Compounds that inhibit protein-protein interaction of CDK4 / 6 with cyclin D

Abstract

The invention provides a compound for inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D. The compound has a result shown as a formula (I), a formula (II), a formula (III), a formula (IV), a formula (V) or a formula (VI), and has application in preparation of drugs for diseases which are benefited by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D or can be treated.

Description

Compounds that inhibit protein-protein interaction between CDK4 / 6 and cyclin D

[0001] Cross-references

[0002] This application claims priority to Chinese Patent Application No. 202311182364.1, filed on September 13, 2023, entitled “Compounds that inhibit protein-protein interaction of CDK4 / 6 and cyclin D”, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present application relates to novel compounds, pharmaceutical compositions comprising the compounds of the present application, a combination, use of the compounds of the present application in the preparation of drugs for diseases that benefit or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D, and methods for treating diseases by using the compounds of the present application. Background Art

[0004] Cyclin-dependent kinases 4 / 6 (CDK4 / 6), members of the serine / tyrosine protein kinase family, are key proteins in regulating the cell cycle. During the G1 phase of the cell cycle, CDK4 / 6 form a CDK4 / 6-CCND complex with cyclin D (CCND). CDK4 / 6 activity is activated, inducing phosphorylation of the tumor suppressor gene retinoblastoma protein (Rb). Rb phosphorylation releases the transcription factor E2F from the Rb-E2F complex, initiating transcription, promoting the transition of the cell cycle from G1 to S phase, and initiating DNA replication. Multiple reports have found that the CDK4 / 6-CCND-Rb pathway is abnormally active in many tumor cells, including breast cancer, renal cancer, pancreatic cancer, liver cancer, brain tumors, and hematologic tumors. Therefore, CDK4 / 6 is an ideal target for the treatment of various tumors.

[0005] Currently, all of the CDK4 / 6 inhibitors on the market target the ATP (adenosine triphosphate) catalytic pocket of CDK4 / 6, such as Palbociclib, Rbociclib, and Abemaciclib. These drugs all have varying degrees of drug resistance and clinical side effects. Therefore, the search for new compounds targeting the CDK4 / 6-CCND-Rb pathway is urgent.

[0006] Summary of the Invention

[0007] In order to solve the above problems, the present application provides a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or (VI), or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof,

[0008] in,

[0009] Ar1, Ar3, Ar4 and Ar7 are each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl;

[0010] Said Ar2, Ar6 and U2 are each independently an 8-10 membered bicyclic heterocycle;

[0011] The U3 is -N(Ar5)2, and the Ar5 are each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl;

[0012] The R1 are each independently hydrogen or C 0-3 alkyl hydroxyl groups;

[0013] The R2 are each independently hydrogen or C 1-6 alkyl;

[0014] R3 is hydrogen, C 2-6 Alkenyl or C 2-6 Alkynyl;

[0015] The R4 is unsubstituted or substituted C 3-8 Cycloalkyl;

[0016] Said X1 is N or C;

[0017] The U1 is -X2-(CH2) n -C(O)-M1;

[0018] Said X2 and X3 are each independently S or O;

[0019] The M1 is -N(R5)(R6);

[0020] The R5 and R6 together with the N to which they are attached form an 8-10 membered bicyclic heterocycle;

[0021] The U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S;

[0022] The R9 and R 10 Together with the attached C, it forms a 5-6 membered aryl or 5-6 membered heteroaryl group;

[0023] The R 11 and R 12 Together with the attached N and C, it forms a 5-6 membered saturated ring;

[0024] n and t are each independently 1, 2 or 3;

[0025] m is 1 or 2.

[0026] The compounds of the present application inhibit the formation of the CDK4 / 6-CCND complex by inhibiting the protein-protein interaction between CDK4 and CCND and / or the protein-protein interaction between CDK6 and CCND, thereby inhibiting the activity of CDK4 / 6 and thus inhibiting the CDK4 / 6-CCND-Rb pathway. The compounds of Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V) and Formula (VI) have good anti-cancer activity. The mechanism of action of the compounds of the present application is different from that of the CDK4 / 6 inhibitors already on the market, and they have reduced drug resistance and clinical side effects.

[0027] The present application provides a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI), or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for a disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D.

[0028] The present application provides a pharmaceutical composition comprising a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) as described herein, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0029] The present application provides a combination comprising a compound of Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V) or Formula (VI) as described herein, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, and one or more therapeutically active agents.

[0030] The present application provides a method for treating a disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D, the method comprising administering to a subject in need of such treatment a therapeutically effective dose of a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) as described herein, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition, or the aforementioned combination.

[0031] It should be understood that the content described in this section is not intended to identify the key or important features of the embodiments of the present application, nor is it intended to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The following provides a brief description of the drawings, which are provided to illustrate exemplary embodiments disclosed herein and not to limit these embodiments.

[0033] Figure 1 shows fluorescence signal diagrams of protein-protein interactions between CDK4 / 6 and CCND1 in Example 1. Specifically shown are fluorescence signal diagrams of VN155-CDK4+VC155-CCND1, VN155-CDK6+VC155-CCND1, and their controls VN155-CDK4+VC155-mCherry and VN155-CDK6+VC155-mCherry, where green signals represent interactions between the corresponding proteins.

[0034] FIG2 is a bar graph showing the protein-protein interaction between CDK4 / 6 and CCND1 blocked by Compounds 1 to 6 in Example 1. DETAILED DESCRIPTION

[0035] Unless otherwise indicated, all numbers used in this specification and claims to represent content, concentration, ratio, weight, particle size, percentage, technical effect, etc. should be understood as being modified by the term "about" or "approximately" under any circumstances. Therefore, unless otherwise indicated, the numerical parameters listed in the following specification and the appended claims are approximate values. Unless otherwise indicated, the terms used herein have the usual understanding meanings for those skilled in the art. For those skilled in the art, it can vary according to the desired properties and effects sought to be obtained through this application, and each numerical parameter should be interpreted according to the number of significant digits and conventional rounding methods or in a manner understood by those skilled in the art.

[0036] Although the numerical ranges and parameters setting forth the broad scope of this application are approximations, the numerical values ​​set forth in the specific examples are provided as precisely as possible. However, any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in its respective testing measurements. Every numerical range given herein will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were expressly written herein.

[0037] In general, the nomenclature used herein and the experimental procedures of organic chemistry, medicinal chemistry, and biology described herein are well known in the art and are commonly used in the art. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as those of ordinary skill in the art to which this application belongs. Where multiple definitions exist for a term used herein, the definitions in this section shall prevail unless otherwise stated.

