Protein degradation agent, and pharmaceutical composition and use thereof

Abstract

A protein degradation agent, and a pharmaceutical composition and the use thereof. The structure as shown in formula (A) is used as an LC3B recruitment compound, and a protein degradation agent is obtained by means of connecting the structure as shown in formula (A), particularly a 2,4-quinazolinedione structure with a ligand of a target protein, which enable the autophagy degradation of the target protein, generate a biological effect, and facilitate the treatment and prevention of related diseases. The protein degradation agent can be used for treating, preventing and / or improving diseases associated with CDK, EZH2, or RAS.

Description

A class of protein degraders and pharmaceutical compositions and uses thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202310710665.0, filed on June 14, 2023, entitled “A class of protein degraders, pharmaceutical compositions and uses thereof”, the entire contents of which are incorporated herein by reference in their entirety to the same extent as if fully set forth herein. Technical Field

[0003] The present invention relates to the field of medical technology, and relates to a class of protein degraders and pharmaceutical compositions thereof, and their use in preparing drugs for treating, preventing and / or improving diseases related to CDK, EZH2 or RAS. Technical Background

[0004] Targeted protein degradation is a highly promising tool for studying biological mechanisms and as a therapeutic approach. Directly degrading the target protein can overcome drug resistance caused by target protein mutations and can also leverage the non-enzymatic clearance of pathogenic proteins that inhibitors lack. Over the past two decades, PROTAC technology has made significant progress, with several PROTAC drug candidates entering clinical trials. However, PROTAC technology still faces challenges, such as the emergence of resistance to E3 ubiquitin ligases and the fact that many substances within the cell are not substrates for proteasome clearance despite relying on the ubiquitin-proteasome system.

[0005] Therefore, there is still a need to develop new protein degradation technologies for degrading target proteins.

[0006] Summary of the Invention

[0007] The inventors used CDK9, which is distributed in cells, as a preliminary screening for degradation fragments and found that by introducing a connecting chain at the 1-position of the structure shown in formula (A) and connecting it to the ligand of CDK9, a series of compounds with degradation effects on the CDK9-cyclin T1 complex were obtained, and it was clarified that it can achieve autophagic degradation of CDK9 by recruiting LC3B. Similarly, the degradation fragments of the structure shown in the relevant representative formula (A) were connected with binding fragments of other targets (such as EZH2, RAS or other CDK inhibitors, etc.), and the related proteins were degraded, indicating that the structure has a broad spectrum of protein degradation. Therefore, the present invention proposes for the first time that the structure shown in formula (A) can be used as an LC3B recruitment compound. By connecting the structure shown in formula (A), especially the 2,4-quinazolinedione structure, to the ligand of the target protein, a protein degrader is obtained, which can achieve autophagic degradation of the target protein, produce biological effects, and treat and prevent related diseases.

[0008] In view of this, one of the objectives of the present invention is to provide a class of protein degradation agents.

[0009] A second object of the present invention is to provide a method for preparing the above-mentioned protein degradation agent.

[0010] A third object of the present invention is to provide a pharmaceutical composition comprising the above-mentioned protein degrading agent.

[0011] A fourth object of the present invention is to provide the use of the above-mentioned protein degrading agent or pharmaceutical composition in the preparation of a drug for treating, preventing and / or ameliorating diseases related to CDK, EZH2, or RAS.

[0012] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0013] In a first aspect, the present invention provides a protein degrading agent or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, wherein the protein degrading agent comprises:

[0014] At least one portion that can bind to LC3 (LC3Binding Moiety, hereinafter sometimes referred to as LCBM portion);

[0015] At least one protein of interest ligand (hereinafter sometimes referred to as POIL) portion that can bind to CDK, EZH2, or RAS; and

[0016] A linker moiety is used to covalently link the LCBM moiety to the POIL moiety independently.

[0017] In this application, LC3 refers to microtubule-associated protein 1A / 1B-light chain 3 (LC3), a soluble protein with a molecular weight of approximately 17 kDa. LC3 is ubiquitous in mammalian tissues and cultured cells and is a key component of autophagy, the recycling system of eukaryotic cells. It is incorporated into the inner and outer membranes of autophagosomes during autophagosome biogenesis. Therefore, LC3 is a specific marker for autophagy, particularly autophagosome formation.

[0018] CDK refers to the cyclin-dependent kinase family of serine / threonine kinases.

[0019] EZH2 refers to Enhancer of zeste homologue 2.

[0020] RAS refers to the RAS protein.

[0021] In a specific embodiment, the present invention provides a protein degradation agent represented by formula (1) or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof,

[0022] LCBM-Linker-POIL(1);

[0023] in:

[0024] LCBM indicates the portion that can bind to LC3;

[0025] Linker represents a covalently linked moiety;

[0026] POIL represents the target protein ligand portion that can bind to CDK, EZH2, or RAS.

[0027] The LCBM moiety and the POIL moiety can each be independently selected from small molecule compounds.

[0028] In some embodiments, the molecular weight of the LCBM portion and the POIL portion are each independently about 100 to about 2000 Da, preferably about 100 to about 1000 Da, for example, about 100 to about 900 Da, about 100 to about 800 Da, about 100 to about 700 Da, about 100 to about 600 Da, about 100 to about 500 Da.

[0029] An LCBM moiety can be linked to one or more POIL moieties, and vice versa. When more than one POIL moiety is present, each POIL moiety can be independently selected and can be the same or different from one another. In some embodiments, multiple POIL moieties are present that target the same target. Even when targeting the same target, multiple identical or different POIL moieties can be used in a single molecule as needed. When more than one POIL moiety is used, the linker used can also be independently selected.

[0030] LCBM part

[0031] This portion indicates a portion that can bind to LC3 and refers to a portion that has affinity for the LC3 protein.

[0032] In some embodiments, the LCBM moiety is of formula (A),

[0033] in:

[0034] Y 1 and Y 2 Each independently selected from O or S;

[0035] Ar ring is selected from C6-C10 aryl or 5-10 membered heteroaryl;

[0036] R 1 is n substituents on the Ar ring, where n is an integer selected from 0 to 4, such as 0, 1, 2, 3, 4;

[0037] Each R 1 Each is independently selected from halogen, cyano (-CN), hydroxyl (-OH), amino (-NH2), nitro (-NO2), carboxyl (-COOH), unsubstituted or substituted C1-C20 alkyl, unsubstituted or substituted C1-C20 alkoxy, C1-C20 alkyl-NH-, (C1-C20 alkyl)(C1-C20 alkyl)N-, C1-C20 alkoxycarbonyl-NH-, unsubstituted or substituted C3-C16 cycloalkyl, unsubstituted or substituted 3-16 membered heterocyclyl, unsubstituted or substituted C6-C14 aryl, unsubstituted or substituted 5-15 membered heteroaryl, wherein the substitution means that one or more hydrogens in the defined group are replaced by one or more substituents selected from halogen, amino (-NH2) and hydroxyl (-OH);

[0038] R 2 Selected from hydrogen, unsubstituted or substituted C1-C20 alkyl, amino C1-C20 alkyl, (C1-C6 alkyl) (C1-C6 alkyl) N-C1-C16 alkyl, C1-C6 alkyl-NH-C1-C16 alkyl, unsubstituted or substituted C3-C16 cycloalkyl, unsubstituted or substituted C3-C16 cycloalkyl C1-C6 alkyl, unsubstituted or substituted 3-16 membered heterocycloalkyl, unsubstituted or substituted 3-10 membered heterocycloalkyl C1-C6 Alkyl, unsubstituted or substituted C6-C14 aryl, unsubstituted or substituted 5-15 membered heteroaryl, unsubstituted or substituted C6-C14 arylC1-C6 alkyl, unsubstituted or substituted 5-15 membered heteroarylC1-C6 alkyl, wherein the substitution means that one or more hydrogen atoms in the defined group are replaced by one or more substituents selected from halogen, amino, hydroxyl, C1-C3 alkyl (e.g., methyl), C1-C3 alkoxy (e.g., methoxy), and trifluoromethyl;

[0039] Indicates that the link is connected from here.

[0040] In some embodiments, in Formula (A), each R 1Each is independently selected from halogen, cyano (-CN), hydroxyl (-OH), amino (-NH2), nitro (-NO2), carboxyl (-COOH), unsubstituted or substituted C1-C10 alkyl, unsubstituted or substituted C1-C10 alkoxy, C1-C10 alkyl-NH-, (C1-C10 alkyl)(C1-C10 alkyl)N-, C1-C10 alkoxycarbonyl-NH-, unsubstituted or substituted C3-C10 cycloalkyl, unsubstituted or substituted 3-10 membered heterocyclyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, and the substitution means that one or more hydrogens in the defined group are replaced by one or more substituents selected from halogen, amino (-NH2), and hydroxyl (-OH).

[0041] In some embodiments, in Formula (A), R 2 Selected from hydrogen, unsubstituted or substituted C1-C10 alkyl, amino C1-C10 alkyl, (C1-C6 alkyl)(C1-C6 alkyl)N-C1-C10 alkyl, C1-C6 alkyl-NH-C1-C10 alkyl, unsubstituted or substituted C3-C10 cycloalkyl, unsubstituted or substituted C3-C10 cycloalkylC1-C6 alkyl, unsubstituted or substituted 3-10 membered heterocycloalkyl, unsubstituted or substituted 3-10 membered heterocycloalkylC1-C6 Alkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, unsubstituted or substituted C6-C10 arylC1-C6 alkyl, unsubstituted or substituted 5-10 membered heteroarylC1-C6 alkyl, wherein the substitution means that one or more hydrogen atoms in the defined group are replaced by one or more substituents selected from halogen, amino, hydroxyl, C1-C3 alkyl (e.g., methyl), C1-C3 alkoxy (e.g., methoxy), and trifluoromethyl.

[0042] Herein, the heteroaryl and heterocyclic groups may contain 1 to 4, for example 1 to 2, heteroatoms selected from N, O and S.

[0043] In some embodiments, the structure represented by formula (A) is selected from the structure represented by the following formula (A-1):

[0044] wherein the Ar ring is selected from phenyl, pyridyl, pyrimidinyl, and pyrazinyl;

[0045] R 1 is n substituents on the Ar ring, where n is an integer selected from 0 to 2, such as 0, 1, 2;

[0046] Each R 1Each is independently selected from halogen, nitro, carboxyl, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, wherein the substitution means that one or more hydrogen atoms in the defined group are replaced by one or more substituents selected from halogen, amino, and hydroxyl; preferably, n is 0 or 1; R 1 is selected from halogen, nitro, methyl, methoxy;

[0047] R 2 is selected from hydrogen, unsubstituted or substituted C1-C10 alkyl, unsubstituted or substituted C3-C10 cycloalkyl C1-C6 alkyl, unsubstituted or substituted 3-10 membered heterocyclyl C1-C6 alkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, unsubstituted or substituted C6-C10 aryl C1-C6 alkyl, unsubstituted or substituted 5-10 membered heteroaryl C1-C6 alkyl, wherein the substitution means that one or more hydrogens in the defined group are replaced by one or more substituents selected from halogen, amino, hydroxyl, C1-C3 alkyl (e.g., methyl), C1-C3 alkoxy (e.g., methoxy), and trifluoromethyl; preferably, R 2 Selected from hydrogen, ethyl, t-pentyl, cyclohexylmethyl, N-methylpiperidinylmethyl, N,N-dimethylaminoethyl, phenyl, benzyl, benzyl substituted by methoxy or trifluoromethyl, phenethyl, pyridyl, N-methylpyrazolyl.

[0048] In some embodiments, the structure represented by formula (A) is selected from the following structures:

[0049] POIL part

[0050] This portion represents a portion that can bind to CDK or EZH2 or RAS, and refers to a portion that can interact with CDK or EZH2 or RAS, that is, the target of the POIL portion is CDK or EZH2 or RAS.

[0051] The CDKs include CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, CDK14, CDK15, CDK16, CDK17, CDK18, CDK19, CDK20, CDK21 and all subtypes thereof.

[0052] In some embodiments, the POIL moiety is capable of binding to CDK9.

[0053] In some embodiments, the POIL moiety is capable of binding to CDK7.

[0054] In some embodiments, the POIL moiety is capable of binding to CDK2.

[0055] In some embodiments, the POIL moiety is capable of binding to CDK2, CDK4, and CDK6 simultaneously.

[0056] In some embodiments, the POIL moiety is capable of binding to CDK5.

[0057] In some embodiments, the POIL moiety is capable of binding to EZH2.

[0058] In some embodiments, the POIL moiety is capable of binding to EZH2, EED, SUZ12, and EZH1 simultaneously.

[0059] In some embodiments, the POIL moiety is capable of binding to RAS.

[0060] In some embodiments, the POIL portion can be a CDK probe, CDK inhibitor, EZH2 probe, EZH2 inhibitor, RAS probe, or RAS inhibitor that has been marketed or reported in the literature, including but not limited to the following compounds:

[0061] Linker

[0062] Linker is a linking portion used to connect the LCBM portion and the POIL portion, which can be a chemical bond or a group.

[0063] The linker can be rigid or flexible. In some preferred embodiments, the linker is flexible.

[0064] In some embodiments, the linker is a chemical bond. Herein, the linker is a chemical bond, which means that the LCBM portion and the POIL portion are directly connected.

[0065] In other embodiments, Linker is a linear or branched alkylene or saturated cyclic alkyl structure comprising 1-50, preferably 2-16 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16), further preferably 2-8 (e.g., 2, 3, 4, 5, 6, 7, 8) carbon atoms, wherein one or more (e.g., 2, 3, 4, 5, 6) carbon atoms, in particular 1 or 2 carbon atoms are optionally replaced by heteroatoms selected from O, S, NR a PR a , preferably O, S or NR a , more preferably O or NR a , especially O, where R ais H or C1-C3 alkyl; or, wherein one or more carbon atoms, in particular 1-6, more in particular 1 or 2 carbon atoms are optionally replaced by -C(=O)-, -C(=S)-, -S(=O)-, -SO2- or a 3- to 6-membered ring having 0 to 4 heteroatoms selected from O, S, N, and P.

[0066] In some embodiments, the linker is selected from:

[0067] -(CH2) n -、-(CH2CH2O) m -(CH2) n -、-CO-(CH2) n -、

[0068] wherein each n is independently an integer selected from 1-20 (for example, n can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20), preferably an integer from 1 to 16, an integer from 1 to 10, or an integer from 1 to 8;

[0069] Each m is independently an integer of 1-10 (for example, m can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10), preferably an integer of 1-6, an integer of 1-2;

[0070] p is an integer of 0-10, preferably an integer of 0-6, and more preferably an integer of 0-2.

[0071] In some embodiments, the linker is selected from the following structures:

[0072] In some embodiments, the POIL moiety is covalently linked to the linker via a carbon atom or a heteroatom selected from oxygen, sulfur, nitrogen, and phosphorus. Those skilled in the art can select an appropriate reaction for the linker based on the structures of the three moieties.

[0073] In some embodiments, the protein degrader is selected from any one of the following structures:

[0074] wherein each n is independently an integer from 1 to 16 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16), preferably an integer from 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8);

[0075] Each m is independently an integer from 1 to 6 (e.g., 1, 2, 3, 4, 5 or 6), preferably 1 or 2;

[0076] R 1 、R 2 Same definition as above.

[0077] In some embodiments, the protein degrader is a degrader of CDK9, cyclin T1 and / or CDK9-cyclin T1 complex, selected from the following structures:

[0078] In some embodiments, the protein degrader is a degrader of one or more of CDK2, cyclin A2, cyclin E1, CDK4, CDK5, CDK6, cyclin D1, CDK7, CDK9, or cyclin T1, selected from the following structures:

[0079] In some embodiments, the protein degrader is a degrader of one or more of EZH2, EED, SUZ12, EZH1, CDK2, CDK4, CDK6, CDK7, cyclin H, cyclin A2, cyclin E1, cyclin D1, or RAS, and is selected from the following structures:

[0080] The terms used in this invention are defined as follows:

[0081] As used herein, "halogen" may be fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine.

[0082] As used herein, the term "C1-C20 alkyl" alone or as part of a composite group refers to a straight or branched chain alkyl group having 1 to 20 carbon atoms, for example, "C1-C10 alkyl," "C1-C6 alkyl," "C1-C4 alkyl," "C1-C3 alkyl," etc. Specific examples thereof may include methyl, ethyl, propyl, n-propyl, isopropyl, butyl, n-butyl, isobutyl, tert-butyl, 1-methyl-butyl, 1-ethyl-butyl, pentyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, tert-pentyl, hexyl, n-hexyl, 1-methylpentyl, 2-methylpentyl, 4-methyl-2-pentyl, 3,3-dimethylbutyl, 2-ethylbutyl, and the like, but are not limited thereto.

[0083] As used herein, the term "C1-C20 alkoxy" refers to an RO- group, wherein R is a C1-C20 alkyl group as described above, for example, "C1-C10 alkoxy," "C1-C6 alkoxy," "C1-C4 alkoxy," "C1-C3 alkoxy," and the like. Specific examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentoxy, isopentoxy, neopentoxy, n-hexyloxy, isohexyloxy, 3-methylpentoxy, 3,3-dimethylbutoxy, 2-ethylbutoxy, and the like.

[0084] As used herein, the term "C3-C16 cycloalkyl" alone or as part of a composite group refers to a fully saturated cyclic hydrocarbon compound group containing 3 to 16 carbon atoms, such as "C3-C10 cycloalkyl", "C3-C7 cycloalkyl", "C3-C6 cycloalkyl", "C4-C6 cycloalkyl", etc., and specific examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0085] As used herein, the term "3-15 membered heterocyclyl" alone or as part of a composite group refers to a 3-15 membered cycloalkyl group containing 1 to 4, for example 1 to 3, 1 to 2 heteroatoms selected from nitrogen, oxygen, and sulfur in the ring, specific examples of which include ethylene oxide, tetrahydroimidazole, tetrahydrofuran, and the like.

[0086] As used herein, the term "C6-C14 aryl" alone or as part of a composite group refers to a monocyclic or polycyclic aromatic group having 6 to 14 carbon atoms, such as "C6-C10 aryl", examples of which are phenyl and naphthyl, preferably phenyl.

[0087] As used herein, the term "5-15 membered heteroaryl" alone or as part of a composite group refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) aromatic ring or aromatic group having 5-15 atoms in the ring and 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur in the ring, preferably a "5-10 membered heteroaryl ring", "5-6 membered heteroaryl ring", or "4-5 membered heteroaryl". Specific examples include pyrrolyl, furyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrazin-2-yl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothienyl, isobenzothienyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzoxazolyl, benzoxazolyl, benzofuranyl, benzoisofuranyl, benzoimidazolyl, benzoxazolyl, benzoxazolyl ... oxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzothiadiazolyl, indolizinyl, quinolyl, isoquinolyl, purinyl, imidazopyridinyl, imidazopyrimidinyl, imidazopyrazinyl, imidazopyridazinyl, imidazotriazinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, pyrazolopyrazinyl, pyrazolotriazinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrrolopyridazinyl, pyrrolopyrazinyl and pyrrolotriazinyl, etc., preferably pyrazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl.

