Deuterated pyrazole derivatives, pharmaceutical compositions, and applications and preparation methods

Abstract

The present invention provides a deuterated pyrazole derivative, the structure of which is shown in formula (I). In addition, the present invention also discloses a pharmaceutically acceptable salt of the derivative, its stereoisomers, a pharmaceutical composition and its application. The compound of the present invention has significant CDK2 inhibitory activity and is of great practical value.

Description

Deuterated pyrazole derivatives, pharmaceutical compositions, applications, and preparation methods Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a deuterated pyrazole derivative, a pharmaceutically acceptable salt, a stereoisomer, a pharmaceutical composition, and an application and preparation method thereof. Background Art

[0002] 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.

[0003] 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.

[0004] 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.

[0005] There are currently few small molecule inhibitors with CDK2 activity in clinical research. Pfizer has developed PF-4091, which has the following structural formula: However, the dosage of PF-4091 in preclinical animal efficacy models was high and required twice daily administration, indicating that both the activity and metabolism of PF-4091 could be improved. Therefore, in order to better develop new drugs for the treatment of cancer and other related diseases, there is an urgent need to develop CDK2 inhibitors with high activity, more stable metabolism, and the ability to treat multiple cancers, especially small molecule inhibitors that selectively target CDK2, which may have better safety.

[0006] Summary of the Invention

[0007] Based on this, the present invention provides a deuterated pyrazole derivative with high CDK2 inhibitory activity, and a pharmaceutically acceptable salt or stereoisomer thereof.

[0008] The present invention is achieved through the following technical solutions.

[0009] A compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0010] in:

[0011] R1 is selected from linear deuterated alkyl having 1 to 20 C atoms, branched deuterated alkyl having 3 to 20 C atoms, cyclic deuterated alkyl having 3 to 20 C atoms, linear deuterated alkoxy having 1 to 20 C atoms, branched deuterated alkoxy having 3 to 20 C atoms, cyclic deuterated alkoxy having 3 to 20 C atoms, or a combination of these systems;

[0012] R2, R3, and R4 are each 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, a cyclic deuterated alkoxy group having 3 to 20 C atoms, or a combination of these systems.

[0013] In one embodiment, the compound of formula (I) is represented by formula (II):

[0014] In one embodiment, R3 is selected from -H or -D.

[0015] In one embodiment, R1 is selected from a linear deuterated alkyl group having 1 to 10 C atoms, a branched deuterated alkyl group having 3 to 10 C atoms, a cyclic deuterated alkyl group having 3 to 10 C atoms, a linear deuterated alkoxy group having 1 to 10 C atoms, a branched deuterated alkoxy group having 3 to 10 C atoms, a cyclic deuterated alkoxy group having 3 to 10 C atoms, or a combination thereof;

[0016] R2 and R4 are each independently selected from -H, -D, a straight-chain alkyl group having 1 to 10 C atoms, a straight-chain deuterated alkyl group having 1 to 10 C atoms, a branched-chain alkyl group having 3 to 10 C atoms, a branched-chain 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 straight-chain alkoxy group having 1 to 10 C atoms, a straight-chain deuterated alkoxy group having 1 to 10 C atoms, a branched-chain alkoxy group having 3 to 10 C atoms, a branched-chain deuterated alkoxy group having 3 to 10 C atoms, a cyclic alkoxy group having 3 to 10 C atoms, a cyclic deuterated alkoxy group having 3 to 10 C atoms, or a combination of these systems.

[0017] In one embodiment, R1 is selected from a linear deuterated alkyl group having 1 to 5 C atoms, a branched deuterated alkyl group having 3 to 5 C atoms, a cyclic deuterated alkyl group having 3 to 5 C atoms, a linear deuterated alkoxy group having 1 to 5 C atoms, a branched deuterated alkoxy group having 3 to 5 C atoms, a cyclic deuterated alkoxy group having 3 to 5 C atoms, or a combination thereof;

[0018] R2 and R4 are independently selected from linear alkyl groups having 1 to 3 C atoms, linear deuterated alkyl groups having 1 to 3 C atoms, branched alkyl groups having 3 to 5 C atoms, branched deuterated alkyl groups having 3 to 5 C atoms, cyclic alkyl groups having 3 to 5 C atoms, cyclic deuterated alkyl groups having 3 to 5 C atoms, or combinations of these systems.

