Selective CDK2 inhibitor and use thereof

Abstract

The present invention relates to a cyclin-dependent kinase (CDK)-targeting compound, and specifically provides a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. The compound has the substituents and structural features recited in the present application. The present application further recites a pharmaceutical composition comprising the compound of formula (I) or the stereoisomer thereof or the pharmaceutically acceptable salt thereof, and a pharmaceutical use of the compound or the stereoisomer thereof or the pharmaceutically acceptable salt thereof.

Description

Selective CDK2 inhibitors and their applications

[0001] Cross-references to related applications

[0002] This application claims priority and benefits from the following patent applications, the entire contents of which are incorporated herein by reference:

[0003] Chinese Invention Patent Application No. 202410937134.X, filed with the State Intellectual Property Office of the People's Republic of China on July 12, 2024. Technical Field

[0004] This disclosure pertains to the pharmaceutical field and relates to a substituted purine compound or its stereoisomer or a pharmaceutically acceptable salt thereof, pharmaceutical compositions containing the same, and their use as selective CDK inhibitors in the prevention or treatment of related diseases. Background Technology

[0005] The development of tumors is associated with an imbalance of various oncogenes and tumor suppressor genes. The functional effects of almost all oncogenes and tumor suppressor genes ultimately converge on the cell cycle. Therefore, tumors can be considered a type of cell cycle disease (CCD), and regulating or blocking the cell cycle is one approach to treating tumors. Currently, many molecules related to cell cycle regulation have been discovered, among which cyclin-dependent kinases (CDKs) are the core molecules of the cell cycle regulatory network.

[0006] CDKs are a class of serine / threonine protein kinases that drive the cell cycle through chemical action on serine / threonine proteins. Working synergistically with cyclins, they are crucial factors in cell cycle regulation. Dysregulation of CDK-cyclin complex activity leads to loss of cell cycle and transcriptional control in tumor cells. Over the past three decades, significant progress has been made in CDK inhibitor research, with selective CDK4 / 6 inhibitors approved for cancer patients. However, more selective inhibitors targeting other CDK subtypes remain to be developed. Another important CDK isoform is CDK2, which phosphorylates retinoblastoma (Rb) and releases E2 transcription factor (E2F) with a sequence identical to CDK4 / 6. CDK2, together with its typical binding partner cyclin E1 (CCNE1), drives the G1 / S process. Amplification of this protein has been observed in various cancer types and is associated with lower overall survival in patients with breast cancer, ovarian cancer, and other cancers. Furthermore, selective CDK2 inhibitors have the potential to benefit cancer patients who have developed resistance to CDK4 / 6 inhibitors due to CCNE1 amplification. Therefore, this application aims to develop novel selective CDK2 inhibitors. Summary of the Invention

[0007] This disclosure relates to a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0008] in:

[0009] X 1 X 2 Selected from C or N;

[0010] X 3 Selected from CR or N;

[0011] R is selected from H, halogen, C1-C3 alkyl, or C1-C3 haloalkyl;

[0012] R 1 Selected from C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C6 alkyl groups, wherein the C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C6 alkyl group is optionally surrounded by one or more R groups. 1a replace;

[0013] R 1a Selected from halogens, C1-C3 alkyl groups, C1-C3 haloalkyl groups, and C(O)NR. a R b C(O)OR c OC(O)NR a R bOC(O)OR c ,NHC(O)OR c or NHC(O)NR a R b ;

[0014] R a R b R c They are independently selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or C3-C6 cycloalkyl;

[0015] R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-10 membered heterocyclic groups, the C3-C6 cycloalkyl or 4-10 membered heterocyclic group optionally enclosed by one or more R 2a replace;

[0016] R 5 Selected from Or be chosen by R 5a Replacement C1-C 10 Alkyl; R 5a Selected from OH, =O, halogen, C3-C6 cycloalkyl or 4-7 membered heterocyclic groups;

[0017] R 2a The group is selected from =O, OH or C1-C3 alkyl groups, wherein the C1-C3 alkyl group is optionally substituted by one or more groups selected from OH, halogen or C3-C6 cycloalkyl groups;

[0018] R 3 Selected from halogens, C1-C3 alkyl groups, C1-C3 haloalkyl groups, or C1-C3 alkoxy groups; or two R groups when k = 2 or 3. 3 The atoms optionally bonded to it may form a C3-C6 cycloalkyl or a 3-6 membered heterocycloalkyl, wherein the C3-C6 cycloalkyl or 3-6 membered heterocycloalkyl may optionally be bonded to one or more R 3a replace;

[0019] R 3a Selected from OH, =O, halogen, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy;

[0020] k is selected from 0, 1, 2, or 3;

[0021] R 4 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclic alkyl, 5-10 membered heteroaryl or NR 6 R 7The C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 heterocyclic alkyl, and 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 4a replace;

[0022] R 4a Selected from halogens, OH, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkoxy;

[0023] R 6 Selected from H or C1-C3 alkyl groups;

[0024] R 7 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-7-membered heterocyclic alkyl, or 5-10-membered heteroaryl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, 4-7-membered heterocyclic alkyl, or 5-10-membered heteroaryl is optionally surrounded by one or more R 7a replace;

[0025] R 7a Selected from halogens, C1-C3 alkyl groups, OH, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C3-C6 cycloalkyl groups, or 4-7 membered heterocyclic alkyl groups;

[0026] m is selected from 0, 1, or 2;

[0027] n is selected from 0, 1, or 2.

[0028] In some implementation schemes, X 1 X 2 One is N, and the other is C.

[0029] In some implementation schemes, X 1 Let N, X 2 The answer is C.

[0030] In some implementation schemes, X 1 Let C and X be the values ​​of C and X respectively. 2 Let N be the number of elements in the array.

[0031] In some implementation schemes, X 3 It is CR.

[0032] In some implementation schemes, X 3 Let N be the number of elements in the array.

[0033] In some implementation schemes, X 1 X 2 One is N, the other is C, and X 3 Let N be the number of elements in the array.

[0034] In some implementations, R is selected from H or halogens, such as F.

[0035] In some implementation schemes, R1 Selected from C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C3 alkyl, wherein the C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C3 alkyl is optionally surrounded by one or more R 1a replace.

[0036] In some implementation schemes, R 1 Selected from cyclopropyl, cyclopentyl, oxacyclopentyl, azacyclopentyl, or C1-C3 alkyl groups, wherein the cyclopropyl, cyclopentyl, oxacyclopentyl, azacyclopentyl, or C1-C3 alkyl group is optionally surrounded by one or more R groups. 1a replace.

[0037] In some implementation schemes, R 1 Selected from cyclopropyl or C1-C3 alkyl groups, wherein the cyclopropyl or C1-C3 alkyl group is optionally converted by one or more R groups. 1a replace.

[0038] In some implementation schemes, R 1a Selected from halogens, C1-C3 alkyl groups, and C(O)NR a R b OC(O)NR a R b or NHC(O)OR c .

[0039] In some implementation schemes, R a R b R c They are independently selected from H or C1-C3 alkyl groups.

[0040] In some implementation schemes, R 1a Selected from halogens, methyl groups, C(O)N(CH3)2, OC(O)NHCH(CH3)2 or NHC(O)OCH(CH3)2.

[0041] In some implementation schemes, R 1a Selected from halogens such as F.

[0042] In some implementation schemes, R 1 Selected from ethyl, isopropyl, CHF2, Cyclopropyl

[0043] In some implementation schemes, R 1 Selected from ethyl, isopropyl, CHF2 or cyclopropyl.

[0044] In some implementation schemes, R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-7 membered heterocyclic alkyl, the C3-C6 cycloalkyl or 4-7 membered heterocycloalkyl may be optionally surrounded by one or more R 2a replace.

[0045] In some implementation schemes, R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-7 membered heterocyclic alkyl, the Cyclopropyl or piperidinyl is optionally coupled with one or more R 2a replace.

[0046] In some implementation schemes, R 2 Selected from NHR 5 OR 5 , Cyclopropyl or piperidinyl, the Cyclopropyl or piperidinyl is optionally coupled with one or more R 2a Replace; the R 5 Selected from Or be chosen by R 5a Substituted C1-C6 alkyl groups.