[0088] As used herein, "pharmaceutically acceptable salts" include anionic salts and cationic salts of the compound of formula (I), such as salts of the compound of formula (1) with an acid or base; for example, inorganic acid or organic acid salts of the compound of formula (1); preferably, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, carbonic acid, perchloric acid; preferably, the organic acid includes formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, glutamic acid, pamoic acid; or inorganic base or organic base salts of the compound of formula (1); for example, I) alkali metal salts, alkaline earth metal salts, ammonium salts of compounds; preferably, the alkali metal includes sodium, potassium, lithium, cesium, and the alkaline earth metal includes magnesium, calcium, and strontium. For example, the organic base includes trialkylamine, pyridine, quinoline, piperidine, imidazole, picoline, dimethylaminopyridine, dimethylaniline, N-alkylmorpholine, 1,5-diazabicyclo[4.3.0]nonene-5, 1,8-diazabicyclo[5.4.0]undecene-7, 1,4-diazabicyclo[2.2.2]octane; preferably, the trialkylamine includes trimethylamine, triethylamine, N,N-diisopropylethylamine; preferably, the N-alkylmorpholine includes N-methylmorpholine.

[0089] The compounds of the present invention may contain chiral centers and, as such, may exist in different isomeric forms. As used herein, "isomers" refer to different compounds having the same molecular formula but differing in the arrangement and configuration of the atoms. As used herein, "stereoisomers" include diastereomers, enantiomers and racemates, geometric isomers, and conformational isomers (including rotamers and atropisomers).

[0090] In another aspect, the present invention provides a method for preparing a protein degrader, comprising a method for synthesizing an LC3-binding portion and covalently linking the portion that can bind to LC3 with a portion that can bind to CDK, EZH2, and RAS.

[0091] The reaction types for achieving the connection include nucleophilic substitution reaction, Mitsunobu reaction, condensation reaction, etc., but are not limited thereto.

[0092] In some embodiments, a pharmaceutically acceptable salt of a protein degradation agent can be prepared by dissolving the protein degradation agent in a corresponding acid-saturated alcohol solution, ethyl acetate solution, or dioxane solution and reacting the solution. For example, the protein degradation agent can be dissolved in a methanol solution saturated with hydrogen chloride, stirred at room temperature for 30 minutes, and the solvent evaporated to dryness to obtain the hydrochloride salt of the corresponding protein degradation agent. However, the present invention is not limited thereto, and those skilled in the art may employ any suitable salt-forming method based on the properties of the protein degradation agent.

[0093] In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above-mentioned protein degraders or their pharmaceutically acceptable salts, stereoisomers, solvates, polymorphs, tautomers, isotopic compounds, metabolites or prodrugs, and an optional pharmaceutically acceptable carrier.

[0094] The pharmaceutically acceptable carrier refers to a conventional pharmaceutical carrier in the pharmaceutical field, for example: diluents such as water; fillers such as starch and sucrose; binders such as cellulose derivatives, alginates, gelatin, and polyvinyl pyrrolidone; humectants such as glycerol; disintegrants such as agar, calcium carbonate, and sodium bicarbonate; absorption enhancers such as quaternary ammonium compounds; surfactants such as cetyl alcohol; adsorption carriers such as kaolin and bentonite; lubricants such as talc, calcium and magnesium stearate, and polyethylene glycol. In addition, other adjuvants such as flavoring agents and sweeteners may be added to the pharmaceutical composition.

[0095] The protein degradation agent of the present invention or its composition can be administered orally or parenterally to a patient in the form of conventional preparations, such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions and emulsions. Suitable preparations can be prepared by commonly used methods using conventional organic or inorganic additives, such as excipients (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate or calcium carbonate), binders (e.g., cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, sucrose or starch), disintegrants (e.g., starch, carboxymethylcellulose, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, sodium bicarbonate). , calcium phosphate or calcium citrate), lubricants (e.g., magnesium stearate, light anhydrous silicic acid, talc or sodium lauryl sulfate), flavoring agents (e.g., citric acid, menthol, glycine or orange powder), preservatives (e.g., sodium benzoate, sodium bisulfite, methylparaben or propylparaben), stabilizers (e.g., citric acid, sodium citrate or acetic acid), suspending agents (e.g., methylcellulose, polyvinylpyrrolidone or aluminum stearate), dispersants (e.g., hydroxypropyl methylcellulose), diluents (e.g., water) and base wax (e.g., cocoa butter, white petrolatum or polyethylene glycol).

[0096] The dosage regimen can be adjusted to provide the optimal desired response. For example, when administered as an injectable, a single bolus, bolus, and / or continuous infusion can be administered, among others. For example, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the therapeutic situation. For example, several divided doses can be administered over time, or the dose can be proportionally reduced or increased depending on the therapeutic situation. It should be noted that dosage values ​​can vary depending on the type and severity of the condition to be alleviated and can include single or multiple doses. Generally, the dosage for treatment varies, depending on considerations such as: the age, sex, and general health of the patient to be treated; the frequency of treatment and the nature of the desired effect; the extent of tissue damage; the duration of symptoms; and other variables that can be adjusted by the individual physician. It will be further understood that for any particular individual, the specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or overseeing the administration of the composition. The dosage and administration regimen of the pharmaceutical composition can be readily determined by one of ordinary skill in the clinical field. For example, the protein degraders or compositions of the present invention can be administered in divided doses from four times daily to once every seven days, with dosages ranging from 0.01 to 1000 mg per dose. The desired dosage can be administered once or multiple times to achieve the desired outcome. Pharmaceutical compositions according to the present invention can also be provided in unit dosage form.

[0097] Another aspect of the present invention provides the use of the above-mentioned protein degrader or its pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotopic compound, metabolite, or prodrug, or pharmaceutical composition, in the preparation of a product for degrading CDK9, cyclin T1, CDK9-cyclin T1 complex, EZH2, EED, SUZ12, EZH1, CDK2, CDK4, CDK5, CDK6, CDK7, cyclin H, cyclin A2, cyclin E1, cyclin D1, or one or more proteins in RAS, particularly for simultaneously degrading CDK9 and cyclin T1, EZH2, EED, SUZ12 and EZH1, CDK2, CDK4 and CDK6, CDK5, as well as cyclin A2, cyclin E1, and cyclin D1. The product can be used as a drug or in preclinical or laboratory research, for example, as a reagent for related protein degradation or corresponding pathway research.

[0098] Another aspect of the present invention provides use of the above-mentioned protein degrading agent or pharmaceutical composition in the preparation of a medicament for treating, preventing and / or ameliorating CDK, EZH2 or RAS-related diseases or conditions.

[0099] Another aspect of the present invention provides use of the above-mentioned protein degrading agent in treating, preventing and / or improving CDK, EZH2 or RAS-related diseases or conditions.

[0100] The above-mentioned CDK, EZH2 or RAS-related diseases or conditions include but are not limited to the following:

[0101] Inflammatory conditions: such as arthritis, rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, juvenile arthritis, and other arthritic conditions; Skin-related conditions: such as psoriasis, eczema, burns, dermatitis, and neuritis; Allergies, pain, and fever; Lung diseases: such as lung inflammation, adult respiratory distress syndrome, pulmonary sarcoidosis, asthma, silicosis, chronic inflammatory lung disease, and chronic obstructive pulmonary disease (COPD); Cardiovascular diseases: such as arteriosclerosis, myocardial infarction (including post-myocardial infarction syndrome), thrombosis, congestive heart failure, and cardiac reperfusion injury. and complications associated with hypertension and / or heart failure, such as vascular organ damage, restenosis, and cardiomyopathy; stroke, including ischemic and hemorrhagic stroke; reperfusion injury; ischemia, including stroke and cerebral ischemia, as well as ischemia caused by cardiac / coronary artery bypass grafting, neurodegenerative diseases, liver disease, or nephritis; gastrointestinal disorders, such as inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome, ulcerative colitis, ulcerative disease, gastric ulcers, viral and bacterial infections; sepsis, including septic shock and Gram-negative sepsis; malaria; meningitis; and HIV infection. ; opportunistic infections; cachexia secondary to infection or malignancy; cachexia secondary to acquired immunodeficiency syndrome (AIDS), AIDS, ARC (AIDS-related complex); myalgia caused by herpes virus infection; influenza; autoimmune diseases; graft-versus-host reaction and allograft rejection; bone resorption diseases; osteoporosis; multiple sclerosis; cancers: such as leukemia, lymphoma, colorectal cancer, brain cancer, bone cancer, tumors of epithelial cell origin (epithelial carcinoma), basal cell carcinoma, adenocarcinoma, gastrointestinal cancer, lip cancer, oral cancer, esophageal cancer, small Intestinal cancer, stomach cancer, colon cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, cervical cancer, lung cancer, breast cancer, skin cancer, squamous cell and / or basal cell carcinoma, prostate cancer, renal cell carcinoma and other cancers known to affect epithelial cells throughout the body, chronic myeloid leukemia (CML), acute myeloid leukemia (AML) and acute promyelocytic leukemia (APL); Central nervous system diseases: including central nervous system diseases with inflammatory or apoptotic components, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal cord injury and peripheral neuropathies, or B-cell lymphoma, etc.

[0102] In addition, by regulating the amount of intracellular CDK, EZH2, or RAS through LC3-mediated autophagic degradation by the protein degraders of the present application (such as those described herein), the present invention also provides a new paradigm for treating, preventing, or ameliorating diseases or conditions in which CDK, EZH2, or RAS play a role.

[0103] The protein degrader or pharmaceutical composition provided by the present invention can also be used in combination with other therapeutic agents for treating or preventing tumors.

[0104] The present invention has the following beneficial effects:

[0105] The present invention provides a series of small molecules containing 2,4-quinazolinediones that have a recruitment effect on LC3, resulting in a series of protein degraders that have a degrading effect on CDK9, cyclin T1, EZH2, EED, SUZ12, EZH1, CDK2 / 4 / 6, cyclin A2 / E1 / D1, RAS, CDK2, CDK7, and cyclin H. These degraders exhibit excellent anti-tumor activity, and the present invention also provides a means for degrading protein complexes.

[0106] The present invention has been described in detail above, but the above embodiments are merely illustrative in nature and are not intended to limit the present invention. In addition, the present invention is not limited by any theory described in the above prior art or invention summary or the following examples. BRIEF DESCRIPTION OF THE DRAWINGS

[0107] Figure 1 shows the Western Blot experimental results of representative compounds inducing CDK9 and Cyclin T1 protein degradation and downregulating Mcl-1 protein, wherein A represents WSU-DLCL2 cells treated with 100 nM compounds Y33, Y35, Y41, Y44, and Y45, and B represents WSU-DLCL2 cells treated with 1 μM compounds Y1, Y2, Y3, Y4, and Y5.

[0108] Figure 2A shows that representative compounds Y2 and Y3 induce the binding of CDK9 and LC3B, and Figure 2B shows that the degradation effect of compound Y35 can be inhibited by the late autophagy inhibitor bafilomycin A1 (BafA1), indicating the autophagy-lysosomal degradation pathway.

[0109] FIG3 shows the Western Blot experimental results showing that compounds Y44 and Y82-Y86 induce degradation of CDK9 and Cyclin T1 proteins and downregulate Mcl-1 protein at a concentration of 100 nM.

[0110] Figure 4A shows the degradation effect of compound Y53 on CDK2 at a series of concentrations (0.1 μM, 1 μM, 10 μM); Figure 4B shows the degradation effect of compound Y67 on CDK7 and the corresponding cyclin H at a concentration of 10 μM; Figure 4C shows the degradation effect of compound Y62 on CDK2 and its corresponding cyclin A2, cyclin E1 and CDK6 and its corresponding cyclin D1; Figure 4D shows the degradation effect of Y59, Y60, and Y61 on CDK2 / 4 / 6 and the corresponding cyclin A2 / E1 / D1 at a concentration of 10 μM.

[0111] Figure 5A shows the degradation effects of compounds Y47 and Y50 on EZH2, EED, SUZ12 complex and EZH1 at a concentration of 10 μM; Figure 5B shows the degradation effects of compounds Y50, Y87-Y89 on EZH2, EED, SUZ12 complex and EZH1 at a concentration of 10 μM.

[0112] FIG6 shows the degradation effect of compound Y69 on Ras protein, which is time- and concentration-dependent. Compound Y69 can also downregulate the levels of related proteins pERK T202 / Y204 and ERK.

[0113] Figure 7 shows the Western Blot experimental results of representative compounds inducing degradation of CDK2, cyclin A2, cyclin E1, CDK4, CDK5, CDK6, cyclin D1, CDK7, CDK9 and Cyclin T1 proteins, where A represents WSU-DLCL2 cells treated with 100 nM compounds Y83, Y95, and Y96, and B represents WSU-DLCL2 cells treated with 100 nM compounds Y84 and Y100-Y106. DETAILED DESCRIPTION

[0114] The present invention will be further described below with reference to the examples. It should be noted that the following examples are provided for illustrative purposes only and do not constitute a limitation on the scope of protection claimed in the present invention.

[0115] Unless otherwise specified, the raw materials, reagents, methods, etc. used in the examples are conventional raw materials, reagents, and methods in the art.

[0116] In the following examples, H NMR spectra were recorded using a Bruker AMX-400 NMR spectrometer, and the unit of chemical shift δ is ppm. Unless otherwise specified, all reaction solvents were purified according to conventional methods. Silica gel (200-300 mesh) for column chromatography was produced by Qingdao Ocean Chemical Plant. Thin-layer chromatography used GF254 high-efficiency plates, produced by Yantai Chemical Research Institute. Preparative thin-layer chromatography plates were prepared by the Shanghai Institute of Materia Medica, Chinese Academy of Sciences, and the stationary phases were prepared using GF254 (HG / T2354-92) silica gel and sodium carboxymethyl cellulose (800-1200), produced by Qingdao Ocean Chemical Co., Ltd. and China National Pharmaceutical (Group) Shanghai Chemical Reagent Co., Ltd., respectively. Unless otherwise specified, all solvents were analytically pure and were purchased from Sinopharm Chemical Reagent Co., Ltd. Iodine and ultraviolet fluorescence were used for color development. The organic solvent was removed by evaporation under reduced pressure on a rotary evaporator.

[0117] Example 1 Synthesis of Protein Degrader Y1

[0118] first step

[0119] Methyl 2-amino-4-chlorobenzoate (1.0 g, 5.39 mmol) was dissolved in 27 mL of anhydrous acetonitrile, and ethoxycarbonyl isothiocyanate (0.8 mL, 6.47 mmol) was added. The mixture was stirred at room temperature overnight. After the starting material was consumed, benzylamine (0.9 mL, 8.08 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 2.1 g, 10.78 mmol) were added. The mixture was stirred at room temperature for 24 hours. The acetonitrile was removed by vortexing and the mixture was dissolved in ethyl acetate. The ethyl acetate phase was then washed twice with 1 M hydrochloric acid and once with water, dried over anhydrous sodium sulfate, and the ethyl acetate was removed by vortexing. The mixture was slurried with ether to give Y1a (white solid, 1.5 g, yield 77.8%). 1 H NMR(400MHz,Chloroform-d)δ12.90(s,1H),8.12(d,J=8.5Hz,1H),7.58–7.51(m,2H),7.36–7 .24(m,4H),7.20(d,J=1.8Hz,1H),5.46(s,2H),4.27(q,J=7.1Hz,2H),1.39(t,J=7.1Hz,3H).

[0120] Step 2

[0121] Y1a (1.5 g, 4.19 mmol) was dissolved in 51 ml of methanol, and 8 mL of 1 M aqueous sodium hydroxide solution was added. The mixture was refluxed for 2 hours. The methanol was removed by vortexing, and dichloromethane was added. The dichloromethane layer was washed once with water and once with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain Y1b (white solid, 150 mg, yield 12.5%). 1 H NMR(400MHz,Chloroform-d)δ9.27(s,1H),8.09(d,J=8.5Hz,1H),7.53(d,J=7.4Hz,2H ),7.37–7.30(m,3H),7.21(dd,J=8.6,1.8Hz,1H),7.07(d,J=1.8Hz,1H),5.26(s,2H).

[0122] Step 3

[0123] Y1b (50 mg, 0.17 mmol) was dissolved in 1 mL of DMF, and 1,2-dibromoethane (60 μL, 0.70 mmol) and potassium carbonate (48 mg, 0.35 mmol) were added. The mixture was stirred at room temperature overnight. Ethyl acetate and water were added, and the ethyl acetate layer was washed four times with water and once with saturated aqueous sodium chloride solution. The mixture was dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain Y1c (white solid, 35 mg, yield 50.7%).1 H NMR (400MHz, Chloroform-d) δ8.21(d,J=8.4Hz,1H),7.55–7.49(m,2H),7.36–7.23(m,5H),5.26(s,2H),4.47(t,J=7.5Hz,2H),3.61(t,J=7.5Hz,2H).

[0124] Step 4

[0125] Y1c (49 mg, 0.12 mmol) was dissolved in 1 mL of DMF, and SNS-032 (47 mg, 0.12 mmol) and diisopropylethylamine (DIPEA, 62 μL, 0.37 mmol) were added. The mixture was stirred at room temperature overnight. After the starting material was completely consumed, ethyl acetate and water were added. The ethyl acetate layer was washed four times with water and once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain Y1 (white solid, 20 mg, yield 23.2%). 1 H NMR(400MHz,Chloroform-d)δ11.40(s,1H),8.19(d,J=8.5Hz,1H),7.55–7.47( m,2H),7.39(s,1H),7.36–7.30(m,3H),7.28–7.21(m,2H),6.61(s,1H),5.27(s ,2H),4.26(t,J=6.9Hz,2H),3.99(s,2H),3.09(d,J=11.1Hz,2H),2.69(t,J=6. 9Hz,2H),2.45–2.34(m,1H),2.26–2.15(m,2H),1.94–1.80(m,4H),1.26(s,9H).

