitor medicaments.

[0095] According to embodiments of the present invention, the disease is, for example, cancer, including colorectal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, or ovarian cancer.

[0096] Beneficial effects

[0097] The compounds of the present invention have good KIF18A inhibitory activity. These compounds can regulate KIF18A protein alone or by forming a binding complex with microtubules, for the treatment of KIF18A-mediated conditions and / or diseases, such as tumor diseases, and for the preparation of medicaments for such conditions or diseases.

[0098] Through structural optimization, this invention has creatively obtained a class of novel compounds that not only have good KIF18A inhibitory activity and OVCAR-3 in vitro cell activity, but also significantly improved physicochemical properties (solubility, permeability), resulting in a significant improvement in the in vivo efficacy of OVCAR-3. Attached Figure Description

[0099] Figure 1 Line graph showing the effect of compound 9 of this invention on tumor volume and body weight in OVCAR-3 mouse xenograft tumor model mice.

[0100] Terminology Definitions and Explanations

[0101] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0102] Unless otherwise stated, the numerical ranges described in this specification and claims 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.

[0103] The term “optional” (or “optionally”, “optionally”) in the general formula definition of this application means the case of being substituted by zero or one or more substituents. For example, “optionally substituted by one, two or more R” means that it may not be substituted by R (no substitution) or may be substituted by one, two or more R.

[0104] "More than three" means three or more.

[0105] Term "C" 1-12 "alkyl" should be understood to refer to straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C 1-8 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1-6 "Alkyl" means a straight-chain or branched alkyl group 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 their isomers.

[0106] Term "C" 3-12 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) hydrocarbon rings or tricyclic alkanes, 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 refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. The C... 3-12 Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, 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]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0107] Term "C"3-12 "Cycloalkenyl" should be understood as referring to monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring), or tricyclic olefins containing carbon-carbon double bonds, having 3 to 12 carbon atoms, preferably "C". 3-10 "Cycloalkenyl", more preferably "C" 3-8 "Cycloalkenyl" can have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. The C 3-12 Cycloalkenyl groups can be monocyclic hydrocarbon groups, such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl or cyclodecenyl, or dicyclic hydrocarbon groups such as spiro[2.5]oct-5-enyl, spiro[3.5]non-6-enyl, spiro[4.5]dec-7-enyl.

[0108] Term "C" 6-14 "Aryl" should preferably be understood to represent a monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6 to 14 carbon atoms and possessing monovalent aromaticity or partial aromaticity. It can be a monoaromatic ring or a polyaromatic ring fused together, preferably "C". 6-10 Aryl. The term "C" 6-14 "Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms. 6-14 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or rings with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. When the C 6-20 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0109] The term "5-14-membered heteroaryl" should be understood to include monocyclic, bicyclic (e.g., fused, bridged, spirocyclic), 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" should also be understood to include monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, and in each case, may be benzo[a]fused. "Hyperaryl" also refers to a group in which the heteroaryl ring is fused with one or more aryl, alicyclic, or heterocyclic rings, wherein the root or point of the connection is on the heteroaryl ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indazinyl, 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-purineyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinazinyl, 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-naphthidyl, 2, 3, 5, 6, 7 or 8-quinazolinyl, 3, 4, 5, 6, 7 or 8-cenolinyl, 2, 4, 6 or 7-pteridyl, 1, 2, 3, 4, 5, 6, 7 or 8-4aH carbazole, 1, 2, 3, 4, 5, 6, 7 - or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbazolyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthridyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-pyridyl, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinel, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenanthrolinel -Phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenthiazinyl, 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-benzisoquinolinyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazin[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furano[3,2-b]-pyranolyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]-o-azinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]-azolel, 2-, 4- or 5-4H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazolo[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furano[3,4-c]cenolinyl, 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]thiopheneyl, 2-, 4-, 5-, 6- or 7-benzozolyl, 2-, 4-, 5-, 6- or 7-benzimidazinyl, 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-benzoazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzozapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thiophene, 2-, 4-, 5-, 6-, or 7-benzozozolyl, 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 compounds of the present invention, the carbon atom on the 5-14-membered heteroaryl ring may be linked to other groups, or the heteroatom on the 5-14-membered heteroaryl ring may be linked to other groups. When the 5-14-membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution sites; for example, hydrogen atoms bonded to carbon atoms on the heteroaryl ring can be substituted, or hydrogen atoms bonded to heteroatoms on the heteroaryl ring can be substituted.

[0110] Unless otherwise defined, the term "3-14 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 (such as a fused ring, bridged ring, or spirocyclic 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 optionally be oxidized to various oxidation states to form nitrides, -S(O)-, or -S(O)2- states. Preferably, the heterocyclic group may 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 containing at least one heteroatom selected from O, S, and N. The heterocyclic group can be connected to the rest of the molecule via any one of the carbon atoms or a nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. Specifically, the heterocyclic group can include, but is not limited to: 4-membered rings, such as azirrobutyl or oxobutyl; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolealkyl, or pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazineyl, or trithiaalkyl; or 7-membered rings, such as diazacycloheptyl. Optionally, the heterocyclic group can be benzofused. The heterocyclic group can be bicyclic, such as, but not limited to, a 5,5-membered ring, like a hexahydrocyclopentano[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, like a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group can be partially unsaturated, meaning it can contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrroleyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl, or 4H-[1,4]thiazinyl, or it can be benzofused, such as, but not limited to, dihydroisoquinolinyl. When the 3-14-membered heterocyclic group is linked to other groups to form the compounds of the present invention, the carbon atom on the 3-14-membered heterocyclic group can be linked to other groups, or the heterocyclic atom on the 3-14-membered heterocyclic ring can be linked to other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazine, the nitrogen atom on the piperazine group can be attached to other groups. Or when the 3-14 membered heterocyclic group is selected from piperidinium, the nitrogen atom on the piperidinium ring and the carbon atom at its para position can be attached to other groups.

[0111] The term "spirocycle" refers to a ring system in which two rings share a single ring atom.

[0112] The term "fused ring" refers to a ring system in which two rings share two cyclic atoms.

[0113] The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

[0114] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0115] "Halogenation" refers to the replacement of a substance by one or more halogens.

[0116] When ring G is selected When substituted, it has three connection sites, which can typically be connected to A, M, and E in the general formula, respectively. Preferably, with For example, connection position 1 is connected to A, connection position 2 is connected to M, and connection position 3 is connected to E. The same explanation applies when rings G and G1 are selected with other substituents.

[0117] Those skilled in the art will understand that when A is selected from unsubstituted or optionally substituted by one, two or more R... a Replacement At that time, R a It can be located anywhere on the ring containing X1. For example, when X1 is CH, R a It can replace H on X1 to form CR a .

[0118] Those skilled in the art will understand that the compounds shown in formula (I) can exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can also form internal salts.

[0119] The compounds of the present invention may exist as solvates (such as hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, may be stoichiometric or non-stoichiometric.