[0038] Most of the currently available CDK4 / 6 inhibitors target the ATP catalytic pocket of CDK4 / 6, such as Palbociclib, Rbociclib, and Abemaciclib. These drugs have shown varying degrees of drug resistance and clinical side effects. Through continuous research, the applicant has found that the compounds of the present application can inhibit the activity of CDK4 / 6 by inhibiting the formation of the CDK4 / 6-CCND complex, thereby inhibiting the CDK4 / 6-CCND-Rb pathway and having good anti-cancer activity. The mechanism of action of the compounds of the present application is different from that of the CDK4 / 6 inhibitors already on the market, and drug resistance and clinical side effects are reduced to varying degrees.

[0039] Compound

[0040] In a first aspect, the present application provides a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI), or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof,

[0041] in,

[0042] Ar1, Ar3, Ar4 and Ar7 are each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl;

[0043] Said Ar2, Ar6 and U2 are each independently an 8-10 membered bicyclic heterocycle;

[0044] The U3 is -N(Ar5)2, and the Ar5 are each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl;

[0045] The R1 are each independently hydrogen or C 0-3 alkyl hydroxyl groups;

[0046] The R2 are each independently hydrogen or C 1-6 alkyl;

[0047] R3 is hydrogen, C 2-6 Alkenyl or C 2-6 Alkynyl;

[0048] The R4 is unsubstituted or substituted C 3-8 Cycloalkyl;

[0049] Said X1 is N or C;

[0050] The U1 is -X2-(CH2) n -C(O)-M1;

[0051] Said X2 and X3 are each independently S or O;

[0052] The M1 is -N(R5)(R6);

[0053] The R5 and R6 together with the N to which they are attached form an 8-10 membered bicyclic heterocycle;

[0054] The U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S;

[0055] The R9 and R 10 Together with the attached C, it forms a 5-6 membered aryl or 5-6 membered heteroaryl group;

[0056] The R 11 and R 12 Together with the attached N and C, it forms a 5-6 membered saturated ring;

[0057] n and t are each independently 1, 2 or 3;

[0058] m is 1 or 2.

[0059] As used herein, the term "plurality" or "multiplicity" refers to two or two, as well as more than two or two.

[0060] As used herein, the term "aryl" refers to a hydrocarbon group containing one or more aromatic rings, for example, an aryl group is a monocyclic, bicyclic, or tricyclic aromatic group having 6 to 20 carbon atoms.

[0061] As used herein, the term "5-6 membered aryl" refers to a monocyclic aryl group having 5 to 6 carbon atoms. Examples of 5-6 membered aryl groups include, but are not limited to, phenyl.

[0062] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or fused ring group containing one or more ring heteroatoms independently selected from N, O, and S. The heteroaryl group can be bonded via a carbon atom or a heteroatom. For example, the heteroaryl group can be an aromatic monocyclic or fused ring group containing 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, and S. The terms "3-8 membered heteroaryl" and "5-6 membered heteroaryl" should be interpreted accordingly and specifically refer to heteroaryl groups having 3-8 ring atoms and heteroaryl groups having 5-6 ring atoms. Examples of heteroaryl groups include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidinyl, or pyridyl.

[0063] As used herein, the term "bicyclic heterocycle" refers to a bicyclic group with unsaturated groups that contains one or more ring heteroatoms independently selected from N, O and S. The term "8-10 membered bicyclic heterocycle" should be interpreted accordingly. Examples of 8-10 membered bicyclic heterocycles include, but are not limited to, a 5-membered ring fused 5-membered ring, a 5-membered ring fused 6-membered ring, or a 6-membered ring fused 6-membered ring. Wherein, the fused 5-membered ring or 6-membered ring is each independently an aromatic monocycle containing ring heteroatoms, an aromatic monocycle not containing ring heteroatoms, a non-aromatic monocycle containing ring heteroatoms, or a non-aromatic monocycle not containing ring heteroatoms.

[0064] As used herein, the term “C 1-6 "Alkyl" refers to a straight or branched saturated hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having from one to six carbon atoms, and attached to the remainder of the molecule by a single bond. 1-5 Alkyl", "C 1-4 Alkyl", "C 1-3 Alkyl", "C 1-2 "Alkyl" should be interpreted accordingly. C 1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, or 1,1-dimethylethyl (tert-butyl).

[0065] As used herein, the term “C 0-3 "Alkylhydroxy" refers to a C 1-3 One of the hydrogen atoms of the alkyl group is replaced by -OH, or, when C is 0, by -OH. 0-3 Examples of alkylhydroxy groups include, but are not limited to, hydroxy, hydroxymethyl, 2-hydroxyethyl, eth-1-ol, 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, and the like.

[0066] As used herein, the term “C 2-6 "Alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to six carbon atoms, and attached to the remainder of the molecule by a single bond. 2-5 Alkenyl", "C 2-4 Alkenyl", "C 2-3 "Alkenyl" should be interpreted accordingly. 2-6 Examples of alkenyl groups include, but are not limited to, vinyl, prop-1-enyl, but-1-enyl, pent-1-enyl, pent-4-enyl, and penta-1,4-dienyl.

[0067] As used herein, the term “C 2-6 "Alkynyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from two to six carbon atoms, and attached to the remainder of the molecule by a single bond. 2-5 Alkynyl", "C 2-4 Alkynyl", "C 2-3 "Alkynyl" should be interpreted accordingly. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-ynyl, pent-4-ynyl, and pent-1,4-diynyl.

[0068] As used herein, the term "cycloalkyl" refers to a monocyclic or bicyclic saturated hydrocarbon group consisting solely of carbon and hydrogen atoms and attached to the remainder of the molecule by a single bond. 3-8 "Cycloalkyl" should be interpreted accordingly and specifically refers to a cycloalkyl group having 3 to 8 carbon atoms. 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0069] As used herein, the term "unsubstituted or substituted" means that a group or ring may be unsubstituted, or the group or ring may be substituted with one or more substituents as defined herein.

[0070] As used herein, the term "halogen" refers to bromine, chlorine, fluorine, or iodine.

[0071] As used herein, the term "saturated ring" refers to a ring in which all the ring bonds involved in the ring formation are saturated. Specifically, the term "5-6 membered saturated ring" refers to a saturated ring having 5-6 carbon atoms; the term "3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S" refers to a 3-, 4-, 5-, or 6-membered saturated carbon ring in which one or more carbon atom ring members may be independently replaced by a heteroatom selected from N, O or S.

[0072] As used herein, the term "fused" refers to two ring members that are connected by sharing two ring atoms.

[0073] In some embodiments, Ar1 is C 1-6 Alkyl-substituted phenyl.

[0074] In some embodiments, Ar3 is a 5-6 membered heteroaryl substituted by halogen.

[0075] In some embodiments, Ar4 is phenyl substituted by halogen.

[0076] In some embodiments, Ar5 and Ar7 are each independently phenyl or a 5-6 membered heteroaryl group.