[0047] In some implementation schemes, R 2 Selected from NHR 5 Or optionally by one or more R 2a Replacement The R 5 Selected from Or be chosen by R 5a Substituted C1-C6 alkyl groups.

[0048] In some implementation schemes, R 2a =O or hydroxyethyl.

[0049] In some implementation schemes, R 5a Selected from halogens, OH, CF3, or methyl.

[0050] In some implementation schemes, R 5a Selected from halogens or OH.

[0051] In some implementation schemes, R 5 Selected from Isopropyl,

[0052] In some implementation schemes, R 5 Selected from

[0053] In some implementation schemes, R2 Selected from NHR 5 OR 5 , Cyclopropyl or piperidinyl, the Cyclopropyl or piperidinyl is optionally coupled with one or more R 2a Replace; the R 5 Selected from

[0054] In some implementation schemes, R 2 Selected from Or cyclopropyl.

[0055] In some implementation schemes, R 2 Selected from

[0056] In some implementation schemes, R 3 Selected from halogens, C1-C3 alkyl groups, or C1-C3 haloalkyl groups, or two R groups when k=2 or 3. 3 The atoms optionally bonded to it can form a C3-C6 cycloalkyl group, wherein the C3-C6 cycloalkyl group is optionally bonded to one or more R... 3a replace.

[0057] In some implementation schemes, R 3 Selected from halogens or C1-C3 alkyl groups, or two Rs when k=2 or 3. 3 The atoms optionally bonded to it can form a C3-C6 cycloalkyl group, wherein the C3-C6 cycloalkyl group is optionally bonded to one or more R... 3a replace.

[0058] In some implementation schemes, R 3 Selected from halogens or methyl groups, or two R groups when k = 2 or 3. 3 Together with the atoms attached to it, they form cyclopropyl or cyclobutyl groups.

[0059] In some implementations, k is 0.

[0060] In some implementations, k = 1 and (R 3 ) k It is a methyl or halogen such as F, or k = 2 and 2 Rs 3 Together with the atoms attached to it, they form cyclopropyl or cyclobutyl groups.

[0061] In some implementations, n is selected from 1 or 2.

[0062] In some implementations, m is 1 and n is 1.

[0063] In some implementation schemes, Selected from

[0064] In some implementation schemes, Selected from

[0065] In some implementation schemes, R 4 Selected from C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl, indole, or NR 6 R 7 The C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl, or indole group is optionally surrounded by one or more R groups. 4a replace.

[0066] In some implementation schemes, R 4 Selected from C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl or NR 6 R 7 The C1-C3 alkyl, cyclopropyl, aziridine, or pyrazolyl group is optionally surrounded by one or more R groups. 4a replace.

[0067] In some implementation schemes, R 4a Selected from C1-C3 alkyl groups or halogens such as F.

[0068] In some implementation schemes, R 4a Selected from methyl or F.

[0069] In some implementation schemes, R 4 For NR 6 R 7 .

[0070] In some implementation schemes, R 6 Selected from H.

[0071] In some implementation schemes, R 7 Selected from methyl, ethyl, indolyl, or oxecyclopentyl, wherein the methyl, ethyl, indolyl, or oxecyclopentyl group is optionally converted by one or more R... 7a replace.

[0072] In some implementation schemes, R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R... 7a replace.

[0073] In some implementation schemes, R 6 Selected from H, and R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R... 7a replace.

[0074] In some implementation schemes, R 4 For NR 6 R 7 R 6 Selected from H, R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R... 7a replace.

[0075] In some implementation schemes, R 7a It is selected from halogens, NH (C1-C3 alkyl) or 4-7 membered heterocyclic alkyl groups.

[0076] In some implementation schemes, R 7a Selected from halogens or 4-7 membered heterocyclic alkyl groups.

[0077] In some implementation schemes, R 7a Selected from F, NHCH3 or 1,4-dioxane.

[0078] In some implementation schemes, R 7a Selected from F or 1,4-dioxane.

[0079] In some implementation schemes, R 4 Selected from NHCH3, NHCH2CH3, NHCH2CF3, Methyl, CH2CF3, Cyclopropyl

[0080] In some implementation schemes, R 4 Selected from NHCH3, NHCH2CH3, NHCH2CF3, Methyl, CH2CF3, Cyclopropyl

[0081] In some implementation schemes, R 4 Selected from NHCH3, NHCH2CH3, NHCH2CF3,

[0082] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0083] Among them, X 1 X 2 X 3 R 1 R 2 R3 R 4 m, n, and k are as defined above.

[0084] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-2) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0085] Among them, X 1 X 2 X 3 R 1 R 2 R 3 R 4 m, n, and k are as defined above.

[0086] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (II) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0087] Among them, X 3 R 1 R 2 R 3 R 4 m, n, and k are as defined above.

[0088] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (III) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0089] Among them, X 3 R 1 R 2 R 3 R 4 m, n, and k are as defined above.

[0090] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the following compounds or their stereoisomers or pharmaceutically acceptable salts:

[0091] On the other hand, this disclosure provides pharmaceutical compositions comprising a compound of formula (I) of this disclosure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

[0092] On the other hand, this disclosure provides a method for treating CDK2-mediated diseases in mammals, including administering to a mammal, preferably a human, a therapeutically effective amount of a compound of general formula (I) of this disclosure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0093] On the other hand, this disclosure provides a method for treating tumors in mammals, comprising administering to a mammal in need of treatment, preferably a human, a therapeutically effective amount of a compound of general formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0094] On the other hand, this disclosure provides the use of compounds of general formula (I) or their stereoisomers or pharmaceutically acceptable salts, or pharmaceutical compositions thereof, in the preparation of medicaments for the prevention or treatment of CDK2-mediated diseases.

[0095] On the other hand, this disclosure provides the use of compounds of general formula (I) or their stereoisomers or pharmaceutically acceptable salts, or pharmaceutical compositions thereof, in the preparation of medicaments for the prevention or treatment of tumors.

[0096] On the other hand, this disclosure provides the use of compounds of general formula (I) or their stereoisomers or pharmaceutically acceptable salts, or pharmaceutical compositions thereof, in the prevention or treatment of CDK2-mediated diseases.

[0097] On the other hand, this disclosure provides the use of compounds of general formula (I) or their stereoisomers or pharmaceutically acceptable salts or pharmaceutical compositions thereof in the prevention or treatment of tumors.

[0098] On the other hand, this disclosure provides compounds of general formula (I) or stereoisomers thereof or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, for the prevention or treatment of CDK2-mediated diseases.

[0099] On the other hand, this disclosure provides compounds of general formula (I) or specific compounds described above for the prevention or treatment of tumors, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof.

[0100] In some implementations, CDK2-mediated diseases are selected from tumors.

[0101] In some implementations, the tumor is selected from cancer.

[0102] Terminology Definitions and Explanations

[0103] Unless otherwise stated, the terms used in this disclosure have the following meanings: the definitions of groups and terms recorded in this disclosure, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined and combined with each other. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0104] In this article Indicates the connection site.

[0105] In this paper, the circles in the ring system represent delocalized large π bonds, indicating that the ring system is an aromatic ring. For example... Both rings in the ring system are aromatic.

[0106] The diagrammatic representation of racemic or enantiomerically pure compounds in this article is derived from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise specified, wedge bonds and virtual wedge bonds are used. The absolute configuration of a solid center is represented by black solid bonds and imaginary bonds. It indicates the relative configuration of a stereocenter (such as the cis-trans configuration of alicyclic compounds).

[0107] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds of this disclosure can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer typically yield a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form is dominant; while in phenols, the enol form is dominant. This disclosure encompasses all tautomeric forms of the compounds.

[0108] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0109] The compounds disclosed herein may have asymmetric atoms such as carbon, sulfur, nitrogen, and phosphorus atoms, or asymmetric double bonds. Therefore, the compounds disclosed herein may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E- and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof or other mixtures, such as mixtures enriched with enantiomers or diastereomers. All such isomers and mixtures thereof are within the scope of the definition of the compounds disclosed herein. Alkyl groups and other substituents may contain additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms. All such isomers involved in all substituents and mixtures thereof are also included within the scope of the definition of the compounds disclosed herein. The compounds containing asymmetric atoms disclosed herein can be isolated in optically active pure form or in racemic form. The optically active pure form can be separated from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0110] The term "substituted" refers to the substitution of one or more hydrogen atoms on a particular atom by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are substituted; oxo substitution does not occur on the aromatic group.