[0120] Depending on their molecular structure, the compounds of the present invention can be chiral, and therefore may exist in various enantiomeric forms. Thus, these compounds can exist in racemic or optically active forms. The compounds of the present invention encompass isomers of each chiral carbon in the R or S configuration, or mixtures thereof, and racemates. The compounds of the present invention or their intermediates can be isolated as enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are obtained from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid in both R and S forms, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomeric separation can also be advantageously performed using optically active resolving agents (e.g., dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized isobutylene ester polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0121] The corresponding stable isomers can be separated using known methods, such as extraction, filtration, or column chromatography.

[0122] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0123] The term “therapeutic effective amount” refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in individuals susceptible to disease, disorder, or symptom but not yet experiencing or exhibiting the pathology or symptoms of the disease; (2) suppression of disease: e.g., suppression of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., prevention of further development of the pathology and / or symptoms); (3) relief of disease: e.g., relief of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., reversal of the pathology and / or symptoms). Detailed Implementation

[0124] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0125] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0126] Example 1

[0127] 5-{6-azaspiro[2.5]octane-6-yl}-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-inden-4-carboxamide (Compound 1)

[0128]

[0129] The first step is the synthesis of 2-bromo-4-fluoro-5-methoxybenzaldehyde (compounds 1-2):

[0130] At room temperature, potassium bromide (38.60 g, 324.380 mmol, 5 eq) was added to a 20 mL aqueous solution of 10 g (64.876 mmol, 1 eq) of 4-fluoro-3-methoxybenzaldehyde, followed by the addition of elemental bromine (9.98 mL, 194.628 mmol, 3 eq). The mixture was stirred at room temperature for 15 hours. The precipitated solid was collected by filtration and washed with water (3 × 5 mL). The solid was dried under vacuum to give 15 g (99.22%) of 2-bromo-4-fluoro-5-methoxybenzaldehyde.

[0131] 1 H NMR (400MHz, DMSO-d6) δ10.13(d,J=0.8Hz,1H),7.83(dd,J=10.4,1.6Hz,1H),7.57-7.54(dd,J=10.4,1.6Hz,1H),3.92(s,3H).

[0132] The second step is the synthesis of 3-(2-bromo-4-fluoro-5-methoxyphenyl)propionic acid (compounds 1-4):

[0133] Under nitrogen protection, at room temperature, McMurray acid (compounds 1-3) (4947.77 mg, 34.329 mmol, 1 eq) was added to a solution of 2-bromo-4-fluoro-5-methoxybenzaldehyde (8 g, 34.329 mmol, 1 eq) in formic acid (10 mL), followed by the dropwise addition of triethylamine (14.32 mL, 102.987 mmol, 3 eq). The reaction mixture was heated to 100 °C and stirred for 10 hours. After the reaction was complete, the mixture was quenched with water at room temperature, extracted with ethyl acetate (3 × 200 mL), and the combined organic phases were washed with saturated sodium chloride solution (1 × 300 mL) and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated 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)propionic acid (2.8 g, 29.44%).

[0134] MS(ESI, m / z): 275.05 [MH] - ,RT(min):0.886.

[0135] Step 3: Synthesis of 4-bromo-6-fluoro-7-methoxy-2,3-dihydroindanone (compounds 1-5):

[0136] 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)propionic acid (2.0 g, 7.218 mmol, 1 eq) in 1,2-dichloroethane (12 mL) at room temperature. The reaction mixture 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 in petroleum ether / ethyl acetate (10:1) to give 4-bromo-6-fluoro-7-methoxy-2,3-dihydroindene-1-one (1.5 g, 80.22%).

[0137] MS(ESI, m / z): 259.15 [M+H] + ,RT(min):1.030.

[0138] 1 H NMR (400MHz, CDCl3-d) δ7.51 (d, J = 11.2Hz, 1H), 4.09 (s, 3H), 2.99-2.96 (m, 2H), 2.77-2.74 (m, 2H).

[0139] Step 4: Synthesis of 7-bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (compounds 1-6):

[0140] 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-dihydro-indene-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 chromatography. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3 × 60 mL). The combined organic phases were 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 in petroleum ether / ethyl acetate (10:1) to give 7-bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (1.5 g, 79.28%).

[0141] 1 ¹H NMR (400MHz, 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).

[0142] Step 5: Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-ol (compounds 1-7):

[0143] 7-Bromo-5-fluoro-4-methoxy-2,3-dihydro-1H-indene (800 mg, 3.264 mmol, 1 eq) and an acetic acid solution of hydrogen bromide (8.00 mL) were heated and stirred at 100 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (50 mL). The reaction mixture was washed successively with distilled water (1 × 30 mL), saturated sodium bicarbonate (1 × 30 mL), and saturated brine (1 × 30 mL). The organic phase was dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product / without further purification was directly added to the next step.

[0144] MS(ESI, m / z): 231.05 [MH] - RT(min):1.041

[0145] Step 6: Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yltrifluoromethanesulfonate (compounds 1-9):

[0146] 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 stirring was continued for 10 minutes. Then, trifluoromethanesulfonic anhydride (compounds 1-8) (3.29 mL, 19.474 mmol, 1.50 eq) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3 × 80 mL). The combined organic phases were backwashed with saturated brine (1 × 100 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 in petroleum ether / ethyl acetate (10:1) to give 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yltrifluoromethanesulfonate (2 g, 42.42%).

[0147] 1 ¹H NMR (400MHz, deuterated chloroform) δ 7.23 (d, J = 9.2Hz, 1H), 3.18–3.14 (m, 2H), 2.99–2.95 (m, 2H), 2.23–2.16 (m, 2H).

[0148] Step 7: Synthesis of methyl 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylate (compounds 1-10):

[0149] Under nitrogen protection, palladium acetate (74.19 mg, 0.331 mmol, 0.1 eq), bis(diphenylphosphinebutane) (140.94 mg, 0.331 mmol, 0.1 eq), and triethylamine (1148.39 μL, 8.263 mmol, 2.5 eq) were added sequentially to a solution of 7-bromo-5-fluoro-2,3-dihydro-1H-inden-4-yltrifluoromethanesulfonate (1.2 g, 3.305 mmol, 1 eq) in dimethyl sulfoxide / methanol (7.5 mL, 3 / 2, v / v) at room temperature. The reaction mixture was heated to 80 °C and carbon monoxide was bubbled through it for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 10 mL). The combined organic phases were 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 in petroleum ether / ethyl acetate (10:1) to give methyl 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylate (400 mg, 44.32%).

[0150] 1¹H NMR (400MHz, 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).

[0151] Step 8: Synthesis of 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylic acid (compounds 1-11):

[0152] Lithium hydroxide (131.54 mg, 5.493 mmol, 3 eq) was added to a tetrahydrofuran / water (4 mL, 3 / 1, v / v) solution of methyl 7-bromo-5-fluoro-2,3-dihydro-1H-indene-4-carboxylate (500 mg, 1.831 mmol, 1 eq) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water at room temperature and acidified to pH 5–6 with 1 mol / L hydrochloric acid solution. The reaction mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic phases were 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 added to the next step.