[0019] In one embodiment, the compound of formula (I) is the following compound:

[0020] The present invention also provides a method for preparing the compound represented by formula (I), comprising the following steps:

[0021] Compound A is reduced to prepare compound B:

[0022] Compound B and compound C are reacted through nucleophilic substitution to prepare compound D:

[0023] Compound D is subjected to a hydrolysis reaction to prepare compound E:

[0024] Compound E and compound F are reacted through acylation to prepare compound G:

[0025] Compound G is subjected to a substitution reaction to prepare a compound of formula (I):

[0026] Wherein, the definitions of R1, R2, R3, and R4 are consistent with those described above; and X is selected from -F, -Cl, -Br, or -I.

[0027] The present invention also provides use of the above-mentioned compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a medicament for treating and / or preventing diseases associated with or mediated by CDK2 activity.

[0028] In one embodiment, the disease associated with or mediated by CDK2 activity is cancer.

[0029] The present invention also provides a pharmaceutical composition comprising the compound as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0030] Compared with the prior art, the compound of the present invention, or its pharmaceutically acceptable salt, or its stereoisomer has the following beneficial effects:

[0031] After extensive and in-depth research, the inventors unexpectedly discovered a novel deuterated pyrazole derivative. This subtle modification replaces some of the hydrogen elements in the drug molecule with deuterium. This subtle adjustment significantly impacts the drug's activity and metabolism. The deuterated pyrazole derivative exhibits unexpectedly high inhibitory activity against CDK2, significantly enhancing pharmacokinetic efficacy and mitigating side effects. It can be used to treat and / or prevent diseases associated with or mediated by CDK2 activity, including various cancers with elevated Cyclin E expression. It has the potential to be a promising treatment for diseases mediated by CDK2 activity. DETAILED DESCRIPTION

[0032] 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.

[0033] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the features. In the description of the invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined. In the description of the present invention, the meaning of "several" is at least one, such as one, two, etc., unless otherwise clearly and specifically defined.

[0034] The terms "preferably," "more preferably," and the like, used herein refer to embodiments of the present invention that may provide certain benefits under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, nor is it intended to exclude other embodiments from the scope of the present invention.

[0035] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein are understood to include any and all subranges subsumed therein.

[0036] Unless otherwise indicated, all percentages, fractions, and ratios are calculated based on the total weight of the compositions of the present invention. Unless otherwise indicated, all masses relating to listed ingredients are given as active ingredients and therefore do not include solvents or byproducts that may be present in commercially available materials. The term "mass percentage content" may be expressed herein with the symbol "%." All molecular weights herein are weight-average molecular weights expressed in Daltons, unless otherwise indicated. All formulations and testing herein took place at 25°C, unless otherwise indicated. The terms "comprise," "include," "contain," "have," "have," and other variations herein are intended to encompass non-exclusive inclusions, and do not distinguish between these terms. The term "comprising" means that additional steps and ingredients that do not affect the end result may be added. The compositions and methods / processes of the present invention comprise, consist of, and consist essentially of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, steps, or limitations described herein. The terms "efficacy," "performance," "effect," and "efficacy" are not used herein to distinguish between.

[0037] 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.

[0038] 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.

[0039] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as formic acid, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and 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.

[0040] 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.

[0041] 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.

[0042] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0043] 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.

[0044] 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.

[0045] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0046] 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

[0047] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is considered as a substituent to which it is connected), if a wavy line is used between the atom on the double bond and its substituent in the compound, When connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of two isomers of formula (A-1) and formula (A-2); the following formula (B) represents that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of two isomers of formula (B-1) and formula (B-2). The following formula (C) represents that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of two isomers of formula (C-1) and formula (C-2).

[0048] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and readily interconvert into each other. If tautomerism is possible (e.g., in solution), 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 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.

[0049] 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%.

[0050] 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%.

[0051] 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).

[0052] 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.

[0053] "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.

[0054] 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.

[0055] 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.

[0056] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0057] 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.

[0058] 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.

[0059] 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.

[0060] 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.

[0061] 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.

[0062] 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).

[0063] 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.

[0064] 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.

[0065]

[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.

[0066] 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.).

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] "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.

[0074] "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.

[0075] 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.