[0126] Example 2 Synthesis of Protein Degrader Y2

[0127] The synthesis method was the same as that in Example 1, except that 1,2-dibromoethane was replaced with 1,3-dibromopropane to obtain protein degradation agent Y2 (white solid, yield 63.4%). 1H NMR(400MHz,Chloroform-d)δ11.30(s,1H),8.18(d,J=8.5Hz,1H),7.56–7.50(m, 2H),7.44(d,J=1.8Hz,1H),7.36(s,1H),7.35–7.30(m,2H),7.29–7.25(m,1H),7. 22(dd,J=8.5,1.7Hz,1H),6.61(s,1H),5.27(s,2H),4.18(t,J=7.2Hz,2H),3.97( s,2H),3.02(d,J=10.5Hz,2H),2.51–2.39(m,3H),2.03–1.88(m,8H),1.26(s,9H).

[0128] Example 3 Synthesis of Protein Degrader Y3

[0129] The synthesis method was the same as that in Example 1, except that 1,2-dibromoethane was replaced with 1,4-dibromobutane to obtain protein degradation agent Y3 (white solid, yield 38.3%). 1 H NMR(400MHz,Chloroform-d)δ11.57(s,1H),8.19(d,J=8.5Hz,1H),7.54–7.46(m,2H ),7.36(s,1H),7.34–7.25(m,4H),7.22(dd,J=8.4,1.7Hz,1H),6.61(s,1H),5.27(s, 2H),4.18–4.11(m,2H),3.98(s,2H),3.04(d,J=11.1Hz,2H),2.56–2.39(m,3H),2.18 –2.03(m,2H),2.01–1.90(m,4H),1.84–1.73(m,2H),1.72–1.62(m,2H),1.26(s,9H).

[0130] Example 4 Synthesis of Protein Degrader Y4

[0131] The synthesis method was the same as that in Example 1, except that 1,2-dibromoethane was replaced with 1,5-dibromopentane to obtain protein degradation agent Y4 (white solid, yield 47.0%). 1H NMR(400MHz,Chloroform-d)δ11.26(s,1H),8.19(d,J=8.4Hz,1H),7.54–7.49(m,2H),7.36(s, 1H),7.35–7.26(m,3H),7.23(dd,J=8.4,1.7Hz,1H),7.17(d,J=1.8Hz,1H),6.61(s,1H),5.27(s ,2H),4.13–4.06(m,2H),3.98(s,2H),3.03(d,J=11.3Hz,2H),2.47–2.36(m,3H),2.14–2.00(m ,2H),1.98–1.87(m,4H),1.81–1.71(m,2H),1.68–1.56(m,2H),1.51–1.38(m,2H),1.26(s,9H).

[0132] Example 5 Synthesis of Protein Degrader Y5

[0133] The synthesis method was the same as that in Example 1, except that 1,2-dibromoethane was replaced with triethylene glycol bis-p-toluenesulfonate to obtain protein degradation agent Y5 (white solid, yield 69.8%). 1 H NMR(400MHz,Chloroform-d)δ11.64(s,1H),8.15(d,J=8.4Hz,1H),7.53–7.48(m,3H),7.35(s,1H),7 .34–7.24(m,3H),7.20(dd,J=8.5,1.7Hz,1H),6.60(s,1H),5.31–5.30(m,1H),5.26(s,2H),4.30(t, J=5.5Hz,2H),3.97(s,2H),3.82(t,J=5.5Hz,2H),3.64–3.60(m,2H),3.59–3.53(m,4H),3.03–2.96( m,2H),2.56(t,J=5.8Hz,2H),2.42–2.31(m,1H),2.12–2.02(m,2H),1.97–1.79(m,4H),1.25(s,9H).

[0134] Example 6 Synthesis of Protein Degrader Y6

[0135] first step

[0136] 4-Chloroisatoic anhydride (500 mg, 2.53 mmol) and aniline (231 μL, 2.53 mmol) were dissolved in 13 mL of dichloromethane and refluxed for 30 minutes. N,N′-carbonyldiimidazole (CDI, 451 mg, 2.78 mmol) was added and refluxed for 3 hours. The dichloromethane was evaporated and ethanol was added. The mixture was slurried and filtered to obtain Y6a (white solid, 230 mg, yield 33.3%). 1 H NMR (400MHz, DMSO-d6) δ11.70 (s, 1H), 7.94 (d, J = 8.4Hz, 1H), 7.52–7.40 (m, 3H), 7.35–7.22 (m, 4H).

[0137] Step 2

[0138] Y6a (230 mg, 0.84 mmol) and 1,2-dibromoethane (364 μL, 4.22 mmol) were dissolved in 4 mL of DMF, and potassium carbonate (234 mg, 1.69 mmol) was added. The mixture was stirred at room temperature overnight, and ethyl acetate and water were added. The ethyl acetate layer was washed four times with water and once with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain compound Y6b (white solid, 163 mg, yield 50.7%). 1 H NMR (400MHz, Chloroform-d) δ 8.23 ​​(d, J = 8.4Hz, 1H), 7.59–7.46 (m, 3H), 7.37–7.26 (m, 4H), 4.51 (t, J = 7.4Hz, 2H), 3.67 (t, J = 7.4Hz, 2H).

[0139] Step 3

[0140] Y6b (85 mg, 0.22 mmol) was dissolved in 1 mL of DMF, and SNS-032 (85 mg, 0.22 mmol) and DIPEA (111 μL, 0.67 mmol) were added. The mixture was stirred at 60° C. overnight to obtain compound Y6 (white solid, 58 mg, yield 38.1%). 1H NMR(400MHz,Chloroform-d)δ10.98(s,1H),8.05(d,J=8.5,2.9Hz,1H),7.41–7.34(m,2H),7.34–7.28(m,1H),7.26–7.17(m,2H),7.15–7.06(m,4H),6. 44(s,1H),4.19–4.08(m,2H),3.81(s,2H),2.99–2.88(m,2H),2.63–2.52(m ,2H),2.30–2.17(m,1H),2.11–1.98(m,2H),1.80–1.66(m,4H),1.09(s,9H).

[0141] Example 7 Synthesis of Protein Degrader Y7

[0142] The synthesis method was the same as that in Example 6, except that aniline was replaced with phenylethylamine to obtain protein degradation agent Y7 (white solid, yield 35.5%). 1 H NMR(400MHz,Chloroform-d)δ10.75(s,1H),8.19(d,J=8.4Hz,1H),7.38(s,1H),7.36–7.30(m,5H),7.27–7.22(m,2H),6.61(s,1H),4.36–4.20(m ,4H),3.98(s,2H),3.13(d,J=10.9Hz,2H),3.03–2.96(m,2H),2.75–2.64 (m,2H),2.43(s,1H),2.31–2.19(m,2H),2.01–1.86(m,4H),1.27(s,9H).

[0143] Example 8 Synthesis of Protein Degrader Y8

[0144] The synthesis method was the same as that in Example 6, except that aniline was replaced with 4-methoxybenzylamine to obtain protein degradation agent Y8 (white solid, yield 35.1%). 1H NMR(400MHz,Chloroform-d)δ11.36(s,1H),8.18(d,J=8.4Hz,1H),7.53–7.46(m ,2H),7.38(s,1H),7.32–7.30(m,1H),7.22(dd,J=8.4,1.7Hz,1H),6.91–6.83(m, 2H),6.61(s,1H),5.21(s,2H),4.31–4.21(m,2H),3.99(s,2H),3.79(s,3H),3.11(d,J=11.0Hz ,2H),2.73–2.66(m,2H),2.47–2.35(m,1H),2.28–2.16(m,2H),1.96–1.84(m,4H),1.26(s,9H).

[0145] Example 9 Synthesis of Protein Degrader Y9

[0146] The synthesis method was the same as that in Example 6, except that aniline was replaced with 4-(trifluoromethyl)benzylamine to obtain protein degradation agent Y9 (white solid, yield 30.1%). 1 H NMR(400MHz,Chloroform-d)δ11.10(s,1H),8.19(d,J=8.5Hz,1H),7.65–7.57( m,4H),7.36(s,1H),7.34(s,1H),7.26(dd,J=8.5,1.7Hz,1H),6.61(s,1H),5.3 2(s,2H),4.26(t,J=7.0Hz,2H),3.99(s,2H),3.10(d,J=11.1Hz,2H),2.74–2.6 6(m,2H),2.47–2.35(m,1H),2.27–2.15(m,2H),1.95–1.83(m,4H),1.26(s,9H).

[0147] Example 10 Synthesis of protein degradation agent Y10

[0148] The synthesis method was the same as that in Example 6, except that aniline was replaced with 4-methylaminopyridine to obtain protein degradation agent Y10 (white solid, yield 11.0%). 1H NMR (400MHz, Methanol-d4) δ8.53–8.48(m,2H),8.17(d,J=8.5Hz,1H),7.64(d,J=1. 8Hz,1H),7.46–7.42(m,2H),7.36(dd,J=8.5,1.7Hz,1H),7.33(s,1H),6.70(s,1H),5 .30(s,2H),4.37(t,J=6.7Hz,2H),4.01(s,2H),3.13(d,J=11.7Hz,2H),2.72(t,J=6. 7Hz, 2H), 2.54–2.42 (m, 1H), 2.20 (t, J = 11.2Hz, 2H), 1.90–1.72 (m, 4H), 1.25 (s, 9H).

[0149] Example 11 Synthesis of protein degradation agent Y11

[0150] The synthesis method was the same as that in Example 6, except that aniline was replaced with 3-aminomethylpyridine to obtain protein degradation agent Y11 (white solid, yield 11.4%). 1 H NMR (400MHz, Methanol-d4) δ8.69(d,J=2.1Hz,1H),8.46(d,J=4.9Hz,1H),8.18(d,J=8.5Hz,1H),7.96(d,J=7.8Hz,1H),7.61( d,J=1.7Hz,1H),7.44(dd,J=8.0,5.0Hz,1H),7.37–7.32(m,2H),6.69(s,1H),5.29(s,2H),4.36(t,J=6.6Hz,2H),4.01(s,2H), 3.10(d,J=11.4Hz,2H),2.70(t,J=6.6Hz,2H),2.53–2.42(m,1H),2.19(t,J=11.6Hz,2H),1.87–1.81(m,2H),1.79–1.68(m,2H),1.26(s,9H).

[0151] Example 12 Synthesis of Protein Degrader Y12

[0152] The synthesis method was the same as that in Example 6, except that aniline was replaced with 2-aminomethylpyridine to obtain degradation agent Y12 (white solid, yield 21.2%). 1H NMR (400MHz, DMSO-d6) δ12.21(s,1H),8.46–8.41(m,1H),8.06(d,J=8.4Hz,1H),7.75(td,J=7.7,1. 8Hz,1H),7.71(d,J=1.8Hz,1H),7.41–7.35(m,2H),7.31(d,J=7.9Hz,1H),7.26–7.21(m,1H),6.73(s ,1H),5.25(s,2H),4.26(t,J=6.3Hz,2H),4.06(s,2H),2.98(d,J=10.9Hz,2H),2.57(t,J=6.3Hz,2H) ,2.48–2.38(m,1H),2.01(t,J=11.4Hz,2H),1.72(d,J=12.5Hz,2H),1.57–1.45(m,2H),1.17(s,9H).

[0153] Example 13 Synthesis of Protein Degrader Y13

[0154] The synthesis method was the same as that of Example 6, except that aniline was replaced with 4-(aminomethyl)-1-methylpyrazole to obtain degradation agent Y13 (white solid, yield 10.0%).

[0155] Example 14 Synthesis of Protein Degrader Y14

[0156] The synthesis method of Y14a was the same as that of Example 6, except that aniline in the first step was replaced with 2,4-dimethoxybenzylamine to obtain Y14a (white solid, yield 46.4%). 1 H NMR(400MHz,Chloroform-d)δ10.64(s,1H),8.20(d,J=8.4Hz,1H),7.37(s,1H),7.33(d,J=1.7Hz,1H), 7.24(dd,J=8.4,1.7Hz,1H),6.98(d,J=8.4Hz,1H),6.61(s,1H),6.47(d,J=2.4Hz,1H),6.43(dd,J=8.4 ,2.4Hz,1H),5.26(s,2H),4.28(t,J=6.8Hz,2H),3.98(s,2H),3.85(s,3H),3.78(s,3H),3.09(d,J=11. 3Hz, 2H), 2.71 (t, J = 6.8Hz, 2H), 2.44–2.32 (m, 1H), 2.25–2.15 (m, 2H), 1.94–1.81 (m, 4H), 1.26 (s, 9H).

[0157] Y14a (95 mg, 0.13 mmol) was dissolved in 1 mL of trifluoroacetic acid and refluxed for 24 hours. Water was added to precipitate a white solid, which was filtered to obtain Y14 (white solid, 50 mg, yield 65.7%). 1 H NMR (400MHz, DMSO-d6) δ12.48–11.52(m,2H),7.98(d,J=8.4Hz,1H),7.60(d,J =1.8Hz,1H),7.38(s,1H),7.31(dd,J=8.4,1.7Hz,1H),6.73(s,1H),4.16(t,J= 6.4Hz,2H),4.05(s,2H),2.99(d,J=11.0Hz,2H),2.56–2.52(m,2H),2.47–2.40 (m,1H),2.07–1.97(m,2H),1.79–1.70(m,2H),1.61–1.48(m,2H),1.17(s,9H).

[0158] Example 15 Synthesis of protein degradation agent Y15

[0159] The synthesis method was the same as that in Example 6, except that aniline in the first step was replaced with ethylamine (2M, THF) to obtain compound Y15a, and then the second and third steps were continued to obtain degradation agent Y15 (white solid, yield 20.0%). 1 H NMR(400MHz,Chloroform-d)δ11.42(s,1H),8.18(d,J=8.4Hz,1H),7.38(s,1H),7. 35–7.31(m,1H),7.23(dd,J=8.5,1.7Hz,1H),6.61(s,1H),4.30–4.23(m,2H),4.15( q,J=7.0Hz,2H),3.99(s,2H),3.13(d,J=11.1Hz,2H),2.75–2.67(m,2H),2.50–2.3 9(m,1H),2.30–2.18(m,2H),1.97–1.86(m,4H),1.30(t,J=7.1Hz,3H),1.26(s,9H).

[0160] Example 16 Synthesis of Protein Degrader Y16

[0161] The synthesis method was the same as that in Example 6, except that aniline was replaced with tert-amylamine to obtain degradation agent Y16 (white solid, yield 36.2%). 1H NMR(400MHz,Chloroform-d)δ11.50(s,1H),8.17(d,J=8.4Hz,1H),7.38(s,1H) ,7.32(d,J=1.8Hz,1H),7.22(dd,J=8.5,1.7Hz,1H),6.61(s,1H),4.26(t,J=7.0 Hz,2H),4.04(s,2H),3.98(s,2H),3.19–3.08(m,2H),2.69(t,J=6.9Hz,2H),2.4 7–2.37(m,1H),2.28–2.14(m,2H),1.96–1.84(m,4H),1.26(s,9H),0.99(s,9H).

[0162] Example 17 Synthesis of Protein Degrader Y17

[0163] The synthesis method was the same as that in Example 6, except that aniline was replaced with N,N-dimethylethylenediamine to obtain degradation agent Y17 (white solid, yield 19.0%). 1 H NMR (400MHz, Methanol-d4) δ8.15 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 1.7 Hz, 1H),7.35–7.29(m,2H),6.70(s,1H),4.34(t,J=6.9Hz,2H),4.25(t,J=6.8Hz,2H),4.00(s,2H),3.18–3.11(m, 2H),2.71(t,J=6.8Hz,4H),2.55–2.45(m,1H),2.38(s,6H),2.26–2.17(m,2H),1.93–1.75(m,4H),1.26(s,9H).

[0164] Example 18 Synthesis of Protein Degrader Y18

[0165] The synthesis method was the same as that in Example 6, except that aniline was replaced with cyclohexylmethylamine to obtain degradation agent Y18 (white solid, yield 39.2%). 1H NMR(400MHz,Chloroform-d)δ8.18(d,J=8.5Hz,1H),7.38(s,1H),7.32(d,J=1.8H z,1H),7.23(dd,J=8.5,1.7Hz,1H),6.61(s,1H),4.26(t,J=7.0Hz,2H),4.00–3.94 (m,4H),3.12(d,J=11.2Hz,2H),2.70(t,J=7.0Hz,2H),2.47–2.36(m,1H),2.28–2 .19(m,2H),1.96–1.81(m,5H),1.77–1.59(m,6H),1.27(s,9H),1.15–1.03(m,2H).

[0166] Example 19 Synthesis of Protein Degrader Y19

[0167] The synthesis method was the same as that in Example 6, except that aniline was replaced with (1-methyl-4-piperidinyl)methylamine to obtain degradation agent Y19 (white solid, yield 4.0%). 1 H NMR(400MHz,Chloroform-d)δ8.17(d,J=8.4Hz,1H),7.35(s,1H),7.30(d,J=1.7Hz,1H),7.23(dd,J=8.4,1.7Hz,1H),6.60(s,1H),4.26(t, J=6.6Hz,2H),4.04(d,J=6.9Hz,2H),3.97(s,2H),3.14–3.00(m,4H),2.51–2.35(m,4H),2.25–2.13(m,4H),1.96–1.57(m,9H),1.25(s,9H).

[0168] Example 20 Synthesis of Protein Degrader Y20

[0169] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by isatoic anhydride and aniline was replaced by benzylamine to obtain degradation agent Y20 (white solid, yield 24.3%). 1H NMR(400MHz,Chloroform-d)δ11.66(s,1H),8.27(dd,J=7.9,1.6Hz,1H),7.74–7 .66(m,1H),7.54–7.49(m,2H),7.38(s,1H),7.35–7.22(m,5H),6.61(s,1H),5.32 –5.28(m,2H),4.31(t,J=7.2Hz,2H),3.99(s,2H),3.13–3.05(m,2H),2.70(t,J=7 .2Hz,2H),2.47–2.35(m,1H),2.25–2.17(m,2H),1.93–1.82(m,4H),1.25(s,9H).

[0170] Example 21 Synthesis of Protein Degrader Y21

[0171] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by isatoic anhydride, and aniline was replaced by ethylamine (2M, THF) to obtain degradation agent Y21 (white solid, yield 34.2%). 1 H NMR(400MHz,Chloroform-d)δ10.58(s,1H),8.27(dd,J=7.9,1.6Hz,1H),7.73–7.66(m ,1H),7.38(s,1H),7.32–7.24(m,5H),6.61(s,1H),4.31(t,J=7.4Hz,2H),4.18(q,J=7. 0Hz,2H),3.98(s,2H),3.13(d,J=11.4Hz,2H),2.71(t,J=7.4Hz,2H),2.47–2.37(m,1H ), 2.23(dt,J=11.0,6.0Hz,2H),1.98–1.85(m,4H),1.32(t,J=7.0Hz,3H),1.27(s,9H).

