Pyrazole derivative, pharmaceutically acceptable salt, stereoisomer, pharmaceutical composition and application thereof

Abstract

A pyrazole derivative having a structure represented by formula (I), a pharmaceutically acceptable salt, a stereoisomer, a pharmaceutical composition, and applications thereof. The compound has good activity inhibition and selectivity on CDK2. # imgabs0 #

Description

Pyrazole derivatives, pharmaceutically acceptable salts, stereoisomers, pharmaceutical compositions and applications thereof

[0001] This application claims priority to:

[0002] CN2023100596930, January 19, 2023. Technical Field

[0003] The present invention relates to the field of medical technology, and in particular to a pyrazole derivative, a pharmaceutically acceptable salt, a stereoisomer, a pharmaceutical composition and applications thereof. Background Art

[0004] Cyclin-dependent kinases (CDKs) belong to the serine / threonine kinase family. They participate in physiological processes such as cell proliferation and transcription. CDKs can be divided into two major categories based on their functions: one type, including CDK1, CDK2, CDK4, and CDK6, participates in cell cycle regulation; the other type, including CDK7, CDK8, CDK9, CDK12, and CDK13, participates in transcriptional regulation.

[0005] Dysregulated CDK2 activity frequently occurs in various human cancers, making it of particular interest to researchers. CDK2 plays a key role in promoting the G1 / S transition and S phase progression. CDK2 forms a complex with cyclin E, phosphorylating retinoblastoma family members (such as pRb), leading to the release and activation of E2F transcription factors, driving the transition from G1 to S phase of the cell cycle. This in turn activates CDK2 / cyclin A, promoting DNA synthesis and replication in the cell cycle.

[0006] Increased copy number and overexpression of Cyclin E1 have been identified in ovarian cancer, gastric cancer, endometrial cancer, breast cancer, and other cancers, and are positively correlated with poor prognosis in the corresponding tumors. In ER+ breast cancer cells, high expression of Cyclin E2 is often accompanied by resistance to hormone therapy, and amplification or overexpression of Cyclin E is closely related to poor prognosis in breast cancer. In HER2+ breast cancer, amplification of Cyclin E has also been reported to contribute to resistance to trastuzumab. There are also reports that overexpression of Cyclin E plays an important role in the progression of triple-negative breast cancer or inflammatory breast cancer. Therefore, CDK2 may become an important anti-tumor target.

[0007] Summary of the Invention

[0008] Based on this, the present invention provides a pyrazole derivative with good selectivity for inhibiting the activity of CDK2, and a pharmaceutically acceptable salt or stereoisomer thereof.

[0009] The present invention is achieved through the following technical solutions.

[0010] A pyrazole derivative represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0011] in:

[0012] Each occurrence of X is independently selected from N or CR4; R4 is absent or selected from -H, -D, -F, -Cl, -Br, -I, straight-chain alkyl having 1 to 6 C atoms, straight-chain deuterated alkyl having 1 to 6 C atoms, branched-chain alkyl having 3 to 6 C atoms, or an aromatic heterocycle;

[0013] R1 is selected from -H, -D, -F, -Cl, -Br, -I, linear alkyl having 1 to 20 C atoms, linear deuterated alkyl having 1 to 20 C atoms, branched alkyl having 3 to 20 C atoms, branched deuterated alkyl having 3 to 20 C atoms, cyclic alkyl having 3 to 20 C atoms, cyclic deuterated alkyl having 3 to 20 C atoms, linear alkoxy having 1 to 20 C atoms, linear deuterated alkoxy having 1 to 20 C atoms, branched alkoxy having 3 to 20 C atoms, branched deuterated alkoxy having 3 to 20 C atoms, cyclic alkoxy having 3 to 20 C atoms, cyclic deuterated alkoxy having 3 to 20 C atoms;

[0014] R3 is selected from substituted or unsubstituted alkyl having 1 to 6 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 6 C atoms, substituted or unsubstituted cycloalkyl having 3 to 7 C atoms, substituted or unsubstituted heterocyclyl having 4 to 7 ring atoms;

[0015] R2 is selected from -F, -Cl, -OH, -CN, -NR5R6, -CH2-ONR7R8, -CH2-ON=R9, -CH2N(R 10 )OR 11 , substituted or unsubstituted alkyl having 1 to 4 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 4 C atoms, substituted or unsubstituted alkoxy having 1 to 4 C atoms, substituted or unsubstituted fluoroalkoxy having 1 to 4 C atoms, substituted or unsubstituted cycloalkyl having 3 to 8 C atoms;

[0016] R5, R6, R7, R8, R 10 、R 11Each of the following groups is independently selected from -H, -D, a linear alkyl group having 1 to 20 C atoms, a linear deuterated alkyl group having 1 to 20 C atoms, a branched alkyl group having 3 to 20 C atoms, a branched deuterated alkyl group having 3 to 20 C atoms, a cyclic alkyl group having 3 to 20 C atoms, a cyclic deuterated alkyl group having 3 to 20 C atoms, a linear alkoxy group having 1 to 20 C atoms, a linear deuterated alkoxy group having 1 to 20 C atoms, a branched alkoxy group having 3 to 20 C atoms, a branched deuterated alkoxy group having 3 to 20 C atoms, a cyclic alkoxy group having 3 to 20 C atoms, and a cyclic deuterated alkoxy group having 3 to 20 C atoms;

[0017] R9 is selected from linear alkylene having 1 to 20 C atoms, linear deuterated alkylene having 1 to 20 C atoms, branched alkylene having 3 to 20 C atoms, branched deuterated alkylene having 3 to 20 C atoms, cyclic alkylene having 3 to 20 C atoms, cyclic deuterated alkylene having 3 to 20 C atoms, or a combination of these systems.

[0018] In one embodiment, the pyrazole derivative has a structure represented by formula (II):

[0019] In one embodiment, the pyrazole derivative has a structure represented by formula (III):

[0020] In one embodiment, R1 is selected from -H, -D, -F, -Cl, -Br, -I, a linear alkyl group having 1 to 10 C atoms, a linear deuterated alkyl group having 1 to 10 C atoms, a branched alkyl group having 3 to 10 C atoms, a branched deuterated alkyl group having 3 to 10 C atoms, a cyclic alkyl group having 3 to 10 C atoms, a cyclic deuterated alkyl group having 3 to 10 C atoms, or a combination of these systems.

[0021] In one embodiment, R1 is selected from -H, -D, -F, -Cl, -Br, a straight-chain alkyl group having 1 to 6 C atoms, a straight-chain deuterated alkyl group having 1 to 6 C atoms, a branched-chain alkyl group having 3 to 6 C atoms, a branched-chain deuterated alkyl group having 3 to 6 C atoms;

[0022] In one embodiment, R1 is selected from -H, -D, -F, -Cl, -Br, a straight-chain alkyl group having 1 to 3 C atoms, a straight-chain deuterated alkyl group having 1 to 3 C atoms, a branched-chain alkyl group having 3 to 6 C atoms, a branched-chain deuterated alkyl group having 3 to 6 C atoms;

[0023] In one embodiment, R1 is selected from -H, -D, -F, -CH3, -CH2CH3, -CH2CH2CH3 and -CH(CH3)2.

[0024] In one embodiment, R2 is selected from -F, -Cl, -OH, -CN, -NR5R6, -CH2-ONR7R8, -CH2-ON=R9, -CH2N(R 10 )OR 11 , substituted or unsubstituted alkyl having 1 to 4 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 4 C atoms, substituted or unsubstituted alkoxy having 1 to 4 C atoms, substituted or unsubstituted fluoroalkoxy having 1 to 4 C atoms, substituted or unsubstituted cycloalkyl having 3 to 8 C atoms;

[0025] In one embodiment, R3 is selected from substituted or unsubstituted alkyl having 1 to 3 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 3 C atoms, substituted or unsubstituted cycloalkyl having 3 to 7 C atoms, substituted or unsubstituted heterocyclyl having 4 to 7 ring atoms;

[0026] In one embodiment, R3 is selected from substituted or unsubstituted -CH3, substituted or unsubstituted -CH2CH3, substituted or unsubstituted -CH2CH2CH3 and substituted or unsubstituted -CH(CH3)2.

[0027] In one embodiment, R3 is selected from substituted or unsubstituted -CH(CH3)2. In one embodiment, R2 is selected from -F, -Cl, -OH, -CN, -NR5R6, -CH2-ONR7R8, -CH2-ON=R9, -CH2N(R 10 )OR 11 , substituted or unsubstituted -CH3, substituted or unsubstituted -CH2CH3, substituted or unsubstituted -CH2CH2CH3, substituted or unsubstituted -CH(CH3)2, substituted or unsubstituted fluoro-CH3, substituted or unsubstituted fluoro-CH2CH3, substituted or unsubstituted fluoro-CH2CH2CH3, substituted or unsubstituted fluoro-CH(CH3)2.