[0111] The terms “optional” or “optionally” mean that the event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, “optionally” substituted with a halogen means that the ethyl group can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible and / or cannot be synthesized is introduced.

[0112] The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents may be arbitrary on the basis of chemical feasibility.

[0113] The term "substituted" refers to the fact that a specific atom or group can be replaced with a specified other atom or group. For example, the CH2 in -CH2CH2CH2- can be replaced by O, S, or NH to obtain -CH2OCH2-, -OCH2CH2-, -CH2SCH2-, -SCH2CH2-, -CH2NHCH2-, or -NHCH2CH2-, etc.

[0114] When any variable (e.g., R) 3 When (R) appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, when (R) appears more than once in the composition or structure of a compound, its definition is independent in each case. 3 When k is 2 in k, it means that it is controlled by 2 R. 3 Replaced, and each R 3 Each has its own independent options.

[0115] For substituents whose substitution positions are not fixed in this paper, such as R in 3 This indicates that the substitution position can be any site on the ring, as long as that site contains a hydrogen atom that can be substituted, such as... include Wait. When When k is 2 or 3, there are 2 R 3 It can replace the same carbon atom and optionally form a C3-C6 cycloalkyl or a 3-6 membered heterocycloalkyl, wherein the C3-C6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally replaced by one or more of the aforementioned R 3a Replace; or 2 Rs 3 The substitution of carbon atoms at different carbon atoms and optionally further bonded thereto, as well as the bonds between the two carbon atoms (calculated based on the shorter chain length of the bond), forms a C3-C6 cycloalkyl or a 3-6 membered heterocycloalkyl, wherein the C3-C6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally replaced by one or more of the aforementioned R 3a replace.

[0116] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a bond. For example... When n is 0, it means that the structure is actually

[0117] C in this article m -C n It refers to having an integer number of carbon atoms, either mn or in the range m to n. For example, "C1-C 10"" means that the group can have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Similarly, "m-membered" to "n-membered" indicates that the number of ring atoms is m to n. For example, 5-14-membered rings include 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, 12-membered, 13-membered, and 14-membered rings. It also includes any range from n to m. For example, 5-14-membered rings include 6-14-membered, 6-11-membered, 5-10-membered, 6-10-membered, and 6-8-membered rings.

[0118] The term "alkyl" refers to a compound with the general formula C10. n H 2n+1 The hydrocarbon group is a straight-chain or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms. The term "C1-C" is used. 10 "Alkyl" should be understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific examples of alkyl groups include, but are not limited to, 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, etc.; the term "C1-C6 alkyl" can be understood as referring to an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms, specific examples including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C4 alkyl" can be understood as referring to a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, or 4 carbon atoms. The term "C1-C3 alkyl" can be understood as referring to a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, or 3 carbon atoms. The "C1-C6 alkyl"... 10 "alkyl" can include the range of "C1-C6 alkyl", "C1-C4 alkyl" or "C1-C3 alkyl", and "C1-C6 alkyl" can further include "C1-C4 alkyl" or "C1-C3 alkyl", and "C1-C4 alkyl" can further include "C1-C3 alkyl".

[0119] The term "haloalkyl" refers to the group obtained by further substituting the alkyl group with a halogen. For example, "C1-C6 haloalkyl" refers to C1-C6 alkyl groups that have been further substituted with a halogen, and "C1-C3 haloalkyl" refers to C1-C3 alkyl groups that have been further substituted with a halogen. The "C1-C6 haloalkyl" may further include "C1-C3 haloalkyl".

[0120] The term "alkoxy" refers to a monovalent group formed by the loss of a hydrogen atom from a hydroxyl group in straight-chain or branched alcohols; it can be understood as "alkyloxy" or "alkyl-O-", where alkyl is defined as described above. The term "C1-C"... 10 "Alkoxy" can be understood as "C1-C" 10 "alkyloxy" or "C1-C" 10 Alkyl-O-”; the term “C1-C6 alkoxy” can be understood as “C1-C6 alkyloxy” or “C1-C6 alkyl-O-”. “C1-C3 alkoxy” can be understood as “C1-C3 alkyloxy” or “C1-C3 alkyl-O-”. The “C1-C” 10 "Alkoxy" can include the range of "C1-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" can further include "C1-C3 alkoxy".

[0121] The term "cycloalkyl" refers to a fully saturated carbon ring existing in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. The term "C3-C" is also used. 10 "Cycloalkyl" should be understood as indicating a saturated monovalent monocyclic, fused, spirocyclic, or bridged ring having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The term "C3-C8 cycloalkyl" should be understood as indicating a saturated monovalent monocyclic, fused, spirocyclic, or bridged ring having 3, 4, 5, 6, 7, or 8 carbon atoms. The term "C3-C6 cycloalkyl" should be understood as indicating a saturated monovalent monocyclic, fused, spirocyclic, or bridged ring having 3, 4, 5, or 6 carbon atoms, specific examples including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The term "C3-C6 cycloalkyl" may include "C3-C5 cycloalkyl" or "C3-C4 cycloalkyl".

[0122] The term "heterocyclic group" refers to a fully saturated or partially saturated (not aromatic as a whole) monovalent monocyclic, fused, spirocyclic, or bridged ring group containing 1, 2, 3, 4, or 5 heteroatoms or heterogroups (i.e., groups containing heteroatoms). These "heteroatoms or heterogroups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=O)2-, -S(=O)-, and optionally substituted -NH-, -S(=O)(=NH)-, -C(=O)NH-, -C(=NH)-, -S(=O)2NH-, S(=O)NH-, or -NHC(=O)NH-, etc., which typically contain 3 to 20 ring atoms. The term "4-7 membered heterocyclic group" refers to a heterocyclic group with 4, 5, 6, or 7 ring atoms, and whose ring atoms contain 1-2 independently selected heteroatoms or heterogroups as described above. The term "4-14 membered heterocyclic group" refers to a heterocyclic group with 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, and whose ring atoms contain 1-5 independently selected heteroatoms or heterogroups as described above. "4-14 membered heterocyclic group" can include "4-10 membered heterocyclic group," "4-7 membered heterocyclic group," "4-6 membered heterocyclic group," "5-6 membered heterocyclic group," or "6-10 membered heterocyclic group." The term "3-6 membered heterocyclic group" can include "4-6 membered heterocyclic group", "5-6 membered heterocyclic group", "3-5 membered heterocyclic group", or "4-5 membered heterocyclic group". Specific examples of 4 membered heterocyclic groups include, but are not limited to, azacyclobutane or oxacyclobutane; specific examples of 5 membered heterocyclic groups include, but are not limited to, tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-1H-pyrrolyl; specific examples of 6 membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine, trithiaalkyl, tetrahydropyridinyl, or 4H-[1,3,4]thiadiazinyl; specific examples of 7 membered heterocyclic groups include, but are not limited to, diazacyclobutane. Cycloheptyl. The heterocyclic group can also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include, but are not limited to, hexahydrocyclopentano[c]pyrrolo-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group can be a benzofused cyclic group of the above-mentioned 4-7-membered heterocyclic groups, specific examples including, but not limited to, dihydroisoquinolinyl. Although some bicyclic heterocyclic groups in this disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0123] The term "heterocyclic alkyl" refers to a fully saturated cyclic group existing in the form of a monocyclic, fused, bridged, or spirocyclic ring, wherein the ring atoms contain 1-5 heteroatoms or heteroatom groups (i.e., atomic groups containing heteroatoms). These "heteroatoms or heteroatom groups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=O)2-, -S(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH-, or -NHC(=O)NH-. The term "4-7 membered heterocyclic alkyl" refers to a heterocyclic alkyl group with 4, 5, 6, or 7 ring atoms, and its ring atoms contain 1-3 independently selected heteroatoms or heteroatom groups as described above. Specific examples of 4-membered heterocyclic alkyl groups include, but are not limited to, acridinel, oxadiazolyl, or thiobutylcyclol; specific examples of 5-membered heterocyclic alkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, imidazolyl, or tetrahydropyrazolyl; specific examples of 6-membered heterocyclic alkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, morpholinyl, piperazine, 1,4-thiaoxalyl, 1,4-dioxane, thiomorpholinyl, 1,3-dithiaalkyl, or 1,4-dithiaalkyl; specific examples of 7-membered heterocyclic alkyl groups include, but are not limited to, azirheptanyl, oxaheptanyl, or thioheptanyl.