[0076] 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.

[0077] 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.

[0078] 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.

[0079] 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.

[0080] "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.

[0081] 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).

[0082] "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.

[0083] "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.

[0084] 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.

[0085] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0086] The solvents used in the present invention are commercially available. The following abbreviations are used in the present invention: DIBAL-H represents diisobutylaluminum hydride; SiO2 represents silicon dioxide; NaH represents sodium hydride; CD3I represents deuterated iodomethane; NH4Cl represents ammonium chloride; LiOH.H2O represents hydrated lithium hydroxide; DMF represents N,N-dimethylformamide; HATU represents 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DIEA represents diisopropylethylamine; and FA represents formic acid.

[0087] Compounds are named according to the conventional nomenclature in the art or using The software named the commercially available compounds using the supplier's catalog name.

[0088] 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.

[0089] The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0090] in:

[0091] R1 is selected from linear deuterated alkyl having 1 to 20 C atoms, branched deuterated alkyl having 3 to 20 C atoms, cyclic deuterated alkyl having 3 to 20 C atoms, linear deuterated alkoxy having 1 to 20 C atoms, branched deuterated alkoxy having 3 to 20 C atoms, cyclic deuterated alkoxy having 3 to 20 C atoms, or a combination of these systems;

[0092] R2, R3, and R4 are each 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, a cyclic deuterated alkoxy group having 3 to 20 C atoms, or a combination of these systems.

[0093] In a specific example, the compound of formula (I) has the structure shown in formula (II):

[0094] In a specific example, R3 is selected from -H or -D.

[0095] In a specific example, R1 is selected from a linear deuterated alkyl group having 1 to 10 C atoms, a branched deuterated alkyl group having 3 to 10 C atoms, a cyclic deuterated alkyl group having 3 to 10 C atoms, a linear deuterated alkoxy group having 1 to 10 C atoms, a branched deuterated alkoxy group having 3 to 10 C atoms, a cyclic deuterated alkoxy group having 3 to 10 C atoms, or a combination of these systems.

[0096] In a specific example, R1 is selected from a linear deuterated alkyl group having 1 to 5 C atoms, a branched deuterated alkyl group having 3 to 5 C atoms, a cyclic deuterated alkyl group having 3 to 5 C atoms, a linear deuterated alkoxy group having 1 to 5 C atoms, a branched deuterated alkoxy group having 3 to 5 C atoms, a cyclic deuterated alkoxy group having 3 to 5 C atoms, or a combination of these systems.

[0097] More specifically, R1 is selected from R1 is selected from linear deuterated alkyl having 1 to 3 C atoms, branched deuterated alkyl having 3 to 5 C atoms, cyclic deuterated alkyl having 3 to 5 C atoms.

[0098] In a specific example, R2 and R4 are independently selected from -H, -D, a straight-chain alkyl group having 1 to 10 C atoms, a straight-chain deuterated alkyl group having 1 to 10 C atoms, a branched-chain alkyl group having 3 to 10 C atoms, a branched-chain 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 straight-chain alkoxy group having 1 to 10 C atoms, a straight-chain deuterated alkoxy group having 1 to 10 C atoms, a branched-chain alkoxy group having 3 to 10 C atoms, a branched-chain deuterated alkoxy group having 3 to 10 C atoms, a cyclic alkoxy group having 3 to 10 C atoms, a cyclic deuterated alkoxy group having 3 to 10 C atoms, or a combination of these systems.

[0099] In a specific example, R2 and R4 are independently selected from 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 5 C atoms, a branched-chain deuterated alkyl group having 3 to 5 C atoms, a cyclic alkyl group having 3 to 5 C atoms, a cyclic deuterated alkyl group having 3 to 5 C atoms, or a combination of these systems.

[0100] In a specific example, the compound of formula (I) is the following compound:

[0101] In a specific example, the pharmaceutically acceptable salt is an alkyl salt. Further, the pharmaceutically acceptable salt is a formate salt.

[0102] In a specific example, the present invention also provides a method for preparing a compound represented by formula (I), comprising the following steps:

[0103] Compound A is reduced to prepare compound B:

[0104] Compound B and compound C are reacted through nucleophilic substitution to prepare compound D:

[0105] Compound D is subjected to a hydrolysis reaction to prepare compound E:

[0106] Compound E and compound F are reacted through acylation to prepare compound G:

[0107] Compound G is subjected to a substitution reaction to prepare a compound of formula (I):

[0108] Wherein, the definitions of R1, R2, R3, and R4 are consistent with those above; and X is selected from -F, -Cl, -Br, or -I.