[0028] In one embodiment, R5, R6, R7, R8, R 10 、R 11Each of the following groups is independently selected from -H, -D, a linear alkyl group having 1 to 10 C atoms, a linear deuterated alkyl group having 1 to 10 C atoms, a branched alkyl group having 3 to 10 C atoms, a branched deuterated alkyl group having 3 to 10 C atoms, a cyclic alkyl group having 3 to 10 C atoms, a cyclic deuterated alkyl group having 3 to 10 C atoms, a linear alkoxy group having 1 to 10 C atoms, a linear deuterated alkoxy group having 1 to 10 C atoms, a branched alkoxy group having 3 to 10 C atoms, a branched deuterated alkoxy group having 3 to 10 C atoms, a cyclic alkoxy group having 3 to 10 C atoms, and a cyclic deuterated alkoxy group having 3 to 10 C atoms;

[0029] In one embodiment, R5, R6, R7, R8, R 10 、R 11 Each of the following groups is independently selected from -H, -D, a linear alkyl group having 1 to 6 C atoms, a linear deuterated alkyl group having 1 to 6 C atoms, a branched alkyl group having 3 to 6 C atoms, a branched deuterated alkyl group having 3 to 6 C atoms, a cyclic alkyl group having 3 to 6 C atoms, a cyclic deuterated alkyl group having 3 to 6 C atoms, a linear alkoxy group having 1 to 6 C atoms, a linear deuterated alkoxy group having 1 to 6 C atoms, a branched alkoxy group having 3 to 6 C atoms, a branched deuterated alkoxy group having 3 to 6 C atoms, a cyclic alkoxy group having 3 to 6 C atoms, and a cyclic deuterated alkoxy group having 3 to 6 C atoms;

[0030] In one embodiment, R5, R6, R7, R8, R 10 、R 11 Each of the following groups is independently selected from -H, -D, a linear alkyl group having 1 to 3 C atoms, a linear deuterated alkyl group having 1 to 3 C atoms, a branched alkyl group having 3 to 6 C atoms, a branched deuterated alkyl group having 3 to 6 C atoms, a cyclic alkyl group having 3 to 6 C atoms, a cyclic deuterated alkyl group having 3 to 6 C atoms, a linear alkoxy group having 1 to 3 C atoms, a linear deuterated alkoxy group having 1 to 3 C atoms, a branched alkoxy group having 3 to 6 C atoms, a branched deuterated alkoxy group having 3 to 6 C atoms, a cyclic alkoxy group having 3 to 6 C atoms, and a cyclic deuterated alkoxy group having 3 to 6 C atoms;

[0031] In one embodiment, R5, R6, R7, R8, R 10 、R 11 Each is independently selected from -H, -D, -CH3, -CH2CH3, -CH2CH2CH3 and -CH(CH3)2.

[0032] In one embodiment, R9 is selected from a linear alkylene group having 1 to 20 C atoms, a linear deuterated alkylene group having 1 to 20 C atoms, a branched alkylene group having 3 to 20 C atoms, a branched deuterated alkylene group having 3 to 20 C atoms, a cyclic alkylene group having 3 to 20 C atoms, and a cyclic deuterated alkylene group having 3 to 20 C atoms.

[0033] In one embodiment, R9 is selected from a linear alkylene group having 1 to 10 C atoms, a linear deuterated alkylene group having 1 to 10 C atoms, a branched alkylene group having 3 to 10 C atoms, a branched deuterated alkylene group having 3 to 10 C atoms, a cyclic alkylene group having 3 to 10 C atoms, and a cyclic deuterated alkylene group having 3 to 10 C atoms.

[0034] In one embodiment, R9 is selected from a linear alkylene group having 1 to 6 C atoms, a linear deuterated alkylene group having 1 to 6 C atoms, a branched alkylene group having 3 to 6 C atoms, a branched deuterated alkylene group having 3 to 6 C atoms, a cyclic alkylene group having 3 to 6 C atoms, and a cyclic deuterated alkylene group having 3 to 6 C atoms.

[0035] In one embodiment, R9 is selected from a linear alkylene group having 1 to 3 C atoms, a linear deuterated alkylene group having 1 to 3 C atoms, a branched alkylene group having 3 to 6 C atoms, a branched deuterated alkylene group having 3 to 6 C atoms, a cyclic alkylene group having 3 to 6 C atoms, and a cyclic deuterated alkylene group having 3 to 6 C atoms.

[0036] In one embodiment, R9 is selected from =CH2, =CHCH3, =CHCH2CH3 and =C(CH3)2.

[0037] In one embodiment, R2 is selected from any one of the following structures:

[0038] The present application provides the following compounds or pharmaceutically acceptable salts thereof;

[0039] In a specific embodiment, the pharmaceutically acceptable salt is an alkyl salt, and further, the pharmaceutically acceptable salt is a formate salt.

[0040] In a specific embodiment, the pharmaceutically acceptable salt is formate or hydrochloride.

[0041] The present invention also provides a use of the above-mentioned pyrazole derivatives, or pharmaceutically acceptable salts thereof, or stereoisomers thereof in the preparation of drugs for treating and / or preventing diseases related to or mediated by CDK2 activity.

[0042] In a specific embodiment, the disease associated with or mediated by CDK2 activity is cancer.

[0043] In a specific embodiment, the invention relates to one or more of ovarian cancer, gastric cancer, endometrial cancer and breast cancer.

[0044] The present invention also provides a pharmaceutical composition comprising the above-mentioned pyrazole derivative, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0045] It is understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0046] Compared with the prior art, the pyrazole derivatives of the present invention, or pharmaceutically acceptable salts thereof, or stereoisomers thereof have the following beneficial effects:

[0047] After long and in-depth research, the inventors unexpectedly discovered a novel pyrazole derivative. The pyrazole derivative of the present invention has unexpected inhibitory activity and selectivity against CDK2, and can be used to treat various cancers with high Cyclin E expression, especially with excellent therapeutic effects on cancer patients resistant to CDK4 / 6 inhibitors. DETAILED DESCRIPTION

[0048] To facilitate understanding of the present invention, the present invention will be described more fully below with reference to the relevant examples. Preferred embodiments of the present invention are provided in the examples. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and comprehensive understanding of the disclosure of the present invention.

[0049] Definition of terms

[0050] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0051] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0052] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, prepared by reacting a compound having specific substituents discovered by the present invention with a relatively non-toxic acid or base. When the compound of the present invention contains relatively acidic functional groups, base addition salts can be obtained by contacting the compound with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compound of the present invention contains relatively basic functional groups, acid addition salts can be obtained by contacting the compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts and organic acid salts, as well as salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0053] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0054] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0055] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0056] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0057] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0058] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0059] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond and straight dashed key

[0060] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers with different functional groups that are in dynamic equilibrium at room temperature and can quickly convert into each other. For example, in the present invention, and isomers. If tautomers are possible (such as in solution), a chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0061] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0062] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0063] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0064] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0065] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0066] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. The type and number of substituents can be any chemically feasible basis. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.

[0067] When any variable (e.g., R1) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 instances of R1, the group may optionally be substituted with up to two instances of R1, with each instance of R1 being an independent choice. Furthermore, combinations of substituents and / or their variants are permitted only if such combinations result in stable compounds.

[0068] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0069] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0070] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the substituent is listed without specifying which atom it is connected to the substituted group, the substituent can be bonded through any atom of the substituent. For example, a pyridyl substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0071] When the listed linking groups do not specify their linking direction, their linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0072] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0073] Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, a "3-7 membered ring" refers to a "ring" having 3-7 atoms arranged around it.

[0074] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-4 Alkyl groups, which may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, tert-butyl, and sec-butyl).

[0075] Unless otherwise specified, the term “C 1-6 "Alkoxy" means an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 1-3 Alkoxy. C 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0076] Unless otherwise specified, the term “C 1-6 "Alkylamino" means an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an amino group. 1-3 Alkylamino. C 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, and the like.

[0077]

[0046] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0078] Unless otherwise specified, the terms "5-membered heteroaryl ring" and "5-membered heteroaryl" are used interchangeably in the present invention. The term "5-membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). The 5-membered heteroaryl group may be attached to the rest of the molecule via a heteroatom or carbon atom. Examples of the 5-membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2 (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.).

[0079] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0080] Unless otherwise specified, “C 3-7 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 7 carbon atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. 3-7 Cycloalkyl groups include C 3-6 、C 4-6 、C 4-5 、C 5-7 or C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-7 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

[0081] Unless otherwise specified, the term "3-7 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 7 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). This includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings. In addition, with respect to the term "3-7 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The term "3-7 membered heterocycloalkyl" includes 5-7 membered, 3 membered, 4 membered, 5 membered, 6 membered and 7 membered heterocycloalkyls, etc. Examples of 3-7 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, etc.

[0082] Unless otherwise specified, the term "3-7 membered nitrogen-containing heterocycloalkyl" refers to a 3-7 membered heterocycloalkyl group containing at least one nitrogen atom.

[0083] Alicyclic refers to a saturated or partially unsaturated all-carbon ring system. Wherein "partially unsaturated" refers to a ring portion that includes at least one double bond or triple bond. "Partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein. Non-limiting examples include cyclopropyl rings, cyclobutyl rings, cyclopentyl rings, cyclopentenyl rings, cyclohexyl rings, cyclohexenyl rings, cyclohexadienyl rings, cycloheptyl rings, cycloheptatrienyl rings, cyclopentanone rings, cyclopentane-1,3-dione rings, and the like.

[0084] Alicyclic groups are saturated or partially unsaturated alicyclic groups in which one, two or three ring carbon atoms are selected from nitrogen, oxygen or S(O) t (wherein t is an integer from 0 to 2) is substituted by a heteroatom, excluding -OO-, -OS- or -SS- ring moieties, and the remaining ring atoms are carbon. Non-limiting examples include an oxetane ring, an azetidine ring, an oxetane ring, a tetrahydrofuran ring, a tetrahydrothiophene ring, a tetrahydropyrrole ring, a piperidine ring, a pyrroline ring, an oxazolidine ring, a piperazine ring, a dioxolane ring, a dioxane ring, a morpholine ring, a thiomorpholine ring, a thiomorpholine-1,1-dioxide, a tetrahydropyran ring, an azetidine-2-one ring, an oxetane-2-one ring, a pyrrolidine-2-one ring, a pyrrolidine-2,5-dione ring, a piperidin-2-one ring, a dihydrofuran-2(3H)-one ring, a dihydrofuran-2,5-dione ring, a tetrahydro-2H-pyran-2-one ring, a piperazin-2-one ring, and a morpholine-3-one ring. Non-limiting examples of partially unsaturated monocyclic heterocycles include 1,2-dihydroazetidine ring, 1,2-dihydrooxetadiene ring, 2,5-dihydro-1H-pyrrole ring, 2,5-dihydrofuran ring, 2,3-dihydrofuran ring, 2,3-dihydro-1H-pyrrole ring, 3,4-dihydro-2H-pyran ring, 1,2,3,4-tetrahydropyridine ring, 3,6-dihydro-2H-pyran ring, 1,2,3,6-tetrahydropyridine ring, 4,5-dihydro-1H-imidazole ring, 1,4,5,6-tetrahydropyrimidine ring, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazolidine ring, 1,6-dihydropyrimidine ring, 4,5,6,7-tetrahydro-1H-1,3-diazepine ring. Cyclic, 2,5,6,7-tetrahydro-1,3,5-oxadiazepine Ring, etc.