[0124] The term "aryl" refers to an aromatic ring group consisting of a monocyclic or fused polycyclic aromatic ring with a conjugated π-electron system. Aryl groups can have 6-20, 6-14, 6-12, or 6-10 carbon atoms. The term "C6-C"... 10 "Aryl" should be understood as a monovalent aromatic monocyclic or bicyclic hydrocarbon ring having 6 to 10 carbon atoms. Specifically, it refers to a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; 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), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0125] The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic system containing at least one, preferably 1, 2, 3, or 4 ring atoms selected from N, O, and S, with the remaining ring atoms being 5-14 membered aromatic cyclic groups of carbon. The heteroaryl is preferably 5-10 membered, more preferably 5- or 6-membered heteroaryl. The term "5-10 membered heteroaryl" should be understood to include monovalent monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, or 10 ring atoms, particularly 5, 6, 9, or 10 ring atoms, and containing 1, 2, 3, 4, or 5, preferably 1, 2, or 3 heteroatoms independently selected from N, O, and S. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzothiazolyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl or isindolyl; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and their benzo[derivatives], such as quinolinyl, quinazolinyl or isoquinolinyl; or acrylinyl, inazinyl, purinyl and their benzo[derivatives]; or cyclolinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphthidyl, pteridinyl, carbazolyl, acrylinyl, phenazinyl, phenothiazinyl or phenothiazinyl. The term "5-6-membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and comprising 1, 2, or 3, preferably 1 or 2, heteroatoms independently selected from N, O, and S. The term "6-membered heteroaryl" refers to an aromatic ring system having 6 ring atoms, and comprising 1, 2, or 3, preferably 1 or 2, heteroatoms independently selected from N, O, and S. The term "5-10-membered heteroaryl" can include either "5-6-membered heteroaryl" or "6-membered heteroaryl," and the term "5-6-membered heteroaryl" can include "6-membered heteroaryl."

[0126] The term "halogen" or "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0127] The term "hydroxyl group" refers to the -OH group.

[0128] The term "cyano" refers to the -CN group.

[0129] The term "amino" refers to the -NH2 group.

[0130] The term "nitro" refers to the -NO2 group.

[0131] The term “therapeutic effective amount” means the amount of a compound of this disclosure used to treat a particular disease, condition, or symptom; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or symptom; or (iii) to delay the onset of one or more symptoms of a particular disease, condition, or symptom described herein. The amount of a compound of this disclosure constituting a “therapeutic effective amount” varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and this disclosure.

[0132] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0133] The term “pharmaceutically acceptable salt” or “medicinal salt” refers to a pharmaceutically acceptable salt of an acid or base, including salts formed by a compound with an inorganic or organic acid, and salts formed by a compound with an inorganic or organic base.

[0134] The term "pharmaceutical composition" refers to a mixture of one or more compounds of this disclosure or their stereoisomers or pharmaceutically acceptable salts with pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compounds of this disclosure to an organism.

[0135] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0136] The word “comprise” or “include” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0137] This disclosure also includes compounds of this disclosure that are identical to those described herein, but in which one or more atoms are labeled with isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as... 2 H, 3 H, 11 C 13C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, etc.

[0138] Compounds of this disclosure labeled with certain isotopes (e.g., using...) 3 H and 14 C-labeling can be used in the analysis of compound and / or substrate tissue distribution. Tritiumization (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. The isotopically labeled compounds of this disclosure can typically be prepared by replacing the unlabeled reagent with an isotopically labeled reagent using a procedure similar to those disclosed in the schemes and / or examples below.

[0139] The pharmaceutical compositions disclosed herein can be prepared by combining the compounds of this disclosure with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0140] Typical routes of administration of the compounds or stereoisomers thereof or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof disclosed herein include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0141] The pharmaceutical compositions disclosed herein can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, emulsification, freeze drying, etc.

[0142] In some embodiments, the pharmaceutical composition is in an oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of this disclosure to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0143] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, flow aids, or flavoring agents.

[0144] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0145] The dosage of compounds or compositions used in the treatments described in this disclosure will generally vary depending on the severity of the disease, the patient’s weight, and the relative efficacy of the compound. However, as a general guideline, a suitable daily dose of the compound of formula (I) described herein is from 0.01 mg / kg to 1000 mg / kg.

[0146] The compounds disclosed herein can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this disclosure.

[0147] The chemical reactions in the specific embodiments of this disclosure are carried out in a suitable solvent, which must be suitable for the chemical changes and the reagents and materials required by this disclosure. To obtain the compounds of this disclosure, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0148] In some embodiments, compounds of general formula (I) of this application can be prepared by those skilled in the art of organic synthesis via the following route:

[0149] Scheme 1

[0150] Among them, X 1 X 2 X 3 R 1 R 3R 4 R 5 m, n and k are as defined in equation (I). Detailed Implementation

[0151] The following detailed description of specific implementation schemes illustrates the contents of this disclosure, but does not imply any adverse limitation thereof. Various specific implementation schemes of this disclosure have been described in detail herein, and it will be apparent to those skilled in the art that various changes and modifications can be made to these specific implementation schemes without departing from the spirit and scope of this disclosure.

[0152] All reagents used in this disclosure are commercially available and can be used without further purification.

[0153] Unless otherwise stated, proportions expressed for mixed solvents are volume-based. Unless otherwise stated, % refers to wt%.

[0154] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are measured in units of 10⁻¹⁰. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., with tetramethylsilane (TMS) as the internal standard; "IC 50 "Half-inhibitory concentration" refers to the concentration at which half of the maximum inhibitory effect is achieved.

[0155] The eluent described below may be a mixture of two or more solvents, with the ratio being the volume ratio of each solvent.

[0156] Abbreviations:

[0157] Et: Ethyl; DMF: N,N-Dimethylformamide; Boc: Tert-Butyloxycarbonyl; NMP: N-Methylpyrrolidone; tBu-BrettPhos Pd G3: 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl(2-amino-1,1'-biphenyl-2-yl)palladium(II); Pd-PEPPSI-IPent: [1,3-bis(2,6-di-3-pentylphenyl)imidazolium-2-yl](3-chloropyridinyl)palladium(II)dichloride; Pd(dppf)Cl2: 1,1-bis(diphenylphosphine)ferrocene palladium chloride; tBuOH: tert-butanol; tBuONa: sodium tert-butoxide; dioxane: 1,4-dioxane; MeOH: methanol; DIEA / DIPEA: N,N-diisopropylethylamine; DCM: dichloromethane; DMSO: dimethyl sulfoxide; TEA: triethylamine; CatacxiumA Pd G3: Mesylate [n-Butylbis(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II); MeCN / ACN: Acetonitrile; NIS: N-iodosuccinimide; HEPES: 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid; DTT: Dithiothreitol; ATP: Adenosine triphosphate; EDTA: Ethylenediaminetetraacetic acid.

[0158] Example 1: Preparation of (S)-N-ethyl-3-((3-ethyl-5-(((2R,3S)-2-hydroxypentan-3-yl)amino)pyrazolo[1,5-a]pyrimidin-7-yl)amino)pyrrolidine-1-sulfonamide (compound 1)

[0159] Step 1: Preparation of (3S)-3-[(5-chloro-3-ethyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]pyrrolidine-1-carboxylic acid tert-butyl ester

[0160] Add 5,7-dichloro-3-ethylpyrazolo[1,5-a]pyrimidine (300 mg, 1.39 mmol) and (3S)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (310.32 mg, 1.67 mmol) to a mixture of anhydrous acetonitrile (3.0 mL), then add N,N-diisopropylethylamine (538.35 mg, 4.17 mmol, 725.53 μL). Stir the resulting mixture at 100 °C for 0.5 hours under microwave conditions. After the reaction was complete, the crude product was separated by a preparative high performance liquid chromatography column (column: Welch Xtimate C18 column, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95% in 18 minutes) to obtain the title compound (400.0 mg, yield: 78.74%).

[0161] MS m / z (ESI): 366.2 [M+H] + .