[0172] Example 22 Synthesis of Protein Degrader Y22

[0173] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced with 5-chloro-1H-benzo[d][1,3]oxazine-2,4-dione and aniline was replaced with benzylamine to obtain degradation agent Y22 (white solid, yield 13.4%). 1H NMR(400MHz,Chloroform-d)δ10.17(s,1H),7.57–7.51(m,3H),7.37(s,1H),7.36–7.29(m,3H),7.27–7.21(m,2H),6.61(s,1H),5.28(s,2H),4.31( t,J=7.1Hz,2H),3.98(s,2H),3.07(d,J=11.2Hz,2H),2.68(t,J=7.0Hz,2 H),2.42–2.32(m,1H),2.25–2.13(m,2H),1.95–1.77(m,4H),1.27(s,9H).

[0174] Example 23 Synthesis of Protein Degrader Y23

[0175] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 5-chloroisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y23 (white solid, yield 12.6%). 1 H NMR(400MHz,Chloroform-d)δ9.92(s,1H),8.24(d,J=2.5Hz,1H),7.64(dd,J=8.9,2 .6Hz,1H),7.53–7.49(m,2H),7.36(s,1H),7.35–7.31(m,2H),7.27–7.22(m,1H),6.6 1(s,1H),5.28(s,2H),4.32–4.23(m,2H),3.98(s,2H),3.07(d,J=11.1Hz,2H),2.74 –2.63(m,2H),2.41–2.31(m,1H),2.27–2.14(m,2H),1.96–1.77(m,4H),1.27(s,9H).

[0176] Example 24 Synthesis of Protein Degrader Y24

[0177] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 3-chloroisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y24 (white solid, yield 14.2%). 1H NMR (400MHz, DMSO-d6) δ12.15 (s, 1H), 8.10 (dd, J = 7.8, 1.7Hz, 1H), 7.89 (dd, J = 7. 9,1.6Hz,1H),7.38(s,1H),7.37–7.28(m,5H),7.24–7.19(m,1H),6.72(s,1H),5.1 2(s,2H),4.58(t,J=6.2Hz,2H),4.06(s,2H),2.61–2.51(m,2H),2.37–2.25(m,1H ),1.89(t,J=11.3Hz,2H),1.57(d,J=12.6Hz,2H),1.28–1.20(m,2H),1.17(s,9H).

[0178] Example 25 Synthesis of Protein Degrader Y25

[0179] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced with 6-methoxy-1H-benzo[d][1,3]oxazine-2,4-dione and aniline was replaced with benzylamine to obtain degradation agent Y25 (white solid, yield 19.3%). 1 H NMR(400MHz,DMSO-d6)δ12.22(s,1H),7.68(t,J=8.5Hz,1H),7.39(s,1H),7.34–7.28(m,4H), 7.25–7.19(m,1H),7.05(d,J=8.6Hz,1H),6.89(d,J=8.5Hz,1H),6.72(s,1H),5.09(s,2H),4. 24(t,J=6.7Hz,2H),4.06(s,2H),3.85(s,3H),2.97(d,J=10.8Hz,2H),2.56(t,J=6.7Hz,2H), 2.48–2.40(m,1H),2.01(t,J=11.3Hz,2H),1.77–1.68(m,2H),1.62–1.49(m,2H),1.17(s,9H).

[0180] Example 26 Synthesis of Protein Degrader Y26

[0181] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 5-methoxyisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y26 (white solid, yield 31.4%). 1H NMR(400MHz,DMSO-d6)δ12.22(s,1H),7.54–7.47(m,2H),7.43–7.38(m,2H),7.34– 7.30(m,4H),7.27–7.20(m,1H),6.72(s,1H),5.16(s,2H),4.25(t,J=6.6Hz,2H),4 .06(s,2H),3.83(s,3H),2.97(d,J=10.9Hz,2H),2.56(t,J=6.6Hz,2H),2.48–2.39 (m,1H),2.05–1.95(m,2H),1.72(d,J=12.6Hz,2H),1.59–1.48(m,2H),1.17(s,9H).

[0182] Example 27 Synthesis of Protein Degrader Y27

[0183] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced with 7-methoxy-1H-benzo[D][1,3]oxazine-2,4-dione and aniline was replaced with benzylamine to obtain degradation agent Y27 (white solid, yield 28.8%). 1 H NMR(400MHz,DMSO-d6)δ12.22(s,1H),8.00(d,J=9.4Hz,1H),7.39(s,1H),7.35–7.28(m ,4H),7.26–7.19(m,1H),6.95–6.89(m,2H),6.72(s,1H),5.13(s,2H),4.27(t,J=6.5Hz, 2H),4.06(s,2H),3.93(s,3H),2.99(d,J=10.8Hz,2H),2.58(t,J=6.6Hz,2H),2.47–2.39 (m,1H),2.01(t,J=11.2Hz,2H),1.73(d,J=12.6Hz,2H),1.63–1.46(m,2H),1.17(s,9H).

[0184] Example 28 Synthesis of Protein Degrader Y28

[0185] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 3-methoxyisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y28 (white solid, yield 32.1%). 1H NMR (400MHz, DMSO-d6) δ12.19(s,1H),7.71(dd,J=7.8,1.5Hz,1H),7.47(dd,J=8.3,1.5H z,1H),7.39(s,1H),7.35–7.26(m,6H),7.25–7.19(m,1H),6.72(s,1H),5.13(s,2H),4.4 8(t,J=6.7Hz,2H),4.06(s,2H),3.93(s,3H),2.79(d,J=10.9Hz,2H),2.58–2.52(m,2H), 2.44–2.33(m,1H),2.02–1.93(m,2H),1.72–1.64(m,2H),1.50–1.37(m,2H),1.18(s,9H).

[0186] Example 29 Synthesis of Protein Degrader Y29

[0187] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 4-methylisatoic anhydride and aniline was replaced by benzylamine to obtain degradation agent Y29 (white solid, yield 11.3%). 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),7.96(d,J=8.0Hz,1H),7.39(s,1H),7.37(s,1H ),7.34–7.28(m,4H),7.26–7.21(m,1H),7.14(d,J=8.2Hz,1H),6.72(s,1H),5.14(s, 2H),4.28–4.22(m,3H),4.06(s,2H),2.99(d,J=10.9Hz,2H),2.60–2.55(m,2H),2.47 –2.45(m,4H),2.06–1.97(m,2H),1.76–1.69(m,2H),1.59–1.48(m,2H),1.17(s,9H).

[0188] Example 30 Synthesis of Protein Degrader Y30

[0189] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 4-bromoisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y30 (white solid, yield 27.6%). 1H NMR (400MHz, DMSO-d6) δ12.21(s,1H),7.97(d,J=8.4Hz,1H),7.81(d,J=1.7Hz,1H),7.49( dd,J=8.4,1.6Hz,1H),7.39(s,1H),7.35–7.31(m,4H),7.27–7.20(m,1H),6.72(s,1H),5.1 3(s,2H),4.27(t,J=6.2Hz,2H),4.06(s,2H),2.97(d,J=11.1Hz,2H),2.56(t,J=6.1Hz,2H) ,2.47–2.39(m,1H),2.04–1.95(m,2H),1.76–1.67(m,2H),1.59–1.46(m,2H),1.17(s,9H).

[0190] Example 31 Synthesis of Protein Degrader Y31

[0191] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 4-fluoroisatoic anhydride, and aniline was replaced by benzylamine to obtain degradation agent Y31 (white solid, yield 20.1%). 1 H NMR(400MHz,DMSO-d6)δ12.22(s,1H),8.17–8.10(m,1H),7.49–7.43(m,1H),7.39(s ,1H),7.34–7.30(m,4H),7.26–7.21(m,1H),7.19–7.14(m,1H),6.72(s,1H),5.14(s, 2H),4.29–4.20(m,2H),4.06(s,2H),2.98(d,J=11.2Hz,2H),2.59–2.54(m,2H),2.36 –2.31(m,1H),2.05–1.95(m,2H),1.75–1.67(m,2H),1.56–1.45(m,2H),1.17(s,9H).

[0192] Example 32 Synthesis of Protein Degrader Y32

[0193] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced by 4-nitroisatoic anhydride and aniline was replaced by benzylamine to obtain degradation agent Y32 (white solid, yield 25.6%). 1H NMR(400MHz,DMSO-d6)δ12.21(s,1H),8.33–8.28(m,2H),8.06(dd,J=8.7,1.9H z,1H),7.39(s,1H),7.37–7.30(m,4H),7.28–7.21(m,1H),6.72(s,1H),5.16(s, 2H),4.34(t,J=6.2Hz,2H),4.06(s,2H),2.99(d,J=10.8Hz,2H),2.62(t,J=6.1H z,2H),2.47–2.39(m,1H),2.09–1.97(m,2H),1.72(d,J=12.7Hz,2H),1.60–1.45 (m,2H),1.17(s,9H).

[0194] Example 33 Synthesis of Protein Degrader Y33

[0195] The synthesis method was the same as that of Example 6, except that aniline was replaced by ethylamine (2M in THF) and 1,2-dibromoethane was replaced by 1,3-dibromoethane to obtain degradation agent Y33 (white solid, yield 33.4%). 1 H NMR (400MHz, DMSO-d6) δ12.21(s,1H),8.03(d,J=8.4Hz,1H),7.66(d,J=1.8Hz, 1H),7.37(s,1H),7.31(dd,J=8.4,1.6Hz,1H),6.70(s,1H),4.10(t,J=7.0Hz,2H ),4.03(s,2H),3.95(q,J=7.0Hz,2H),2.85(d,J=10.9Hz,2H),2.46–2.38(m,1H ),2.36–2.27(m,2H),1.90–1.69(m,6H),1.65–1.53(m,2H),1.18–1.10(m,12H).

[0196] Example 34 Synthesis of Protein Degrader Y34

[0197] The synthesis method was the same as that in Example 6, except that 1,2-dibromoethane was replaced with 1,3-dibromoethane to obtain degradation agent Y34 (white solid, yield 39.1%). 1H NMR(400MHz, DMSO-d6)δ12.22(s,1H),8.04(d,J=8.3Hz,1H),7.75(s,1H),7.51–7.27(m,6H),6.70(s,1H),4.11(t,J=7.0Hz,2H),4 .03(s,2H),2.88(d,J=10.9Hz,2H),2.46–2.40(m,1H),2.39–2.32(m,2H),1.92–1.72(m,6H),1.65(t,J=11.8Hz,2H),1.15(s,9H).

[0198] Example 35 Synthesis of Protein Degrader Y35

[0199] The synthesis method was the same as that in Example 6, except that aniline was replaced by 4-methylaminopyridine and 1,2-dibromoethane was replaced by 1,3-dibromoethane to obtain degradation agent Y35 (white solid, yield 26.0%). 1 H NMR(400MHz,DMSO-d6)δ12.23(s,1H),8.53–8.45(m,2H),8.07(d,J=8.5Hz,1H) ,7.75(d,J=1.8Hz,1H),7.42–7.35(m,2H),7.32–7.25(m,2H),6.72(s,1H),5.1 5(s,2H),4.14(t,J=6.9Hz,2H),4.06(s,2H),2.87(d,J=10.2Hz,2H),2.47–2.4 0(m,1H),2.37–2.31(m,2H),1.92–1.71(m,6H),1.68–1.54(m,2H),1.17(s,9H).

[0200] Example 36 Synthesis of Protein Degrader Y36

[0201] The synthesis method was the same as that in Example 6, except that 4-chloroisatoic anhydride was replaced with 5-methoxy-[1,3]benzoxazine-2,4-dione, aniline was replaced with 4-methylaminopyridine, and 1,2-dibromoethane was replaced with 1,3-dibromoethane to obtain degradation agent Y36 (white solid, yield 38.9%). 1H NMR (400MHz, DMSO-d6) δ12.24(s,1H),8.51–8.43(m,2H),7.69(t,J=8.5Hz,1H),7.39(s ,1H),7.27–7.22(m,2H),7.14(d,J=8.6Hz,1H),6.92(d,J=8.5Hz,1H),6.73(s,1H),5.1 0(s,2H),4.13(t,J=7.3Hz,2H),4.06(s,2H),3.85(s,3H),2.87(d,J=11.1Hz,2H),2.47 –2.41(m,1H),2.36(t,J=6.8Hz,2H),1.92–1.70(m,6H),1.66–1.53(m,2H),1.18(s,9H).

[0202] Example 37 Synthesis of Protein Degrader Y37

[0203] The synthesis method of Y37a was the same as that of Y6a in Example 6, except that aniline in the first step was replaced with 4-methylaminopyridine to obtain Y37a (white solid, yield 15.9%). 1 H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.53–8.42(m,2H),7.95(d,J=8.5Hz,1H),7.30–7.26(m,3H),7.24(d,J=2.0Hz,1H),5.09(s,2H).

[0204] first step

[0205] Y37a (500 mg, 1.74 mmol), methyl 4-hydroxycyclohexanecarboxylate (550 mg, 3.48 mmol), and triphenylphosphine (912 mg, 3.48 mmol) were dissolved in 10 mL of anhydrous tetrahydrofuran. Diisopropyl azodicarboxylate (DIAD, 0.7 mL, 3.48 mmol) was added dropwise under an ice bath. The mixture was then heated to room temperature and stirred overnight. The tetrahydrofuran was evaporated and the mixture was purified by silica gel column chromatography to obtain 500 mg of a mixture of Y37b and triphenylphosphine as a colorless oil.

[0206] Step 2

[0207] The mixture obtained in the previous step was dissolved in 3 mL of methanol, and 3 mL of water and lithium hydroxide monohydrate (245 mg, 5.84 mmol) were added. The mixture was stirred at room temperature overnight to obtain 200 mg of Y37c. 1H NMR (400MHz, DMSO-d6) δ12.09(s,1H),8.54–8.41(m,2H),8.09(d,J=8.5Hz,1H),7.54(d,J=2.0Hz,1H),7.43(dd,J=8.5,2.1Hz,1H ),7.28–7.19(m,2H),5.38–5.31(m,1H),5.24(s,2H),2.32–2.20(m,1H),1.86–1.77(m,2H),1.69–1.54(m,4H),1.43–1.27(m,2H).

[0208] Step 3

[0209] SNS-032 (50 mg, 0.13 mmol), Y37c (60 mg, 0.14 mmol), -(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 75 mg, 0.20 mmol), and DIPEA (70 μL, 0.39 mmol) were dissolved in 2 mL of dichloromethane, stirred at room temperature for 2 hours, and purified by silica gel column chromatography to obtain degradation agent Y37 (white solid, 80 mg, yield 85.7%). 1 H NMR (400MHz, DMSO-d6) δ12.33(s,1H),8.53–8.47(m,2H),8.09(d,J=8.5Hz,1H),7.54(d,J=2.0Hz,1H) ,7.43(dd,J=8.5,2.1Hz,1H),7.40(s,1H),7.29–7.26(m,2H),6.72(s,1H),5.49–5.39(m,1H),5.27–5. 19(m,2H),4.42(d,J=12.9Hz,1H),4.06(s,2H),3.99(d,J=13.6Hz,1H),3.04(t,J=12.7Hz,1H),2.82– 2.64(m,2H),2.63–2.53(m,1H),1.98–1.78(m,4H),1.76–1.62(m,2H),1.57–1.32(m,6H),1.18(s,9H).

[0210] Example 38 Synthesis of Protein Degrader Y38

[0211] first step

[0212] 3-Aminopyridine-2-carboxylic acid (5.0 g, 36.20 mmol) and benzylamine (5.9 mL, 54.30 mmol) were dissolved in 90 mL of dichloromethane, and DIPEA (18.9 mL, 108.60 mmol) and HATU (17.9 g, 47.06 mmol) were added. The mixture was stirred at room temperature overnight and purified by silica gel column chromatography to obtain Y38a (pale yellow oil, 5.6 g, yield 68.6%).

[0213] Step 2

[0214] Y38a (5.6 g, 24.82 mmol) was dissolved in 62 mL of acetonitrile, and Boc2O (6.5 g, 29.8 mmol) and p-dimethylaminopyridine (DMAP, 303 mg, 2.48 mmol) were added. The mixture was stirred at room temperature overnight. After Y38a was completely consumed, Y38b (3.2 g, white solid, yield 51.3%) was obtained by filtration. 1 H NMR (400MHz, DMSO-d6) δ11.64(s,1H),8.51(dd,J=4.3,1.4Hz,1H),7.71–7.59(m,2H),7.37–7.21(m,5H),5.11(s,2H).

[0215] Step 3

[0216] Y38b (300 mg, 1.18 mmol) was dissolved in 4 mL of DMF, and 1,3-dibromopropane (0.7 mL, 7.11 mmol) and potassium carbonate (327 mg, 2.37 mmol) were added. The mixture was stirred at room temperature overnight. Ethyl acetate and water were added. The ethyl acetate layer was washed four times with water and once with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain Y38c (white solid, 392 mg, yield 88.4%). 1 H NMR (400MHz, DMSO-d6) δ8.57(d,J=4.3Hz,1H),8.03(d,J=8.6Hz,1H),7.79(dd,J=8.7,4.4Hz,1H ),7.39–7.22(m,5H),5.16(s,2H),4.22(t,J=7.2Hz,2H),3.67–3.63(m,2H),2.21–2.12(m,2H).

[0217] Step 4

[0218] Y38c (100 mg, 0.27 mmol) was dissolved in 2 mL of DMF, and SNS-032 (91.5 mg, 0.24 mmol) and DIPEA (140 μL, 0.80 mmol) were added. The mixture was stirred at 60° C. overnight. Ethyl acetate and water were added. The ethyl acetate layer was washed four times with water and once with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain Y38 (white solid, 112 mg, yield 63.5%). 1 H NMR(400MHz,DMSO-d6)δ12.22(s,1H),8.62–8.47(m,1H),8.25–8.03(m,1H) ,7.88–7.67(m,1H),7.49–7.19(m,6H),6.81–6.66(m,1H),5.17(s,2H),4.1 4(t,J=7.2Hz,2H),4.08–4.04(m,2H),2.82(d,J=10.9Hz,2H),2.47–2.38(m ,1H),2.38–2.31(m,2H),1.89–1.68(m,6H),1.63–1.45(m,2H),1.18(s,9H).

[0219] Example 39 Synthesis of Protein Degrader Y39

[0220] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 4-aminopyridine-3-carboxylic acid to obtain degradation agent Y39 (white solid, yield 27.2%). 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),9.10(s,1H),8.73(d,J=5.9Hz,1H),7.58(d,J=6.1Hz,1H),7.47–7.13(m,6H),6.73(s,1H),5. 14(s,2H),4.17–4.09(m,2H),4.06(s,2H),2.86–2.76(m,2H),2.45–2.30(m,3H),1.89–1.66(m,6H),1.59–1.42(m,2H),1.18(s,9H).