[0153] MS(ESI, m / z): 257.10 [MH] - RT(min):1.053

[0154] 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 (compounds 1-13):

[0155] Under nitrogen protection, tetramethylchloromethane hexafluorophosphate (2166.02 mg, 7.720 mmol, 4 eq) and N-methylimidazolium (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 (compounds 1-12) (660.76 mg, 2.895 mmol, 1.5 eq) (660.76 mg, 2.895 mmol, 1.5 eq) at room temperature. The mixture was stirred 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 residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (10:1) to give 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%).

[0156] MS(ESI, m / z): 469.25 [M+H] + ,RT(min):1.455.

[0157] Step 10: Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-2,3-dihydro-1H-indene-4-carboxamide (compounds 1-15):

[0158] 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 (compounds 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 reacted 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 combined organic phases were 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 residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (10:1) to give 5-{6-azaspiro[2.5]octane-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%).

[0159] MS(ESI,m / z):560.60[M+H]+,RT(min):1.494.

[0160] 1 ¹H NMR (400MHz, deuterated chloroform) δ 12.69 (s, 1H), 7.46 (s, 1H), 7.27 (s, 1H), 3.99 (d, J = 7.2Hz, 4H), 3.47 (t, J = 7.6Hz, 2H), 3.03 (t, J = 5.4Hz, 4H), 2.93 (t, J = 7.6Hz, 2H), 2.37 (s, 3H), 2.10 (q, J = 7.6Hz, 2H), 1.99 (s, 4H), 1.55 (s, 4H), 0.38 (s, 4H).

[0161] Step 11: Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (compound 1):

[0162] Under nitrogen protection, at room temperature, dicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine (Ephos, 8.59 mg, 0.016 mmol, 0.2 eq) was added to a solution of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-2,3-dihydro-1H-indene-4-carboxamide (45 mg, 0.080 mmol, 1 eq) in 1,4-dioxane (2 mL). 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 mixture was heated to 100 °C and heated for 1 hour. The reaction mixture 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 combined organic phases were 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 high-performance liquid chromatography (HPLC) under the following conditions (column specification: XBridge). Prep OBD C18 Column, 30*150mm, 5μm; Mobile phase A: water (10mmol / L ammonium bicarbonate), Mobile phase B: acetonitrile; Flow rate: 60mL / min; Elution gradient: 48% B to 80% B, 80% B over 8 min; Detection wavelength: UV 220nm; Retention time (min): 7.88. Compound 1 was obtained (21.93mg, 44.81%).

[0163] MS(ES,m / z):605.45[M+H] + ,RT(min):1.894.

[0164] 1 H NMR: (400MHz, DMSO-d6) δ12.92(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=6.5Hz,2H),3 .31(s,2H),3.22(t,J=7.5Hz,2H),2.94(s,4H),2.85(t,J=7.5Hz,2H),2.31(s,3H),2.05-1.86(m,6H),1.65(s,4H),0.36(s,4H).

[0165] Example 2

[0166] 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-inden-4-carboxamide (Compound 9)

[0167]

[0168] 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-inden-4-carboxamide (compound 9-2):

[0169] Under nitrogen protection, at room temperature, N-methylimidazolium (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'-tetramethylchloromethanesulfonamide 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). 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 combined organic phases were 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 in petroleum ether / ethyl acetate (5:1) to give 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%).

[0170] MS:(ESI,m / z):468.30[M+H] + ,RT(min):1.397.

[0171] The second step is the synthesis of 5-{6-azaspiro[2.5]octane-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):

[0172] 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 combined organic phases were 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 residue was purified by preparative chromatography using petroleum ether / ethyl acetate (1:1) to give 5-{6-azaspiro[2.5]octane-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%).

[0173] MS:(ESI,m / z):559.45[M+H] + ,RT(min):1.590.

[0174] Step 3: Synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (compound 9):

[0175] Under nitrogen protection, at room temperature, 1 mmol of 5-{6-azaspiro[2.5]octane-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) were applied. Add dicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine (8.6 mg, 0.016 mmol, 0.2 eq) and (methanesulfonic acid {dicyclohexyl(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 mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture 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 high performance liquid chromatography to give 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (8.8 mg, 17.98%).

[0176] MS:(ESI,m / z):604.40[M+H] + ,RT(min):1.861.

[0177] 1 H NMR: (400MHz, DMSO-d6) δ12.16(s,1H),9.31(s,1H),7.46(s,1H),7.22(s,1 H),6.53(s,1H),4.96(s,1H),3.77(t,J=6.5Hz,2H),3.71(t,J=5.8Hz,4H),3 .33(s,1H),3.29(s,1H),3.21(t,J=7.5Hz,2H),2.94(t,J=5.3Hz,4H),2.85 (t,J=7.5Hz,2H),2.26(s,3H),2.05-1.88(m,6H),1.63(s,4H),0.34(s,4H).

[0178] Example 3

[0179] 5-(6-azaspiro[2.5]octane-6-yl)-N-[6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-inden-4-carboxamide (Compound 8)

[0180]

[0181] The first step was the synthesis of 5-(6-azaspiro[2.5]octane-6-yl)-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (compound 8-1):

[0182] 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 1 mL of N-methylpyrrolidone 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 mixture 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 in petroleum ether / ethyl acetate (3:1) to give 5-(6-azaspiro[2.5]octane-6-yl)-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (30 mg, 53.11%).

[0183] MS:(ESI,m / z):377.00[M+H] + RT(min): 0.827

[0184] The second step is the 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):

[0185] Under nitrogen protection, at room temperature, 6-(4,4-difluoropiperidin-1-yl)pyridine-2-amine (9.13 mg, 0.044 mmol, 1.5 eq), tetramethylchlorourea hexafluorophosphate (64.09 mg, 0.228 mmol, 4 eq), and N-methylimidazolium (46.88 mg, 0.570 mmol, 10 eq) were added to a solution of 5-(6-azaspiro[2.5]octane-6-yl)-7-bromo-2,3-dihydro-1H-indene-4-carboxylic acid (20 mg, 0.057 mmol, 1 eq) in dichloromethane (1 mL). The reaction mixture was heated to 80 °C and reacted 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 residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (3:1) to give 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 (30 mg, 96.32%).

[0186] MS:(ESI,m / z):365.90[M+H] + RT(min):1.140

[0187] The third step is the synthesis of 5-(6-azaspiro[2.5]octane-6-yl)-N-[6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (compound 8):

[0188] Under nitrogen protection, at room temperature, a solution 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 (25 mg, 0.046 mmol, 1 eq) in 1,4-dioxane (1 mL, 11.804 mmol, 257.55 eq) was prepared with dicyclohexyl(3-isopropoxy-2′,4′,6′-triisopropyl-[1,1′-biphenyl]-2-yl)phosphonane (4.90 mg, 0.009 mmol, 0.2 eq), (methanesulfonic acid {dicyclohexyl(3-isopropoxy-2′,4′,6′-triisopropyl-[1,1′- The reaction mixture consisted of: [biphenyl-2-yl]phosphonane}(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 mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3 × 10 mL). The combined organic phases were 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 high-performance liquid chromatography (HPLC) under the following conditions (Kinete 5 μm column). EVO C18, 30mm*150mm, mobile phase A: water (10mmol / L ammonium bicarbonate), mobile phase B: acetonitrile, flow rate: 60mL / min, elution gradient: 40% B to 75% B in 8min; 254nm; Rt: 6.9min), yielded 5-(6-azaspiro[2.5]octane-6-yl)-N-[6-(4,4-difluoropiperidin-1-yl)pyridin-2-yl]-7-(2-hydroxyethanesulfonamide)-2,3-dihydro-1H-indene-4-carboxamide (2.79mg, 10.29%).