[0085] "Aryl" and "aromatic ring" are used interchangeably to refer to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, which may be fused to a cycloalkyl ring, a heterocycloalkyl ring, a cycloalkenyl ring, a heterocycloalkenyl ring, or a heteroaryl ring. 6-10 The term "aryl" refers to a monocyclic or bicyclic aromatic group having 6 to 10 carbon atoms. Non-limiting examples of aryl include phenyl, naphthyl, and the like.

[0086] "Heteroaryl" and "heteroaryl ring" are used interchangeably and refer to a group of a monocyclic, bicyclic or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic arrangement) having ring carbon atoms and ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur. In the present invention, heteroaryl also includes a ring system in which the above-mentioned heteroaryl ring is fused to one or more cycloalkyl rings, heterocycloalkyl rings, cycloalkenyl rings, heterocycloalkenyl rings or aromatic rings. The heteroaryl ring may be optionally substituted. "5 to 10 membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl group having 5 to 10 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include thienyl, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrolyl, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and tetrazinyl. "8- to 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 8 to 10 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms, non-limiting examples of which include indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, The term "heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as valence permits. Heteroaryl bicyclic ring systems may include one or more heteroatoms in one or both rings.

[0087] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; acyloxy groups such as acetoxy and trifluoroacetoxy, etc.

[0088] The term "protecting group" includes, but is not limited to, an "amino protecting group," a "hydroxy protecting group," or a "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions at the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0089] Unless otherwise defined, the "substituents independently selected from..." described in the present invention means that when more than one hydrogen on a group is replaced by a substituent, the substituents may be the same or different, and the substituents selected are independently of each other.

[0090] Generally, the compound of the present invention or its pharmaceutically acceptable salt, or its stereoisomer can be formed into a suitable dosage form for administration with one or more pharmaceutical carriers. These dosage forms are suitable for oral, rectal, topical, oral and other parenteral administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, dosage forms suitable for oral administration include capsules, tablets, granules and syrups. The compound of the present invention contained in these preparations can be a solid powder or granules; a solution or suspension in an aqueous or non-aqueous liquid; an oil-in-water or water-in-oil emulsion, etc. The above dosage forms can be made from the active compound and one or more carriers or excipients through common pharmaceutical methods. The above carriers need to be compatible with the active compound or other excipients. For solid preparations, commonly used non-toxic carriers include but are not limited to mannitol, lactose, starch, magnesium stearate, cellulose, glucose, sucrose, etc. Carriers for liquid preparations include water, physiological saline, aqueous glucose solution, ethylene glycol and polyethylene glycol, etc. The active compound can form a solution or suspension with the above carriers.

[0091] The compositions of the present invention are formulated, dosed and administered in a manner consistent with standard medical practice. The "therapeutically effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0092] "Therapeutically effective amount" refers to the amount of the compound of the present invention that will elicit a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0093] The therapeutically effective amount of the compound of the present invention or its pharmaceutically acceptable salt, or its stereoisomer contained in the pharmaceutical composition or pharmaceutical use composition of the present invention is preferably 0.1 mg-5 g / kg (body weight).

[0094] "Patient" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

[0095] "Treatment" refers to alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0096] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0097] The structures of the compounds of the present invention can be confirmed using conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed using conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data from a cultured single crystal using a Bruker D8venture diffractometer, using CuKα radiation as the light source and φ scanning mode. After collecting relevant data, the crystal structure can be further analyzed using a direct method (Shelxs97) to confirm the absolute configuration.

[0098] The solvents used in the present invention are commercially available. The present invention uses the following abbreviations: N2 represents nitrogen; DMSO represents dimethyl sulfoxide; Pd(dppf)Cl2 represents 1,1'-bis(diphenylphosphinoferrocene)palladium dichloride; DIEA represents N,N-diisopropylethylamine; THF represents tetrahydrofuran; EtOAc represents ethyl acetate; FA represents formic acid; TFA represents trifluoroacetic acid; Select-F represents selective fluorine reagent; tBuOK represents potassium tert-butoxide; HCl represents hydrochloric acid; and MeOH represents methanol.

[0099] Compounds are named according to the conventional nomenclature in the art or using Software naming, commercially available compounds use supplier catalog names.

[0100] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. The terms used herein in the specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items. BRIEF DESCRIPTION OF THE DRAWINGS

[0101] FIG1 is a graph showing the results of Experimental Example 7.

[0102] DETAILED DESCRIPTION

[0103] The pyrazole derivatives and their preparation methods of the present invention are further described in detail below with reference to specific examples. Unless otherwise specified, the raw materials used in the following examples are all commercially available products.

[0104] Example 1

[0105] This embodiment provides compound 1, whose structural formula is as follows:

[0106] The reaction route is as follows:

[0107] Step 1:

[0108] Compound 1-2 (186 g, 1.77 mol, 192 mL) was dissolved in tetrahydrofuran (700 mL) at room temperature and cooled to 0°C. Under nitrogen, trimethylaluminum (2.00 M, 883 mL) was added. After stirring at 25°C for 1 hour, the reaction system was cooled to 0°C and compound 1-1 (250 g, 1.18 mol) was dissolved in tetrahydrofuran (500 mL) and added. The air was replaced with nitrogen three times, and the reaction was continued at 25°C under nitrogen for 4 hours. The temperature was then lowered to 0°C, and methanol (1.00 L) was slowly added dropwise to the reaction system. After reacting at 20°C for 2 hours, the trimethylaluminum was completely quenched. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain compound 1-3. MS (ESI) m / z = 239.9 [M+H] + .

[0109] Step 2:

[0110] At room temperature, compound 1-3 (260 g, 958 mmol) was dissolved in water (500 mL), cooled to 0°C, and hydrochloric acid (12 M, 479 mL) was added. The mixture was reacted at 25°C for 4 hours. The mixture was filtered and the filter cake was dried to obtain compound 1-4. MS (ESI) m / z = 226.1 [M+H] + Step 3:

[0111] Compound 1-4 (50.0 g, 222 mmol) was dissolved in 400 g, 4.16 mol, 296 mL of methanesulfonic acid at room temperature and allowed to react at 25°C for 10 hours. The reaction solution was slowly poured into saturated sodium bicarbonate solution (500 mL) and extracted six times with ethyl acetate (200 mL x 6). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 1-5. MS (ESI) m / z = 207.7 [M+H] + Step 4:

[0112] At room temperature, compound 1-5 (11.0 g, 53.1 mmol) and N,N-diisopropylethylamine (6.86 g, 53.1 mmol, 9.25 mL) were dissolved in toluene (100 mL). The temperature was lowered to 0°C. Under nitrogen, phosphorus oxychloride (16.3 g, 106 mmol, 9.87 mL) was added and stirred at 120°C for 10 hours. The mixture was concentrated, excess toluene was removed, and the mixture was extracted three times with ethyl acetate (100 mL x 3). The organic phases were combined, washed twice with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain compound 1-6. MS (ESI) m / z = 225.7 [M+H] + . 1 H NMR:DMSO-d6,,400MHz,δppm 8.94 (dd, J=4.8, 0.80Hz, 1H), 7.92 (d, J=4.75Hz, 1H), 7.46 (d, J=0.80Hz, 1H), 4.39 (q, J=7.2Hz, 2H), 1.35 (t, J=7.13Hz, 3H).

[0113] Step 5:

[0114] Compound 1-6 was dissolved in tetrahydrofuran (100 mL) and the air was replaced with nitrogen three times. The system was cooled to -68°C and diisobutylaluminum hydride (1M, 75.3 mL) was slowly added under nitrogen. The reaction was allowed to react at 25°C for 1 hour. Methanol (40.0 mL) was added to the reaction solution at 0°C. The reaction was stirred at 25°C for 2 hours, then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-7. MS (ESI) m / z = 183.7 [M+H] + . 1 H NMR: DMSO-d6, 400MHz δppm 8.77 (d, J = 4.8 Hz, 1H), 7.72 (d, J = 4.4 Hz, 1H), 6.92 (s, 1H), 4.70 (d, J = 6.0 Hz, 2H).

[0115] Step 6:

[0116] Compound 1-7 was dissolved in N,N-dimethylformamide (50.0 mL), the air was replaced with nitrogen three times, and the temperature was lowered to -5°C. Under nitrogen, iodomethane (24.5 g, 173 mmol, 10.7 mL) and sodium hydride (1.27 g, 31.7 mmol, 60% purity) were added to the reaction solution in sequence and reacted at -5°C for 1 hour. The reaction solution was slowly poured into saturated ammonium chloride solution (30.0 mL) and extracted three times with ethyl acetate (20.0 mL x 3). The organic phases were combined, washed once with saturated brine (20.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-8. MS (ESI) m / z = 197.7 [M+H] + . 1 H NMR: DMSO-d6, 400MHz δppm 8.81 (d, J = 4.8 Hz, 1H), 7.75 (d, J = 4.4 Hz, 1H), 6.99 (s, 1H), 4.64 (s, 2H), 3.34 (s, 3H).