[0221] Example 40 Synthesis of Protein Degrader Y40

[0222] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 3-aminoisonicotinic acid to obtain degradation agent Y40 (white solid, yield 16.9%). 1H NMR(400MHz,DMSO-d6)δ12.22(s,1H),9.07(s,1H),8.53(d,J=5.0Hz,1H),7 .93(d,J=5.0Hz,1H),7.39(s,1H),7.37–7.22(m,5H),6.73(s,1H),5.16(s,2 H),4.24(t,J=7.1Hz,2H),4.06(s,2H),2.82(d,J=10.9Hz,2H),2.46–2.32(m ,3H),1.89–1.79(m,4H),1.75–1.67(m,2H),1.59–1.45(m,2H),1.18(s,9H).

[0223] Example 41 Synthesis of Protein Degrader Y41

[0224] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 2-aminonicotinic acid to obtain degradation agent Y41 (white solid, yield 23.4%). 1 H NMR(400MHz, DMSO-d6)δ12.20(s,1H),8.77(dd,J=4.8,1.9Hz,1H),8.43(dd,J=7.8,1.9Hz,1H),7.41–7.22(m,7H),6.72(s,1H),5.15(s,2H),4.3 2(t,J=7.2Hz,2H),4.06(s,2H),2.81(d,J=10.9Hz,2H),2.45–2.29(m,3H ),1.89–1.74(m,4H),1.72–1.65(m,2H),1.52–1.39(m,2H),1.18(s,9H).

[0225] Example 42 Synthesis of Protein Degrader Y42

[0226] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 4-aminopyrimidine-5-carboxylic acid to obtain degradation agent Y42 (white solid, yield 7.9%). 1H NMR(400MHz, DMSO-d6)δ12.20(s,1H),9.22(d,J=7.1Hz,2H),7.41–7.36(m,3H),7.34–7.23(m,3H),6.73(s,1H),5.13(s,2H),4.26(t,J=7.1Hz,2H), 4.06(s,2H),2.79(d,J=10.9Hz,2H),2.44–2.30(m,3H),1.88–1.73(m,4H),1.71–1.63(m,2H),1.47–1.33(m,2H),1.18(s,9H).

[0227] Example 43 Synthesis of Protein Degrader Y43

[0228] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 3-aminopyrazine-2-carboxylic acid to obtain degradation agent Y43 (white solid, yield 6.1%). 1 H NMR (400MHz, DMSO-d6) δ12.21 (s, 1H), 8.81 (d, J = 2.4Hz, 1H), 8.64 (d, J = 2. 3Hz,1H),7.42–7.36(m,3H),7.34–7.23(m,3H),6.72(s,1H),5.17(s,2H), 4.27(t,J=7.2Hz,2H),4.06(s,2H),2.81(d,J=10.9Hz,2H),2.44–2.30(m, 3H),1.88–1.76(m,4H),1.72–1.65(m,2H),1.52–1.39(m,2H),1.18(s,9H).

[0229] Example 44 Synthesis of Protein Degrader Y44

[0230] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 2-aminonicotinic acid and benzylamine was replaced with ethylamine hydrochloride to obtain degradation agent Y44 (white solid, yield 4.6%). 1H NMR (400MHz, DMSO-d6) δ12.20(s,1H),8.75(dd,J=4.8,1.8Hz,1H),8.41(dd,J=7.8 ,1.9Hz,1H),7.38(s,1H),7.35(dd,J=7.7,4.8Hz,1H),6.73(s,1H),4.32(t,J=7.2 Hz,2H),4.06(s,2H),4.00(q,J=7.0Hz,2H),2.84(d,J=10.9Hz,2H),2.46–2.33(m, 3H),1.89–1.77(m,4H),1.76–1.65(m,2H),1.54–1.39(m,2H),1.23–1.16(m,12H).

[0231] Example 45 Synthesis of Protein Degrader Y45

[0232] The synthesis method was similar to that of Example 38, except that 3-aminopyridine-2-carboxylic acid was replaced with 2-aminonicotinic acid and benzylamine was replaced with 4-methylaminopyridine to obtain degradation agent Y45 (white solid, yield 6.7%). 1 H NMR(400MHz, DMSO-d6)δ12.20(s,1H),8.80(dd,J=4.8,1.9Hz,1H),8.51–8.49(m,2H),8.44(dd,J=7.7,1.9Hz,1 H),7.42–7.37(m,2H),7.35–7.31(m,2H),6.73(s,1H),5.17(s,2H),4.33(t,J=7.2Hz,2H),4.06(s,2H),2.83(d, J=11.0Hz,2H),2.47–2.32(m,3H),1.90–1.77(m,4H),1.74–1.67(m,2H),1.55–1.42(m,2H),1.19(s,9H).

[0233] Example 46 Synthesis of Protein Degrader Y46

[0234] The synthesis of Y46a was carried out with reference to Y38a. In the first step, 3-aminopyridine-2-carboxylic acid was replaced with 2-aminonicotinic acid, and benzylamine was replaced with ethylamine hydrochloride to obtain Y46a (white solid, yield 66.0%). 1H NMR (400MHz, DMSO-d6) δ11.95(s,1H),8.63(dd,J=4.7,1.9Hz,1H),8.31(dd,J=7.8,1 .9Hz,1H),7.28(dd,J=7.8,4.8Hz,1H),3.93(q,J=7.0Hz,2H),1.16(t,J=7.0Hz,3H).

[0235] first step

[0236] Y46a (500 mg, 2.62 mmol), 1,2-ethylene glycol (974 mg, 15.69 mmol), and triphenylphosphine (1.4 g, 5.23 mmol) were dissolved in 10 mL of anhydrous tetrahydrofuran. DIAD was added dropwise under ice-cooling. After the addition was complete, the mixture was warmed to room temperature and stirred overnight. The mixture was then purified by silica gel column chromatography to obtain a mixture of Y46b and triphenylphosphine (881 mg, colorless oil).

[0237] Step 2

[0238] Y46b obtained in the previous step was dissolved in 12 mL of dichloromethane, and Dess-Martin periodinane (1.44 g, 3.40 mmol) was added under ice-cooling. The mixture was stirred at 0°C for 5 hours and purified by silica gel column chromatography to obtain Y46c (white solid, 305 mg, yield 34.9%). 1 H NMR (400MHz, DMSO-d6) δ9.68(s,1H),8.68(dd,J=4.8,1.9Hz,1H),8.45(dd,J=7.8,1.9Hz,1 H),7.39(dd,J=7.8,4.8Hz,1H),5.12(s,2H),3.99(q,J=7.1Hz,2H),1.18(t,J=7.1Hz,3H).

[0239] Step 3

[0240] Y46c (13 mg, 0.06 mmol) and C24a (20 mg, 0.04 mmol) were dissolved in 1 mL of dichloroethane (DCE), and one drop of acetic acid was added. After stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (24 mg, 0.11 mmol) was added and stirred at room temperature for 30 minutes. Dichloromethane and water were added, and the dichloromethane layer was washed twice with water and once with saturated sodium chloride aqueous solution. The mixture was dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain degradation agent Y46 (white solid, 17 mg, yield 60.6%). 1H NMR(400MHz,Chloroform-d)δ8.67(dd,J=4.7,1.9Hz,1H),8.52–8.47(m,2H),8.39(s,1H),7.83–7.75(m,2H),7.72–7.70(m,1H),7.61(s,1H) ),7.22(dd,J=7.7,4.7Hz,1H),6.72(d,J=8.8Hz,1H),6.03(s,1H),4.96–4.85(m,1H),4.65(d,J=5.9Hz,2H),4.60(t,J=6.8Hz,2H),4.17(q, J=7.0Hz,2H),3.62–3.56(m,4H),2.82(t,J=6.8Hz,2H),2.79–2.74(m,4H ),2.25(s,3H),2.08(s,3H),1.62(d,J=6.6Hz,6H),1.31(t,J=7.0Hz,3H).

[0241] Example 47 Synthesis of Protein Degrader Y47

[0242] The synthesis method was similar to that of Example 46, except that 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y47 (white solid, yield 56.7%). 1 H NMR (400MHz, Chloroform-d) δ12.25(s,1H),8.68–8.65(m,1H),8.50–8.46(m,2H),8.41(s,1H),7.97(t,J=5. 9Hz,1H),7.78(dd,J=8.8,2.6Hz,1H),7.73(s,1H),7.60(s,1H),7.24–7.19(m,1H),6.69(d,J=8.9Hz,1H),5.9 4(s,1H),4.96–4.83(m,1H),4.66(d,J=5.8Hz,2H),4.49–4.42(m,2H),4.23–4.10(m,2H),3.61–3.54(m,4H), 2.62–2.54(m,6H),2.43(s,3H),2.18(s,3H),2.05–1.98(m,2H),1.61(d,J=6.6Hz,6H),1.32(t,J=7.0Hz,3H).

[0243] Example 48 Synthesis of Protein Degrader Y48

[0244] The synthesis method was similar to that of Example 46, except that 1,2-ethylene glycol was replaced with 1,4-butanediol to obtain degradation agent Y48 (white solid, yield 33.5%). 1 H NMR(400MHz,Chloroform-d)δ12.44(s,1H),8.66(dd,J=4.8,2.0Hz,1H),8.53–8.46(m,2H),8.42(s,1H),8.00(t,J=5.8Hz,1H) ,7.79(dd,J=8.8,2.6Hz,1H),7.74(d,J=1.3Hz,1H),7.60(s,1H),7.21(dd,J=7.8,4.8Hz,1H),6.70(d,J=8.9Hz,1H),5.95(s,1 H),4.96–4.81(m,1H),4.66(d,J=5.8Hz,2H),4.46–4.35(m,2H),4.21–4.12(m,2H),3.66–3.57(m,4H),2.64–2.58(m,4H),2.51 (t,J=7.6Hz,2H),2.43(s,3H),2.18(s,3H),1.87–1.77(m,2H),1.74–1.63(m,2H),1.60(d,J=6.6Hz,6H),1.31(t,J=7.1Hz,3H).

[0245] Example 49 Synthesis of Protein Degrader Y49

[0246] The synthesis method was similar to that of Example 46, except that 1,2-ethylene glycol was replaced with 1,5-pentanediol to obtain degradation agent Y49 (white solid, yield 60.0%). 1H NMR(400MHz,Chloroform-d)δ8.66(dd,J=4.8,1.9Hz,1H),8.51(d,J=2.5Hz,1H),8.48(dd,J=7.8,1.9Hz,1H),8.40(s,1H),7.84(t,J =6.0Hz,1H),7.80(dd,J=8.9,2.5Hz,1H),7.72(d,J=1.2Hz,1H),7.60(s,1H),7.21(dd,J=7.7,4.8Hz,1H),6.72(d,J=8.8Hz,1H),6.0 1(s,1H),4.99–4.83(m,1H),4.65(d,J=5.8Hz,2H),4.43–4.29(m,2H),4.16(q,J=7.0Hz,2H),3.69–3.61(m,4H),2.69–2.62(m,4H),2 .51–2.44(m,5H),2.23(s,3H),1.84–1.74(m,2H),1.72–1.64(m,2H),1.61(d,J=6.6Hz,6H),1.53–1.43(m,2H),1.31(t,J=7.1Hz,3H).

[0247] Example 50 Synthesis of Protein Degrader Y50

[0248] The synthesis method was similar to that of Example 46, except that 1,2-ethylene glycol was replaced with diethylene glycol to obtain degradation agent Y50 (white solid, yield 65.7%). 1 H NMR(400MHz,Chloroform-d)δ8.65(dd,J=4.8,2.0Hz,1H),8.51–8.46(m,2H),8.41(s,1H),7.91(t,J=5.9Hz,1H),7 .80(dd,J=8.8,2.6Hz,1H),7.73(d,J=1.2Hz,1H),7.60(s,1H),7.22(dd,J=7.8,4.7Hz,1H),6.69(d,J=8.9Hz,1H), 5.99(s,1H),4.97–4.83(m,1H),4.70–4.56(m,4H),4.16(q,J=7.0Hz,2H),3.84(t,J=5.9Hz,2H),3.73(t,J=5.4Hz, 2H),3.61–3.55(m,4H),2.66–2.56(m,6H),2.45(s,3H),2.21(s,3H),1.61(d,J=6.6Hz,6H),1.30(t,J=7.0Hz,3H).

[0249] Example 51 Synthesis of Protein Degrader Y51

[0250] The synthesis method was similar to that of Example 46, except that Y46a was replaced with 7-chloro-3-ethylquinazoline-2,4(1H,3H)-dione and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y51 (white solid, yield 30.0%). 1 H NMR(400MHz,Chloroform-d)δ8.51(d,J=2.5Hz,1H),8.42(s,1H),8.18(d,J=8.4Hz,1H),7.91(t, J=6.0Hz,1H),7.81(dd,J=8.8,2.6Hz,1H),7.74(s,1H),7.61(s,1H),7.47(d,J=1.7Hz,1H),7.23 (dd,J=8.5,1.6Hz,1H),6.73(d,J=8.8Hz,1H),5.99(s,1H),4.98–4.81(m, 1H),4.67(d,J=5.9Hz,2H),4.23(t,J=7.1Hz,2H),4.16(q,J=7.1Hz,2H),3. 75–3.66(m,4H),2.72–2.65(m,4H),2.59(t,J=6.6Hz,2H),2.46(s,3H),2. 22(s,3H),2.06–2.00(m,2H),1.62(d,J=6.6Hz,6H),1.31(t,J=7.0Hz,3H).

[0251] Example 52 Synthesis of Protein Degrader Y52

[0252] The synthesis method was similar to that of Example 46, except that Y46a was replaced with 7-chloro-3-(pyridin-4-ylmethyl)quinazoline-2,4(1H,3H)-dione and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y52 (white solid, yield 34.5%). 1H NMR(400MHz,Chloroform-d)δ8.62–8.55(m,2H),8.51(s,1H),8.43(s,1H),8.20(d,J=8.5Hz,1H), 7.89–7.77(m,2H),7.71(s,1H),7.61(s,1H),7.51(s,1H),7.40–7.34(m,2H),6.74(d,J=8.9Hz,1H) ,5.99(s,1H),5.27(s,2H),4.97–4.83(m,1H),4.66(d,J=5.8Hz,2H),4.25(t,J=7.1Hz,2H),3.80–3 .65(m,4H),2.79–2.55(m,6H),2.46(s,3H),2.24(s,3H),2.10–2.01(m,2H),1.62(d,J=6.6Hz,6H).

[0253] Example 53 Synthesis of Protein Degrader Y53

[0254] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5609a and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y53 (white solid, yield 70.7%). 1 H NMR(400MHz,Chloroform-d)δ11.86(d,J=21.9Hz,1H),8.73–8.41(m,4H),8.06(d,J=26.7Hz,1H),7.66–7.41(m ,1H),7.23–7.11(m,1H),6.48–6.21(m,1H),4.57–4.37(m,2H),4.33–4.23(m,1H),4.20–4.10(m,2H),2.76–2.42 (m,4H),2.17–2.07(m,6H),2.03–1.56(m,6H),1.35–1.21(m,5H).

[0255] Example 54 Synthesis of Protein Degrader Y54

[0256] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422a and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y54 (white solid, yield 72.6%). 1H NMR(400MHz,Chloroform-d)δ8.71–8.62(m,2H),8.53–8.46(m,2H),8.29(d,J=8.6Hz,1H),8.24(d,J=2.3H z,1H),7.89(t,J=1.6Hz,1H),7.70(dd,J=10.8,1.8Hz,1H),7.56(dd,J=8.7,2.4Hz,1H),7.21(dd,J=7.8,4. 7Hz,1H),4.99–4.86(m,1H),4.67(s,2H),4.46(t,J=7.3Hz,2H),4.17(q,J=7.0Hz,2H),3.09(d,J=11.0Hz, 2H),2.65–2.43(m,3H),2.15–1.97(m,4H),1.88–1.69(m,4H),1.61(d,J=7.0Hz,6H),1.31(t,J=7.0Hz,3H).

[0257] Example 55 Synthesis of Protein Degrader Y55

[0258] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422a and 1,2-ethylene glycol was replaced with 1,5-pentanediol to obtain degradation agent Y55 (white solid, yield 60.7%). 1 H NMR(400MHz,Chloroform-d)δ8.71(s,1H),8.67(dd,J=4.8,2.0Hz,1H),8.51–8.45(m,2H),8.29(d,J=8.6Hz,1H),8.27(d,J=2. 3Hz,1H),7.89(t,J=1.6Hz,1H),7.70(dd,J=10.8,1.8Hz,1H),7.59(dd,J=8.7,2.4Hz,1H),7.21(dd,J=7.7,4.8Hz,1H),4.98–4 .87(m,1H),4.67(s,2H),4.42–4.33(m,2H),4.16(q,J=7.0Hz,2H),3.14(d,J=11.1Hz,2H),2.60–2.52(m,1H),2.46(t,J=7.9Hz ,2H),2.17–2.07(m,2H),1.92–1.75(m,6H),1.72–1.65(m,2H),1.61(d,J=7.0Hz,6H),1.52–1.42(m,2H),1.31(t,J=7.1Hz,3H).

[0259] Example 56 Synthesis of Protein Degrader Y56

[0260] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422a and 1,2-ethylene glycol was replaced with diethylene glycol to obtain degradation agent Y56 (white solid, yield 52.8%). 1 H NMR(400MHz,Chloroform-d)δ8.68–8.61(m,2H),8.51–8.44(m,2H),8.29(d,J=8.6Hz,1H),8.24(d,J=2.3Hz,1H),7.90(t,J =1.6Hz,1H),7.70(dd,J=10.9,1.9Hz,1H),7.56(dd,J=8.6,2.4Hz,1H),7.21(dd,J=7.7,4.8Hz,1H),4.99–4.86(m,1H),4.67 (s,2H),4.64(t,J=5.9Hz,2H),4.16(q,J=7.1Hz,2H),3.85(t,J=5.9Hz,2H),3.73(t,J=5.6Hz,2H),3.07(d,J=11.2Hz,2H),2 .62(t,J=5.7Hz,2H),2.53–2.43(m,1H),2.16–2.07(m,2H),1.84–1.75(m,4H),1.61(d,J=7.0Hz,6H),1.31(t,J=7.0Hz,3H).