[0109] The present invention also provides a use of the above compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a drug for treating and / or preventing diseases related to or mediated by CDK2 activity.

[0110] In one specific example, the disease associated with or mediated by CDK2 activity is cancer.

[0111] In a specific example, the cancer is one or more of ovarian cancer, gastric cancer, endometrial cancer, and breast cancer.

[0112] The present invention also provides a pharmaceutical composition comprising the above compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0113] 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.

[0114] The deuterated 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.

[0115] Example 1

[0116] This embodiment provides compound 1, whose structural formula is as follows:

[0117] The reaction scheme is as follows:

[0118] The preparation process is as follows:

[0119] Step 1:

[0120] At a temperature of -60 ° C, compound 1-1 (2.00 g, 8.84 mmol, 1.00 eq) was dissolved in tetrahydrofuran (15.0 mL), and DIBAL-H (1.00 M, 17.6 mL, 2.00 eq) was slowly added dropwise. After completion of the addition, the temperature was raised to -20 ° C, and the reaction was allowed to react for 3 hours. Methanol (20.0 mL) was added to the reaction system at 0 ° C to quench the reaction. The reaction was stirred at 20 ° C for 2 hours, and then concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to obtain compound 1-2. 1 HNMR: (400MHz, CDCl3) δ6.82 (s, 1H), 4.67 (s, 2H), 4.39-4.32 (m, 2H), 4.15 (s, 3H), 2.02 (br s, 1H), 1.38 (t, J = 7.2Hz, 3H).

[0121] Step 2:

[0122] At 0°C, NaH (52.1 mg, 1.30 mmol, 60% purity, 1.20 eq) was dissolved in tetrahydrofuran (1.00 mL). Compound 1-2 (200 mg, 1.09 mmol, 1.00 eq) was dissolved in tetrahydrofuran (1.00 mL) and added dropwise to the reaction mixture. After reacting at 25°C for 1 hour, the temperature was lowered to 0°C and CD3I (770 mg, 5.43 mmol, 330 μL, 5.00 eq) was added dropwise. After reacting at 25°C for 9 hours, 2.0 mL of NH4Cl solution was slowly added at 0°C and extracted three times with ethyl acetate (3.00 mL). The organic phases were combined, washed three times with saturated sodium chloride (4.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to yield compound 1-3. LCMS: RT=0.310min, MS(ESI)m / z=167.0[M-34]+ 1 HNMR: (400MHz, CDCl3) δ = 6.84 (s, 1H), 4.44 (s, 2H), 4.34 (q, J = 7.2Hz, 2H), 4.16 (s, 3H), 1.37 (t, J = 7.2Hz, 3H).

[0123] Step 3:

[0124] Compound 1-3 (200 mg, 993 μmol, 1.00 eq) was dissolved in tetrahydrofuran (1.00 mL) and water (1.00 mL) at 25°C. LiOH.H2O (43.7 mg, 1.04 mmol, 1.05 eq) was added and reacted at 25°C for 10 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, adjusted to pH 1.0 with hydrochloric acid, and extracted three times with ethyl acetate (2.00 mL). The organic phases were combined, washed three times with saturated aqueous sodium chloride solution (3.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-4. LCMS: MS (ESI) m / z = 139.0 [M-34] + 1 HNMR: (400MHz, CDCl3) δ = 6.97 (s, 1H), 4.48 (s, 2H), 4.19 (s, 3H).

[0125] Step 4:

[0126] At 25°C, compound 1-4 (168 mg, 972 μmol, 1.00 eq) and compound 1-5 (300 mg, 972 μmol, 1.00 eq) were dissolved in DMF (3.00 mL). HATU (406 mg, 1.07 mmol, 1.10 eq) and DIEA (377 mg, 2.92 mmol, 508 μL, 3.00 eq) were added. After stirring at 25°C for 10 hours, the reaction solution was diluted with water (9.00 mL) and extracted three times with ethyl acetate (10.0 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (15.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-6. LCMS: m / z = 464.4 (M+H) + 1 HNMR: EC6454-331-P1N1 (400MHz, DMSO-d6) δ = 9.92 (s, 1H), 7.95 (s, 1H), 7.00 (s, 1H), 6.02 (s, 1H), 5.22 (s, 1H), 4.70 (s, 1 H),4.36(s,1H),4.03(s,3H),3.57(brdd,J=6.4,13.2Hz,1H),3.05-2.94(m,1H),1.81-1.60(m,6H),1.51(s,9H),1.03(br d,J=6.4Hz,6H).