[0189] MS:(ESI,m / z):590.05[M+H] + RT(min):1.874

[0190] 1H-NMR: (400MHz, DMSO-d6) δ12.30(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=6.5Hz,2H),3 .72(t,J=5.7Hz,4H),3.30(s,2H),3.22(t,J=7.5Hz,2H),3.03–2.89(m, 4H), 2.83 (t, J = 7.5Hz, 2H), 2.09–1.90 (m, 6H), 1.64 (s, 4H), 0.35 (s, 4H).

[0191] Example 4

[0192] 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethanesulfonamide)-2,3-dihydro-1H-indene-4-carboxamide (Compound 13)

[0193]

[0194] The first step was the synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-N-[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (compound 13-2):

[0195] Under nitrogen protection, at room temperature, N,N,N,N-tetramethylchloromethane hexafluorophosphate (80.11 mg, 0.284 mmol, 4 eq) and N-methylimidazolium (58.60 mg, 0.710 mmol, 10 eq) were added to a solution of 5-{6-azaspiro[2.5]octane-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). 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. The filtrate was filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (3:1) to give 5-{6-azaspiro[2.5]octane-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%).

[0196] MS:(ESI,m / z):548.05[M+H] + RT(min):1.255

[0197] The second step is the synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (compound 13):

[0198] Under nitrogen protection, at room temperature, dicyclohexyl (3-isopropoxy-2′,4′,6′-triisopropyl-[1,1′)) was added to a 1,4-dioxane solution (5 mL) 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) containing 1,4-dioxane. [-biphenyl]-2-yl)phosphonane (2.45 mg, 0.005 mmol, 0.1 eq) and methanesulfonic acid {dicyclohexyl[3-isopropoxy-2',4',6'-triisopropyl-(1,1'-biphenyl)-2-yl]phosphonane}(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) were added. The reaction mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture 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. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (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: 254 nm / 220 nm; retention time (min): 7.55, yielding 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)pyrazin-2-yl]-7-(2-hydroxyethanesulfonamide)-2,3-dihydro-1H-indene-4-carboxamide (2.23 mg, 8.17%).

[0199] MS:(ESI,m / z):591.20[M+H] + RT(min): 1.754

[0200] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),8.76(s,1H),8.16(s,1H),7.24(s,1H),5.00(s,1H),3.77(d,J=6.5Hz,6H) ,3.24(m,4H),2.95(d,J=5.3Hz,4H),2.80(t,J=7.6Hz,2H),2.05(m,4H),1.95(m,2H),1.64(s,4H),0.36(s,4H).

[0201] Example 5

[0202] 5-(6-azaspiro[2.5]octane-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)

[0203]

[0204] The first step was the synthesis of 5-(6-azaspiro[2.5]octane-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):

[0205] Under nitrogen protection, at room temperature, 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]octane-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-fluoropyridine-4-amine (18.15 mg, 0.078 mmol, 1.1 eq) in dichloromethane (2 mL). The mixture was heated to 60 °C and stirred for 1 h. The reaction mixture was diluted with water (10 mL). 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 silica gel column chromatography in petroleum ether / ethyl acetate (10:1) to give 5-(6-azaspiro[2.5]octane-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%).

[0206] MS:(ESI,m / z):563.15[M+H]+ RT(min):1.184

[0207] The second step was the synthesis of 5-(6-azaspiro[2.5]octane-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):

[0208] Under nitrogen protection at room temperature, cesium carbonate (52.04 mg, 0.159 mmol, 3 eq) and bicyclohexyl(3-isopropoxy-2′,4′,6′-triisopropyl-[1,1′-biphenyl]-2-yl)phosphonane (5.69 mg, 0.011 mmol) were added to a 1,4-dioxane solution (2 mL) of 5-(6-azaspiro[2.5]octane-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) at 1,4-dioxane (2 mL). 0.2 mmol (ol, 0.2 eq) and (methanesulfonic acid {bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphonane}(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 the 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. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions: column specification: XBridge BEH ShieldRP18 5μm, 30mm*150mm; Mobile phase A: water (10mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL / min; elution gradient: 50% B to 75% B in 8min; 254nm; Rt: 6.9min. 5-(6-azaspiro[2.5]octane-6-yl)-N-[2-(4,4-difluoropiperidin-1-yl)-3-fluoropyridin-4-yl]-7-(2-hydroxyethylsulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (17.1mg, 52.75%) was obtained.

[0209] MS:(ESI,m / z):608.45[M+H] + RT(min): 1.736

[0210] 1H NMR(400MHz,DMSO-d6)δ11.50(s,1H),9.33(s,1H),8.00–7.85(m,2H),7.1 8(s,1H),4.99(s,1H),3.78(t,J=6.5Hz,2H),3.54(t,J=5.7Hz,4H),3.30( d,J=6.4Hz,2H),3.10(t,J=7.5Hz,2H),2.94(t,J=5.2Hz,4H),2.85(t,J=7 .5Hz,2H),2.14–2.01(m,4H),2.01–1.91(m,2H),1.46(s,4H),0.32(s,4H).

[0211] Example 6

[0212] 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (Compound 28)

[0213]

[0214] The first step was the synthesis of 5-{6-azaspiro[2.5]octane-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):

[0215] Under nitrogen protection, 6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridine-2-amine (23.77 mg, 0.103 mmol, 1.2 eq), tetramethylchloromethane hexafluorophosphate (96.13 mg, 0.344 mmol, 4 eq), and N-methylimidazolium (70.33 mg, 0.860 mmol, 10 eq) were added to a solution of 5-{6-azaspiro[2.5]octane-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 h. 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 in petroleum ether / ethyl acetate (8:1) to give 5-{6-azaspiro[2.5]octane-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%).