[0117] Step 7:

[0118] Tetrabutylammonium fluoride (21.7 g, 83.3 mmol) was dissolved in N-methylpyrrolidone (25 mL) at room temperature and heated to 130°C for 1 hour. Compound 1-8 (5.49 g, 27.7 mmol) was stirred at 130°C for 3 hours under N2 protection. The reaction solution was poured into ice water (200 mL) and extracted three times with ethyl acetate (50 mL x 3). The organic phases were combined, washed twice with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain compound 1-9.

[0119] MS (ESI) m / z = 181.7 [M+H] + . 1 H NMR: CDCl3, 400MHz δppm 8.24 (d, J = 4.8 Hz, 1H), 7.41 (d, J = 4.4 Hz, 1H), 6.88 (s, 1H), 4.72 (s, 2H), 3.48 (s, 3H).

[0120] Step 8:

[0121] Compound 1-9 (1.00 g, 5.52 mmol) and N-iodosuccinimide (2.48 g, 11.0 mmol) were dissolved in acetonitrile (10.0 mL) at room temperature and stirred at 25°C for 20 hours. The mixture was poured into water (10.0 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-10. MS (ESI) m / z = 307.8 [M+H] + .

[0122] Step 9:

[0123] Compound 1-10 (3.20 g, 10.4 mmol) was dissolved in dioxane (20.0 mL). 50% trimethylboroxine in tetrahydrofuran (3.5 M, 29.7 mL) and potassium carbonate (4.32 g, 31.2 mmol) were added sequentially. The air was replaced with nitrogen three times. Pd(dppf)Cl2 (762 mg, 1.04 mmol) was added under nitrogen. The reaction was incubated at 90°C for 10 hours. The mixture was poured into water (20.0 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (10.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain compound 1-12. MS (ESI) m / z = 196.1 [M+H] + .

[0124] Step 10:

[0125] Compound 1-13 (250 g, 1.76 mol) was dissolved in anhydrous methanol (1.00 L). Trimethyl orthoformate (1.31 kg, 12.3 mol, 1.35 L) and p-toluenesulfonic acid (6.06 g, 35.2 mmol) were added sequentially. The mixture was allowed to react at 25°C for 30 hours. The reaction solution was slowly poured into saturated aqueous sodium bicarbonate (300 mL). The mixture was concentrated, excess methanol was removed, and the mixture was extracted twice with ethyl acetate (200 mL x 2). The organic phases were combined, washed twice with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-15.

[0126] Step 11:

[0127] Acetonitrile (139 g, 3.40 mol, 179 mL) was dissolved in tetrahydrofuran (700 mL) at room temperature and cooled to -68°C. Under nitrogen, a 2.5 M solution of n-butyllithium in n-hexane (2.50 M, 1.36 L) was added. The mixture was stirred at -68°C for 1 hour. Compound 1-15 (320 g, 1.70 mol) was dissolved in tetrahydrofuran (300 mL) and added. The air was replaced with nitrogen three times. The mixture was reacted at -68°C under nitrogen for 1 hour. The reaction mixture was slowly poured into ice water (2.00 L), the pH was adjusted to 7 with 1 M aqueous hydrochloric acid, and the mixture was extracted three times with ethyl acetate (1.50 L x 3). The organic phases were combined, washed twice with saturated sodium chloride (700 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 1-16. 1 H NMR: CDCl3, 400MHz δppm 3.52 (s, 2H), 3.20 (d, J = 2.40Hz, 6H), 2.08 (d, J = 8.40Hz, 3H), 1.97-2.04 (m, 1H), 1.83-1.92 (m, 3H).

[0128] Step 12:

[0129] At room temperature, sodium hydroxide (73.0 g, 1.83 mol) was dissolved in ethanol (500 mL). Under N2 protection, tert-butylhydrazine salt (227 g, 1.83 mol) was added. The mixture was stirred at 25°C for 1 hour. Compound 1-16 (320 g, 1.70 mol) was dissolved in ethanol (250 mL) and added. The air was replaced with N2 three times. The reaction was continued at 75°C under N2 conditions for 15 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain compound 1-18.

[0130] Step 13:

[0131] Compound 1-18 (425 g, 1.59 mol) was dissolved in acetonitrile (1.00 L) at room temperature. Benzyl chloroformate (542 g, 3.18 mol, 452 mL) was added under N2 protection and stirred at 25°C for 2 hours. Sodium bicarbonate (401 g, 4.77 mol, 185 mL) was added to the reaction system. The air was replaced with N2 three times and the reaction was continued at 25°C under N2 conditions for 11 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain compound 1-19. MS (ESI) m / z = 356.1 [M+H] + .

[0132] Step 14:

[0133] At room temperature, compound 1-19 (758 g, 1.89 mol) and p-toluenesulfonic acid (39.0 g, 226 mmol) were dissolved in water (1.00 L) and acetone (1.00 L). The air was replaced with N2 three times and stirred at 60°C for 10 hours. The mixture was concentrated, excess acetone was removed, and the mixture was extracted three times with dichloromethane (300 mL x 3). The organic phases were combined, washed twice with saturated brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain compound 1-20. MS (ESI) m / z = 356.0 [M+H] + .

[0134] Step 15:

[0135] At room temperature, compound 1-20 (177 g, 497 mmol) was dissolved in tetrahydrofuran (900 mL). The temperature was lowered to -68°C. Under nitrogen, a 1M solution of lithium triethylborohydride in tetrahydrofuran (996 mL) was added and stirred at -68°C for 1.5 hours. The temperature was then lowered to -30°C, and saturated aqueous sodium bicarbonate (900 mL) was slowly added dropwise to the reaction system. The temperature was then lowered to -10°C, and hydrogen peroxide (362 g, 3.20 mol, 307 mL, 30% purity) was slowly added dropwise to the reaction system. After reacting at 10°C for 1 hour, the lithium triethylborohydride and 1M solution of tetrahydrofuran were completely quenched. After the reaction was complete, the mixture was extracted with ethyl acetate (1000 mL x 3). The combined organic phases were then slowly poured into a saturated sodium sulfite solution under stirring, paying attention to the exothermic reaction. The oxidizing property was tested using moistened starch potassium iodide test paper to ensure that the test paper did not turn blue. The organic phase was then washed twice with aqueous sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was extracted three times with ethyl acetate (1.00 L x 3). The organic phases were combined, washed twice with saturated aqueous sodium chloride solution (700 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain the crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 1:100) to obtain the desired product. The product was separated by SFC (column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 μm); mobile phase: [CO₂-MeOH]; B%: 35%, isocratic elution) to obtain compound 1-21.

[0136] Compound 1-21: SFC (column: Chiralpak AD-3 50×4.6 mm ID, 3 μm; mobile phase: phase A: CO 2 , phase B: MeOH (0.05% DEA); elution gradient: 5% to 40% B in A; flow rate: 3 mL / min; detector: PDA; column temperature: 35°C; back pressure: 100 Bar), RT = 1.158 min;

[0137] MS (ESI) m / z = 358.7 [M+H] + . 1H NMR:DMSO-d6,400MHzδppm 9.06(s,1H),7.27-7.46(m,5H),5.92(s,1H),5.12(s,2H),4.56(d,J=4.4Hz,1H),4.10-4.19(m,1H),2.89(t,J=8.0Hz ,1H),2.14-2.23(m,1H),1.80-1.91(m,1H),1.65-1.78(m,2H),1.52-1.62(m,2H),1.48(s,9H),1.04(d,J=6.2Hz,1H).

[0138] Step 16:

[0139] Compound 1-21 (43.0 g, 120 mmol) was dissolved in tetrahydrofuran (100 mL), and triethylamine (36.5 g, 360 mmol, 50.2 mL) and isopropyl isocyanate (40.9 g, 481 mmol, 47.2 mL) were added in sequence. The mixture was reacted at 80°C for 10 hours. The reaction solution was slowly poured into water (100 mL) and extracted twice with dichloromethane (100 mL x 2). The organic phases were combined, washed twice with saturated brine (50.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (100-200 mesh silica gel, petroleum ether:ethyl acetate = 100:1 to 2:1). Compound 1-23 was obtained. MS (ESI) m / z = 443.8

[0140] [M+H] + .

[0141] 1 H NMR:DMSO-d6,400MHzδppm 9.07(s,1H),7.23-7.46(m,5H),6.92(d,J=8.0Hz,1H),5.93(s,1H),5.11(s,2H),3.55-3.62(m,1H),2.90-2.99(m,1H),2.36 (t,J=16,8.0Hz,1H),1.89-1.97(m,1H),1.84(td,J=8.0,4.4Hz,1H),1.56-1.75(m,4H),1.47(s,9H),1.02(d,J=6.6Hz,6H).

[0142] Step 17:

[0143] Compound 1-23 (32.0 g, 72.3 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL). The atmosphere was replaced with nitrogen three times, and wet palladium on carbon (4.00 g, 50% purity) was added. The nitrogen atmosphere was replaced with hydrogen (15 psi) three times. The reaction was continued at 25°C under hydrogen (15 psi) for 2 hours. The reaction solution was filtered and the filtrate was concentrated to obtain compound 1-24. MS (ESI) m / z = 309.3 [M+H] + .

[0144] 1 H NMR:CDCl 3, 400MHzδppm 6.90(d,J=8.0Hz,1H),5.22(s,1H),4.70(s,2H),3.52-3.60(m,1H),2.78(s,1 H), 2.25-2.33 (m, 1H), 1.51-1.89 (m, 6H), 1.47 (s, 9H), 1.03 (d, J = 6.4Hz, 6H).