[0162] Step 2: Preparation of 5-chloro-3-ethyl-N-[(3S)-pyrrolidine-3-yl]pyrazolo[1,5-a]pyrimidine-7-amine hydrochloride

[0163] (3S)-3-[(5-chloro-3-ethyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]pyrrolidine-1-carboxylic acid tert-butyl ester (200 mg, 546.66 μmol) was added to methanol (1 mL), followed by HCl solution of dioxane (4 M, 3 mL). The reaction was stirred at 25 °C for 2.0 h. After the reaction was complete, the solution was concentrated under reduced pressure to obtain the crude product, the title compound (140 mg, yield: 96.37%). No purification was required, and the product was used directly in the next step.

[0164] MS m / z(ESI): 266.1 [M+H] + .

[0165] Step 3: Preparation of (3S)-3-[(5-chloro-3-ethyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-N-ethyl-pyrrolidine-1-sulfonamide

[0166] 5-Chloro-3-ethyl-N-[(3S)-pyrrolidine-3-yl]pyrazolo[1,5-a]pyrimidine-7-amine hydrochloride (140 mg, 463.27 μmol) was added to a solution of dichloromethane (5 mL), followed by N,N-diisopropylethylamine (544.71 mg, 4.21 mmol, 734.11 μL). N-ethylsulfonyl chloride (113.47 mg, 790.24 μmol) in dichloromethane (2 mL) was slowly added dropwise at -78 °C. The reaction was stirred at -78 °C for 1 hour. After the reaction was complete, saturated ammonium chloride aqueous solution was added under ice bath to quench the reaction. The solution was extracted with dichloromethane and concentrated under reduced pressure to obtain the crude product. The crude product was then subjected to high-performance liquid chromatography (HPLC) with a Welch Xtimate C18 column (150 mm length, 30 mm inner diameter, 5 μm particle size); mobile phase A: water (containing 0.225%). Separation was performed using NH3 (mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95% over 18 minutes) to give the title compound (120.0 mg, yield: 61.09%).

[0167] MS m / z(ESI): 373.1 [M+H] + .

[0168] Step 4: Preparation of (S)-N-ethyl-3-((3-ethyl-5-(((2R,3S)-2-hydroxypentan-3-yl)amino)pyrazolo[1,5-a]pyrimidin-7-yl)amino)pyrrolidine-1-sulfonamide (compound 1)

[0169] (3S)-3-[(5-chloro-3-ethyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-N-ethyl-pyrrolidine-1-sulfonamide (60 mg, 160.91 μmol), (2R,3S)-3-aminopentan-2-ol (33.20 mg, 321.83 μmol), Pd-PEPPSI-IPent (12.74 mg, 16.09 μmol), and sodium tert-butoxide (46.39 mg, 482.74 μmol) were added to 1,4-dioxane (2 mL), and nitrogen was purged. The resulting mixture under nitrogen atmosphere was heated to 90 °C and stirred for 2.0 h under nitrogen protection. After the reaction was complete, the mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then separated by a preparative high-performance liquid chromatography column (column: Welch Xtimate C18 column, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95% in 18 minutes) to obtain the title compound, compound 1 (24 mg, yield: 35.34%).

[0170] MS m / z (ESI): 440.3 [M+H] +.

[0171] 1 H NMR (400MHz, DMSO-d6) δ7.60(s,1H),7.05(d,J=6.8Hz,1H),6.46(d,J=8.1Hz,1H),5.40(s,1H),5.06(s,1H),4. 19–4.09(m,1H),3.82(s,1H),3.65(p,J=6.1Hz,1H),3.55(dd,J=10.0,6.4Hz,1H),3.40(s,2H),3.26–3.19(m,2 H),2.96(q,J=7.2Hz,2H),2.47(t,J=7.5Hz,2H),2.32–2.22(m,1H),2.15(ddt,J=12.5,7.8,6.1Hz,1H),1.66(d tt,J=14.5,7.2,3.6Hz,1H),1.46–1.34(m,1H),1.16(t,J=7.6Hz,3H),1.08–1.02(m,6H),0.88(t,J=7.4Hz,3H).

[0172] Example 2: Preparation of (S)-N-ethyl-3-((2-(((2R,3S)-2-hydroxypentan-3-yl)amino)-8-isopropylpyrazolo[1,5-a][1,3,5]triazine-4-yl)amino)pyrrolidine-1-sulfonamide (compound 2)

[0173] Step 1: Preparation of (S)-3-((2-chloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0174] 2,4-Dichloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazine (60.0 mg, 0.26 mmol) and (3S)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (48.4 mg, 0.26 mmol) were dissolved in anhydrous acetonitrile (2.6 mL), and triethylamine (79 mg, 0.78 mmol) was added at 0 °C. The reaction was carried out at 0 °C for 10 min. After the reaction was completed, the organic phase was evaporated to dryness, and the residue was purified by normal column chromatography (dichloromethane:methanol = 96:04) to give the title compound (95 mg, yield 96%).

[0175] m / z(ESI): 381.3 [M+H] + .

[0176] Step 2: Preparation of (S)-2-chloro-8-isopropyl-N-(pyrrolidine-3-yl)pyrazolo[1,5-a][1,3,5]triazine-4-amine hydrochloride

[0177] (S)-3-((2-chloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (95 mg, 0.25 mmol) was dissolved in dichloromethane (2.5 mL), and dioxane hydrochloride solution (0.5 mL, 4 M) was added. The reaction was carried out at 25 °C for 2 hours, and the reaction was completed. The organic phase was concentrated to dryness under reduced pressure to obtain the title compound (crude product), which was used directly in the next step.

[0178] m / z(ESI): 281.3 [M+H] + .

[0179] Step 3: Preparation of (S)-3-((2-chloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazine-4-yl)amino)-N-ethylpyrrolidine-1-sulfonamide

[0180] The compound (S)-2-chloro-8-isopropyl-N-(pyrrolidine-3-yl)pyrazolo[1,5-a][1,3,5]triazine-4-amine hydrochloride obtained in the previous step was dissolved in dichloromethane (2.5 ml), and the solution was transferred to -78 °C. Triethylamine (101 mg, 1.01 mmol) and ethylaminosulfonyl chloride (72 mg, 0.50 mmol) were added, and the reaction was stirred at -78 °C for 10 minutes. After the reaction was completed, the reaction solution was purified by normal column chromatography (dichloromethane:methanol = 96:04) to give the title compound (90 mg, yield 93%).

[0181] m / z(ESI): 388.4 [M+H] + .

[0182] Step 4: Preparation of (S)-N-ethyl-3-((2-(((2R,3S)-2-hydroxypentan-3-yl)amino)-8-isopropylpyrazolo[1,5-a][1,3,5]triazine-4-yl)amino)pyrrolidine-1-sulfonamide (compound 3)

[0183] Compound (S)-3-((2-chloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino)-N-ethylpyrrolidine-1-sulfonamide (45 mg, 0.12 mmol), sodium tert-butoxide (38.4 mg, 0.40 mmol), and [1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-yl](3-chloropyridinyl)palladium dichloride (9.5 mg, 0.012 mmol) were dissolved in 1,4-dioxane (1.0 mL). The reaction was carried out at 100 °C for 4.0 h under a nitrogen atmosphere, and the reaction was completed. The organic phase obtained by filtration of the reaction solution was concentrated to dryness under reduced pressure. The reaction solution was purified by reverse-phase C18 column (mobile phase: acetonitrile-water (the aqueous phase contains 0.025% ammonia), acetonitrile ratio 10-90%, gradient elution for 8 column volumes) to obtain the title compound, compound 2 (12.0 mg, yield 27%).

[0184] m / z (ESI): 455.4 [M+H] + .

[0185] 1 H NMR(400MHz,DMSO-d6)δ7.77(s,1H),7.05(brs,1H),7.09(s,1H),4.95(brs,1H),4.79–4.65(m,1H),4. 45–4.42(m,1H),4.00(q,J=7.2Hz,2H),3.55(dd,J=9.7,6.7Hz,1H),3.40–3.36(m,2H),3.30–3.29(m,1H ),3.25–3.19(m,1H),3.09(dd,J=9.8,6.1Hz,1H),3.03–2.88(m,2H),2.81(t,J=13.0Hz,1H),2.21(td, J=12.8,6.8Hz,1H),1.90–1.82(m,1H),1.69–1.55(m,6H),1.35(t,J=7.2Hz,4H),1.06(t,J=7.2Hz,3H).