[0216] MS:(ESI,m / z):562.65[M+H] + RT(min):1.806

[0217] The second step was the synthesis of 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (compound 28):

[0218] Under nitrogen protection, at room temperature, add (8.15 mg, 0.009 mmol, 0.1 eq) of methanesulfonic acid {bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphonane}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (8.15 mg, 0.009 mmol, 0.1 eq) and (3-isopropoxy-2'-[6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-2,3-dihydro-1H-indene-4-carboxamide (50 mg, 0.089 mmol, 1 eq) to a solution of 1,4-dioxane (2 mL) 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) and (2 mL) of bicyclohexyl(3-isopropoxy-2'-[6,4-difluoropiperidin-1-yl)palladium(II) (8.15 mg, 0.009 mmol, 0.1 eq)) to a solution of 1,4-dioxane (2 mL) 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) at 4′,6′-triisopropyl-[1,1′-biphenyl]-2-yl)phosphonane (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 mixture 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 high-performance liquid chromatography (HPLC) under the following conditions: column specification: YMC. Triart C18EXRs 5μm, 30mm*150mm; Mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL / min; elution gradient: 60% B to 85% B in 8min; detection wavelength: 220nm; retention time (min): 7.8. 5-{6-azaspiro[2.5]octane-6-yl}-N-[6-(4,4-difluoropiperidin-1-yl)-5-fluoropyridin-2-yl]-7-(2-hydroxyethanesulfonamido)-2,3-dihydro-1H-indene-4-carboxamide (10.08mg, 18.69%) was obtained.

[0219] MS:(ESI,m / z):608.45[M+H] + RT(min):1.863

[0220] 1 H NMR: (400MHz, DMSO-d6) δ12.43(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).

[0221] Example 7

[0222] 7-{6-azaspiro[2.5]octane-6-yl}-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethanesulfonamide)imidazo[1,2-a]pyridine-8-carboxamide (compound 29)

[0223]

[0224] The first step was the synthesis of methyl 5,7-dichloroimidozop[1,2-a]pyridine-8-carboxylate (compound 29-3):

[0225] Under nitrogen protection, a solution of 2-bromo-1,1-dimethoxyethane (3.2 g, 19.001 mmol, 2.1 eq) in 20 mL of hydrobromic acid was stirred for 30 minutes at room temperature. Ethanol (15 mL) was then added, followed by 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). The reaction mixture was heated to 60 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature, quenched with water, and extracted with ethyl acetate (3 × 80 mL). The combined organic phases were 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 in petroleum ether / ethyl acetate (3:1) to give methyl 5,7-dichloroimidazolo[1,2-a]pyridine-8-carboxylate (1.5 g, 67.65%).

[0226] MS:(ESI,m / z):244.08[M+H] + RT(min): 0.886

[0227] The second step is the synthesis of 5,7-dichloroimidazolo[1,2-a]pyridine-8-carboxylic acid (compound 29-4):

[0228] 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).

[0229] MS:(ESI,m / z):230.90[M+H] + RT(min): 0.498

[0230] 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):

[0231] Under nitrogen protection, at room temperature, 6-(4,4-dichloroimidizolo[1,2-a]pyridine-8-carboxylic acid (800 mg, 3.463 mmol, 1 eq) in dichloromethane (4 mL) was added to a solution of 5,7-dichloroimidizolo[1,2-a]pyridine-8-carboxylic acid (800 mg, 3.463 mmol, 1 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). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phases were 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 residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (5:1) to give 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%).

[0232] MS:(ESI,m / z):441.25[M+H] + RT(min):1.198

[0233] Step 4: Synthesis of ethyl acetate 2-[(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl)aminosulfonyl] (compound 29-7):

[0234] Under nitrogen protection, at room temperature, ethyl 2-aminosulfonyl acetate (170.49 mg, 1.020 mmol, 1.5 eq), (methanesulfonic acid {bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphonane}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (62.45 mg, 0.068 mmol, 0.1 eq), and (bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphonane) (2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (62.45 mg, 0.068 mmol, 0.1 eq) 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) and (bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphonane) at room temperature. 2-(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl)aminosulfonyl]ethyl acetate (150 mg, 38.57%). The reaction mixture was heated to 100 °C and reacted for 1 hour. The reaction mixture was cooled to room temperature and 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. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (5:1) to give ethyl acetate 2-[(7-chloro-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl)aminosulfonyl] (150 mg, 38.57%).

[0235] MS:(ESI,m / z):572.45[M+H] + RT(min):1.094

[0236] Step 5: Synthesis of 2-[(7-{6-azaspiro[2.5]octane-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl]aminosulfonyl]ethyl acetate (compound 29-8):

[0237] Under nitrogen protection, at room temperature, 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 acetate (120 mg, 0.210 mmol, 1 eq) in tert-butanol (1 mL). 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 in petroleum ether / ethyl acetate (1:1) to give ethyl acetate 2-[(7-{6-azaspiro[2.5]octane-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl]aminosulfonyl] (60 mg, 44.22%).

[0238] MS:(ESI,m / z):647.45[M+H] + RT(min):1.240

[0239] Step 6: Synthesis of 7-{6-azaspiro[2.5]octane-6-yl}-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethanesulfonamide)imidazo[1,2-a]pyridine-8-carboxamide (compound 29):

[0240] Under nitrogen protection, lithium aluminum hydride (5.28 mg, 0.140 mmol, 1.5 eq) was added to a solution of ethyl 2-[(7-{6-azaspiro[2.5]octane-6-yl}-8-{[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]carbamoyl}imidazo[1,2-a]pyridin-5-yl]aminosulfonyl]ethyl acetate (60 mg, 0.093 mmol, 1 eq) in tetrahydrofuran (3.00 mL). The mixture was stirred at room temperature for 30 minutes. The reaction solution was quenched with sodium sulfate decahydrate at 0 °C. The mixture was filtered and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (HPLC) under the following conditions (Column specifications: XBridge BEH Shield RP18). 5 μm, 30 mm * 150 mm; Mobile phase A: water (10 mmol / L sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; elution gradient: 40% B to 60% B in 10 min; detection wavelength: 254 nm / 220 nm; retention time (min): 8.33). 7-{6-azaspiro[2.5]octane-6-yl}-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-5-(2-hydroxyethanesulfonamide)imidazo[1,2-a]pyridine-8-carboxamide (1.8 mg, 3.21%) was obtained.

[0241] MS:(ES,m / z):605.45[M+H] + ,RT(min):1.559.

[0242] 1 H NMR: (400MHz, DMSO-d6) δ12.31(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).

[0243] Example 8

[0244] N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-(2-hydroxyethanesulfonamido)-7-{6-azaspiro[2.5]octane-6-yl}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (compound 30)

[0245]

[0246] The first step is the synthesis of methyl 4,6-dichloro-2-[[(E)-(hydroxyamino)methylene]amino]pyridine-3-carboxylate (compound 30-2):

[0247] Under nitrogen protection, at room temperature, N,N-dimethylformamide dimethyl acetal (2.18 mL, 16.286 mmol, 1.20 eq) was added to a solution of methyl 2-amino-4,6-dichloropyridine-3-carboxylate (3 g, 13.572 mmol, 1 eq) in isopropanol (60.00 mL). The mixture was heated to 70 °C and stirred for 3 hours. After cooling to room temperature, hydroxylamine hydrochloride (1.13 g, 16.286 mmol, 1.2 eq) was added to the reaction solution, and the mixture was stirred at room temperature for 3 hours. The mixture was filtered, and the filter cake was collected and washed with isopropanol (3 × 50 mL). The product methyl 4,6-dichloro-2-[[(E)-(hydroxyamino)methylene]amino]pyridine-3-carboxylate (1.5 g, 41.85%) was obtained.