[0145] Step 18:

[0146] At room temperature, compound 1-24 (1.10 g, 3.57 mmol) was dissolved in tetrahydrofuran (10.0 mL), the temperature was controlled at -20°C, and lithium bis(trimethylsilyl)amide (1 M, 10.7 mL, 3.00 eq) was added to the mixture in a 1.0 M tetrahydrofuran solution (1 M, 10.7 mL, 3.00 eq) under N2 protection. The mixture was stirred at -20°C for 1 hour. Compound 1-12 (730 mg, 3.74 mmol) was dissolved in tetrahydrofuran (5.00 mL) and added. The air was replaced with N2 three times, and the mixture was reacted at -20°C under N2 conditions for 1 hour. The reaction solution was poured into saturated aqueous ammonium chloride (10.0 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 1-25. MS (ESI) m / z = 484.2 [M+H] + .

[0147] Step 19:

[0148] Compound 1-25 (1.00 g, 2.07 mmol) was dissolved in anhydrous formic acid (5.00 mL). The mixture was reacted at 100°C for 0.5 hours. The reaction solution was concentrated to remove formic acid to obtain compound 1-26. MS (ESI) m / z = 428.2 [M+H] + .

[0149] Step 20:

[0150] Compound 1-26 (160 mg, 374 μmol) was dissolved in dichloromethane (2.00 mL) at room temperature and the temperature was controlled at -70°C. Boron tribromide (187 mg, 748 μmol, 72.1 μL) was added under N2 protection and stirred at 20°C for 10 hours. The reaction solution was poured into an ice-water solution (5.00 mL) and extracted three times with dichloromethane (3.00 mL x 3). The organic phases were combined, washed twice with saturated brine (3.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-27. MS (ESI) m / z = 478.0 [M+H] + Step 21

[0151] At room temperature, DIEA (157 mg, 1.22 mmol, 212 μL) was added to a solution of compound 1-27 (44.7 mg, 93.9 μmol) and compound 1-28 (44.1 mg, 939 μmol) in THF (1.00 mL) and stirred at 50°C for 5 hours. The reaction system was diluted with 5.0 mL of H2O and extracted three times with 15.0 mL (5.0 mL*3) of EtOAc. The mixture was then washed three times with 15.0 mL (5.0 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The product was purified by high performance liquid chromatography (FA conditions: column: Phenomenexluna C18 150*25.0 mm*10.0 μm; mobile phase: [water (FA) ACN]; gradient: 46.0%-76.0% B over 10.0 min) to obtain the formate salt of compound 1. MS (ESI) m / z = 443.1 [M+H] + . 1 H NMR: DMSO-d6,400MHzδ=12.28-12.13(m,1H),11.72-11.52(m,1H),8.27(br s,1H),8.07-7.98(m,1H),7.34(dd,J=5.6,11.8Hz,1H),7.31-7.30(m,1H), 7.05-7.00(m,1H),6.83-6.75(m,1H),5.97-5.88(m,1H),5.12-4.99(m,2H), 4.14-4.12(m,1H),4.04(d,J=6.4Hz,1H),3.71-3.57(m,2H),3.41(s,4H),3. 21-3.09(m,2H),2.60(s,3H),2.13-2.08(m,1H),2.00-1.95(m,1H),1.71(br s, 4H), 1.12-1.07 (m, 10H).

[0152] Example 2

[0153] This embodiment provides compound 2, whose structural formula is as follows:

[0154] The reaction route is as follows:

[0155] Step 1:

[0156] Compound 2-1 (245 mg, 2.52 mmol) and N,N-diisopropylethylamine (406 mg, 3.15 mmol, 548 μL) were dissolved in tetrahydrofuran (2.00 mL). The air was replaced with nitrogen three times. Compound 1-27 (150 mg, 314 μmol) was dissolved in tetrahydrofuran (1.00 mL) and added. The mixture was reacted at 25°C for 10 hours and extracted three times with ethyl acetate (10.0 mL x 2). The organic phases were combined, washed twice with saturated brine (10.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by reverse-phase HPLC (0.1%, formic acid conditions) to obtain the formate salt of compound 2. MS (ESI) m / z = 457.1 [M+H] + . 1 H NMR:DMSO-d6,400MHzδppm12.18(br s,1H),11.59(s,1H),7.99(d,J=4.8Hz,1H),7.29(dd,J=5.2,16.8Hz,1H),7.01-6.91(m,1H),5.88(s,1H),5.01(br s,1H),3.93(s,1H),3.84(s,1H),3.58(br d,J=6.8Hz,1H),3.31(s,3H),3.09(brdd,J=1.6,6.4Hz,1H),2.56(s,3H),2.53(br s,3H),2.08-2.02(m,1H),1.95-1.86(m,1H),1.75-1.64(m,4H),1.05-1.02(m,6H).

[0157] Example 3

[0158] This example provides compound 3, whose structural formula is as follows:

[0159] The reaction route is as follows:

[0160] Step 1:

[0161] Compound 1-27 (350 mg, 657 μmol) was dissolved in formic acid (3.00 mL) at room temperature. The reaction was incubated at 90°C for 30 minutes. The reaction solution was concentrated to obtain a crude product. The crude product was purified by HPLC (preparative column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (TFA)-ACN]; gradient: 24%-54% B over 10 min) to obtain compound 3-1. MS (ESI) m / z = 478.0 (M+H) + .

[0162] Step 2:

[0163] At room temperature, compound 3-2 (154 mg, 1.50 mmol) was dissolved in tetrahydrofuran (1.00 mL), cooled to 0°C, and sodium hydride (10.5 mg, 262 μmol, 60% purity) was added and stirred at 0°C for 30 minutes. Compound 3-1 (50.0 mg, 52 μmol, TFA) was then dissolved in tetrahydrofuran (0.50 mL) and slowly added dropwise to the reaction mixture at 0°C. After the addition was complete, the mixture was heated to 25°C and allowed to react for 30 minutes. The reaction mixture was poured into ice water (1.00 mL) and extracted three times with ethyl acetate (1.00 mL x 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (1.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain the crude product. The crude product was purified by HPLC (preparative column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (TFA)-ACN]; gradient: 27%-57% B over 10min) to obtain compound 3-3. MS (ESI) m / z = 543.7 (M+H) + .

[0164] Step 3:

[0165] Compound 3-3 (28.0 mg, 42.6 μmol, TFA) was dissolved in HCl / dioxane (1.00 mL) at room temperature. The reaction was allowed to proceed at 25°C for 30 minutes. The reaction solution was directly concentrated, dissolved in methanol, and dried to obtain the hydrochloride salt of compound 3. m / z = 443.1 (M+H) + . 1H NMR: (400MHz METHANOL-d4)δ=8.23(d,J=5.6Hz,1H),7.49(d,J=5.6Hz,1H),6.40(s,1H),5.45(s,2H),5.12(br s,1H),3.76-3.65(m,1H),3.29-3.22(m,1H),3.06(s,3H),2.78(s,3H),2. 69-2.57(m,1H),2.29-2.14(m,1H),2.06-1.77(m,4H),1.17-1.08(m,6H).

[0166] Example 4

[0167] This example provides compound 4, whose structural formula is as follows:

[0168] The reaction route is as follows:

[0169] Step 1:

[0170] Compound 1-9 (500 mg, 2.76 mmol) and Select-F (2.93 g, 8.28 mmol) were dissolved in acetonitrile (10.0 mL) at room temperature and stirred at 25°C for 20 hours. The mixture was poured into water (10.0 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4-1. MS (ESI) m / z = 199.9 [M+H] + .

[0171] Step 2:

[0172] At room temperature, compound 1-24 (800 mg, 2.59 mmol) was dissolved in tetrahydrofuran (8.00 mL), the temperature was controlled at -20°C, and lithium bis(trimethylsilyl)amide, a 1.0 M solution in tetrahydrofuran (7.78 mL) was added under N2 protection and stirred at -20°C for 1 hour. Compound 4-1 (516 mg, 2.59 mmol) was dissolved in tetrahydrofuran (5.00 mL) and added. The air was replaced with N2 three times, and the mixture was reacted at -20°C under N2 conditions for 1 hour. The reaction solution was poured into saturated aqueous ammonium chloride (10.0 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4-2. MS (ESI) m / z = 488.1 [M+H] + .

[0173] Step 3:

[0174] Compound 4-2 (1.12 g, 2.30 mmol) was dissolved in anhydrous formic acid (5.00 mL). The mixture was reacted at 100°C for 0.5 h. The reaction solution was concentrated to remove formic acid to obtain a crude product. The crude product was purified by reverse-phase HPLC (0.1% formic acid) to obtain compound 4-3. MS (ESI) m / z = 432.0 [M+H] + .

[0175] Step 4:

[0176] At room temperature, compound 4-3 (315 mg, 730 μmol) was dissolved in dichloromethane (3.00 mL), the temperature was controlled to -70°C, and boron tribromide (365 mg, 1.46 mmol, 140 μL) was added under N2 protection and stirred at 20°C for 10 hours. The reaction solution was poured into an ice-water solution (5.00 mL) and extracted three times with dichloromethane (3.00 mL x 3). The organic phases were combined, washed twice with saturated brine (3.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4-4. MS (ESI) m / z = 481.9 [M+H] + .

[0177] Step 5:

[0178] To a solution of compound 4-4 (150 mg, 312 μmol) and compound 4-5 (415 mg, 3.12 mmol) in THF (1.00 mL) was added tBuOK (105 mg, 936 μmol) and stirred at 0°C for 10 min. The reaction system was diluted with 10.0 mL of H2O and extracted three times with 15.0 mL (5.0 mL*3) of EtOAc. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 4-6. MS (ESI) m / z = 533.2 [M+H] + .

[0179] Step 6:

[0180] Compound 4-6 (160 mg, 300 μmol) was stirred in HCl / ethyl acetate (1.00 mL) for 1 hour. 10.0 mL of H2O was added to the reaction system for dilution, and the mixture was extracted three times with 15.0 mL (5.0 mL*3) of EtOAc. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, compound 4-7. MS (ESI) m / z = 433.0 [M+H] + .