[0186] Example 3: Preparation of (S)-N-ethyl-3-((3-ethyl-6-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-sulfonamide (compound 3)

[0187] Step 1: Preparation of 8-bromo-6-chloro-3-ioimidazole[1,2-b]pyridazine

[0188] 8-Bromo-6-chloroimidazolo[1,2-b]pyridazine (0.5 g, 2.2 mmol) was dissolved in N,N-dimethylformamide (7 mL), and N-iodosuccinimide (0.6 g, 2.6 mmol) was added. The resulting mixture was stirred at 80 °C for 2 hours. After the reaction solution cooled to room temperature, it was directly passed through a high-performance liquid chromatography column (column: Welch Xtimate C18, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95%, elution for 8 column volumes) to give the title compound (0.4 g, yield 52.0%).

[0189] m / z(ESI): 358.8 [M+H] + .

[0190] Step 2: Preparation of (S)-3-((6-chloro-3-iodoimidazole[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0191] (S)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (0.2 g, 1.0 mmol) was dissolved in N,N-dimethylformamide (1 mL), and 8-bromo-6-chloro-3-iodoimidazole[1,2-b]pyridazine (0.4 g, 1.0 mmol) was added. The air in the solution was replaced three times with argon, and the mixture was stirred at 80 °C for 1 hour. After the reaction solution was cooled to room temperature, it was directly passed through a high-performance liquid chromatography column (column: Welch Xtimate C18 column, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95%, elution for 8 column volumes) to give the title compound (0.3 g, yield 64.4%).

[0192] m / z (ESI): 464.7 [M+H] + .

[0193] Step 3: Preparation of (S)-3-((6-chloro-3-vinylimidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0194] Potassium trifluoroborate (36 mg, 0.27 mmol), cesium carbonate (0.3 g, 0.9 mmol), (S)-3-((6-chloro-3-iodomizo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (0.14 g, 0.3 mmol), and methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (43 mg, 60 μmol) were dissolved in 1,4-dioxane / water (1.5 mL / 0.5 mL). The mixture was purged three times with argon, and reacted at 100 °C for 1 hour. After the system cooled to room temperature, the reaction solution was filtered and evaporated to dryness, and then subjected to high performance liquid chromatography (HPLC) column (column: Welch Xtimate C18 column, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95%, elution for 8 column volumes) to obtain the title compound (0.1 g, yield 91.0%).

[0195] m / z(ESI): 364.2 [M+H] + .

[0196] Step 4: Preparation of (S)-3-((6-chloro-3-ethylimidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0197] (S)-3-((6-chloro-3-vinylimidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (80 mg, 0.22 mmol) was dissolved in methanol (5 mL), followed by the addition of platinum dioxide (40 mg, 0.18 mmol). The air was replaced three times with hydrogen, and the reaction was carried out at room temperature for 10 minutes. The reaction solution was filtered and evaporated to dryness to obtain the crude title compound (53 mg, yield 65.9%), which was used directly in the next step.

[0198] m / z(ESI): 366.2 [M+H] + .

[0199] Step 5: Preparation of (S)-3-((3-ethyl-6-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0200] (S)-3-((6-chloro-3-ethylimidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (53 mg, 0.14 mmol), cesium carbonate (0.14 g, 0.43 mmol), (2R,3S)-3-aminopentan-2-ol (37 mg, 0.36 mmol), and 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl(2-amino-1,1'-biphenyl-2-yl)palladium(II) (24 mg, 29 μmol) were dissolved in anhydrous tert-butanol (3 mL), the air in the solution was replaced three times with argon, and the solution was stirred at 90 °C for 1 hour. The reaction solution was filtered and evaporated to dryness, and then subjected to high performance liquid chromatography (HPLC) column (column: Welch Xtimate C18 column, 150 mm long, 30 mm inner diameter, 5 μm particle size; mobile phase A: water (containing 0.225% NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95%, elution for 8 column volumes) to obtain the title compound (20 mg, yield 31.9%).

[0201] m / z(ESI): 433.3 [M+H] + .

[0202] Step 6: Preparation of (2R,3S)-3-((3-ethyl-8-(((S)-pyrrolidine-3-yl)amino)imidazo[1,2-b]pyridazin-6-yl)amino)pentane-2-ol hydrochloride

[0203] (S)-3-((3-ethyl-6-((((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (20 mg, 46 μmol) was added to anhydrous dichloromethane (3 mL), followed by the addition of 4M dioxane hydrochloride solution (1 mL). The resulting mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and 1,4-dioxane (10 mL) was added. The mixture was then concentrated again under reduced pressure and dried to obtain (2R,3S)-3-((3-ethyl-8-((((S)-pyrrolidine-3-yl)amino)imidazo[1,2-b]pyridazin-6-yl)amino)pentan-2-ol hydrochloride (16 mg crude), which was directly used in the next step of the reaction.

[0204] m / z(ESI): 333.3 [M+H] + .

[0205] Step 7: Preparation of (S)-N-ethyl-3-((3-ethyl-6-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[1,2-b]pyridazin-8-yl)amino)pyrrolidine-1-sulfonamide (compound 3)

[0206] Add 3 mL of dichloromethane to (2R,3S)-3-((3-ethyl-8-(((S)-pyrrolidine-3-yl)amino)imidazo[1,2-b]pyridazin-6-yl)amino)pentane-2-ol hydrochloride (crude product 16 mg), stir and react at -78 °C for 10 minutes, then add triethylamine (26 mg, 0.26 mmol), stir for 10 minutes, add ethylaminosulfonyl chloride (6.6 mg, 46 μmol) dropwise, continue reaction at -78 °C for 10 minutes, quench with saturated sodium bicarbonate (4 mL), extract three times with dichloromethane (3 mL), combine organic phases and evaporate to dryness, concentrate is subjected to high performance liquid chromatography (HPLC) column (column: Welch Xtimate C18 column, length 150 mm, inner diameter 30 mm, particle size 5 μm; mobile phase A: water (containing 0.225%) (NH3), mobile phase B: acetonitrile; gradient: mobile phase B from 5% to 95%, elution for 8 column volumes) to give title compound 3 (5 mg, two-step yield 23.9%).

[0207] m / z (ESI): 440.5 [M+H] + .

[0208] 1 H NMR (400MHz, DMSO-d6) δ7.12(t,J=5.7Hz,1H),6.99(s,1H),6.69(d,J=6.6Hz,1H),5.93(d,J=8.3Hz,1H) ,5.61(s,1H),4.60(d,J=5.2Hz,1H),4.17–4.06(m,1H),3.74–3.58(m,2H),3.56–3.50(m,1H),3.27–3.1 9(m,1H),3.19–3.12(m,1H),2.99–2.91(m,2H),2.71(q,J=7.5Hz,2H),2.30–2.20(m,1H),2.13–2.05(m, 1H),1.77–1.66(m,1H),1.46–1.35(m,1H),1.29–1.19(m,4H),1.12–1.02(m,6H),0.88(t,J=7.3Hz,3H).

[0209] Example 4: Preparation of (S)-N-ethyl-3-((7-ethyl-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-sulfonamide (compound 4)

[0210] Step 1: Preparation of (S)-3-((7-bromo-2-chloroimidozono[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0211] 7-Bromo-2,4-dichloroimidazolo[2,1-f][1,2,4]triazine (50 mg, 0.188 mmol) and (3S)-3-aminopyrrolidine-1-carboxylic acid tert-butyl ester (52.14 mg, 0.279 mmol) were added to a mixture of dichloromethane (2.0 mL), followed by the addition of N,N-diisopropylethylamine (72.36 mg, 0.560 mmol). The resulting solution was stirred at room temperature for 4 hours. After the reaction was complete, the dichloromethane was evaporated to dryness, and the crude product was subjected to reversed-phase column chromatography (column: [column information missing]). Rapid silica column chromatography; mobile phase: acetonitrile / water, acetonitrile ratio 5%-95%, 18 min) to separate the title compound (65.0 mg, yield: 83.38%).

[0212] MS m / z (ESI): 484.1 [M+H] + .