[0077] In some embodiments, Ar2 is an 8-membered bicyclic heterocycle. In some embodiments, the 8-membered bicyclic heterocycle contains one or more N atoms.

[0078] In some embodiments, the Ar6 is a 9-membered bicyclic heterocycle. In some embodiments, the 9-membered bicyclic heterocycle contains one or more N atoms.

[0079] In some embodiments, U2 is a 9-membered bicyclic heterocycle. In some embodiments, the 9-membered bicyclic heterocycle contains one or more N atoms.

[0080] In some embodiments, the R1 are each independently C 0-3 Alkyl hydroxyl groups.

[0081] In some embodiments, the R2 are each independently C 1-3 alkyl.

[0082] In some embodiments, R3 is C 2-6 Alkynyl.

[0083] In some embodiments, the R4 is unsubstituted C 3-8 Cycloalkyl.

[0084] In some embodiments, X1 is N.

[0085] In some embodiments, X2 is S, and X3 is O.

[0086] In some embodiments, said R5 and R6 together with the N to which they are attached form a 10-membered bicyclic heterocycle.

[0087] In some embodiments, the R7 and R8 together with the attached C form a 5-membered saturated ring. In some embodiments, the 5-membered saturated ring contains one or more O atoms.

[0088] In some embodiments, the R9 and R 10 Together with the attached C, it forms a phenyl group or a 6-membered heteroaryl group.

[0089] In some embodiments, the R 11 and R 12 Together with the attached N and C, it forms a 5-membered saturated ring.

[0090] In some embodiments, the compound of formula (I) is in,

[0091] The Ar1 is an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl; the Ar2 is an 8-10 membered bicyclic heterocycle.

[0092] In some embodiments, in the compound of formula (I), Ar2 is an 8-membered bicyclic heterocycle, a 9-membered bicyclic heterocycle, or a 10-membered bicyclic heterocycle.

[0093] In some embodiments, in the compound of formula (I), Ar2 is an 8-membered bicyclic heterocycle containing one or more N atoms.

[0094] In some embodiments, in the compound of formula (I), Ar2 is a 5-membered ring fused to a 5-membered ring. In some embodiments, Ar2 is a 5-membered unsaturated heterocyclic ring fused to a 5-membered saturated carbocyclic ring. In some embodiments, Ar2 is a 5-membered unsaturated heterocyclic ring fused to a 5-membered saturated carbocyclic ring, and Ar2 contains two nitrogen atoms.

[0095] In some embodiments, in the compound of formula (I), Ar2 is or In some embodiments, in the compound of formula (I), Ar2 is

[0096] In some embodiments, the compound of formula (I) has the structure of formula (I-1):

[0097] Wherein, Ar1 is an unsubstituted or substituted 5-6 membered aryl group or 3-8 membered heteroaryl group.

[0098] In some embodiments, in the compound of formula (I) or formula (I-1), Ar1 is unsubstituted phenyl or C 1-6 In some embodiments, in the compound of formula (I) or formula (I-1), Ar1 is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C1-3 Alkyl, C 1-2 Alkyl or methyl substituted phenyl.

[0099] In some embodiments, in the compounds of formula (I) or formula (I-1), Ar1 is phenyl substituted with a methyl group. In some embodiments, Ar1 is phenyl substituted with 1, 2, 3, 4, or 5 methyl groups. In some embodiments, the methyl substitution may be ortho, meta, or para relative to the linker.

[0100] In some embodiments, the compound of formula (I) or formula (I-1) is selected from

[0101] In some embodiments, the compound of formula (II) is:

[0102] wherein each of the R1s is independently hydrogen or C 0-3 Alkylhydroxyl; said R2 are each independently hydrogen or C 1-6 Alkyl; R3 is hydrogen, C 2-6 Alkenyl or C 2-6 Alkynyl.

[0103] In some embodiments, in the compound of formula (II), each of the R1 is independently C 0-3 In some embodiments, in the compound of formula (II), each of the R1 groups is independently hydroxy, hydroxymethyl, 2-hydroxyethyl, ethanol, 1-hydroxypropyl, 2-hydroxypropyl, or 3-hydroxypropyl. In some embodiments, in the compound of formula (II), each of the R1 groups is hydroxy.

[0104] In some embodiments, in the compound of formula (II), each of the R2 is independently C 1-6 In some embodiments, in the compound of formula (II), each of the R2 is independently C 1-5 Alkyl, C 1-4 Alkyl, C 1- 3 alkyl, C 1-2 Alkyl or methyl.

[0105] In some embodiments, in the compound of formula (II), R3 is C 2-6 In some embodiments, in the compound of formula (II), R3 is C 2-5 Alkenyl, C 2-4 Alkenyl, C 2-3 alkenyl or vinyl.

[0106] In some embodiments, in the compound of formula (II), R3 is C2-6 In some embodiments, in the compound of formula (II), R3 is C 2-5 Alkynyl, C 2-4 Alkynyl, C 2-3 Alkynyl or ethynyl.

[0107] In some embodiments, in the compound of formula (II), R1 is hydroxyl; R2 is methyl; and R3 is ethynyl.

[0108] In some embodiments, the compound of formula (II) is selected from

[0109] In some embodiments, the compound of formula (III) is:

[0110] wherein Ar3 is an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl; and R2 is independently hydrogen or C 1-6 Alkyl; said R4 is unsubstituted or substituted C 3-8 Cycloalkyl; said X1 is N or C.

[0111] In some embodiments, in the compound of formula (III), Ar3 is an unsubstituted 5-6 membered heteroaryl or a substituted 5-6 membered heteroaryl. In some embodiments, in the compound of formula (III), Ar3 is a substituted 5-6 membered heteroaryl. In some embodiments, in the compound of formula (III), Ar3 is a 5-6 membered heteroaryl substituted by halogen. In some embodiments, in the compound of formula (III), Ar3 is a pyridyl, pyrazolyl, imidazolyl, pyrrolyl, pyridazinyl, pyrimidinyl or pyrazinyl substituted by halogen. In some embodiments, in the compound of formula (III), Ar3 is a pyridyl substituted by halogen.

[0112] In some embodiments, in the compound of formula (III), Ar3 is In some embodiments, in the compound of formula (III), Ar3 is

[0113] In some embodiments, in the compound of formula (III), each of the R2 is independently C 1-6 In some embodiments, in the compound of formula (III), each of the R2 is independently C 1-5 Alkyl, C 1-4 Alkyl, C 1- 3 alkyl, C 1-2In some embodiments, in the compound of formula (III), R2 is methyl.

[0114] In some embodiments, in the compound of formula (III), X1 is N.