[0127] Step 5:

[0128] Compound 1-6 (273 mg, 588 μmol, 1.00 eq) was dissolved in formic acid (2.50 mL) at room temperature and reacted at 100°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove formic acid to obtain a crude product, which was purified by prep-HPLC (FA conditions) to obtain compound 1. LCMS: m / z = 408.3 (M+H) + 1 H NMR: (400MHz, DMSO-d6) δ12.22(br s,1H),10.72(s,1H),7.11(s,1H),6.95(br d,J=7.2Hz,1H),6.41(br s,1H),5.01(br s,1H),4.33(s,2H),4.05(s,3H),3.58(qd,J=6.8,13.6Hz,1H),3.14-3.03(m,1H),2.10-1.55(m,6H),1.03(d,J=6.4Hz,6H).

[0129] In vitro activity test

[0130] Experimental Example 1: In vitro CDK2 / CyclinE enzyme activity test

[0131] Experimental Materials:

[0132] 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).

[0133] Experimental methods:

[0134] 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.

[0135] Preparation of stop solution: Use 100 μL of 1M EDTA stock solution, add 0.625 μL of 1X detection buffer and mix with 1725 μL of distilled water to prepare the stop solution.

[0136] 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 degrees 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 degrees 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).

[0137] Data Analysis:

[0138] 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.

[0139] Experimental Example 2: In vitro CDK1 / CyclinB1 enzyme activity test

[0140] Experimental Materials:

[0141] 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).

[0142] Experimental methods:

[0143] 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.

[0144] Preparation of stop solution: Use 100 μL of 1M EDTA stock solution, add 0.625 μL of 1X detection buffer and mix with 1725 μL of distilled water to prepare the stop solution.

[0145] Dilute the enzyme, Ulight-4E-BP1 peptide, ATP, and inhibitors in kinase buffer.

[0146] Dilute Eu-anti-phospho-tyrosine antibody to 8 nM / L in assay buffer.

[0147] 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 degrees 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 degrees 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).

[0148] Data Analysis:

[0149] 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.

[0150] Experimental Example 3: In vitro GSK3β enzyme activity test

[0151] Experimental Materials:

[0152] GSK3βActive was purchased from SignalChem; GSK3 Substrate was purchased from SignalChem; ADP-Glo ​​Kinase Assay was purchased from Promega; Kinase assay buffer III was purchased from SignalChem; Nivo multilabel analyzer (PerkinElmer).

[0153] Experimental methods:

[0154] The test compound was diluted to 100 μM in 100% DMSO as the first concentration. The compound was then diluted five-fold using a pipette to the eighth concentration, from 100 μM to 0.0013 μM. Each compound concentration point was diluted 20-fold in 1X kinase buffer to prepare a compound working solution containing 5% DMSO. 1 μL of each compound concentration gradient working solution was added to a microplate, with duplicate wells. 2 μL of GSK3β enzyme (1 ng) and 2 μL of a mixture of substrate and ATP (62.5 μM ATP, 0.5 μg / μL GSK3 substrate) were added to the microplate, resulting in a final compound concentration gradient of 1 μM to 0.013 nM, with final ATP and substrate concentrations of 25 μM and 0.2 μg / μL, respectively. The reaction system was incubated at 25°C for 60 minutes. After the reaction, 5 μl of ADP-Glo ​​reagent was added to each well and the reaction was continued at 25°C for 40 minutes. After the reaction was completed, 10 μL of kinase detection reagent was added to each well and the reaction was continued at 25°C for 30 minutes. Chemiluminescence was read using a PerkinElmer Nivo multi-label analyzer with an integration time of 0.5 seconds.