[0213] Step 2: Preparation of (S)-3-((7-bromo-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazine-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0214] Add (S)-3-((7-bromo-2-chloroimidazolo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (65 mg, 155.62 μmol), (2R,3S)-3-aminopentan-2-ol (64.22 mg, 622.47 μmol), N,N-diisopropylethylamine (100.56 mg, 778.09 μmol), and N-methylpyrrolidone (0.5 mL) to a sealed tube. Stir the resulting solution at 150 °C for 24 hours. After the reaction is complete, cool to room temperature. The crude product is then subjected to reversed-phase column chromatography (column: [column information missing]). Rapid silica column chromatography; mobile phase: acetonitrile / water, acetonitrile ratio 5%-95%, 18 min) to separate the title compound (61.0 mg, yield: 80.92%).

[0215] MS m / z(ESI): 485.2 [M+H]+

[0216] Step 3: Preparation of (S)-3-((7-ethyl-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0217] (S)-3-((7-bromo-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (61 mg, 125.93 μmol), potassium vinyltrizoborate (50.61 mg, 377.80 μmol), 1,1-bis(diphenylphosphine)ferrocene palladium chloride (9.21 mg, 12.59 μmol), and sodium carbonate (40.04 mg, 377.80 μmol) were added to 1,4-dioxane (2 mL) and water (0.5 mL), and nitrogen was purged. The resulting mixture under nitrogen atmosphere was heated to 90 °C and stirred for 2.0 hours. After the reaction was complete, the mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in methanol (2 mL), and palladium on carbon (15 mg, 14.09 μmol) was added. Hydrogen was purged three times, and the mixture was stirred in a hydrogen atmosphere for 2 hours. The solid was filtered off, and the methanol was evaporated to dryness to obtain the crude product of the title compound (50.0 mg, yield: 91.58%). The crude product was directly used in the next reaction.

[0218] MS m / z (ESI): 434.3 [M+H] + .

[0219] Step 4: Preparation of (S)-N-ethyl-3-((7-ethyl-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-sulfonamide (compound 4)

[0220] Dissolve (S)-3-((7-ethyl-2-(((2R,3S)-2-hydroxypentan-3-yl)amino)imidazo[2,1-f][1,2,4]triazin-4-yl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester (50 mg, 115.33 μmol) in methanol (1 mL), and add 4 M A solution of 1,4-dioxane in HCl was added, and the resulting solution was stirred for 2 hours. The solvent was then removed by rotary evaporation. Dichloromethane (1 mL) and triethylamine (35.01 mg, 345.98 μmol) were added to the crude product. The mixture was cooled to -70°C, and then N-ethylsulfonyl chloride (33.12 mg, 230.66 μmol) in dichloromethane (1 mL) was slowly added dropwise. The reaction was stirred at -70°C for 1 hour. After the reaction was completed, saturated NaHCO3 aqueous solution was added under ice bath to quench the reaction. The product was extracted with dichloromethane and concentrated under reduced pressure to obtain the crude product. The crude product was then subjected to high-performance liquid chromatography (HPLC) using a Welch Xtimate column. The C18 column was 150 mm long, 30 mm inner diameter, and 5 μm particle size. Mobile phase A was water (containing 0.225% NH3), and mobile phase B was acetonitrile. Gradient separation was performed using mobile phase B from 5% to 95% in 18 minutes to obtain the title compound, compound 4 (3.0 mg, yield: 5.90%).

[0221] MS m / z (ESI): 441.4 [M+H] + .

[0222] 1 H NMR (400MHz, DMSO-d6) δ8.35(d,J=5.7Hz,1H),7.17–7.06(m,2H),6.00(d,J=8.6Hz,1H),4.66(dd,J=13. 5,6.8Hz,1H),4.55(d,J=4.5Hz,1H),3.65–3.59(m,2H),3.55(dd,J=9.9,6.9Hz,1H),3.26–3.19(m,2H), 3.15(dd,J=9.8,6.3Hz,1H),2.99–2.91(m,2H),2.76–2.68(m,2H),2.25–2.21(m,1H),2.18–2.05(m,1H) ,1.80–1.66(m,1H),1.47–1.36(m,1H),1.24(t,J=7.5Hz,3H),1.15–1.02(m,6H),0.87(t,J=7.4Hz,3H).

[0223] Test Example 1: Detection of the inhibitory effect of the compound of the present invention on CDK2 kinase

[0224] The in vitro activity of CDK2 was determined by detecting the phosphorylation level of the substrate in the kinase reaction using the Perkin Elmer Lance Ultra TR-FRET kinase assay kit. The reaction buffer contained the following components: 50 mM HEPES (pH 7.5), 10 mM MgCl2, 2 mM DTT, and 0.01% Tween 20; human recombinant CDK2 / CyclinE1 protein (Carna Biosciences, 04-165) diluted to 6 nM with the reaction buffer; the substrate reaction solution consisted of ULight-4E-BP1 kinase substrate (PerkinElmer, TRF0128) diluted to 100 nM with the reaction buffer and 400 μM ATP; the assay buffer consisted of Europium-anti-phospho-4E-BP1 antibody (PerkinElmer, TRF0216) diluted to 2 nM with 1× assay buffer (PerkinElmer, CR97-100) and 20 mM EDTA.

[0225] Using an Echo 650 automated workstation (Beckman Coulter Life Sciences), 100 nmol of different concentrations of compound or DMSO was added to a 384-well assay plate (Perkin Elmer, 6007299), followed by 5 μL of CDK2 kinase solution. After thorough mixing, the mixture was incubated on ice for 30 minutes. Then, 5 μL of substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 60 minutes. In the reaction system, the final compound concentration started at 3000 nM and was serially diluted 4-fold, ranging from 3000 nM to 0.01 nM. The final concentration of DMSO in the system was 1%. Subsequently, 10 μL of assay buffer was added to each well, thoroughly mixed, and incubated at room temperature for 60 minutes. Signal values ​​were then detected at 615 nm and 665 nm using an Envision microplate reader (Perkin Elmer). The signal values ​​(absorbance at 665 nm / absorbance at 615 nm) were positively correlated with the phosphorylation level of the substrate, thus reflecting the kinase activity of CDK2 and the inhibitory effect of the compound.

[0226] The percentage of CDK2 activity inhibition by a compound can be calculated using the following formula: Inhibition percentage (%) = 100 - 100 * (signal value) 化合物 -Signal value 阴性对照 ) / (signal value) 阳性对照 -Signal value 阴性对照 ).

[0227] Where: signal value 化合物 Signal value refers to the signal value of a pore containing a compound, enzyme, substrate, and ATP; signal value 阴性对照Signal value refers to the signal value of a well containing only 1% DMSO, substrate, and ATP; signal value 阳性对照 The signal value refers to the pore containing 1% DMSO, enzyme, substrate, and ATP.

[0228] The inhibition curve was fitted using a four-parameter model using XLfit (ID Business Solutions Ltd., UK) software, and the half-maximal inhibitory concentration (IC50) was calculated. 50 ).

[0229] Experimental results:

[0230] Although specific embodiments of this disclosure have been illustrated and described, those skilled in the art will recognize that, where feasible, the technical features described in one embodiment may be applied to another embodiment or combined with the technical features described in another embodiment. Therefore, those skilled in the art can make various changes and modifications to the embodiments of this disclosure without departing from the spirit and scope of this disclosure.