[0115] In some embodiments, the compound of formula (III) has the structure of formula (III-1):

[0116] Wherein, said R4 is unsubstituted or substituted C 3-8 Cycloalkyl.

[0117] In some embodiments, in the compound of formula (III), R4 is unsubstituted C 3-8 Cycloalkyl or C 1-6 Alkyl substituted C 3-8 In some embodiments, in the compound of formula (III), R4 is unsubstituted C 3-8 In some embodiments, in the compound of formula (III), R4 is cyclopropyl.

[0118] In some embodiments, in the compound of formula (III), Ar3 is The R2 is a methyl group; the R4 is a cyclopropyl group; and the X1 is N.

[0119] In some embodiments, the compound of formula (III) is selected from:

[0120] In some embodiments, the compound of formula (IV) is:

[0121] wherein Ar4 is an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl; and U1 is -X2-(CH2) n -C(O)-M1; wherein M1 is -N(R5)(R6), wherein R5 and R6 together with the N to which they are connected form an 8-10 membered bicyclic heterocyclic ring; wherein R2 is hydrogen or C 1-6 Alkyl; X2 and X3 are each independently S or O; n is 1, 2 or 3.

[0122] In some embodiments, in the compound of formula (IV), Ar4 is unsubstituted phenyl or substituted phenyl. In some embodiments, in the compound of formula (IV), Ar4 is substituted by halogen or C 1-6 Alkyl-substituted phenyl.

[0123] In some embodiments, in the compound of formula (IV), Ar4 is phenyl substituted by halogen. In some embodiments, the halogen is F. In some specific embodiments, the F substitution can be ortho-, meta-, or para-substituted relative to the connecting bond.

[0124] In some embodiments, in the compound of formula (IV), the R5 and R6 form an 8-, 9- or 10-membered bicyclic heterocycle together with the connected N. In some embodiments, in the compound of formula (IV), the R5 and R6 form a 10-membered bicyclic heterocycle together with the connected N. In some embodiments, in the compound of formula (IV), the R5 and R6 form a 6-membered ring fused 6-membered ring together with the connected N. In some embodiments, the R5 and R6 form a 6-membered aromatic ring fused 6-membered saturated heterocycle together with the connected N. In some embodiments, the R5 and R6 form a phenyl-fused hexahydropyridyl, phenyl-fused hexahydropyridazinyl, phenyl-fused hexahydropyrimidinyl or phenyl-fused hexahydropyrazinyl together with the connected N.

[0125] In some embodiments, in the compound of formula (IV), each of the R2 is independently C 1-6 In some embodiments, in the compound of formula (III), each of the R2 is independently C 1-5 Alkyl, C 1-4 Alkyl, C 1- 3 alkyl, C 1-2 In some embodiments, in the compound of formula (III), R2 is methyl.

[0126] In some embodiments, in the compound of formula (IV), n is 1.

[0127] In some embodiments, in the compound of formula (IV), R2 is methyl; and n is 1.

[0128] In some embodiments, in the compound of formula (IV), X2 is S, and X3 is O.

[0129] In some embodiments, in the compound of formula (IV), Ar4 is a phenyl group substituted by a halogen; R5 and R6 together with the attached N form a phenyl-fused hexahydropyridinyl group; R2 is a methyl group; n is 1; X2 is S; and X3 is O.

[0130] In some embodiments, the compound of formula (IV) is selected from:

[0131] In some embodiments, the compound of formula (V) is:

[0132] wherein Ar6 and U2 are each independently an 8-10 membered bicyclic heterocycle; U3 is -N(Ar5)2, and Ar5 is each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl.

[0133] In some embodiments, in the compound of formula (V), the Ar6 is an 8-, 9- or 10-membered bicyclic heterocycle. In some embodiments, in the compound of formula (V), the Ar6 is a 9-membered bicyclic heterocycle, and the 9-membered bicyclic heterocycle contains one or more N atoms. In some embodiments, in the compound of formula (V), the Ar6 is a 6-membered ring fused to a 5-membered ring, and the 6-membered ring fused to a 5-membered ring contains 1, 2, 3 or 4 nitrogen atoms. In some embodiments, in the compound of formula (V), the Ar6 is a 6-membered aromatic ring fused to a 5-membered heteroaromatic ring, and the 5-membered heteroaromatic ring contains 1, 2, 3 or 4 nitrogen atoms. In some embodiments, the Ar6 is a phenyl-fused imidazolyl, a phenyl-fused pyrazolyl, a phenyl-fused pyrrolyl, a phenyl-fused triazolyl or a phenyl-fused tetrazolyl. In some embodiments, the Ar6 is a phenyl-fused triazolyl, optionally, the Ar6 is In some embodiments, the Ar6 can also be

[0134] In some embodiments, the compound of formula (V) has the structure of formula (V-1);

[0135] wherein U2 is an 8-10 membered bicyclic heterocycle; U3 is -N(Ar5)2, and each Ar5 is independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl.

[0136] In some embodiments, in the compound of formula (V), the Ar5 is each independently unsubstituted phenyl or 5-6 membered heteroaryl, or is substituted phenyl or 5-6 membered heteroaryl. In some embodiments, the Ar5 is each independently unsubstituted phenyl, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidinyl or pyridyl. In some embodiments, the Ar5 is phenyl.

[0137] In some embodiments, in the compound of formula (V), U2 is an 8-, 9-, or 10-membered bicyclic heterocycle. In some embodiments, in the compound of formula (V), U2 is a 9-membered bicyclic heterocycle, and the 9-membered bicyclic heterocycle contains one or more N atoms. In some embodiments, in the compound of formula (V), U2 is a 6-membered ring fused to a 5-membered ring, and the 6-membered ring fused to the 5-membered ring contains 1, 2, 3, or 4 nitrogen atoms. In some embodiments, in the compound of formula (V), U2 is a 6-membered aromatic ring fused to a 5-membered heteroaromatic ring, and the 5-membered heteroaromatic ring contains 1, 2, 3, or 4 nitrogen atoms. In some embodiments, U2 is a phenyl-fused imidazolyl, a phenyl-fused pyrazolyl, a phenyl-fused pyrrolyl, a phenyl-fused triazolyl, or a phenyl-fused tetrazolyl. In some embodiments, U2 is a phenyl-fused triazolyl, optionally, U2 is In some embodiments, U2 can also be

[0138] In some embodiments, the compound of formula (V) is selected from:

[0139] In some embodiments, the compound of formula (VI) is:

[0140] wherein Ar7 is an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl;

[0141] The U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S;

[0142] The R9 and R 10 Together with the attached C, it forms a 5-6 membered aryl or 5-6 membered heteroaryl group;

[0143] The R 11 and R 12 Together with the attached N and C, it forms a 5-6 membered saturated ring;

[0144] t is 1, 2, or 3;

[0145] m is 1 or 2.