[0248] MS:(ESI,m / z):264.10[M+H] + ,RT(miN):0.949.

[0249] The second step is the synthesis of methyl 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (compound 30-3):

[0250] Under nitrogen protection, a solution of trifluoroacetic anhydride (1.79 g, 8.521 mmol, 1.5 eq) in acetonitrile (10 mL) was added dropwise 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 (5 mL) at 0 °C. The mixture was heated to room temperature and stirred for 4 hours. The reaction solution was concentrated under reduced pressure. The solution was neutralized to pH 7 with saturated sodium bicarbonate solution. The solution was extracted with dichloromethane (3 × 20 mL). The combined organic phases were 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 in petroleum ether / ethyl acetate (1:2) to give methyl 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (1.1 g, 77.13%).

[0251] MS:(ESI,m / z):246.20[M+H] + ,RT(min):0.828.

[0252] The third step is the synthesis of methyl 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (compound 30-5):

[0253] Under nitrogen protection, triethylamine (740.29 mg, 7.317 mmol, 3 eq) was added to a solution of methyl 5,7-dichloro-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (600 mg, 2.439 mmol, 1 eq) in tert-butanol (6 mL) at room temperature. After stirring for 2 minutes, 3,4-dimethoxybenzylamine (16.31 mg, 0.097 mmol, 1.2 eq) was added dropwise at room temperature. The mixture was heated to 60 °C and stirred for 2 hours. The reaction solution was cooled to room temperature. It was diluted with water (20 mL) 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 in petroleum ether / ethyl acetate (1:3) to give methyl 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (700 mg, 68.56%).

[0254] MS:(ESI,m / z):377.35[M+H] + ,RT(min):1.062.

[0255] 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):

[0256] Under nitrogen protection at room temperature, lithium hydroxide (114.41 mg, 4.776 mmol, 3 eq) was added in portions to a solution of methyl 7-chloro-5-{[(2,4-dimethoxyphenyl)methyl]amino}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (600 mg, 1.592 mmol, 1 eq) in methanol (2 mL), tetrahydrofuran (2 mL), and water (2 mL). The mixture was heated to 60 °C and stirred for 1 hour. The reaction mixture was acidified to pH 4 with 1 N hydrochloric acid solution. A white solid precipitated; the solid was filtered, and the filter cake was collected to give 1 g of crude product.

[0257] MS:(ESI,m / z):363.60[M+H] + ,RT(min):1.000.

[0258] 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):

[0259] Under nitrogen protection at room temperature, N,N,N',N'-tetramethylchloroformamide hexafluorophosphate (1.5 g, 5.512 mmol, 4 eq) and N-methylimidazolium (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-methylpyridine-2-amine (375.87 mg, 1.654 mmol, 1.2 eq) in 10 mL of N,N-dimethylformamide (10 mL). The mixture was heated to 60 °C and 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 residue was purified by silica gel column chromatography in petroleum ether / ethyl acetate (3:1) to give 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%).

[0260] MS:(ESI,m / z):572.50[M+H] + ,RT(min):1.370.

[0261] Step 6: Synthesis of 7-{6-azaspiro[2.5]octane-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):

[0262] Under nitrogen protection at room temperature, 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). The mixture was heated to 120 °C and stirred for 2 hours. The reaction mixture was cooled to room temperature and diluted with water (10 mL). It was 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 silica gel column chromatography in dichloromethane / methanol (10:1) to give compound 30-8 (445 mg, 62.32%).

[0263] MS:(ESI,m / z):647.60[M+H] + ,RT(min):1.172.

[0264] Step 7: Synthesis of 5-amino-7-{6-azaspiro[2.5]octane-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):

[0265] Under nitrogen protection, the reaction mixture of compound 30-8 (410 mg, 0.634 mmol, 1 eq) in trifluoroacetic acid (5 mL, 67.315 mmol, 106.18 eq) was stirred for 1 hour at 60 °C. The reaction mixture was cooled to room temperature, and the resulting residue was concentrated under reduced pressure. The solution was alkalized to pH 8 with saturated sodium bicarbonate solution. Extraction was performed with ethyl acetate (3 × 20 mL). The combined organic phases were 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 reversed-phase column chromatography under the following conditions: column specification (C18 column), mobile phase: water and acetonitrile, gradient from 0% to 80% for 10 min, UV 254 nm detector. Compound 30-9 (310 mg, 88.63%) was obtained.

[0266] MS:(ESI,m / z):497.30[M+H] + ,RT(min):1.410.

[0267] Step 8: Synthesis of 2-[(7-{6-azaspiro[2.5]octane-6-yl}-8-{[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]carbamoyl}-[1,2,4]triazolo[1,5-a]pyridin-5-yl)aminosulfonyl]ethyl acetate (compound 30-11):

[0268] Under nitrogen protection, at 0°C, 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). The mixture was heated to room temperature and stirred for 2 hours. The solution was 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 reversed-phase column chromatography under the following conditions: C18 column, mobile phase: water and acetonitrile, gradient from 10% to 70% for 10 min, UV 254 nm detector. Compound 30-11 (80 mg, 61.43%) was obtained.

[0269] MS:(ESI,m / z):647.30[M+H] + ,RT(min):1.100.

[0270] Step 9: Synthesis of N-[6-(4,4-difluoropiperidin-1-yl)-4-methylpyridin-2-yl]-5-(2-hydroxyethanesulfonamido)-7-{6-azaspiro[2.5]octane-6-yl}-[1,2,4]triazolo[1,5-a]pyridine-8-carboxamide (compound 30):

[0271] Under nitrogen protection, at 0°C, a tetrahydrofuran solution of lithium aluminum hydride (6.16 mg, 0.162 mmol, 1.5 eq) was added dropwise to a tetrahydrofuran solution of compound 30-11 (70 mg, 0.108 mmol, 1 eq) (3 mL). After the addition was complete, the system was stirred at 0°C for 1 hour. 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. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high-performance liquid chromatography (HPLC) under the following conditions: column specifications: SuNfire C18 5μm, 30mm*150mm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 60mL / min; elution gradient: 35% B to 68% B in 8min; 254nm / 220nm; Rt: 7.42min. Compound 30 (23.73mg, 35.42%) was obtained.

[0272] MS:(ESI,m / z):605.20[M+H] + ,RT(min):1.720.

[0273] 1 H NMR (400MHz, DMSO-d6) δ14.04(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. 6Hz,2H),3.72(t,J=6.1Hz,4H),3.21(t,J=6.7Hz,2H),3.10(s,4H),2.27(s,3H),1.97(t,J=15.6Hz,4H),1.72(s,4H),0.39(s,4H).

[0274] Example 9

[0275] 5-{6-azaspiro[2.5]octane-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)

[0276]

[0277] The first step is the synthesis of methyl 4-bromo-2,6-difluoro-3-nitrobenzene (compound 31-2):

[0278] 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. The reaction mixture 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 to obtain crude methyl 4-bromo-2,6-difluoro-3-nitrobenzene (6.01 g).