[0261] Example 57 Synthesis of Protein Degrader Y57

[0262] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422a and 1,2-ethylene glycol was replaced with triethylene glycol to obtain degradation agent Y57 (white solid, yield 61.8%). 1H NMR(400MHz,Chloroform-d)δ8.72(s,1H),8.65–8.61(m,1H),8.50–8.43(m,2H),8.29(d,J=8.6Hz,1H),8.26–8.23(m ,1H),7.89(s,1H),7.74–7.67(m,1H),7.62–7.55(m,1H),7.21(dd,J=7.7,4.7Hz,1H),4.99–4.86(m,1H),4.67(s,2H), 4.62(t,J=6.2Hz,2H),4.15(q,J=7.1Hz,2H),3.85(t,J=6.2Hz,2H),3.77–3.55(m,6H),3.14(d,J=11.3Hz,2H),2.67(t ,J=5.9Hz,2H),2.58–2.47(m,1H),2.26–2.17(m,2H),1.93–1.78(m,4H),1.61(d,J=7.0Hz,6H),1.30(t,J=7.1Hz,3H).

[0263] Example 58 Synthesis of Protein Degrader Y58

[0264] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422a and 1,2-ethylene glycol was replaced with 1,8-octanediol to obtain degradation agent Y58 (white solid, yield 61.8%). 1 H NMR(400MHz,Chloroform-d)δ8.76(s,1H),8.66(dd,J=4.7,2.0Hz,1H),8.51–8.44(m,2H),8.33–8.25(m,2H),7. 89(t,J=1.6Hz,1H),7.70(dd,J=10.8,1.8Hz,1H),7.59(dd,J=8.7,2.4Hz,1H),7.20(dd,J=7.7,4.7Hz,1H),4.98– 4.85(m,1H),4.67(s,2H),4.40–4.29(m,2H),4.16(q,J=7.0Hz,2H),3.17(d,J=11.2Hz,2H),2.61–2.50(m,1H),2. 49–2.41(m,2H),2.20–2.10(m,2H),1.97–1.84(m,4H),1.79–1.69(m,2H),1.64–1.55(m,8H),1.46–1.24(m,11H).

[0265] Example 59 Synthesis of Protein Degrader Y59

[0266] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422b and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y59 (white solid, yield 41.1%). 1 H NMR(400MHz,Chloroform-d)δ8.66(dd,J=4.8,1.9Hz,1H),8.57(s,1H),8.48(dd,J=7.8,1.9Hz,1H),8.39(d,J=3.8Hz,1 H),8.33(d,J=8.7Hz,1H),8.23(d,J=2.3Hz,1H),7.55(dd,J=8.7,2.4Hz,1H),7.47(s,1H),7.31(dd,J=11.3,1.8Hz,1H) ,7.21(dd,J=7.7,4.7Hz,1H),4.45(t,J=7.3Hz,2H),4.38(t,J=4.4Hz,2H),4.25–4.13(m,3H),3.33(t,J=4.4Hz,2H),3. 14–3.03(m,2H),2.62–2.43(m,3H),2.13–1.96(m,4H),1.88–1.67(m,4H),1.31(t,J=7.0Hz,3H),1.26(d,J=6.5Hz,6H).

[0267] Example 60 Synthesis of Protein Degrader Y60

[0268] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422b and 1,2-ethylene glycol was replaced with 1,5-pentanediol to obtain degradation agent Y60 (white solid, yield 64.6%). 1 H NMR(400MHz,Chloroform-d)δ8.68(dd,J=4.8,1.9Hz,1H),8.49(dd,J=7.7,1.9Hz,1H) ,8.41–8.33(m,2H),8.26–8.08(m,2H),7.61(d,J=8.7Hz,1H),7.48(s,1H),7.34–7.30 (m,1H),7.22(dd,J=7.8,4.8Hz,1H),4.44–4.34(m,4H),4.28–4.09(m,3H),3.49–3.08 (m,3H),2.84–2.18(m,4H),2.06–1.58(m,8H),1.56–1.41(m,2H),1.35–1.25(m,11H).

[0269] Example 61 Synthesis of Protein Degrader Y61

[0270] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422b and 1,2-ethylene glycol was replaced with diethylene glycol to obtain degradation agent Y61 (white solid, yield 59.5%). 1 H NMR(400MHz,Chloroform-d)δ8.70–8.64(m,1H),8.49(d,J=7.5Hz,1H),8.37(d,J=3.8Hz,1H),8.34(d,J=8.7Hz,1H),8 .20–8.17(m,1H),8.10–8.03(m,1H),7.56(d,J=8.9Hz,1H),7.48(s,1H),7.32(d,J=11.7Hz,1H),7.22(dd,J=7.9,4.8H z,1H),4.65(t,J=5.9Hz,2H),4.40(t,J=4.4Hz,2H),4.29–4.09(m,3H),3.86(t,J=5.9Hz,2H),3.82–3.70(m,1H),3.34 (t,J=4.4Hz,2H),3.18–3.00(m,1H),2.57(d,J=51.1Hz,2H),2.24–2.01(m,2H),1.94–1.59(m,4H),1.36–1.15(m,12H).

[0271] Example 62 Synthesis of Protein Degrader Y62

[0272] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422b and 1,2-ethylene glycol was replaced with triethylene glycol to obtain degradation agent Y62 (white solid, yield 65.2%). 1H NMR(400MHz, DMSO-d6)δ9.92(s,1H),8.76–8.72(m,1H),8.62(d,J=3.9Hz,1H),8.40(dd,J=7.8,1.9Hz,1H),8.20–8.17(m, 1H),8.12(d,J=8.7Hz,1H),7.61(d,J=8.7Hz,1H),7.49(s,1H),7.36(dd,J=7.7,4.8Hz,1H),7.19(d,J=12.0Hz,1H),4.44(t ,J=6.5Hz,2H),4.31(t,J=4.2Hz,2H),4.20–4.11(m,1H),3.98(q,J=7.4Hz,2H),3.69(t,J=6.5Hz,2H),3.60–3.54(m,2H), 3.52–3.44(m,4H),3.33–3.29(m,2H),2.96(s,1H),2.46(s,1H),2.10–1.97(m,2H),1.82–1.56(m,3H),1.23–1.12(m,13H).

[0273] Example 63 Synthesis of Protein Degrader Y63

[0274] The synthesis method was similar to that of Example 46, except that C24a was replaced with NUV-422b and 1,2-ethylene glycol was replaced with 1,8-octanediol to obtain degradation agent Y63 (white solid, yield 60.9%). 1 H NMR(400MHz,Chloroform-d)δ9.26(s,1H),8.65(dd,J=4.8,1.9Hz,1H),8.45(dd,J=7.8,1.9Hz,1H),8.42(d,J=3 .8Hz,1H),8.37–8.31(m,2H),7.57(dd,J=8.6,2.4Hz,1H),7.44(s,1H),7.32–7.24(m,1H),7.18(dd,J=7.8,4.7H z,1H),4.39–4.27(m,4H),4.23–4.07(m,3H),3.34–3.22(m,4H),2.67–2.48(m,3H),2.38–2.24(m,2H),2.14–1.9 9(m,2H),1.95–1.84(m,2H),1.80–1.59(m,4H),1.43–1.32(m,8H),1.29(t,J=7.1Hz,3H),1.25(d,J=6.5Hz,6H).

[0275] Example 64 Synthesis of Protein Degrader Y64

[0276] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5102 and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain degradation agent Y64 (white solid, yield 79.9%). 1 H NMR(400MHz,Chloroform-d)δ11.64(s,1H),8.80–8.53(m,3H),8.45(d,J=7.6Hz,1H),7.83(d,J=33.5Hz,1H),7.27–7.09(m,2H),6.37(d ,J=58.6Hz,1H),4.56–4.22(m,3H),4.14(q,J=7.0Hz,2H),2.73–2.23(m,12H),2.02–1.91(m,2H),1.89–1.58(m,4H),1.33–1.22(m,3H).

[0277] Example 65 Synthesis of Protein Degrader Y65

[0278] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5102 and 1,2-ethylene glycol was replaced with 1,5-pentanediol to obtain degradation agent Y65 (white solid, yield 55.7%). 1 H NMR(400MHz,Chloroform-d)δ11.13(s,1H),8.76–8.53(m,3H),8.46(dd,J=7.8,1.9Hz,1H),7.88(d,J=27.1Hz,1H),7.27–7.15(m,2 H),6.15(s,1H),4.41–4.28(m,3H),4.21–4.08(m,3H),2.59–2.53(m,3H),2.46–2.39(m,3H),1.94–1.40(m,11H),1.33–1.22(m,7H).

[0279] Example 66 Synthesis of Protein Degrader Y66

[0280] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5102 and 1,2-ethylene glycol was replaced with diethylene glycol to obtain degradation agent Y66 (white solid, yield 6.3%). 1H NMR(400MHz,Chloroform-d)δ10.36(s,1H),8.79–8.50(m,3H),8.43(s,1H),7.91(d,J=29.9Hz,1H),7.27–7.24(m,1H ),7.20–7.12(m,1H),6.15–6.05(m,1H),4.66–4.57(m,2H),4.30–4.20(m,1H),4.19–4.06(m,2H),3.89–3.76(m,2H), 3.74–3.61(m,2H),2.78–2.52(m,8H),2.45(s,3H),2.40–2.32(m,1H),1.84–1.64(m,3H),1.62–1.54(m,1H),1.30–1.25(m,3H).

[0281] Example 67 Synthesis of Protein Degrader Y67

[0282] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5102 and 1,2-ethylene glycol was replaced with triethylene glycol to obtain degradation agent Y67 (white solid, yield 41.2%). 1 H NMR(400MHz,Chloroform-d)δ10.96(s,1H),8.75–8.51(m,3H),8.45(d,J=7.9Hz,1H),7 .87(d,J=31.6Hz,1H),7.25(d,J=8.0Hz,1H),7.18(dd,J=7.7,4.8Hz,1H),6.18(s,1H), 4.59(t,J=6.0Hz,2H),4.29(s,1H),4.18–4.08(m,2H),3.82(t,J=6.2Hz,2H),3.72–3.6 5(m,3H),3.62–3.52(m,3H),2.81–2.37(m,12H),1.88–1.58(m,4H),1.34–1.22(m,3H).

[0283] Example 68 Synthesis of Protein Degrader Y68

[0284] The synthesis method was similar to that of Example 46, except that C24a was replaced with SY-5102 and 1,2-ethylene glycol was replaced with 1,8-octanediol to obtain degradation agent Y68 (white solid, yield 31.6%). 1H NMR(400MHz,Chloroform-d)δ10.99(s,1H),8.77–8.53(m,3H),8.47(dd,J=7.8, 1.8Hz,1H),7.88(d,J=33.3Hz,1H),7.25(d,J=8.2Hz,1H),7.19(dd,J=7.8,4.8Hz ,1H),6.12(s,1H),4.33(t,J=7.8Hz,3H),4.16(q,J=7.1Hz,2H),2.82–2.28(m,12 H),1.88–1.79(m,2H),1.77–1.64(m,4H),1.55–1.44(m,2H),1.42–1.24(m,11H).

[0285] Example 69 Synthesis of Protein Degrader Y69

[0286] first step

[0287] To 7-chloro-8-fluoropyrido[4,3-d]pyrimidine-2,4(1H,3H)-dione (500 mg, 2.32 mmol) was added N,N-diisopropylethylamine (1.80 mL, 11.60 mmol) under nitrogen protection. Phosphorus oxychloride (4.96 mL, 53.35 mmol) was added under ice bath, and the mixture was reacted at 50 °C for 3 h. After the reaction, the phosphorus oxychloride was evaporated at 70 °C. The product was diluted with dichloromethane, washed once with ice water, and dried to give Y69a (brown block, 585.56 mg, 2.32 mmol). It was directly used for the next step without further purification.

[0288] Step 2

[0289] To a solution of Y69a in 10 mL of dichloromethane was added dropwise N,N-diisopropylethylamine (DIEA, 2.71 mL, 15.54 mmol) at -40°C, followed by a solution of tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (492 mg, 2.32 mmol) in dichloromethane. The mixture was allowed to react for 0.5 h after the addition was complete. The reaction was monitored to be complete by TLC. The mixture was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and purified by column chromatography to give Y69b (yellow solid, 695 mg, two-step yield 70%). 1 H NMR (400MHz, Chloroform-d) δ 8.84 (s, 1H), 4.54 (d, 2H), 4.40 (d, 2H), 3.73 (m, 2H), 1.99 (d, J = 5.9Hz, 2H), 1.67 (d, J = 7.7Hz, 2H), 1.52 (s, 9H).

[0290] Step 3

[0291] To 2-aminonicotinic acid (10 g, 72.40 mmol) was added 120 mL of dichloromethane, followed by HATU (35.78 g, 94.12 mmol) and DIEA (31.53 mL, 180.99 mmol). After 15 min, ethylamine hydrochloride (7.08 g, 86.88 mmol) was added. The mixture was reacted at room temperature for 16 hours, and the mixture was dried and used for the next step.

[0292] Step 4

[0293] The product from the previous step was added to a single-necked flask with 90 mL of 1,4-dioxane, followed by the addition of N,N'-carbonyldiimidazole (17.61 g, 108.60 mmol) and DIEA (25.22 mL, 144.80 mmol). The reaction was refluxed at 120°C for 16 h. The reaction was complete as monitored by TLC. The product was allowed to cool to room temperature to precipitate crystals, which were filtered and washed with DCM. The filter cake was dried to give the product Y46a (white solid, 7.20 g, yield 52%). 1 H NMR (400MHz, DMSO-d6) δ11.93 (s, 1H), 8.61 (dd, J = 4.8, 1.9Hz, 1H), 8.30 (dd,J=7.8,1.9Hz,1H),7.27(dd,J=7.8,4.8Hz,1H),3.91(q,J=7.0Hz,2H),1.14(t,J=7.0Hz,3H).

[0294] Step 5

[0295] Y69c (150 mg, 784.56 μmol) and potassium carbonate (110.60 mg, 800.25 μmol) were dissolved in 5 mL of N,N-dimethylformamide. After 5 min, 2-bromoethanol (111 μL, 1.58 mmol) was added. After reacting at room temperature for 16 h, water and ethyl acetate were added to the system for extraction. The ethyl acetate was washed with saturated sodium chloride aqueous solution and dried over anhydrous sodium sulfate. The product Y69d (white solid, 103 mg, yield 56%) was obtained by column chromatography. 1 H NMR(400MHz, DMSO-d6)δ8.72(dd,J=4.8,2.0Hz,1H),8.39(dd,J=7.7,1.9Hz,1H),7.40–7.28(m,1H),4.79( t,J=6.0Hz,1H),4.34(t,J=6.7Hz,2H),3.97(q,J=7.0Hz,2H),3.63(q,J=6.5Hz,2H),1.17(t,J=7.0Hz,3H).

[0296] Step 6

[0297] Under nitrogen protection, Y69d (85.68 mg, 364.24 μmol) was dissolved in 5 mL of dry tetrahydrofuran, and then sodium hydride (14.57 mg, wt 60%, 364.24 μmol) was slowly added under ice bath. After 10 min, 5 mL of a tetrahydrofuran solution of Y69b (130 mg, 303.53 μmol) was added. The reaction was moved to room temperature for 16 h. TLC monitored the complete consumption of Y69b. Water and ethyl acetate were added to the reaction system for extraction, and the mixture was washed with saturated sodium chloride aqueous solution and dried over anhydrous sodium sulfate. The product Y69e (light yellow solid, 109 mg, yield 57%) was obtained by column chromatography. 1 H NMR (400MHz, DMSO-d6) δ8.86(s,1H),8.61(dd,J=4.8,1.8Hz,1H),8.33(dd,J=7.8,1.9Hz,1H),7.29(dd,J=7.7,4.8Hz,1H),4.71(dd,J=13.9,4.9Hz,4H),4 .42(d,J=12.7Hz,2H),4.20(s,2H),3.89(q,J=7.0Hz,2H),3.57(d,J=12.6Hz ,2H),1.76(s,2H),1.55(d,J=7.7Hz,2H),1.46(s,9H),1.05(t,J=7.0Hz,3H).

[0298] Step 7

[0299] Under argon protection, Y69e was dissolved in 3 mL of toluene, and 500 μL of water was added. Potassium phosphate (101 mg, 478.41 μmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (90 mg, 175.42 μmol) were added, and finally methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II)) (13.4 mg, 15.95 μmol) was added. The mixture was ventilated three times and the reaction was monitored by TLC. The product was directly dried and purified by column chromatography to give compound Y69f (52 mg). It was not further isolated and directly used in the next step.

[0300] Step 8

[0301] Under nitrogen protection, Y69f (52 mg) was dissolved in dry N,N-dimethylformamide, and then cesium fluoride was added. The reaction was allowed to react at room temperature for 1 h and monitored by TLC. Water and ethyl acetate were added and extracted three times. The ethyl acetate layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The organic layer was spin-dried and then a dioxane hydrochloride solution was added thereto in an ice bath under nitrogen protection. The reaction was monitored by TLC. The reaction was separated and prepared to give compound Y69 (yellow solid, 8 mg). 1 H NMR(400MHz, Methanol-d4)δ8.97(s,1H),8.58(dd,J=4.8,1.9Hz,1H),8.40(dd,J=7.8, 1.9Hz,1H),7.85(dd,J=9.2,5.7Hz,1H),7.35–7.33(m,1H),7.33–7.19(m,3H),4.84(s,3 H),4.62(d,J=16.6Hz,2H),4.01(q,J=7.0Hz,2H),3.82(s,2H),3.74(d,J=12.2Hz,2H), 3.34(d,J=1.0Hz,1H),3.33(s,2H),1.89(dd,J=22.0,8.9Hz,4H),1.15(t,J=7.0Hz,3H).

[0302] Example 70 Synthesis of Protein Degrader Y82

[0303] first step

[0304] Y46a (1.0 g, 5.23 mmol), methyl 3-hydroxypropionate (816.8 mg, 7.85 mmol), and triphenylphosphine (2.7 g, 10.46 mmol) were dissolved in 15 mL of anhydrous tetrahydrofuran. DIAD (2.0 mL, 10.46 mmol) was added dropwise under ice-bath. After the addition was complete, the temperature was raised to room temperature overnight. The tetrahydrofuran was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography to give Y82b (white solid, 1.58 g, 100% yield). 1 H NMR (400MHz, DMSO-d6) δ8.74(dd,J=4.8,1.9Hz,1H),8.41(dd,J=7.7,1.9Hz,1H),7.37(dd,J=7.7,4.8 Hz,1H),4.55–4.43(m,2H),3.97(q,J=7.0Hz,2H),3.59(s,3H),2.75–2.64(m,2H),1.17–1.13(m,3H).