[0146] In some embodiments, in the compound of formula (VI), the Ar7 is unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl. In some embodiments, in the compound of formula (VI), the Ar7 is phenyl or 5-6 membered heteroaryl. In some embodiments, in the compound of formula (VI), the Ar7 is phenyl, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidinyl or pyridyl.

[0147] In some embodiments, in the compound of formula (VI), t is 1, 2 or 3.

[0148] In some embodiments, in the compound of formula (VI), Ar7 is phenyl; and t is 1.

[0149] In some embodiments, in the compound of formula (VI), said R9 and R 10 Together with the attached C, they form a 5-6 membered aryl or 5-6 membered heteroaryl. In some embodiments, in the compound of formula (VI), the R9 and R 10 Together with the attached C, they form a phenyl group or a 5-6 membered heteroaryl group. In some embodiments, in the compound of formula (VI), the R9 and R 10 Together with the attached C, it forms a phenyl group, a furyl group, a pyrrolyl group, a thienyl group, a pyrazolyl group, an imidazolyl group, a thiazolyl group, an isothiazolyl group, an oxazolyl group, an isoxazolyl group, a triazolyl group, a tetrazolyl group, a pyrazinyl group, a pyridazinyl group, a pyrimidinyl group or a pyridyl group.

[0150] In some embodiments, in the compound of formula (VI), the R 11 and R 12 Together with the attached N and C, they form a 5-6 membered saturated ring. In some embodiments, in the compound of formula (VI), the R 11 and R 12 Together with the attached N and C, a 5-membered saturated ring is formed. In some embodiments, in the compound of formula (VI), the 5-membered saturated ring comprises 1 N atom, and optionally further comprises 1, 2 or 3 heteroatoms selected from N, O or S.

[0151] In some embodiments, in the compound of formula (VI), said R9 and R 10 Together with the connected C, a phenyl group is formed; the R 11 and R 12 Together with the attached N and C, it forms a 5-membered saturated ring.

[0152] In some embodiments, the compound of formula (VI) has the structure of formula (VI-1);

[0153] Among them, the U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S, and m is 1 or 2.

[0154] In some embodiments, in the compound of formula (VI), the R7 and R8 together with the C to which they are attached form a 3-6 membered saturated ring, and the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S. In some embodiments, in the compound of formula (VI), the R7 and R8 together with the C to which they are attached form a 5-membered saturated ring, and the 5-membered saturated ring contains one or more heteroatoms selected from N, O or S. In some embodiments, in the compound of formula (VI), the R7 and R8 together with the C to which they are attached form a 5-membered saturated ring, and the 5-membered saturated ring contains one or more O atoms. In some embodiments, in the compound of formula (VI), the R7 and R8 together with the C to which they are attached form a 5-membered saturated ring, and the 5-membered saturated ring contains 1, 2, 3 or 4 O atoms. In some embodiments, in the compound of formula (VI), the R7 and R8 together with the C to which they are attached form a 5-membered saturated ring, and the 5-membered saturated ring contains 2 O atoms, optionally, the 5-membered saturated ring is selected In some embodiments, the 5-membered saturated ring can also be selected from

[0155] In some embodiments, in the compound of formula (VI), m is 1.

[0156] In some embodiments, the compound of formula (VI) is selected from

[0157] In some embodiments, unless otherwise indicated, the compounds of Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V) or (VI) of the present application may be in one of the following forms: possible stereoisomers, rotamers, atropisomers, tautomers or mixtures thereof, solvates, isotopic derivatives or pharmaceutically acceptable salts. For example, as substantially pure geometric (cis or trans) stereoisomers, diastereomers, optical isomers (enantiomers), racemates or mixtures thereof.

[0158] As used herein, the term "solvate" refers to the association of one or more solvent molecules with the compound of the present application or a salt thereof. Solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, ethyl acetate, or acetic acid.

[0159] As used herein, the term "isotopic derivative" refers to a structure depicted by the chemical formula given herein in which one or more atoms are replaced by an isotopic atom having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the present invention include, for example, isotopes of hydrogen. In some specific embodiments, the incorporation of certain isotopes, particularly deuterium (i.e., 2 H or D) may offer certain therapeutic advantages by providing greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements or improved therapeutic index or tolerability.

[0160] In some embodiments, isotopes that can be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, chlorine, fluorine, or iodine, such as 3 H. 11 C. 13 C. 1 4 C. 15 N. 18 F. 35 S. 36 Cl, 1 23 I. 124 I or 125 I, etc. In some embodiments, the present application includes compounds incorporating one or more of the above isotopes, for example, compounds incorporating radioactive isotopes 3 H and / or 14 C, or also including the incorporation of non-radioactive isotopes 2 H and / or 13 Part C.

[0161] In some embodiments, the compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) of the present application may exist in a free form or in the form of a salt thereof.

[0162] As used herein, the term "pharmaceutically acceptable salt" refers to salts that retain the desired biological activity of the compounds described herein and exhibit reduced undesirable toxicological effects. These pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in its free acid or free base form with a suitable base or acid, respectively.

[0163] In some embodiments, pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. The inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, or phosphoric acid. The organic acids include acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, or sulfosalicylic acid.

[0164] In some embodiments, pharmaceutically acceptable base addition salts can be formed with inorganic bases and organic bases. The inorganic bases include, for example, ammonium salts and metals from columns I to XII of the periodic table. The organic bases include, for example, primary, secondary, and tertiary amines; substituted amines (including naturally occurring substituted amines); cyclic amines; basic ion exchange resins, and the like. In some specific embodiments, the organic bases include isopropylamine, benzathine, choline salts, diethanolamine, diethylamine, lysine, meglumine, piperazine, or tromethamine, and the like.

[0165] Pharmaceutical composition

[0166] In a second aspect, the present application provides a pharmaceutical composition comprising a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) as described in the first aspect of the present application, or an enantiomer, diastereomer, racemate, solvate, isotope derivative or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0167] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable raw material, component, or vehicle that imparts form or consistency to the pharmaceutical composition. When combined, each carrier is compatible with the other ingredients of the pharmaceutical composition to avoid significantly reducing the efficacy of the compound of the present invention upon administration to a subject and to avoid interactions with the drug components that would render them pharmaceutically unacceptable.

[0168] In some embodiments, the pharmaceutically acceptable carrier includes, but is not limited to, a diluent, a filler, a binder, a disintegrant, a lubricant, a glidant, a granulating agent, a coating agent, a wetting agent, a solvent, a co-solvent, a suspending agent, an emulsifier, a sweetener, a flavoring agent, a taste masking agent, a coloring agent, an anti-caking agent, a humectant, a chelating agent, a plasticizer, a viscosity increasing agent, an antioxidant, a preservative, a stabilizer, a surfactant, or a buffer, etc. Those skilled in the art may selectively use the above-mentioned pharmaceutically acceptable carriers according to the dosage form of the pharmaceutical composition.