[0181] Step 7:

[0182] Paraformaldehyde (90.2 mg, 3.01 mmol, 82.8 μL) was added to a solution of compound 4-7 (130 mg, 300 μmol) in MeOH (0.80 mL) and H2O (0.20 mL) and stirred at 25°C for 1 hour. 10.0 mL of H2O was added to the reaction system for dilution, and the mixture was extracted three times with 15.0 mL (5.00 mL*3) of EtOAc. The mixture was then washed three times with 15.0 mL (5.00 mL*3) of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The product was purified by HPLC (FA conditions: column: Phenomenexluna C18 150*25 mm*10 μm; mobile phase: [water(FA)-ACN]; gradient: 21.0%-51.0% B over 10 min) to obtain compound 4. MS (ESI) m / z = 445.0 [M+H] + . 1 H NMR: DMSO-d6, 400MHzδ=12.21-12.09(m,1H),8.02-7.95(m,1H),7.42-7.34(m,1H),7.15-7.07(m,1H),7.02-6.90(m,1H),6.65(br d,J=7.2Hz,1H),5.28-5.18(m,2H),5.17-4.95(m,2H),4.92-4.92(m,1H),3.63-3.52(m,1H),3.30(br s,1H),3.13-3.04(m,2H),2.08-2.01(m,1H),1.94-1.87(m,1H),1.79-1.60(m,4H),1.03(br d,J=5.2Hz,6H).

[0183] Example 5

[0184] This example provides compound 5, whose structural formula is as follows:

[0185] The reaction route is as follows:

[0186] Step 1:

[0187] Compound 5-1 (139 mg, 1.67 mmol, 126 μL) and N,N-diisopropylethylamine (565 mg, 4.37 mmol, 761 μL) were dissolved in tetrahydrofuran (2.00 mL). The air was replaced with nitrogen three times. Compound 4-4 (100 mg, 208 μmol) was dissolved in tetrahydrofuran (1.00 mL) and added. The mixture was reacted at 25°C for 10 hours. The reaction solution was poured into ice water (5.00 mL) to quench the mixture and extracted three times with ethyl acetate (5.00 mL x 2). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by reverse-phase HPLC (0.1%, formic acid) to obtain compound 5. MS (ESI) m / z = 447.2 [M+H] + . 1 H NMR: DMSO-d6,400MHzδppm12.24-12.12(m,1H),8.61-8.43(m,1H),7.96(br d,J=7.0Hz,1H),7.39-7.28(m,1H),7.00-6.93(m,1H),6.91-6.88(m,1H),6.53-6.42(m,1H),5.01(br s,1H),4.09(d,J=6.0Hz,2H),3.61-3.54(m,1H),3.36(s,3H),3.11-3.04(m,1H ),2.07-2.00(m,1H),1.95-1.86(m,1H),1.80-1.68(m,4H),1.05-1.01(m,6H).

[0188] Example 6

[0189] This example provides compound 6, whose structural formula is as follows:

[0190] The reaction route is as follows:

[0191] Step 1:

[0192] Compound 2-1 (162 mg, 1.67 mmol) and N,N-diisopropylethylamine (269 mg, 2.08 mmol, 362 μL) were dissolved in tetrahydrofuran (2.00 mL). The air was replaced with nitrogen three times. Compound 4-4 (100 mg, 208 μmol) was dissolved in tetrahydrofuran (1.00 mL) and added. The mixture was reacted at 25°C for 10 hours. The reaction solution was poured into ice water (5.00 mL) to quench the mixture and extracted three times with ethyl acetate (5.00 mL x 2). The organic phases were combined, washed twice with saturated brine (5.00 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by reverse-phase HPLC (0.1%, formic acid conditions) to obtain compound 6. MS (ESI) m / z = 461.1 [M+H] + . 1 H NMR:DMSO-d6,400MHzδppm 12.24-12.09(m,1H),8.59-8.45(m,1H),8.02-7.92(m,1H),7.34(br d,J=4.8Hz,1H),6.96(br d,J=8.4Hz,1H),6.49(br s,1H),5.07-4.92(m,1H),3.98(s,2H),3.58(br dd,J=6.8,12.0Hz,1H),3.33-3.33(m,3H),3.10(br s,1H),2.60-2.55(m,3H),2.11-1.99(m,1H),1.96-1.85(m,1H),1.80-1.60(m,4H),1.03(dd,J=2.4,6.4Hz,6H).

[0193] Example 7

[0194] This example provides compound 7, whose structural formula is as follows:

[0195] The reaction route is as follows:

[0196] Step 1:

[0197] At room temperature, compound 3-2 (367 mg, 2.50 mmol) was dissolved in tetrahydrofuran (1.00 mL). The atmosphere was replaced with nitrogen three times, the temperature was lowered to 0°C, sodium hydride (14.9 mg, 374 μmol, 60% purity) was added, and the mixture was stirred for 30 minutes. Compound 4-4 (60.0 mg, 124 μmol) was then dissolved in tetrahydrofuran (1.00 mL) and slowly added dropwise to the reaction mixture. The temperature was raised to 20°C and the reaction mixture was allowed to react for 30 minutes. The reaction mixture was poured into ice water (1.00 mL) and extracted three times with ethyl acetate (1.00 mL x 3). The organic phases were combined, washed with saturated sodium chloride aqueous solution (1.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by HPLC (preparative column: Welch Xtimate C18 150*25mm*5.0μm; mobile phase: [water(FA)-ACN]; gradient: 32.0%-62.0% B over 10min) to obtain compound 7-1. m / z=547.5 (M+H) + .

[0198] Step 2:

[0199] Compound 7-1 (35.0 mg, 59.0 μmol) was dissolved in HCl / EtOAc (0.50 mL) at room temperature. The reaction was allowed to proceed at 25°C for 30 minutes. The reaction solution was directly concentrated and dissolved in methanol, diluted with deionized water, and lyophilized to obtain the hydrochloride salt of compound 7. m / z = 447.0 (M+H) + . 1 H NMR: (400MHz METHANOL-d4)δ8.17(d,J=4.8Hz,1H),7.52(d,J=5.6Hz,1H),6.34(s,1H),5.40(s,2H),5.12(br s,1H),3.75-3.65(m,1H),3.03(s,3H),2.68-2.57(m,1H),2.25-2.16(m,1H),2.11-1.88(m,4H),1.82(br d,J=8.4Hz,1H),1.14-1.10(m,6H).

[0200] In vitro activity test

[0201] Experimental Example 1: In vitro CDK2 / CyclinE enzyme activity test

[0202] Experimental Materials:

[0203] CDK2 / Cyclin E1 was purchased from Syngenecon. Ulight-4E-BP1 peptide, Eu-anti-phospho-tyrosine antibody, and 1X detection buffer were purchased from PerkinElmer. High-purity ATP was purchased from Promega. EDTA was purchased from Sigma. Nivo Multilabel Analyzer was used (PerkinElmer).

[0204] Experimental methods:

[0205] Preparation of kinase buffer: Kinase buffer contains 50mM HEPES, 1mM EDTA, 10mM MgCl2, 0.01% Brij-35, pH 7.4. Add 2.38g HEPES, 58mg EDTA, 406mg MgCl2, 20mg Brij-35 to 200ml buffer and adjust the pH to 7.4.

[0206] Preparation of stop solution:

[0207] Prepare the stop solution by adding 100 μL of 1M EDTA stock solution, 0.625 μL of 1X detection buffer, and 1725 μL of distilled water.

[0208] Use kinase buffer to dilute the enzyme, Ulight-4E-BP1 peptide, ATP, and inhibitor. Use detection buffer to dilute the Eu-anti-phospho-tyrosine antibody to a concentration of 8nM / L. Use a pipette to dilute the test compound 5-fold to the eighth concentration, that is, from 4μM to 0.0512nM, with a final DMSO concentration of 4%, and set up a duplicate well experiment. Add 2.5μL of each inhibitor concentration gradient, 5μL of LCDK2 / CyclinE 1 enzyme (10ng), and 2.5μL of a mixture of substrate and ATP (4mM ATP, 100nM Ulight-4E-BP1 peptide) to the microplate. At this time, the final compound concentration gradient is diluted from 1μM to 0.0128Nm, and the final concentrations of ATP and substrate are 1mM and 25nM. The reaction system is placed at 25 degrees for 120 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution was added to each well. After the reaction was completed at 25°C for 60 minutes, data were collected using the PerkinElmer Nivo multi-label analyzer in TR-FRET mode (excitation wavelength was 320 nm, emission wavelengths were 615 nm and 665 nm).

[0209] Data Analysis:

[0210] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (derived by using the log(inhibitor) vs. response--Variable slope mode in GraphPad Prism). Table 1 provides the inhibitory activities of the compounds of the present invention on CDK2 / CyclinE 1 enzyme.

[0211] Experimental Example 2: In vitro CDK1 / CyclinB1 enzyme activity test

[0212] Experimental Materials:

[0213] CDK1 / Cyclin B1 was purchased from CARNA. Ulight-4E-BP1 peptide, Eu-anti-phospho-tyrosine antibody, and 1X detection buffer were purchased from PerkinElmer. High-purity ATP was purchased from Promega. EDTA was purchased from Sigma. Nivo Multilabel Analyzer was used (PerkinElmer).

[0214] Experimental methods:

[0215] Preparation of kinase buffer: Kinase buffer contains 50mM HEPES, 1mM EDTA, 10mM MgCl2, 0.01% Brij-35, pH 7.4. Add 2.38g HEPES, 58mg EDTA, 406mg MgCl2, 20mg Brij-35 to 200ml buffer and adjust the pH to 7.4.

[0216] Preparation of stop solution:

[0217] Prepare the stop solution by adding 100 μL of 1M EDTA stock solution, 0.625 μL of 1X detection buffer, and 1725 μL of distilled water.

[0218] Dilute the enzyme, Ulight-4E-BP1 peptide, ATP, and inhibitors in kinase buffer.