Claims

1. The compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt, in: X 1 X 2 Selected from C or N; X 3 Selected from CR or N; R is selected from H, halogen, C1-C3 alkyl, or C1-C3 haloalkyl; R 1 Selected from C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C6 alkyl groups, wherein the C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C6 alkyl group is optionally surrounded by one or more R groups. 1a replace; R 1a Selected from halogens, C1-C3 alkyl groups, C1-C3 haloalkyl groups, and C(O)NR. a R b C(O)OR c OC(O)NR a R b OC(O)OR c ,NHC(O)OR c or NHC(O)NR a R b ; R a R b R c They are independently selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or C3-C6 cycloalkyl; R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-10 membered heterocyclic groups, the C3-C6 cycloalkyl or 4-10 membered heterocyclic group optionally enclosed by one or more R 2a replace; R 5 Selected from Or be chosen by R 5a Replacement C1-C 10 Alkyl; R 5a Selected from OH, =O, halogen, C3-C6 cycloalkyl or 4-7 membered heterocyclic groups; R 2a The group is selected from =O, OH or C1-C3 alkyl groups, wherein the C1-C3 alkyl group is optionally substituted by one or more groups selected from OH, halogen or C3-C6 cycloalkyl groups; R 3 Selected from halogens, C1-C3 alkyl groups, C1-C3 haloalkyl groups, or C1-C3 alkoxy groups; or two R groups when k = 2 or 3. 3 The atoms optionally bonded to it may form a C3-C6 cycloalkyl or a 3-6 membered heterocycloalkyl, wherein the C3-C6 cycloalkyl or 3-6 membered heterocycloalkyl may optionally be bonded to one or more R 3a replace; R 3a Selected from OH, =O, halogen, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy; k is selected from 0, 1, 2, or 3; R 4 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclic alkyl, 5-10 membered heteroaryl or NR 6 R 7 The C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 heterocyclic alkyl, and 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 4a replace; R 4a Selected from halogens, OH, C1-C3 alkyl, C1-C3 haloalkyl, or C1-C3 alkoxy; R 6 Selected from H or C1-C3 alkyl groups; R 7 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-7-membered heterocyclic alkyl, or 5-10-membered heteroaryl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, 4-7-membered heterocyclic alkyl, or 5-10-membered heteroaryl is optionally surrounded by one or more R 7a replace; R 7a Selected from halogens, C1-C3 alkyl groups, OH, NH2, NH(C1-C3 alkyl), N(C1-C3 alkyl)2, C3-C6 cycloalkyl groups, or 4-7 membered heterocyclic alkyl groups; m is selected from 0, 1, or 2; n is selected from 0, 1, or 2.

2. The compound of formula (I) according to claim 1, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, X 1 X 2 One is N, and the other is C; or, X 1 Let N, X 2 For C; or, X 1 Let C and X be the values ​​of C and X respectively. 2 For N; or, X 3 For CR; or, X 3 For CR, R is selected from H or halogens, such as F; or, X 3 For N; or, X 1 X 2 One is N, the other is C, and X 3 Let N be the number of elements in the array.

3. The compound of formula (I) according to any one of claims 1-2, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 Selected from C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C3 alkyl, wherein the C3-C6 cycloalkyl, 4-7 membered heterocyclic, or C1-C3 alkyl is optionally surrounded by one or more R 1a Replace; R 1 Selected from cyclopropyl, cyclopentyl, oxacyclopentyl, azacyclopentyl, or C1-C3 alkyl groups, wherein the cyclopropyl, cyclopentyl, oxacyclopentyl, azacyclopentyl, or C1-C3 alkyl group is optionally surrounded by one or more R groups. 1a Replace; R 1 Selected from cyclopropyl or C1-C3 alkyl groups, wherein the cyclopropyl or C1-C3 alkyl group is optionally converted by one or more R groups. 1a replace.

4. The compound of formula (I) according to any one of claims 1-3, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1a Selected from halogens, C1-C3 alkyl groups, and C(O)NR a R b OC(O)NR a R b or NHC(O)OR c .

5. The compound of formula (I) according to any one of claims 1-4, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R a R b R c They are independently selected from H or C1-C3 alkyl groups.

6. The compound of formula (I) according to any one of claims 1-5, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1a Selected from halogens, methyl groups, C(O)N(CH3)2, OC(O)NHCH(CH3)2, or NHC(O)OCH(CH3)2; R 1a Selected from halogens such as F.

7. The compound of formula (I) according to any one of claims 1-6, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-7 membered heterocyclic alkyl, the C3-C6 cycloalkyl or 4-7 membered heterocycloalkyl may be optionally surrounded by one or more R 2a Replace; or, R 2 Selected from NHR 5 OR 5 , C3-C6 cycloalkyl or 4-7 membered heterocyclic alkyl, the Cyclopropyl or piperidinyl is optionally coupled with one or more R 2a Replace; or, R 2 Selected from NHR 5 OR 5 , Cyclopropyl or piperidinyl, the Cyclopropyl or piperidinyl is optionally coupled with one or more R 2a Instead, the R 5 Selected from Or be chosen by R 5a Substituted C1-C6 alkyl; or, R 2 Selected from NHR 5 Or optionally by one or more R 2a Replacement The R 5 Selected from Or be chosen by R 5a Substituted C1-C6 alkyl groups.

8. The compound of formula (I) according to any one of claims 1-7, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 2a =O or hydroxyethyl.

9. The compound of formula (I) according to any one of claims 1-8, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 5a Selected from halogens, OH, CF3, or methyl; or, R 5a Selected from halogens or OH.

10. The compound of formula (I) according to any one of claims 1-9, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 3 Selected from halogens, C1-C3 alkyl groups, or C1-C3 haloalkyl groups, or two R groups when k=2 or 3. 3 The atoms optionally bonded to it can form a C3-C6 cycloalkyl group, wherein the C3-C6 cycloalkyl group is optionally bonded to one or more R... 3a Replace; R 3 Selected from halogens or C1-C3 alkyl groups, or two Rs when k=2 or 3. 3 The atoms optionally bonded to it can form a C3-C6 cycloalkyl group, wherein the C3-C6 cycloalkyl group is optionally bonded to one or more R... 3a Replace; R 3 Selected from halogens or methyl groups, or two R groups when k = 2 or 3. 3 Together with the atoms attached to it, they form cyclopropyl or cyclobutyl groups.

11. The compound of formula (I) according to any one of claims 1-10, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, k is 0; or k = 1 and (R 3 ) k It is a methyl or halogen such as F, or k = 2 and 2 Rs 3 Together with the atoms attached to it, they form cyclopropyl or cyclobutyl groups.

12. The compound of formula (I) according to any one of claims 1-11, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, n is selected from 1 or 2; or m is 1 and n is 2.

13. The compound of formula (I) according to any one of claims 1-12, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 4 Selected from C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl, indole, or NR 6 R 7 The C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl, or indole group is optionally surrounded by one or more R groups. 4a Replace; or, R 4 Selected from C1-C3 alkyl, cyclopropyl, aziridine, pyrazolyl or NR 6 R 7 The C1-C3 alkyl, cyclopropyl, aziridine, or pyrazolyl group is optionally surrounded by one or more R groups. 4a Replace; or, R 4 For NR 6 R 7 R 6 Selected from H, R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R... 7a replace.

14. The compound of formula (I) according to any one of claims 1-13, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 4a Selected from C1-C3 alkyl groups or halogens such as F; or, R 4a Selected from methyl or F.

15. The compound of formula (I) according to any one of claims 1-14, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 7 Selected from methyl, ethyl, indolyl, or oxecyclopentyl, wherein the methyl, ethyl, indolyl, or oxecyclopentyl group is optionally converted by one or more R... 7a Replace; or, R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R groups. 7a Replace; or, R 6 Selected from H, and R 7 Selected from methyl, ethyl, or oxecyclopentyl, wherein the methyl, ethyl, or oxecyclopentyl group is optionally converted by one or more R groups. 7a replace.

16. The compound of formula (I) according to any one of claims 1-15, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 7a Selected from halogens, NH (C1-C3 alkyl) or 4-7 membered heterocyclic alkyl groups; or, R 7a Selected from halogens or 4-7 membered heterocyclic alkyl groups.

17. The compound of formula (I) according to any one of claims 1-16, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound of formula (II) or its stereoisomer or its pharmaceutically acceptable salt. Among them, X 3 R 1 R 2 R 3 R 4 m, n and k are as defined in any one of claims 1-16.

18. The compound of formula (I) according to any one of claims 1-16, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound of formula (III) or its stereoisomer or its pharmaceutically acceptable salt. Among them, X 3 R 1 R 2 R 3 R 4 m, n and k are as defined in any one of claims 1-16.

19. The compound of formula (I) according to any one of claims 1-18, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt thereof is selected from the following compounds or their stereoisomers or pharmaceutically acceptable salts:

20. A pharmaceutical composition comprising the compound of any one of claims 1-19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

21. A method for treating a CDK2-mediated disease in a mammal, comprising administering to a mammal, preferably a human, a therapeutically effective amount of the compound of any one of claims 1-19 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 20.

22. A method of treating tumors in mammals, comprising administering to a mammal in need of treatment, preferably a human, a therapeutically effective amount of the compound of any one of claims 1-19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 20.

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