[0169] In some embodiments, the compound of the present application and one or more pharmaceutically acceptable carriers are formulated into a dosage form suitable for administration to a subject via a desired route of administration. In some embodiments, the dosage form of the pharmaceutical composition includes but is not limited to tablets, capsules, caplets, pills, lozenges, powders, syrups, brews, suspensions, solutions, emulsions, transdermal patches, suppositories, inhalants, creams, ointments, lotions, pastes, sprays, injections, or gels.

[0170] A combination

[0171] In a third aspect, the present application provides a combination comprising a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) described in the first aspect of the present application, or an enantiomer, diastereomer, racemate, solvate, isotope derivative or pharmaceutically acceptable salt thereof, and one or more therapeutically active agents.

[0172] In some embodiments, the therapeutically active agent includes but is not limited to other anticancer agents, antiallergic agents, antiemetics, or analgesics.

[0173] In some embodiments, other anticancer agents can be selected for use with the compounds of the present invention based on the disease or condition being treated. In some specific embodiments, for the treatment of breast cancer, the other anticancer agents may include but are not limited to trastuzumab, pertuzumab, lapatinib, tamoxifen, everolimus, paclitaxel or sipatinib; for the treatment of cervical cancer, the other anticancer agents may include but are not limited to bleomycin, cisplatin or topotecan; for the treatment of liver cancer, the other anticancer agents may include but are not limited to sorafenib, lenvatinib mesylate, fluorouracil or oxaliplatin; for the treatment of prostate cancer, the other anticancer agents may include but are not limited to flutamide, goserelin, docetaxel, cabazitaxel, mitoxantrone or estradiol nitrogen mustard; for the treatment of lung cancer, the other anticancer agents may include but are not limited to methotrexate, paclitaxel, maleic acid Afatinib, pemetrexed disodium, bevacizumab, carboplatin, cisplatin, crizotinib, erlotinib hydrochloride, gefitinib or gemcitabine hydrochloride, etc.; for the treatment of leukemia, other anticancer agents may include but are not limited to bosutinib, cyclophosphamide, cytarabine, dasatinib, imatinib, ponatinib mesylate, nilotinib or homoharringtonine, etc.; for the treatment of ovarian cancer, the other anticancer agents may include but are not limited to doxorubicin hydrochloride, carboplatin, cyclophosphamide, cisplatin, doxorubicin hydrochloride liposome, gemcitabine hydrochloride, topotecan hydrochloride or paclitaxel, etc.; for the treatment of colon cancer, the other anticancer agents may include but are not limited to oxaliplatin, fluorouracil, S-1, bevacizumab or cetuximab, etc.

[0174] In some embodiments, for possible allergic reactions, the compounds of the present application can be used in combination with antiallergic agents. In some embodiments, the antiallergic agents include but are not limited to dexamethasone, beclomethasone, hydrocortisone, prednisone, prednisolone, diphenhydramine, cyproheptadine, salbutamol or terbutaline.

[0175] In some embodiments, the compounds of the present application can be used in combination with an antiemetic to address possible vomiting. In some embodiments, the antiemetic includes but is not limited to aprepitant, ondansetron, granisetron, lorazepam, dexamethasone, prochlorperazine, or casopitant.

[0176] In some embodiments, in order to relieve the pain of the subject, the compound of the present application can be used in combination with an analgesic. In some embodiments, the analgesic includes but is not limited to morphine, oxycodone, fentanyl or oxymorphone hydrochloride.

[0177] use

[0178] In a fourth aspect, the present application provides a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) as described in the first aspect of the present application, or its enantiomer, diastereomer, racemate, solvate, isotope derivative or pharmaceutically acceptable salt, for the preparation of a drug for a disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D.

[0179] As used herein, the term "disease" refers to any change in the state of the body or some organ that interrupts or interferes with the performance of functions and / or causes symptoms (such as discomfort, dysfunction, adverse stress or even death) in the person suffering from the disease or those who come into contact with it.

[0180] As used herein, the term "treat," ...

[0181] As used herein, the term "cyclin D (CCND)" is an important member of the cyclin family that regulates G1-S transition. Cyclin D has three isoforms: cyclin D1 (CCND1), cyclin D2 (CCND2), and cyclin D3 (CCND3), all of which have similar expression patterns.

[0182] In some embodiments, the inhibitory activity IC of the compounds of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) provided herein on CDK4 / 6 kinases is 50 Not exceeding 1000μM, 800μM, 600μM, 500μM, 400μM, 300μM, 200μM, 100μM, 50μM.

[0183] In some embodiments, the disease that benefits from or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D includes cancer. Alternatively, the disease that benefits from or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D is selected from breast cancer, glioma, liver cancer, prostate cancer, cervical cancer, lung cancer, leukemia, gastric cancer, ovarian cancer or colon cancer.

[0184] In some embodiments, the toxicity IC of the compounds of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) provided herein to human breast cancer cells MCF-7 is 50 Not exceeding 1000μM, 800μM, 600μM, 500μM, 400μM, 300μM, 200μM, 100μM, 50μM.

[0185] In some embodiments, the toxicity IC of the compounds of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) provided herein to human breast cancer cells HCC1937 is 50 Not exceeding 1000μM, 800μM, 600μM, 500μM, 400μM, 300μM, 200μM, 100μM, 50μM.

[0186] In some embodiments, the toxicity IC of the compounds of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) provided herein to cervical cancer cells HeLa is 50 Not exceeding 1000μM, 800μM, 600μM, 500μM, 400μM, 300μM, 200μM, 100μM, 50μM.

[0187] In some embodiments, the disease that can benefit from or be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D includes breast cancer and / or cervical cancer.

[0188] Treatment

[0189] In a fifth aspect, the present application provides a method for treating a disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D, the method comprising administering to a subject in need of such treatment a therapeutically effective dose of a compound of formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) as described in the first aspect of the present application, or an enantiomer, diastereomer, racemate, solvate, isotope derivative or pharmaceutically acceptable salt thereof.

[0190] As used herein, the term "subject" refers to primates (e.g., humans), dogs, rabbits, guinea pigs, pigs, rats, and mice. In some embodiments, the subject is a primate. In some specific embodiments, the subject is a human.

[0191] As used herein, a subject is "in need of" a treatment if the subject would benefit biologically, medically, or in quality of life from such treatment.