[0219] Dilute Eu-anti-phospho-tyrosine antibody to 8 nM / L in assay buffer.

[0220] The test compound was diluted five-fold using a pipette to the eighth concentration, from 4 μM to 0.0512 nM, with a final DMSO concentration of 4%. A duplicate assay was performed. To the microplate, 2.5 μL of each inhibitor concentration gradient, 5 μL of 0.5 ng of LCDK1 / CyclinB1 enzyme, and 2.5 μL of a substrate and ATP mixture (4 mM ATP, 200 nM Ulight-4E-BP1 peptide) were added. The final compound concentration gradient was 1 μM to 0.0128 nM, with final ATP and substrate concentrations of 1 mM and 50 nM, respectively. The reaction system was incubated at 25°C for 60 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution was added to each well. After the reaction was completed at 25°C for 60 minutes, data were collected using the PerkinElmer Nivo multi-label analyzer in TR-FRET mode (excitation wavelength was 320 nm, emission wavelengths were 615 nm and 665 nm).

[0221] Data Analysis:

[0222] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (derived by using the log(inhibitor) vs. response--Variable slope mode in GraphPad Prism). Table 1 provides the inhibitory activities of the compounds of the present invention on CDK1 / CyclinB 1 enzymes.

[0223] Experimental Example 3: In vitro OVCAR3 cell activity test

[0224] Experimental Materials:

[0225] 1640 culture medium, fetal bovine serum, and penicillin / streptomycin antibiotics were purchased from Vicente. CellTiter-Glo (a chemiluminescent cell viability assay) was purchased from Promega. The OVCAR3 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. The Envision multi-label analyzer was purchased from PerkinElmer.

[0226] Experimental methods:

[0227] OVCAR3 cells were seeded in a white 384-well plate, with 40 μL of cell suspension per well, containing 300 OVCAR3 cells. The plate was incubated in a CO2 incubator overnight.

[0228] The compound to be tested was diluted 5-fold to the 8th concentration, that is, from 2000 μM to 0.00512 μM, using a dispenser, and a double-well experiment was set up. 78 μL of culture medium was added to the middle plate, and then 2 μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position. After mixing, 10 μL of each well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 10 μM to 0.026 nM. The cell plate was placed in a carbon dioxide incubator and cultured for 7 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. Add 10 μL of cell viability chemiluminescence detection reagent to each well of this cell plate and incubate at room temperature for 10 minutes to stabilize the luminescence signal. Read using a multi-label analyzer.

[0229] Add 10 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Read the results using a multi-label analyzer.

[0230] Data Analysis:

[0231] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value of can be obtained by four-parameter curve fitting (obtained in the "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 1 provides the inhibitory activity of the compounds of the present invention on OVCAR3 cell proliferation.

[0232] Experimental Example 4: In vitro T47D cell activity test

[0233] Experimental Materials:

[0234] 1640 culture medium, fetal bovine serum, and penicillin / streptomycin antibiotics were purchased from Vicente. CellTiter-Glo (a chemiluminescent cell viability assay) was purchased from Promega. T-47D cell lines were purchased from Nanjing Kebai Biotechnology Co., Ltd. Envision multi-label analyzer was purchased from PerkinElmer.

[0235] Experimental methods:

[0236] T-47D cells were seeded in a white 384-well plate, with 40 μL of cell suspension per well, containing 300 T-47D cells. The plate was incubated overnight in a CO2 incubator.

[0237] The compound to be tested was diluted 5-fold to the 8th concentration, that is, from 2000 μM to 0.00512 μM, using a dispenser, and a double-well experiment was set up. 78 μL of culture medium was added to the middle plate, and then 2 μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position. After mixing, 10 μL of each well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 10 μM to 0.026 nM. The cell plate was placed in a carbon dioxide incubator and cultured for 7 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. Add 10 μL of cell viability chemiluminescence detection reagent to each well of this cell plate and incubate at room temperature for 10 minutes to stabilize the luminescence signal. Read using a multi-label analyzer.

[0238] Add 10 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Read the results using a multi-label analyzer.

[0239] Data Analysis:

[0240] The raw data were converted into inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)*100%. 50 The value of can be obtained by four-parameter curve fitting (obtained in the "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 1 provides the inhibitory activity of the compounds of the present invention on T-47D cell proliferation.

[0241] Table 1

[0242] Results and conclusions: The compounds of the present invention have good in vitro activity and good selectivity for CDK2 over CDK1. Specifically, the IC value of the formate salt of compound 1 for CDK1 is 50 (nM) is 1908, the IC of CDK1 of the formate salt of compound 2 50 (nM) is 4084, and the IC of CDK1 of the hydrochloride salt of compound 3 50 (nM) is 1354, and the IC of compound 5CDK1 50 (nM) is 1501, and the IC of compound 6CDK1 50 (nM) is 3493, and the IC of CDK1 of the hydrochloride salt of compound 7 is 50 (nM)>10000, better than PF-07104091.

[0243] In vivo pharmacokinetic studies

[0244] Experimental Example 5: Pharmacokinetic study of the test compound after oral and intravenous administration in mice

[0245] Purpose of the experiment:

[0246] In this study, ICR male mice were used as test animals. LC / MS / MS was used to quantitatively determine the drug concentrations in the plasma of mice at different time points after intravenous or oral administration of the test compound and the reference compound to evaluate the pharmacokinetic characteristics of the test drug in mice.

[0247] Experimental Materials:

[0248] ICR mice (male, 20-30 g, 6-8 weeks old, purchased from Beijing Weitonglihua).

[0249] Experimental operation:

[0250] The clear solution of the test compound was injected into ICR mice via the tail vein (fasted or fed overnight) or administered orally to ICR mice (fasted or fed overnight). For intravenous administration, 50 μL of blood was collected from the cheek puncture at 0 h (before administration) and 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 24 h after administration, and placed in an anticoagulant tube with sodium heparin. The mixture was thoroughly vortexed and centrifuged at 6000 rpm for 3 minutes at 4 ° C. For oral administration, blood was collected from the cheek puncture at 0 h (before administration) and 0.083, 0.25, 1, 2, 4, 6, 8, 24 h after administration, and placed in an anticoagulant tube with sodium heparin. The mixture was thoroughly vortexed and centrifuged at 6000 rpm for 3 minutes. Plasma drug concentrations were determined by LC-MS / MS, and relevant pharmacokinetic parameters were calculated using the non-compartmental linear-log trapezoidal method using Phoenix WinNonlin 8.2.0 pharmacokinetic software. The experimental results are shown in Table 2.

[0251] Table 2

[0252] Results and Conclusions: The compound of the present invention exhibits excellent bioavailability. The peak time and AUC at the same dose are significantly better than those of PF-07104091.

[0253] In vivo efficacy studies

[0254] Experimental Example 6: In vivo pharmacodynamic study of a subcutaneous xenograft tumor model of human ovarian cancer OVCAR-3 cells in CB17-SCID mice

[0255] Experimental procedures:

[0256] Cell Culture: OVCAR-3 cells (human ovarian cancer cell line, obtained from the Xinxin Cell Bank) were cultured in RPMI-1640 (ATCC modified, Gibco) supplemented with 20% FBS (Gibco) and 1% polysaccharide (PS) at 37°C, 5% CO₂, and passaged twice weekly. Exponentially phase OVCAR-3 cells were harvested and resuspended in a mixture of PBS and Matrigel (Corning) (1:1 volume ratio of PBS to Matrigel) to a concentration of 5 × 10⁷ cells / mL for inoculation into mice.

[0257] Animals: Female CB17-SCID mice, 6-8 weeks old, weighing 18-21 g. Forty mice were inoculated. Provided by Shanghai Lingchang Biotechnology Co., Ltd.

[0258] Tumor inoculation: Each mouse was subcutaneously inoculated with 0.2 mL of OVCAR-3 tumor cells (10×10 6 ).

[0259] Experimental Indicators: These indicators assess whether tumor growth is inhibited, delayed, or cured. Tumor diameter is measured twice weekly with a vernier caliper. Tumor volume is calculated using the formula: V = 0.5a × b², where a and b represent the major and minor diameters of the tumor, respectively.

[0260] Antitumor index (TGI) (%): reflects the rate of tumor growth inhibition. Calculation of TGI (%): TGI (%) = [(1 - (average tumor volume at the end of treatment in a given treatment group - average tumor volume at the start of treatment in that treatment group)) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the start of treatment in the solvent control group)] × 100%.

[0261] Experimental results: The experimental results are shown in Table 3

[0262] Table 3

[0263] The compounds of the present application exhibited unexpected tumor reduction effects in the OVCAR3 in vivo efficacy model and were well tolerated by animals.

[0264] In vivo PD studies

[0265] Experimental Example 7: In vivo PD study in the human ovarian cancer OVCAR-3 mouse model

[0266] Experimental procedures:

[0267] Cell Culture: Human ovarian cancer OVCAR-3 cells (ATCC, Manassas, cat#HTB-161) were cultured as monolayers in RPMI 1640 medium supplemented with 20% fetal bovine serum, 0.01 mg / ml bovine insulin, and 1% anti-antibody in a 37°C, 5% CO2 incubator. Cells were routinely digested and passaged twice weekly using trypsin-EDTA. When cell saturation reached 80%-90% and the desired number of cells was reached, cells were harvested, counted, and plated.

[0268] Animals: Female BALB / c nude mice, 7-9 weeks old, weighing 18-23 g. A total of 100 mice were inoculated. These mice were provided by the Experimental Animal Management Department of the Shanghai Institute of Family Planning Science (formerly Shanghai Xipu-Bikai).

[0269] Tumor inoculation: 0.2 mL (10 × 106 cells) of OVCAR-3 cells (with Matrigel, volume ratio of 1:1) were subcutaneously inoculated into the right back of each mouse. When the average tumor volume reached 171 mm3, the mice were divided into groups for administration, with 6 mice per group for a total of 10 groups.