[0305] Step 2

[0306] Y82b (1.58 g, 5.69 mmol) was dissolved in 10 mL of methanol, and 10 mL of water and lithium hydroxide monohydrate (716.4 mg, 17.07 mmol) were added. The mixture was stirred at room temperature overnight. The methanol was removed by rotary evaporation and the pH was adjusted to 4 with 1N dilute hydrochloric acid to precipitate a white solid. Y82c (white solid, 514.1 mg, yield 34.7%) was obtained by filtration. 1 H NMR (400MHz, DMSO-d6) δ8.75(dd,J=4.8,1.9Hz,1H),8.40(dd,J=7.8,1.9Hz,1H),7.36(dd,J=7.7,4 .8Hz,1H),4.45(dd,J=8.7,7.0Hz,2H),3.97(q,J=7.0Hz,2H),2.68–2.55(m,2H),1.17–1.13(m,3H).

[0307] Step 3

[0308] KI-ARV-03 (20.0 mg, 0.08 mmol) was dissolved in 1 mL of dichloromethane, and HATU (44.0 mg, 0.12 mmol), DIPEA (0.04 mL, 0.23 mmol), and Y82c (24.4 mg, 0.09 mmol) were added. The mixture was stirred at room temperature for 3 hours, and Y82 was obtained by silica gel column chromatography (colorless oil, 8 mg, yield 17.6%). 1 H NMR(400MHz,Chloroform-d)δ8.68(dd,J=4.8,1.9Hz,1H),8.50(dd,J=7.8,1.9Hz,1H),7.95(d,J=2.2Hz,1H ),7.25(dd,J=7.8,4.8Hz,1H),6.45–6.41(m,2H),6.26(d,J=6.9Hz,1H),5.82(s,1H),4.68(t,J=7.1Hz,2H), 4.50–4.38(m,1H),4.26–4.12(m,3H),2.81–2.68(m,4H),2.45–2.36(m,1H),2.34–2.25(m,1H),2.23–2.15(m ,1H),2.14–2.04(m,1H),1.88–1.72(m,3H),1.70–1.55(m,1H),1.31(t,J=7.1Hz,3H),1.03(t,J=7.4Hz,3H).

[0309] Example 71 Synthesis of Protein Degrader Y83

[0310] The synthesis method was as follows: with reference to Example 70, except that KI-ARV-03 was replaced with compound 754 to obtain compound Y83 (white solid, yield 79.8%). 1 H NMR(400MHz, DMSO-d6)δ9.07(s,1H),8.75(dd,J=4.8,1.9Hz,1H),8.41(dd,J=7.7,1.9Hz,1H),8.23(s,1H),7.97(s,1H),7.84 (d,J=7.6Hz,1H),7.68(s,1H),7.43(d,J=7.6Hz,1H),7.36(dd,J=7.7,4.8Hz,1H),4.49–4.43(m,2H),4.02–3.92(m,4H),3.57– 3.39(m,2H),2.88(s,2H),2.46(t,J=7.5Hz,2H),1.89(s,2H),1.76(s,2H),1.27(s,6H),1.26–1.15(m,7H).

[0311] Example 72 Synthesis of Protein Degrader Y84

[0312] The synthesis method was as described in Example 46, except that C24a was replaced with compound 754, and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain compound Y84 (colorless oil, yield 22.9%). 1 H NMR(400MHz,Chloroform-d)δ8.95(s,1H),8.80–8.67(m,1H),8.63(dd,J=4.8,2.0Hz,1H),8.45(dd ,J=7.8,1.9Hz,1H),8.11(s,1H),7.97(s,1H),7.21(dd,J=7.7,4.8Hz,1H),7.17(s,1H),4.46(t,J=6 .6Hz,2H),4.12(q,J=7.0Hz,2H),3.93(s,2H),3.80–3.65(m,1H),2.95(t,J=7.2Hz,2H),2.92(s,2H) ,2.89–2.81(m,1H),2.29–2.11(m,6H),1.63–1.49(m,2H),1.40–1.29(m,8H),1.27(t,J=7.0Hz,3H).

[0313] Example 73 Synthesis of Protein Degrader Y85

[0314] The synthesis method was similar to that of Example 70, except that KI-ARV-03 was replaced with compound 627 to obtain compound Y85 (white solid, yield 57.3%). 1 H NMR (400MHz, DMSO-d6) δ9.17(s,1H),8.74(dd,J=4.8,1.9Hz,1H),8.40(dd,J=7.7,1.9Hz,1H),8.27(s,1H) ,7.82(d,J=7.6Hz,1H),7.52(s,1H),7.44–7.32(m,2H),7.22(dd,J=8.4,6.7Hz,1H),7.08(dd,J=11.5,2.4 Hz,1H),6.89(td,J=8.4,2.5Hz,1H),4.45(t,J=7.6Hz,2H),3.98(q,J=7.0Hz,2H),3.77(s,3H),3.58–3.47 (m,1H),3.46–3.36(m,1H),2.45(t,J=7.7Hz,2H),1.95–1.84(m,2H),1.80–1.66(m,2H),1.28–1.11(m,7H).

[0315] Example 74 Synthesis of Protein Degrader Y86

[0316] The synthesis method was as described in Example 46, except that C24a was replaced with compound 627, and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain compound Y86 (white solid, yield 20.2%). 1 H NMR(400MHz,Chloroform-d)δ8.89(s,1H),8.67(dd,J=4.9,1.9Hz,1H),8.50(dd,J=7.7,1.9Hz,1H),8.21(s,1H),7 .68(s,1H),7.25(dd,J=7.8,4.9Hz,1H),7.13(dd,J=8.3,6.6Hz,1H),6.80–6.71(m,3H),4.49(t,J=6.6Hz,2H),4.16 (q,J=7.0Hz,2H),3.84–3.68(m,4H),2.85(t,J=6.7Hz,2H),2.79–2.68(m,1H),2.22–2.09(m,6H),1.46–1.34(m,4H),1.30(t,J=7.4Hz,3H).

[0317] Example 75 Synthesis of Protein Degrader Y87

[0318] first step

[0319] 2-Aminonicotinic acid (2.0 g, 14.48 mmol) was dissolved in 36 mL of DCM, HATU (7.2 g, 18.82 mmol) and DIPEA (5.0 mL, 180.99 mmol) were added, and 4-methylaminopyridine (1.9 g, 17.38 mmol) was added after 15 minutes. The mixture was stirred at room temperature for 16 hours, and the reaction was complete after TLC monitoring. Y87a was obtained, which was spin-dried and used in the next step.

[0320] Step 2

[0321] The product from the previous step was placed in a single-necked flask and added with 36 mL of 1,4-dioxane, followed by CDI (2.4 g, 14.50 mmol) and DIPEA (2.5 mL, 14.50 mmol). The mixture was refluxed at 110°C for 16 hours, and the reaction was monitored for completion by TLC. After cooling to room temperature, a solid precipitated. The solid was filtered, and the filter cake was washed with DCM and dried to give crude Y87b (white solid, 4.4 g). 1 H NMR (400MHz, DMSO-d6) δ12.14(s,1H),8.66(dd,J=4.8,1.8Hz,1H),8.49(d,2H),8.33(dd,J=7.8,1.9Hz,1H),7.33–7.29(m,3H),5.10(s,2H).

[0322] Step 3

[0323] Y87b (1.0 g, 3.90 mmol) was dissolved in 20 mL of DMF, and KCO (1.1 g, 7.90 mmol) and 2,2'-dibromodiethyl ether (3.0 mL, 23.60 mmol) were added sequentially. The reaction was allowed to react at room temperature for approximately 5 hours. After the reaction was complete, water and ethyl acetate were added. The ethyl acetate layer was washed three times with water, then with saturated brine, and dried over anhydrous sodium sulfate. Silica gel column chromatography afforded compound Y87c (oily liquid, 67.0 mg, yield 10.6%). 1 H NMR(400MHz, DMSO-d6)δ8.78(dd,J=4.8,1.9Hz,1H),8.50–8.48(m,2H),8.43(dd,J=7.7,1.9Hz,1H),7.40(dd,J= 7.8, 4.8Hz, 1H), 7.33–7.30 (m, 2H), 5.16 (s, 2H), 4.47 (t, J = 6.3Hz, 2H), 3.78–3.72 (m, 4H), 3.49 (t, J = 5.6Hz, 2H).

[0324] Step 4

[0325] Y87c (136.0 mg, 0.34 mmol) was dissolved in 2 mL of DMF, and C24a (168.0 mg, 0.34 mmol) and KCO (93.0 mg, 0.67 mmol) were added. The mixture was reacted at 60°C for approximately 4 hours. After completion of the reaction as monitored by TLC, the protein degrader Y87 was isolated and purified by silica gel column chromatography (white solid, 7.0 mg, 2.6% yield). 1 H NMR(400MHz,Chloroform-d)δ12.30(s,1H),8.67(dd,J=4.8,1.9Hz,1H),8.56–8.52(m,2H),8.48(td,J=3.6,1.9Hz,2H), 8.40(s,1H),7.95(t,J=5.8Hz,1H),7.76(dd,J=8.8,2.6Hz,1H),7.71(s,1H),7.58(s,1H) ,7.36–7.33(m,2H),7.23(dd,J=7.8,4.8Hz,1H),6.65(d,J=8.9Hz,1H),5.91(s,1H),5.24( s,2H),4.87(p,J=6.6Hz,1H),4.67–4.60(m,4H),3.82(t,J=5.8Hz,2H),3.69(t,J=5.6Hz,2 H),3.58–3.51(m,4H),2.60–2.50(m,6H),2.41(s,3H),2.15(s,3H),1.58(d,J=6.6Hz,6H).

[0326] Example 76 Synthesis of Protein Degrader Y88

[0327] The synthesis method was as described in Example 75, except that 2-aminonicotinic acid was replaced with 4-aminopyrimidine-5-carboxylic acid, and 4-methylaminopyridine was replaced with ethylamine hydrochloride to obtain compound Y88 (white solid, yield 36.7%). 1H NMR(400MHz,Chloroform-d)δ9.28(s,1H),9.11(s,1H),8.47(d,J=2.6Hz,1H),8.40(s,1H),8.02(t,J=5.8Hz,1H), 7.76(dd,J=8.9,2.6Hz,1H),7.73(s,1H),7.58(s,1H),6.65(d,J=8.9Hz,1H),5.93(s,1H),4.88(p,J=6.6Hz,1H),4 .64(d,J=5.8Hz,2H),4.56(t,J=5.7Hz,2H),4.12(q,J=7.1Hz,2H),3.82(t,J=5.7Hz,2H),3.69(t,J=5.5Hz,2H),3. 55(t,J=5.0Hz,4H),2.59–2.53(m,6H),2.41(s,3H),2.15(s,3H),1.59(s,3H),1.57(s,3H),1.28(t,J=7.0Hz,3H).

[0328] Example 77 Synthesis of Protein Degrader Y89

[0329] The synthesis method was as shown in Example 75, except that 2-aminonicotinic acid was replaced with 4-aminopyrimidine-5-carboxylic acid and 4-methylaminopyridine was replaced with benzylamine to obtain compound Y89 (white solid, yield 28.5%). 1 H NMR(400MHz,Chloroform-d)δ9.31(s,1H),9.12(s,1H),8.49(d,J=2.5Hz,1H),8.41(s,1H),8.01(t,J=5.8Hz,1H),7.78(d d,J=8.9,2.6Hz,1H),7.74(d,J=1.3Hz,1H),7.60(s,1H),7.54–7.51(m,1H),7.51–7.50(m,1H),7.35–7.29(m,3H),6.66(d, J=8.8Hz,1H),5.94(s,1H),5.24(s,2H),4.89(p,J=6.7Hz,1H),4.66(d,J=5.8Hz,2H),4.58(t,J=5.7Hz,2H),3.82(t,J=5. 7Hz,2H),3.68(t,J=5.6Hz,2H),3.57–3.52(m,4H),2.56–2.50(m,6H),2.43(s,3H),2.16(s,3H),1.61(s,3H),1.59(s,3H).

[0330] Example 78 Synthesis of Protein Degrader Y90

[0331] The synthesis method was similar to that of Example 46, except that ethylene glycol was replaced with diethylene glycol and C24a was replaced with EPZ6438a to obtain compound Y90 (white solid, yield 60.1%). 1 H NMR(400MHz,Chloroform-d)δ8.62(dd,J=4.8,1.9Hz,1H),8.44(dd,J=7.8,2.0Hz,1H),7.48–7.42(m,2H),7.32(d,J=7.9Hz,3H),7 .29(d,J=1.8Hz,1H),7.22–7.15(m,2H),5.91(s,1H),4.60(t,J=5.9Hz,2H),4.56(d,J=5.8Hz,2H),4.13(q,J=7.1Hz,2H),3.98–3. 92(m,2H),3.80(t,J=5.9Hz,2H),3.66(t,J=5.6Hz,2H),3.52(s,2H),3.32(td,J=11.3,3.1Hz,2H),3.10(q,J=7.0Hz,2H),3.06–2. 98(m,1H),2.57–2.46(m,8H),2.40(s,3H),2.35(s,3H),2.12(s,3H),1.76–1.63(m,4H),1.34–1.20(m,5H),0.90(t,J=6.9Hz,3H).

[0332] Example 79 Synthesis of Protein Degrader Y91

[0333] The synthesis method was as described in Example 46, except that ethylene glycol was replaced with 1,5-pentanediol and C24a was replaced with EPZ6438a to obtain compound Y91 (white solid, yield 60.0%). 1H NMR(400MHz,Chloroform-d)δ12.57(s,1H),8.63(dd,J=4.8,2.0Hz,1H),8.44(dd,J=7.7,2.0Hz,1H),7.46–7.40(m,2H),7.33–7.2 9(m,3H),7.28(d,1H),7.22(t,J=5.8Hz,1H),7.18(dd,J=7.7,4.8Hz,1H),5.89(s,1H),4.54(d,J=5.8Hz,2H),4.36–4.28(m,2H),4. 17–4.11(m,2H),3.96–3.90(m,2H),3.52(s,2H),3.35–3.26(m,2H),3.08(q,J=7.0Hz,2H),3.04–2.95(m,1H),2.54(s,6H),2.46–2 .39(m,2H),2.38(s,3H),2.33(s,3H),2.09(s,3H),1.78–1.55(m,8H),1.45–1.36(m,2H),1.30–1.23(m,5H),0.88(t,J=6.9Hz,3H).

[0334] Example 80 Synthesis of Protein Degrader Y92

[0335] The synthesis method was as described in Example 46, except that ethylene glycol was replaced with 1,4-butanediol and C24a was replaced with EPZ6438a to obtain compound Y92 (white solid, yield 63.6%). 1H NMR(400MHz,Chloroform-d)δ12.66(s,1H),8.62(dd,J=4.7,1.9Hz,1H),8.44(dd,J=7.8,2.0Hz,1H),7.42(d,J=8.2Hz,2H),7.33–7.29( m,3H),7.28–7.26(m,1H),7.21(t,J=5.8Hz,1H),7.17(dd,J=7.7,4.8Hz,1H),5.89(s,1H),4.55(d,J=5.8Hz,2H),4.38–4.31(m,2H),4.1 2(t,J=6.9Hz,2H),3.97–3.90(m,2H),3.50(s,2H),3.30(td,J=11.3,3.0Hz,2H),3.09(q,J=7.0Hz,2H),3.04–2.96(m,1H),2.49(s,6H), 2.41(d,J=7.7Hz,2H),2.38(s,3H),2.33(s,3H),2.09(s,3H),1.78–1.55(m,8H),1.28–1.26(m,2H),1.25(s,3H),0.88(t,J=6.9Hz,3H).

[0336] Example 81 Synthesis of Protein Degrader Y93

[0337] The synthesis method was as described in Example 46, except that ethylene glycol was replaced with 1,3-propylene glycol and C24a was replaced with EPZ6438a to obtain compound Y93 (white solid, yield 59.2%). 1H NMR(400MHz,Chloroform-d)δ8.62(dd,J=4.7,2.0Hz,1H),8.45(dd,J=7.7,2.0Hz,1H),7.44(d,J=8.0Hz,2H),7.36–7.31(m, 3H),7.29(d,J=2.1Hz,1H),7.20–7.16(m,2H),5.90(s,1H),4.56(d,J=5.8Hz,2H),4.41(t,J=7.3Hz,2H),4.14(q,J=7.1Hz,2 H),3.98–3.92(m,2H),3.53(s,2H),3.32(td,J=11.2,3.0Hz,2H),3.10(q,J=7.0Hz,2H),3.05–2.97(m,1H),2.60–2.45(m,8H ),2.40(s,3H),2.35(s,3H),2.11(s,3H),1.97(p,J=7.2Hz,2H),1.75–1.64(m,4H),1.32–1.24(m,5H),0.89(t,J=6.9Hz,3H).

[0338] Example 82 Synthesis of Protein Degrader Y94

[0339] The synthesis method was similar to that of Example 46, except that C24a was replaced with EPZ6438a to give compound Y94 (white solid, yield 71.9%). 1H NMR(400MHz,Chloroform-d)δ12.66(s,1H),8.61(dd,J=4.8,2.0Hz,1H),8.44(dd,J=7.8,2.0Hz,1H),7.43(d,J=8.2Hz,2H),7.34–7.30( m,3H),7.28(d,1H),7.23(t,J=5.8Hz,1H),7.18(dd,J=7.7,4.8Hz,1H),5.90(s,1H),4.55(d,J=5.8Hz,2H),4.50(t,J=7.0Hz,2H),4.13( q,J=7.0Hz,2H),3.96–3.90(m,2H),3.51(s,2H),3.31(td,J=11.3,3.0Hz,2H),3.09(q,J=7.0Hz,2H),3.04–2.96(m,1H),2.70(t,J=7.0H z,2H),2.63(s,4H),2.47(s,4H),2.39(s,3H),2.34(s,3H),2.09(s,3H),1.72–1.60(m,4H),1.28(t,J=7.0Hz,3H),0.89(t,J=7.0Hz,3H).

[0340] Example 83 Synthesis of Protein Degrader Y95

[0341] Y46a (300.0 mg, 1.57 mmol), methyl 4-hydroxycyclohexanecarboxylate (496.5 mg, 3.14 mmol), and triphenylphosphine (823.1 mg, 3.14 mmol) were dissolved in 4 mL of tetrahydrofuran under nitrogen protection. DIAD (0.6 mL, 3.14 mmol) was added dropwise at 0°C. After the addition was complete, the mixture was allowed to stand at room temperature overnight. The tetrahydrofuran was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography to give Y95a (white solid, 455.4 mg, yield 87.6%).