[0192] As used herein, the term "therapeutically effective dose" refers to an amount that results in a benefit or treatment of a disease compared to a corresponding subject not receiving that amount, but which is sufficiently low within the scope of sound medical judgment to avoid serious side effects. The therapeutically effective dose of a compound will vary with the specific compound selected (e.g., taking into account the potency, efficacy, and half-life of the compound); the route of administration selected; the disease being treated; the severity of the disease being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient being treated; the duration of treatment; the nature of concurrent treatment; the desired therapeutic effect, and the like, but can still be determined in a routine manner by those skilled in the art.

[0193] In some embodiments, the disease that benefits from or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D includes cancer. Alternatively, the disease that benefits from or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D is selected from breast cancer, glioma, liver cancer, prostate cancer, cervical cancer, lung cancer, leukemia, gastric cancer, ovarian cancer or colon cancer.

[0194] The various embodiments and preferences for the compounds, pharmaceutical compositions or combinations of the present application described above can be combined with each other (as long as they are not inherently contradictory to each other) and are applicable to the use of the present application. The various embodiments formed by such combinations are considered to be part of the present application.

[0195] The technical solutions of the present application will be more clearly and specifically described below in conjunction with examples in an illustrative manner. It should be understood that these examples are for illustrative purposes only and are in no way intended to limit the scope of protection of the present application. The scope of protection of the present application is defined solely by the claims.

[0196] Example

[0197] Unless otherwise stated, all reagents and instruments used in the following examples are commercially available conventional products. Unless otherwise stated, experiments were performed under conventional conditions or conditions recommended by the manufacturer.

[0198] Compound

[0199] Compound 1: purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0200] Compound 2: purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0201] Compound 3: purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0202] Compound 4: purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0203] Compound 5: purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0204] Compound 6 was purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0205] Biological experiments

[0206] Example 1: Inhibition of protein-protein interaction between CDK4 / 6 and CCND

[0207] Experimental methods

[0208] The protein-protein interaction experiment of inhibiting CDK4 / 6 and CCND was performed based on bimolecular fluorescence complementation (BiFC). The specific steps are as follows:

[0209] (1) VN155-CDK4, VN155-CDK6, and VC155-CCND1 vectors (CDK4: NCBI No. NM_000075.4; CDK6: NCBI No. NM_001145306.2; CCNDI: NCBI No. NM_053056.3) were constructed and confirmed by sequencing. HEK293T cells (from Zhejiang University Laboratory) plated in 24-well plates were co-transfected with VN155-CDK4+VC155-CCND1 and VN155-CDK6+VC155-CCND1, as well as the negative control plasmid VC155-mCherry (0.3 μg of each plasmid per well). After incubation for 18 hours, the fluorescence intensity of intracellular protein-protein interactions was detected using a fluorescence microscope (Olympus, IX73).

[0210] (2) HEK293T cells were plated in a 96-well plate. 12 hours later, the cells were co-transfected with VN155-CDK4+VC155-CCND1 and VN155-CDK6+VC155-CCND1 (50 ng / well for each plasmid). Four hours after transfection, the compound of the present application was added at a final concentration of 10 μM, and the cells were cultured in a 37°C cell culture incubator for 18 hours. Fluorescence signals representing intracellular protein-protein interactions were detected using a fluorescence microscope (Olympus, IX73) (detection channel: FITC), and the fluorescence intensity of the cells was quantitatively measured using a high-content system (MD, IXM-C). The co-transfected mCherry signal intensity was used as a reference signal to normalize the protein-protein interaction signal.

[0211] Experimental results

[0212] As shown in Figure 1, 293T cells co-transfected with VN155-CDK4+VC155-CCND1 and VN155-CDK6+VC155-CCND1 have strong green fluorescence signals, while the negative control plasmid has no fluorescence signal due to the lack of interaction. This confirms that the constructed system for detecting protein-protein interaction between CDK4 / 6 and CCND can work normally.

[0213] As shown in Figure 2, compounds 1 to 5 can block the interaction of CDK4 / CCND1, and compounds 4-6 can block the interaction of CDK6 / CCND1.

[0214] Example 2: Experiment on inhibition of CDK4 / 6 kinase biological activity

[0215] Experimental methods

[0216] Using 293T cells as the basic reaction system, CDK4-CCND1-RB1CT (RB1 protein C-terminus, containing the S795 site) and CDK6-CCND1-RB1CT were overexpressed. After treatment with different concentrations of compounds (200, 100, 50, 25, 12.5, 6.25, 3.125, and 1.5625 μM) for 20 hours, cells were collected and the phosphorylation level of RB1CT S795 was detected by western blotting (S795 antibody purchased from Abclonal), and the corresponding IC was calculated. 50 The kinase inhibitory activity of the compound was indirectly measured.

[0217] The experimental results are shown in the following table:

[0218] The enzyme inhibitory activity (IC 50 , unit: μM)

[0219] Compounds 1-6 showed varying degrees of inhibitory activity against CDK4 kinase, with Compounds 1 and 6 showing excellent inhibitory activity against CDK4 kinase, and Compound 5 showing good inhibitory activity against CDK4 kinase. Compounds 4-6 showed good inhibitory activity against CDK6 kinase, with Compounds 5 and 6 showing excellent inhibitory activity against CDK6 kinase.

[0220] Example 3: Experiment on inhibiting proliferation of various cancer cells

[0221] Experimental methods

[0222] The CCK-8 assay was used to test the inhibitory effect of the compounds of the present invention on the proliferation of human breast cancer (MCF-7, HCC1937) cell lines and cervical cancer (HeLa) cell lines. The specific method is as follows:

[0223] (1) Healthy and vigorous tumor cells were cultured at a rate of 1×10 5 Cells / well were seeded in 96-well plates and cultured at 37°C, 5% CO2 for more than 12 h;

[0224] (2) Add the culture medium solution of the compound of the present application diluted in the required concentration gradient (200, 100, 50, 25, 12.5, 6.25, 3.125 and 1.5625 μM) to the culture plate and incubate for 48 hours, setting up 6 replicate wells;

[0225] (3) One hour before the end of incubation, 10 μL of 11xCCK-8 solution (Taoshu, TP11970) was added to each well. After the incubation, OD450 was measured with a microplate reader, and the IC of the compound of the present application was obtained by fitting. 50 .

[0226] The experimental results are shown in the following table:

[0227] The cytotoxicity of the compound (IC 50 , μM)

[0228] Compounds 1-6 showed good cytotoxicity to MCF-7 cells, HCC1937 cells and HeLa cells. Compound 6 showed excellent cytotoxicity to MCF-7 cells, and compound 1 showed excellent cytotoxicity to HCC1937 cells and HeLa cells.

[0229] Although specific embodiments have been described, it is possible that alternatives, modifications, variations, improvements, and substantial equivalents of the above embodiments may exist or are not currently foreseeable to the applicant or other skilled in the art. Therefore, the appended claims, as filed and as they may be amended, are intended to cover all such alternatives, modifications, variations, improvements, and substantial equivalents.