[0270] Tumor sampling: Tumor samples were collected after 21 consecutive days of administration.

[0271] Protein extraction and quantification:

[0272] 1) Take out the quick-frozen tissue sample from a -80℃ refrigerator or liquid nitrogen tank.

[0273] 2) Working on dry ice, remove a portion of tissue (approximately 30-100 mg) and place it in a 2 mL centrifuge tube containing steel beads. Add 400 μL of RIPA cell lysis buffer (to which 1% protease inhibitors and phosphatase inhibitors have been freshly added).

[0274] 3) Crush the tissue using a cryo-grinder at 50 Hz for 5 minutes.

[0275] 4) Place the tissue lysis solution on ice for 30 minutes.

[0276] 5) Centrifuge at 15,000 rpm, 4°C for 10 minutes and transfer the supernatant into a new 1.5 mL centrifuge tube.

[0277] 6) Protein quantification was performed using a BCA quantification kit.

[0278] 7) Based on the quantitative results, prepare the protein sample for loading, unify the sample protein concentration to 4 μg / μL, add LDS loading buffer (4X) and sample reducing agent (10X), and heat the sample at 100°C for 10 minutes.

[0279] 8) Western blotting, or store the denatured samples in a -80°C refrigerator.

[0280] Immunoblotting:

[0281] 1) Sample loading: Thaw the sample and load 10 μL into each well of Bis-Tris gel.

[0282] 2) Electrophoresis: 1X MES electrophoresis buffer, 80 V, 30 min, then 120 V, 90 min.

[0283] 3) Transfer: Use the iBlot2 transfer kit and transfer apparatus at 20 volts for 7 minutes.

[0284] 4) After transfer, cut the membrane according to the molecular weight of the protein to be detected and wash the membrane three times with 10 mL of 1xTBS for 5 minutes each time at room temperature with shaking.

[0285] 5) Blocking: Place the membrane in 10 mL of blocking solution ( Block in blocking solution) at room temperature with slow shaking for 1 hour.

[0286] 6) Incubate with primary antibody: Add 10 mL of primary antibody of appropriate dilution (using 0.1% tween-20 The cells were incubated with blocking buffer at 4°C with slow shaking. The pRb (T821) antibody was diluted 1:1000 and incubated for 3 days. The GAPDH antibody was diluted 1:2000 and incubated overnight.

[0287] 7) Wash the membrane with 10 mL of 1xTBST three times for 10 minutes each time, shaking at room temperature.

[0288] 8) Incubate with secondary antibody: Add 10 mL of secondary antibody of appropriate dilution (using 0.1% tween-20 Blocking solution was diluted at a ratio of 1:10,000) and shaken slowly at room temperature for 1 hour.

[0289] 9) Wash the membrane with 10 mL of 1xTBST three times for 10 minutes each time, shaking at room temperature.

[0290] 10) Wash the membrane twice with 10 mL of 1X TBS to remove residual Tween-20, 5 minutes each time, shaking at room temperature.

[0291] 11) Fluorescence signal values ​​were detected using Odyssey Clx and its software Image Studio Ver 5.2.

[0292] Expression quantification:

[0293] Image Studio Lite Ver 5.2 software was used to perform relative quantification of immunoblot fluorescence bands.

[0294] Data Analysis:

[0295] Based on the band density intensity quantified by the software, the expression ratio of the target protein relative to the internal reference (GAPDH) was calculated. The normalized expression fold of the target protein in the treatment group was then calculated using the blank control group as the benchmark. The normalized expression fold of the protein in each group was displayed in GraphPad Prism 6.02.

[0296] Experimental results: The experimental results are shown in Figure 1.

[0297] Conclusion: Immunoblotting assays demonstrated that the compounds of this application effectively inhibited CDK2 enzyme activity and reduced pRb protein levels, thereby blocking the transition from the G1 to S phase of the cell cycle and inhibiting tumor cell proliferation. At the same dose, the compounds of this application demonstrated superior inhibitory activity against T821 phosphorylation of pRb, a downstream target of CDK2, compared to PF-07104091.

[0298] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0299] The embodiments described above only express several implementation methods of the present invention, which are convenient for understanding the technical solutions of the present invention in a specific and detailed manner, but they cannot be understood as limiting the scope of protection of the invention patent. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present invention, several variations and improvements can be made, which all fall within the scope of protection of the present invention. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided by the present invention are all within the scope of protection of the claims attached to the present invention. Therefore, the scope of protection of the patent of the present invention shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. A pyrazole derivative represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in: X is independently selected from N or CR4 at each occurrence; R4 is absent or selected from -H, -D, -F, -Cl, -Br, -I, straight chain alkyl having 1 to 6 C atoms, straight chain deuterated alkyl having 1 to 6 C atoms, branched alkyl having 3 to 6 C atoms or aromatic heterocycle; R1 is selected from -H, -D, -F, -Cl, -Br, -I, linear alkyl having 1 to 20 C atoms, linear deuterated alkyl having 1 to 20 C atoms, branched alkyl having 3 to 20 C atoms, branched deuterated alkyl having 3 to 20 C atoms, cyclic alkyl having 3 to 20 C atoms, cyclic deuterated alkyl having 3 to 20 C atoms, linear alkoxy having 1 to 20 C atoms, linear deuterated alkoxy having 1 to 20 C atoms, branched alkoxy having 3 to 20 C atoms, branched deuterated alkoxy having 3 to 20 C atoms, cyclic alkoxy having 3 to 20 C atoms, cyclic deuterated alkoxy having 3 to 20 C atoms; R3 is selected from substituted or unsubstituted alkyl having 1 to 6 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 6 C atoms, substituted or unsubstituted cycloalkyl having 3 to 7 C atoms, substituted or unsubstituted heterocyclyl having 4 to 7 ring atoms; R2 is selected from -F, -Cl, -OH, -CN, -NR5R6, -CH2-ONR7R8, -CH2-ON=R9, -CH2N(R 10 )OR 11 , substituted or unsubstituted alkyl having 1 to 4 C atoms, substituted or unsubstituted fluoroalkyl having 1 to 4 C atoms, substituted or unsubstituted alkoxy having 1 to 4 C atoms, substituted or unsubstituted fluoroalkoxy having 1 to 4 C atoms, substituted or unsubstituted cycloalkyl having 3 to 8 C atoms; R5, R6, R7, R8, R 10 , R 11 Each of the following is independently selected from -H, -D, a linear alkyl group having 1 to 20 C atoms, a linear deuterated alkyl group having 1 to 20 C atoms, a branched alkyl group having 3 to 20 C atoms, a branched deuterated alkyl group having 3 to 20 C atoms, a cyclic alkyl group having 3 to 20 C atoms, a cyclic deuterated alkyl group having 3 to 20 C atoms, a linear alkoxy group having 1 to 20 C atoms, a linear deuterated alkoxy group having 1 to 20 C atoms, a branched alkoxy group having 3 to 20 C atoms, a branched deuterated alkoxy group having 3 to 20 C atoms, a cyclic alkoxy group having 3 to 20 C atoms, and a cyclic deuterated alkoxy group having 3 to 20 C atoms; R9 is selected from linear alkylene having 1 to 20 C atoms, linear deuterated alkylene having 1 to 20 C atoms, branched alkylene having 3 to 20 C atoms, branched deuterated alkylene having 3 to 20 C atoms, cyclic alkylene having 3 to 20 C atoms, and cyclic deuterated alkylene having 3 to 20 C atoms.

2. The pyrazole derivative, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, characterized in that: The pyrazole derivative is a structure shown in formula (II):

3. The pyrazole derivative according to claim 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The pyrazole derivative is a structure shown in formula (III):

4. The pyrazole derivative, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, characterized in that: R1 is selected from -H, -D, -F, -Cl, -Br, -I, straight-chain alkyl having 1 to 10 C atoms, straight-chain deuterated alkyl having 1 to 10 C atoms, branched-chain alkyl having 3 to 10 C atoms, branched-chain deuterated alkyl having 3 to 10 C atoms, cyclic alkyl having 3 to 10 C atoms, cyclic deuterated alkyl having 3 to 10 C atoms.

5. The pyrazole derivative, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, characterized in that: R5, R6, R7, R8, R 10 , R 11 Each of the following is independently selected from -H, -D, a linear alkyl group having 1 to 10 C atoms, a linear deuterated alkyl group having 1 to 10 C atoms, a branched alkyl group having 3 to 10 C atoms, a branched deuterated alkyl group having 3 to 10 C atoms, a cyclic alkyl group having 3 to 10 C atoms, a cyclic deuterated alkyl group having 3 to 10 C atoms, a linear alkoxy group having 1 to 10 C atoms, a linear deuterated alkoxy group having 1 to 10 C atoms, a branched alkoxy group having 3 to 10 C atoms, a branched deuterated alkoxy group having 3 to 10 C atoms, a cyclic alkoxy group having 3 to 10 C atoms, and a cyclic deuterated alkoxy group having 3 to 10 C atoms; R9 is selected from linear alkylene having 1 to 20 C atoms, linear deuterated alkylene having 1 to 20 C atoms, branched alkylene having 3 to 20 C atoms, branched deuterated alkylene having 3 to 20 C atoms, cyclic alkylene having 3 to 20 C atoms, cyclic deuterated alkylene having 3 to 20 C atoms, or a combination of these systems.

6. The pyrazole derivative, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, characterized in that: R2 is selected from any one of the following structures:

7. The pyrazole derivative according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The pyrazole derivative is any of the following compounds:

8. Use of the pyrazole derivative according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a medicament for treating and / or preventing a disease associated with or mediated by CDK2 activity.

9. The use according to claim 8, characterized in that: The disease associated with CDK2 activity or mediated by CDK2 activity is cancer.

10. A pharmaceutical composition, characterized in that The invention comprises the pyrazole derivative according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

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