[0155] Data Analysis:

[0156] The raw data were converted to inhibition using the equation (Sample-Min) / (Max-Min)*100%. The IC50 value was then obtained by four-parameter curve fitting (using the log(inhibitor) vs. response - Variable slope mode in GraphPad Prism). Table 1 provides the GSK3β enzymatic inhibitory activity of the compounds of the present invention.

[0157] Max well: positive control well reading value, blank well without enzyme;

[0158] Min wells: Negative control wells contain 1% DMSO solvent.

[0159] Experimental Example 4: In vitro OVCAR3 cell activity test

[0160] Experimental Materials:

[0161] 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.

[0162] Experimental methods:

[0163] 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.

[0164] 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.

[0165] 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.

[0166] Data Analysis:

[0167] 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.

[0168] Experimental Example 5: In vitro T47D cell activity test

[0169] Experimental Materials:

[0170] 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.

[0171] Experimental methods:

[0172] 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.

[0173] 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.

[0174] 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.

[0175] Data Analysis:

[0176] 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.

[0177] Table 1

[0178] 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.

[0179] 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 compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in: R1 is selected from linear deuterated alkyl having 1 to 20 C atoms, branched deuterated alkyl having 3 to 20 C atoms, cyclic deuterated alkyl having 3 to 20 C atoms, linear deuterated alkoxy having 1 to 20 C atoms, branched deuterated alkoxy having 3 to 20 C atoms, cyclic deuterated alkoxy having 3 to 20 C atoms, or a combination of these systems; R2, R3, and R4 are each 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, a cyclic deuterated alkoxy group having 3 to 20 C atoms, or a combination of these systems.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The compound of formula (I) has the structure shown in formula (II):

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R3 is selected from -H or -D.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R1 is selected from linear deuterated alkyl having 1 to 10 C atoms, branched deuterated alkyl having 3 to 10 C atoms, cyclic deuterated alkyl having 3 to 10 C atoms, linear deuterated alkoxy having 1 to 10 C atoms, branched deuterated alkoxy having 3 to 10 C atoms, cyclic deuterated alkoxy having 3 to 10 C atoms, or a combination of these systems; R2 and R4 are each independently selected from -H, -D, a straight-chain alkyl group having 1 to 10 C atoms, a straight-chain deuterated alkyl group having 1 to 10 C atoms, a branched-chain alkyl group having 3 to 10 C atoms, a branched-chain 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 straight-chain alkoxy group having 1 to 10 C atoms, a straight-chain deuterated alkoxy group having 1 to 10 C atoms, a branched-chain alkoxy group having 3 to 10 C atoms, a branched-chain deuterated alkoxy group having 3 to 10 C atoms, a cyclic alkoxy group having 3 to 10 C atoms, a cyclic deuterated alkoxy group having 3 to 10 C atoms, or a combination of these systems.

5. The compound according to claim 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R1 is selected from linear deuterated alkyl having 1 to 5 C atoms, branched deuterated alkyl having 3 to 5 C atoms, cyclic deuterated alkyl having 3 to 5 C atoms, linear deuterated alkoxy having 1 to 5 C atoms, branched deuterated alkoxy having 3 to 5 C atoms, cyclic deuterated alkoxy having 3 to 5 C atoms, or a combination of these systems; R2 and R4 are independently selected from linear alkyl groups having 1 to 3 C atoms, linear deuterated alkyl groups having 1 to 3 C atoms, branched alkyl groups having 3 to 5 C atoms, branched deuterated alkyl groups having 3 to 5 C atoms, cyclic alkyl groups having 3 to 5 C atoms, cyclic deuterated alkyl groups having 3 to 5 C atoms, or combinations of these systems.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The compounds of formula (I) are the following compounds:

7. A method for preparing the compound of formula (I), characterized in that: The steps include: Compound A is reduced to prepare compound B: Compound B and compound C are reacted through nucleophilic substitution to prepare compound D: Compound D is subjected to a hydrolysis reaction to prepare compound E: Compound E and compound F are reacted through acylation to prepare compound G: Compound G is subjected to a substitution reaction to prepare a compound of formula (I): Wherein, the definitions of R1, R2, R3, and R4 are consistent with those described in any one of claims 1 to 6; and X is selected from -F, -Cl, -Br, or -I.

8. Use of the compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a medicament for treating and / or preventing diseases 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 compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

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