Claims

1. A compound, or an enantiomer, diastereomer, racemate, solvate, isotope derivative or pharmaceutically acceptable salt thereof, wherein the compound is represented by formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI), in, Ar1, Ar3, Ar4 and Ar7 are each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl; Ar2, Ar6 and U2 are each independently an 8-10 membered bicyclic heterocycle; The U3 is -N(Ar5)2, and the Ar5 is independently an unsubstituted or substituted 5-6-membered aryl or 3-8-membered heteroaryl; The R1 are each independently hydrogen or C 0-3 Alkyl hydroxyl; The R2 are each independently hydrogen or C 1-6 alkyl; R3 is hydrogen, C 2-6 Alkenyl or C 2-6 Alkynyl; The R4 is unsubstituted or substituted C 3-8 Cycloalkyl; X1 is N or C; The U1 is -X2-(CH2) n -C(O)-M1; Said X2 and X3 are each independently S or O; The M1 is -N(R5)(R6); The R5 and R6 together with the N to which they are attached form an 8-10 membered bicyclic heterocycle; The U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S; The R9 and R 10 Together with the attached C, it forms a 5-6 membered aryl or 5-6 membered heteroaryl; The R 11 and R 12 Together with the attached N and C, it forms a 5-6 membered saturated ring; n and t are each independently 1, 2 or 3; m is 1 or 2.

2. The compound according to claim 1, wherein Ar1 is C 1-6 Alkyl substituted phenyl, Ar2 is an 8-membered bicyclic heterocycle, the 8-membered bicyclic heterocycle contains one or more N atoms, U2 and Ar6 are each independently a 9-membered bicyclic heterocycle, the 9-membered bicyclic heterocycle contains one or more N atoms; Ar3 is a 5-6 membered heteroaryl group substituted by halogen, Ar4 is a phenyl group substituted by halogen, Ar5 and Ar7 are each independently phenyl or a 5-6 membered heteroaryl group; The R1 are each independently C 0-3 Alkyl hydroxyl; The R2 are each independently C 1-3 alkyl; R3 is C 2-6 Alkynyl; The R4 is an unsubstituted C 3-8 Cycloalkyl; X1 is N; X2 is S, and X3 is O; The R5 and R6 together with the N to which they are attached form a 10-membered bicyclic heterocycle; The R7 and R8 together with the connected C form a 5-membered saturated ring, and the 5-membered saturated ring contains one or more O atoms; The R9 and R 10 Together with the attached C, it forms a phenyl group or a 6-membered heteroaryl group; The R 11 and R 12 Together with the attached N and C, it forms a 5-membered saturated ring.

3. The compound according to claim 1 or 2, wherein The compound satisfies at least one of the following: (1) In the compound of formula (I), Ar2 is a 5-membered unsaturated heterocyclic ring fused to a 5-membered saturated carbon ring, wherein Ar2 contains two N atoms; alternatively, Ar2 is (2) In the compound of formula (III), Ar3 is pyridine substituted by halogen; alternatively, Ar3 is The R2 is a methyl group; (3) In the compound of formula (V), Ar6 is a phenyl-fused triazole group; (4) In the compound of formula (VI), R9 and R 10 Together with the connected C, a phenyl group is formed; the R 11 and R 12 Together with the connected N and C, a 5-membered saturated ring is formed; Ar7 is phenyl, and t is 1.

4. The compound according to any one of claims 1 to 3, wherein The compound of formula (I) has the structure of formula (I-1): Wherein, Ar1 is an unsubstituted or substituted 5-6-membered aryl or 3-8-membered heteroaryl; The compound of formula (III) has the structure of formula (III-1): Wherein, said R4 is unsubstituted or substituted C 3-8 Cycloalkyl; The compound of formula (V) has the structure of formula (V-1): The U2 is an 8-10 membered bicyclic heterocyclic ring; the U3 is -N(Ar5)2, and the Ar5 is each independently an unsubstituted or substituted 5-6 membered aryl or 3-8 membered heteroaryl; The compound of formula (VI) has the structure of formula (VI-1): Among them, the U4 is The R7 and R8 together with the connected C form a 3-6 membered saturated ring, wherein the 3-6 membered saturated ring contains one or more heteroatoms selected from N, O or S; m is 1 or 2.

5. The compound according to any one of claims 1 to 4, wherein The compound satisfies at least one of the following conditions: (1) In the compound of formula (I) or formula (I-1), Ar1 is a phenyl group substituted with a methyl group; (2) In the compound of formula (II), R1 is hydroxyl; R2 is methyl; R3 is ethynyl; (3) In the structure of formula (III) or formula (III-1), R4 is C 3-6 Cycloalkyl; (4) In the compound of formula (IV), Ar4 is a phenyl group substituted by a halogen, and the halogen is F; (5) In the compound of formula (IV), R5 and R6 together with the N to which they are attached form a phenyl-fused hexahydropyridinyl group; (6) In the compound of formula (IV), R2 is methyl and n is 1; (7) In the compound of formula (V), U2 is a phenyl-fused triazole group, and optionally, U2 is (8) In the compound of formula (V), Ar5 is phenyl; (9) In the compound of formula (VI), R7 and R8 together with the connected C form a 5-membered saturated ring, the 5-membered saturated ring contains two O atoms, and optionally, the 5-membered saturated ring is selected from The m is 2.

6. The compound according to any one of claims 1 to 5, wherein The compound of formula (I) or formula (I-1) is selected from: The compound of formula (II) is selected from: The compound of formula (III) or formula (III-1) is selected from: The compound of formula (IV) is selected from: The compound of formula (V) is selected from: The compound of formula (VI) is selected from:

7. A pharmaceutical composition comprising a compound as described in any one of claims 1 to 6, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

8. A combination comprising a compound as claimed in any one of claims 1 to 6, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, and one or more therapeutically active agents.

9. Use of a compound according to any one of claims 1 to 6, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7, or a combination according to claim 8, in the preparation of a medicament for a disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D.

10. A method for treating a disease that benefits or can be treated by inhibiting the protein-protein interaction of CDK4 / 6 and cyclin D, the method comprising administering to a subject in need of such treatment a therapeutically effective dose of a compound as described in any one of claims 1 to 6, or an enantiomer, diastereomer, racemate, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 7, or a combination as described in claim 8.

11. The use according to claim 9 or the method according to claim 10, wherein The disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D includes cancer. Optionally, the disease that benefits or can be treated by inhibiting the protein-protein interaction between CDK4 / 6 and cyclin D includes one or more of breast cancer, glioma, liver cancer, prostate cancer, cervical cancer, lung cancer, leukemia, gastric cancer, ovarian cancer and colon cancer.

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