[0279] 1 H NMR: (400MHz, DMSO-d6) δ8.11 (dd, J=9.5, 2.1Hz, 1H), 3.93 (s, 3H).

[0280] The second step involves the synthesis of methyl 4-bromo-6-fluoro-2-methylamino-3-nitrobenzene (compound 31-3):

[0281] Under nitrogen protection, methylamine (472 mg, 15 mmol, 1.5 eq) was added to a methanol solution of methyl 4-bromo-2,6-difluoro-3-nitrobenzene (3 g, 10 mmol, 1 eq) in 30 mL at room temperature. The reaction mixture was stirred at 60 °C for 16 hours. The reaction mixture 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). The mixture was 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 give methyl 4-bromo-6-fluoro-2-methylamino-3-nitrobenzene (2.21 g, 71%).

[0282] MS:(ESI,m / z):306.85[M+H] + RT(min):1.180

[0283] The third step was the synthesis of methyl 6-{6-azaspiro[2.5]octane-6-yl}-4-bromo-2-methylamino-3-nitrobenzene (compound 31-4):

[0284] Under nitrogen protection, methyl 4-bromo-6-fluoro-2-methylamino-3-nitrobenzene (500 mg, 1.6 mmol, 1 eq) was added to a tert-butanol (15 mL) solution in which 6-azaspiro[2.5]octane hydrochloride (217 mg, 2 mmol, 1.2 eq) and triethylamine (824 mg, 8 mmol, 5 eq) were added. The reaction mixture was heated to 100 °C and stirred for 16 hours. The reaction mixture 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). The mixture was 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 give methyl 6-{6-azaspiro[2.5]octane-6-yl}-4-bromo-2-methylamino-3-nitrobenzene (460 mg, 71%).

[0285] MS:(ESI,m / z):398.35[M+H] + RT(min):1.380

[0286] Step 4: Synthesis of methyl 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylate (compound 31-6):

[0287] Under nitrogen protection, at room temperature, 3 mL of an ethanol solution (440 mg, 1.1 mmol, 1 eq) of methyl 6-{6-azaspiro[2.5]octane-6-yl}-4-bromo-2-methylamino-3-nitrobenzene was added, along with 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). After the addition was complete, the reaction was stirred at room temperature for 1 hour. Quenching with water, extraction with ethyl acetate (3 × 30 mL), combining the organic phases, backwashing with saturated brine (2 × 30 mL), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the residue by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give methyl 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylate (190 mg, 45.5%).

[0288] MS:(ESI,m / z):378.30[M+H] + RT(min):1.193

[0289] 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):

[0290] 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 (100 mg, 0.3 mmol, 1 eq) of methyl 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylate (2 mL) at room temperature. The reaction solution 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 1M dilute hydrochloric acid, and extracted with ethyl acetate (2 × 10 mL). The organic phases were combined and backwashed with saturated brine (2 × 10 mL). The solution was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid (101 mg, 105%).

[0291] MS:(ESI,m / z):364.30[M+H] + RT(min): 0.763

[0292] 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):

[0293] Under nitrogen protection, at room temperature, 6-(4,4-difluoropiperidin-1-yl)-4-methylpyridine-2-amine (112 mg, 0.5 mmol, 3 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (185 mg, 0.7 mmol, 4 eq), and N-methylimidazolium (135 mg, 1.7 mmol, 10 eq) were added to a dichloromethane solution (4 mL) containing 5-{6-azaspiro[2.5]octane-6-yl}-7-bromo-3-methyl-1,3-benzodiazole-4-carboxylic acid (60 mg, 0.16 mmol, 1 eq), and the reaction mixture was heated to 60 °C and stirred for 1 hour. The reaction mixture 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. The filtrate was filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give 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 (72 mg, 76%).

[0294] MS:(ESI,m / z):573.30[M+H] +RT(min):1.392

[0295] Step 7: Synthesis of 5-{6-azaspiro[2.5]octane-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):

[0296] Under nitrogen protection, at room temperature, 2-hydroxyethanesulfonamide (26 mg, 0.2 mmol, 1.2 eq) and methanesulfonic acid {bicyclohexyl[3-isopropoxy-2',4',6'-triisopropoxy-2'-triisopropoxy-2'-trioxane-2' ... The reaction mixture was prepared by heating propyl-(1,1′-biphenyl)-2-yl]phosphonane}(2'-methylamino-1,1′-biphenyl-2-yl)palladium(II) (16.02 mg, 0.017 mmol, 0.1 eq), bicyclohexyl(3-isopropoxy-2′,4′,6′-triisopropyl-[1,1′-biphenyl]-2-yl)phosphonane (18.65 mg, 0.035 mmol, 0.2 eq), and cesium carbonate (170.44 mg, 0.522 mmol, 3 eq) to 100 °C and stirring 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). The solution was 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]octane-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%).

[0297] MS:(ESI,m / z):618.70[M+H] + RT(min):1.903

[0298] 1H NMR: (400MHz, DMSO-d6) δ10.73(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=6.6Hz,2H),3.8 0(d,J=3.9Hz,3H),3.69(t,J=5.5Hz,4H),3.45(t,J=6.6Hz,2H),2.96(t, J=5.2Hz,4H),2.29(s,3H),2.05–1.85(m,4H),1.45(s,4H),0.29(s,4H).

[0299] Biological evaluation

[0300] Test Example 1

[0301] Test Name: Imaging-Based Nuclear Counting Analysis (NCA) in OVCAR-3 Cells

[0302] Day 0: Compound dilution and treatment

[0303] a) The final test concentrations of AM-5308 were: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, and 0.5 nM.

[0304] b) The final test concentrations of the compounds to be tested were: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, and 0.5 nM.

[0305] c) The cells were cultured for 4 days in an incubator at 37°C and 5% CO2.

[0306] d) The DMSO concentration is 0.1%.

[0307] On day 1, cells were seeded into 384-well cell culture plates.

[0308] a) When the cell confluence reaches 80%-90%, process the cells.

[0309] b) Resuspend the cells in the culture medium, and then count and dilute the cells at the desired density.

[0310] c) Add 30 μL / well of cell suspension containing appropriate cells to a 384-well plate: 600 cells / well.

[0311] Day 4 testing

[0312] a) Add 30 μL of 8% fixative (final concentration 4%) and incubate the plate at room temperature for 30 minutes.

[0313] b) Centrifuge plate, 1000 RPM, 30 s.

[0314] c) Wash twice with 60 μl / well of PBS.

[0315] d) After fixation, the cells were permeabilized and stained in 60 μL of wash buffer containing 2 μg / mL Hoechst 33342 DNA dye (1% BSA, 0.2% Triton X-100, 1X PBS).

[0316] e) Seal the plate and incubate it at room temperature in the dark for 1 hour.

[0317] f) Wash 3 times with PBS.

[0318] g) Add 50 μL PBS per well and scan the plate using HCS.

[0319] h) Data Acquisition and Detection

[0320] Data Analysis

[0321] Inhibition rate (%) = 100 - (Compound well reading - Low reading control well reading) / (High reading control well reading - Low reading control well reading) * 100

[0322] High reading control wells: cells with 30 nL DMSO; low reading control wells: 10 μM AM-5308.

[0323] Calculate IC using GraphPad Prism 8 software 50 (nM) and plot the effect-dose curve of the compound.

[0324] Table 1. Biological activity data of the compounds in this application.

[0325]

[0326] Test Example 2

[0327] Test Name: ADP-Glo TM kinase assay

[0328] Operating steps:

[0329] 1) Prepare 1× reaction buffer.

[0330] 2) Transfer 100 nmL of the diluted compound stock solution to each well of the reaction plate using an Echo 655. The final concentration of DMSO is 1%.

[0331] 3) Seal the reaction plate with a sealing film and centrifuge at 1000g for 1 minute.

[0332] 4) Prepare 2× enzyme solution using 1× reaction buffer.

[0333] 5) Add 5 μL of 2× enzyme solution to each well of the reaction plate. Seal the plate with sealing film, centrifuge at 1000g for 1 minute, and incubate at room temperature for 15 minutes.

[0334] 6) Prepare a 2×ATP solution using 1× reaction buffer.

[0335] 7) Add 5 μL of 2×ATP solution to the reaction plate, centrifuge at 1000g for 1 minute to start the reaction.

[0336] 8) React at room temperature for 60 minutes.

[0337] 9) Add 10 μL of ADP Glo reagent. Centrifuge at 1000g for 1 minute and incubate at room temperature for 60 minutes.

[0338] 10) Add 20 μL of kinase detection reagent. Centrifuge at 1000g for 1 minute and incubate at room temperature for 60 minutes.

[0339] 11) Centrifuge at 1000g for 1 minute.

[0340] 12) Read the light emission signal on Envision 2104.

[0341] Data Analysis:

[0342] The inhibition percentage is calculated as follows:

[0343] %inhibition = 100 - (Signal) cmpd -Signal Ave_PC ) / (Signal Ave_VC -Signal Ave_PC )×100

[0344] Signal cmpd : The average value of the compounds tested on the reaction plate.

[0345] Signal Ave_PC : The average value of the positive control on the reaction plate.

[0346] Signal Ave_VC : The average value of the negative control on the reaction plate.

[0347] Computing IC 50 And the dose-response curve of the fitted compound:

[0348] Using GraphPad 8.0, the IC of the compound is obtained using a nonlinear fitting formula. 50 .

[0349] 3) Quality Control

[0350] Z factor > 0.5; S / B > 2.

[0351] Table 2 Enzyme activity data of the compounds in this application

[0352]

[0353] Test Example 3: Effect of the compound of the present invention on tumor regression

[0354] To demonstrate the effect of KIF18A inhibitors on tumor regression, OVCAR-3 cells (ATCC) were selected for the experiment. Approximately 0.1 mL (5 × 10⁶ cells) was subcutaneously injected into the right side of female nude mice. 6 OVCAR-3 cells. Based on tumor volume (average tumor volume 150 mm). 3 Animals were randomly divided into 6 groups (solvent group, compound 9 group with three doses of 10, 20, or 30 mg / kg, and AMG650 group with two doses of 15 or 30 mg / kg), with 10 animals in each group. Oral administration was administered once daily, starting on day 28 post-tumor inoculation. Tumor volume was measured using electronic calipers (Chengdu Sanhe Measuring Instruments Co., Ltd.). Tumor volume and animal weight were measured twice weekly (from day 28 at the start of the study to day 46 at the end of the study), and the tumor growth inhibition rate of the compound was calculated.

[0355] The formula for calculating tumor volume is: 1 / 2 × a × b 2 , where a and b are the length and width of the tumor, respectively.

[0356] The formula for calculating the 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.

[0357] Data was plotted using GraphPad Prism software (V9.5.0). Tumor volume and weight data are expressed as the mean plus or minus the standard error of the mean. Figure 1 As can be seen, the TGI of compound 9 at doses of 10, 20, and 30 mg / kg were 47.35%, 106.60%, and 115.58%, respectively; the TGI of AMG650 at doses of 15 and 30 mg / kg were 72.58% and 113.15%, respectively. All dose groups of compound 9 had no significant effect on the change in animal body weight relative to the solvent group, and no obvious toxicity was observed.

[0358] The above results indicate that compound 9 of this invention can induce tumor regression in a female nude mouse xenograft model induced by human hyperserous ovarian cancer OVCAR-3 cells (TP53 mutation, CCNE1 amplification). Compound 9 significantly inhibited tumor growth in a mouse subcutaneous xenograft model carrying OVCAR-3 cells at doses of 20 and 30 mg / kg. The inhibitory effect of compound 9 at 20 mg / kg on mouse subcutaneous xenografts was comparable to that of AMG650 at 30 mg / kg.

[0359] As shown in Table 3, compound 9 has a better tumor-to-blood ratio (the ratio of tumor tissue exposure to plasma exposure) than AMG650, indicating that a higher proportion of the drug enters the tumor tissue to exert its effect. Furthermore, compared to AMG650, the lower absolute exposure of compound 9 may reduce the potential for cumulative toxicity.

[0360] Table 3. Exposed levels of mouse plasma and tumor tissue

[0361]

[0362] The embodiments of the technical solution of the present invention have been described above by way of example. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the protection scope of the claims of this application.

Claims

1. A compound of formula (I), its racemate, or a pharmaceutically acceptable salt thereof: （I） in, The compound represented by formula (I) has the following structure: (IIB-2) Where A is selected from ; X1, X2, and X3 may be the same or different, and are independently selected from N or CR0; R0 is selected from H, halogens, and C. 1-6 Alkyl; and when X1 is N and X2 is CR0, X3 is N or CR0; when X1 is N or CR0 and X2 is N, X3 is CR0; B is selected from ; And the condition is that the following compounds are excluded: 。 2. The compound of formula (I) according to claim 1, its racemate, or a pharmaceutically acceptable salt thereof, characterized in that, A is selected from , , , ,or .

3. The compound of formula (I) according to claim 1, its racemate, or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from the following structures: 、 、 、 、 、 。 4. A method for preparing the compound represented by formula (I) according to any one of claims 1-3, its racemate, or a pharmaceutically acceptable salt thereof, comprising the following steps: (1) Compound a reacts with compound A-NH2 to give compound b; (2) Compound b reacts with compound MH to give compound c; (3) Compound c reacts with compound EH to give the compound shown in formula (I); Wherein, the compound represented by formula (I) has the definition in any one of claims 1-3; L is selected from halogens; Q is selected from halogens.

5. A method for preparing the compound represented by formula (I) according to claim 4, its racemate, or a pharmaceutically acceptable salt thereof, characterized in that, L is selected from Cl, Br, and I; Q is selected from F, Cl, and Br.

6. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of any one of claims 1-3, their racemates, or pharmaceutically acceptable salts thereof.

7. Use of at least one of the compounds according to any one of claims 1-3, their racemates, or pharmaceutically acceptable salts thereof in the preparation of a medicament; The intended use is in the preparation of a medicament for treating ovarian cancer.

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