[0342] Y95a (455.4 mg, 1.37 mmol) was dissolved in 2.5 mL of methanol and 2.5 mL of water, and lithium hydroxide (173.0 mg, 4.12 mmol) was added. The mixture was stirred overnight at room temperature, and the organic solvent was removed by rotary evaporation. Water was added to dissolve the mixture, and the pH was adjusted to 4-5 with 1N hydrochloric acid. A white solid precipitated and was filtered to obtain Y95b (white solid, 315.6 mg, yield 72.4%). 1H NMR(400MHz,DMSO-d6)δ12.19(s,1H),8.83–8.65(m,1H),8.52–8.24(m,1H),7.48–7.24(m,1H),5.48–5.1 7(m,1H),4.05–3.85(m,2H),2.69–2.54(m,2H),2.27–1.99(m,2H),1.80–1.41(m,4H),1.21–1.08(m,3H).

[0343] Y95b (50.0 mg, 0.16 mmol), compound 754 (63.5 mg, 0.16 mmol), HATU (71.9 mg, 0.19 mmol) and DIPEA (0.04 mL, 0.24 mmol) were dissolved in 1 mL of dichloromethane, stirred at room temperature overnight, and purified by silica gel column chromatography to give Y95 (white solid, 57.0 mg, yield 51.5%). 1 H NMR(400MHz,Chloroform-d)δ8.71–8.57(m,2H),8.51–8.43(m,1H),8.34–8.14(m,2H ),7.97(s,1H),7.25–7.14(m,1H),7.04–6.90(m,1H),5.79–5.51(m,1H),4.19–4.09(m ,2H),4.05–3.94(m,3H),3.87–3.67(m,1H),3.25–3.12(m,1H),3.01–2.81(m,3H),2.2 2–2.12(m,2H),1.88–1.60(m,6H),1.53–1.37(m,8H),1.35(s,6H),1.32–1.26(m,3H).

[0344] Example 84 Synthesis of Protein Degrader Y96

[0345] The synthesis method was as follows: Referring to Example 83, 4-hydroxycyclohexanecarboxylic acid methyl ester was replaced with cis-3-hydroxycyclobutylcarboxylic acid methyl ester to obtain compound Y96 (white solid, yield 72.5%). 1H NMR (400MHz, DMSO-d6) δ9.09(s,1H),8.74(d,J=4.7Hz,1H),8.39(d,J=7.8Hz,1H),8.23(s ,1H),7.97(s,1H),7.76(d,J=7.7Hz,1H),7.67(s,1H),7.53–7.44(m,1H),7.41–7.30(m,1 H),6.23–6.09(m,1H),4.04–3.89(m,4H),3.67–3.44(m,2H),3.21–3.05(m,3H),2.88(s,2 H),2.36(t,J=10.1Hz,2H),2.00–1.78(m,4H),1.34–1.22(m,10H),1.18(t,J=7.0Hz,3H).

[0346] Example 85 Synthesis of Protein Degrader Y97

[0347] The synthesis method was as follows: Referring to Example 83, 4-hydroxycyclohexanecarboxylic acid methyl ester was replaced with trans-3-hydroxycyclobutylcarboxylic acid methyl ester to obtain compound Y97 (white solid, yield 63.7%). 1 H NMR(400MHz,DMSO-d6)δ9.08(s,1H),8.69(dd,J=4.7,1.9Hz,1H),8.38(dd,J=7.7,2.0Hz,1 H),8.22(s,1H),7.96(s,1H),7.67(s,1H),7.63(d,J=7.8Hz,1H),7.45(d,J=7.6Hz,1H),7.3 4(dd,J=7.8,4.8Hz,1H),5.39–5.25(m,1H),4.01–3.90(m,4H),3.64–3.42(m,2H),3.01–2. 84(m,4H),2.79–2.66(m,1H),2.00–1.73(m,4H),1.33–1.22(m,10H),1.17(t,J=7.0Hz,3H).

[0348] Example 86 Synthesis of Protein Degrader Y98

[0349] The synthesis method was as described in Example 83, except that Y46a was replaced with Y15a, and 4-hydroxycyclohexanecarboxylic acid methyl ester was replaced with cis-3-hydroxycyclobutylcarboxylic acid methyl ester to give compound Y98 (white solid, yield 82.9%). 1H NMR (400MHz, DMSO-d6) δ9.09(s,1H),8.24(s,1H),8.03(d,J=8.5Hz,1H),7.97(s,1H),7.83(d,J=7 .7Hz,1H),7.67(s,1H),7.52(d,J=2.0Hz,1H),7.47(d,J=7.7Hz,1H),7.39(dd,J=8.4,2.0Hz,1H),5 .53–5.43(m,1H),4.09–4.00(m,2H),3.95(s,2H),3.70–3.44(m,3H),3.20–2.97(m,2H),2.89(s,2H ),2.62–2.53(m,2H),2.46–2.36(m,2H),1.98–1.89(m,2H),1.87–1.80(m,2H),1.32–1.18(m,11H).

[0350] Example 87 Synthesis of Protein Degrader Y99

[0351] The synthesis method was as described in Example 83, except that Y46a was replaced with Y37a, and 4-hydroxycyclohexanecarboxylic acid methyl ester was replaced with cis-3-hydroxycyclobutylcarboxylic acid methyl ester to obtain compound Y99 (white solid, yield 76.1%). 1 H NMR(400MHz,DMSO-d6)δ9.09(s,1H),8.57–8.50(m,2H),8.23(s,1H),8.07(d,J=8.5Hz,1H),7.97(s,1H ),7.78(d,J=7.8Hz,1H),7.67(s,1H),7.57(d,J=2.1Hz,1H),7.47(d,J=8.2Hz,1H),7.43(dd,J=8.5,2.1 Hz,1H),7.32–7.27(m,2H),5.49–5.38(m,1H),5.23(s,2H),3.94(s,2H),3.66–3.42(m,2H),2.96–2.82( m,3H),2.50–2.45(m,2H),2.29–2.18(m,2H),1.97–1.88(m,2H),1.85–1.76(m,2H),1.33–1.17(m,10H).

[0352] Example 88 Synthesis of Protein Degrader Y100

[0353] The synthesis method was similar to that of Example 46, except that compound C24a was replaced by compound 754, 1,2-ethylene glycol was replaced by 4-hydroxycyclohexanone, and the second step of Example 46, i.e., the step of converting the hydroxyl group to an aldehyde group, was omitted to obtain compound Y100 (white solid, yield 6.1%). 1 H NMR(400MHz,Chloroform-d)δ8.69–8.57(m,1H),8.52–8.41(m,1H),8.15(s,1H),8.06 (s,1H),7.49–7.33(m,1H),7.24–7.15(m,1H),5.79–5.36(m,1H),4.17–4.07(m,2H),3 .96(s,2H),3.88–3.74(m,1H),3.31–3.14(m,1H),3.03(s,2H),2.87–2.69(m,1H),2.5 0–2.17(m,6H),2.06–1.62(m,8H),1.49–1.39(m,2H),1.36(s,6H),1.32–1.24(m,3H).

[0354] Example 89 Synthesis of Protein Degrader Y101

[0355] The synthesis method was similar to that of Example 46, except that C24a was replaced with compound 754, 1,2-ethylene glycol was replaced with 4-hydroxymethylcyclohexanone, and the second step of Example 46, i.e., the step of converting the hydroxyl group to an aldehyde group, was omitted to obtain compound Y101 (white solid, yield 34.8%). 1 H NMR (400MHz, DMSO-d6) δ9.12(d,J=2.6Hz,1H),8.73(d,J=4.8Hz,1H),8.41(dd,J=7.7,1.9Hz,1H),8.21(d ,J=2.1Hz,1H),7.97(s,1H),7.70(d,J=2.6Hz,1H),7.54–7.46(m,1H),7.40–7.31(m,1H),4.28(d,J=7.2Hz ,1H),4.14(d,J=7.1Hz,1H),4.05–3.96(m,2H),3.96(s,2H),3.54–3.42(m,2H),3.18–2.98(m,2H),2.88(s ,2H),2.19–1.67(m,10H),1.65–1.55(m,1H),1.52–1.36(m,3H),1.34–1.22(m,8H),1.17(t,J=7.0Hz,3H).

[0356] Example 90 Synthesis of Protein Degrader Y102

[0357] The synthesis method was as described in Example 46, except that C24a was replaced with compound 754, 1,2-ethylene glycol was replaced with 3-(hydroxymethyl)cyclobutan-1-one, and the second step of Example 46, i.e., the step of converting the hydroxyl group to an aldehyde group, was omitted to obtain compound Y102 (white solid, yield 19.3%). 1 H NMR(400MHz,Chloroform-d)δ8.95–8.72(m,1H),8.67–8.59(m,1H),8.52–8.43(m,1H),8.16(d,J=3 .5Hz,1H),8.00(d,J=4.4Hz,1H),7.25–7.14(m,1H),7.09–6.90(m,1H),4.54(d,J=8.0Hz,1H),4.45 (d,J=6.3Hz,1H),4.21–4.09(m,2H),3.97(s,2H),3.81–3.67(m,1H),3.40–3.24(m,1H),2.93(s,2H ),2.83–2.63(m,1H),2.55–2.39(m,2H),2.30–2.07(m,7H),1.71–1.43(m,3H),1.41–1.22(m,12H).

[0358] Example 91 Synthesis of Protein Degrader Y103

[0359] The synthesis method was as described in Example 46, except that C24a was replaced with compound 754 and 1,2-ethylene glycol was replaced with diethylene glycol to obtain compound Y103 (white solid, yield 43.7%). 1 H NMR(400MHz,Chloroform-d)δ8.95(s,1H),8.76–8.66(m,1H),8.63(dd,J=4.8,1.9Hz,1H),8.46( dd,J=7.7,1.9Hz,1H),8.13(s,1H),7.96(s,1H),7.21(dd,J=7.7,4.8Hz,1H),7.11(s,1H),4.59( t,J=5.8Hz,2H),4.13(q,J=7.0Hz,2H),3.94(s,2H),3.83(t,J=5.8Hz,2H),3.76(t,J=5.0Hz,2H) ,3.73–3.67(m,1H),2.95–2.88(m,4H),2.75–2.65(m,1H),2.19–2.02(m,4H),1.48–1.21(m,13H).

[0360] Example 92 Synthesis of Protein Degrader Y104

[0361] The synthesis method was similar to that of Example 46, except that C24a was replaced with compound 754 to obtain compound Y104 (white solid, yield 68.6%). 1 H NMR(400MHz,DMSO-d6)δ9.13(s,1H),8.75(d,J=4.6Hz,1H),8.45(d,J=7.7Hz,1H),8 .22(s,1H),7.96(s,1H),7.66(s,1H),7.54(s,1H),7.45–7.34(m,1H),4.55–4.40(m, 2H),4.04–3.96(m,2H),3.94(s,2H),3.20–2.95(m,4H),2.88(s,2H),2.07–1.88(m,4H),1.34–1.12(m,13H).

[0362] Example 93 Synthesis of Protein Degrader Y105

[0363] first step

[0364] Y46a (1.0 g, 5.23 mmol), cis-4-(Boc-amino)cyclohexanol (1.4 g, 6.28 mmol), and triphenylphosphine (2.7 g, 10.46 mmol) were dissolved in 13 mL of anhydrous tetrahydrofuran. DIAD (2 mL, 10.46 mmol) was added dropwise under ice-bath. After the addition was complete, the mixture was allowed to stand at room temperature overnight. The organic solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography to give crude Y105a (white solid, 2.7 g). 1 H NMR (400MHz, DMSO-d6) δ8.75 (dd, J=4.7, 2.0Hz, 1H), 8.40 (dd, J=7.7, 1.9Hz, 1 H),7.35(dd,J=7.7,4.7Hz,1H),6.79(d,J=8.1Hz,1H),5.37–5.21(m,1H),4.9 3–4.85(m,1H),3.96(q,J=7.0Hz,2H),2.64–2.53(m,2H),1.89(d,J=12.5Hz,2 H),1.66(d,J=12.3Hz,2H),1.40(s,9H),1.36–1.29(m,2H),1.24–1.21(m,3H).

[0365] Step 2

[0366] Y105a (2.7 g, 7.04 mmol) was dissolved in 18 mL of dichloromethane, and 8.8 mL of 4 M HCl in ethyl acetate was added. The mixture was stirred at room temperature overnight to obtain Y105b (white solid, 428.3 mg). 1 H NMR (400MHz, DMSO-d6) δ8.74(dd,J=4.7,2.0Hz,1H),8.40(dd,J=7.7,1.9Hz,1H),7.35(dd,J=7.7,4.7Hz,1H),5.41–5.22(m,1 H),3.96(q,J=7.0Hz,2H),2.69–2.53(m,3H),1.95–1.80(m,2H),1.71–1.55(m,2H),1.24–1.18(m,2H),1.16(t,J=7.1Hz,3H).

[0367] Step 3

[0368] 5-Chloro-4-iodopyridin-2-amine (375.0 mg, 1.47 mmol) was dissolved in anhydrous dichloromethane. A dichloromethane solution of triphosgene (148.7 mg, 0.50 mmol) was added dropwise under an ice bath. Triethylamine (0.4 mL, 2.95 mmol) was then added dropwise. The reaction was allowed to react at room temperature until the starting material was completely consumed. The solvent was then dried, and toluene, Y105b (425.0 mg, 1.47 mmol), and triethylamine (0.4 mL, 2.95 mmol) were added. The reaction was continued at 100°C for 4 hours. Water was added, the mixture was extracted with ethyl acetate, dried, and purified by silica gel column chromatography to afford Y105c (white solid, 550.0 mg, 65.6% yield). 1 H NMR(400MHz,DMSO-d6)δ10.15(s,1H),9.18(s,1H),8.76(dd,J=4.7,2.0Hz,1H), 8.41(dd,J=7.7,2.0Hz,1H),8.31(s,1H),8.26(s,1H),7.36(dd,J=7.7,4.8Hz,1 H),5.49–5.26(m,1H),3.97(q,J=6.9Hz,2H),3.62–3.48(m,1H),2.74–2.57(m,2 H),2.09–1.99(m,2H),1.76–1.67(m,2H),1.47–1.30(m,2H),1.25–1.10(m,3H).

[0369] Step 4

[0370] Y105c (200.0 mg, 0.35 mmol), 5,5-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydro-4H-pyrrolo[1,2-b]pyrazole (110.6 mg, 0.42 mmol), Pd(dppf)Cl2 (14.4 mg, 0.02 mmol), and potassium carbonate (721.5 mg, 0.88 mmol) were dissolved in 2 mL of dioxane and 2 mL of water, and stirred at 60°C overnight. After completion of the reaction, water was added to the reaction system, and the mixture was extracted with ethyl acetate and purified by silica gel column chromatography to obtain Y105 (white solid, 46.7 mg, yield 23.0%). 1 H NMR (400MHz, DMSO-d6) δ9.12(s,1H),8.76(dd,J=4.8,2.0Hz,1H),8.41(dd,J=7.7,2.0Hz,1H ),8.24(s,1H),7.98(s,1H),7.70(s,1H),7.51(d,J=7.5Hz,1H),7.36(dd,J=7.7,4.7Hz,1H) ,5.45–5.26(m,1H),4.04–3.90(m,4H),3.67–3.54(m,1H),2.90(s,2H),2.76–2.61(m,2H),2 .08–2.01(m,2H),1.77–1.67(m,2H),1.46–1.33(m,2H),1.28(s,6H),1.17(t,J=7.0Hz,3H).

[0371] Example 94 Synthesis of Protein Degrader Y106

[0372] The synthesis method was as described in Example 46, except that C24a was replaced with compound 754a, and 1,2-ethylene glycol was replaced with 1,3-propylene glycol to obtain compound Y106 (white solid, yield 35.7%). 1H NMR(400MHz,Chloroform-d)δ8.83(s,1H),8.65(dd,J=4.8,1.9Hz,1H),8.46(dd,J=7.7, 1.9Hz,1H),8.24(s,1H),8.02(s,1H),7.21(dd,J=7.7,4.8Hz,1H),4.49(t,J=6.5Hz,2H), 4.17–4.07(m,3H),3.96(s,2H),3.00–2.91(m,4H),2.90–2.83(m,1H),2.31–2.21(m,2H), 2.01–1.90(m,2H),1.84–1.71(m,2H),1.69–1.54(m,2H),1.34(s,6H),1.29–1.25(m,5H).

[0373] Biological activity test examples

[0374] Test Example 1: Cell IC 50 Determination

[0375] In each well of a 96-well plate, 10,000 WSU-DLCL2 cells (human diffuse large B-cell lymphoma cells) were inoculated with 100 μl of fresh culture medium containing different concentrations (the highest concentration was 10 μM, 3-fold dilution, a total of 8 gradient concentrations) of the compound of the present invention or the positive control drug. After incubation of the cells with the compound for 72 hours, 10 μL of Cell Counting Kit-8 (CCK8, purchased from Li Ji Biotechnology) was added to each well. After incubation at 37°C for 2 hours, the absorbance of each well was measured at 450 nm using SpectraMAX190 (Molecular Devices), and the IC was calculated using SoftMax Pro. 50 The results are shown in Table 1.

[0376] The positive control compounds used were SNS-032 and THAL-SNS-032, and their structures are shown below.

[0377] Table 1 Inhibitory activity of compounds on WSU-DLCL2 cells

[0378] Test Example 2: Western Blot Experiment

[0379] CDK can combine with cyclin (cell cycle protein) to form a heterodimer, in which CDK is the catalytic subunit and cyclin is the regulatory subunit. Different cyclin-CDK complexes catalyze the phosphorylation of different substrates through CDK activity, thereby achieving the promotion and transformation of different phases of the cell cycle.

[0380] Polycomb repressive complex 2 (PRC2) is a histone methyltransferase complex that includes three core subunits: EZH2 (or EZH1), EED, and SUZ12. Dysregulation of PRC2 is associated with the development, progression, and poor prognosis of hematological and solid tumors. EZH2 is generally considered to be the main catalytic subunit of the PRC2 complex, catalyzing the trimethylation modification of histone H3 lysine 27 (H3K27me3) through its SET domain, thereby maintaining the silencing of downstream target genes. EZH2 is overexpressed or has gain-of-function mutations in various cancers, and the role of EZH2 inhibitors in cancer treatment has been clinically validated.

[0381] The above proteins and Ras proteins can all be used as targets for tumor therapy. Therefore, this test case studied the degradation effects of the degraders of the present invention on CDK9, CDK2, CDK2 / 4 / 6, CDK5, CDK7, EZH2, EED, SUZ12, EZH1, Ras, and their related proteins. The specific method is as follows:

[0382] Appropriate numbers of WSU-DLCL2, U-2932, or GP2D cells (human colon cancer cells) were plated in 6-well plates and treated with drugs (see Figures 1-7 for drug and concentration). Cells were then harvested by centrifugation for subsequent experiments. Depending on the cell population, the appropriate amount of 1X loading buffer (50 mM Tris-HCl (pH 6.8), 2% (w / v) SDS, 0.1% (w / v) BPB (bromophenol blue), 10% (v / v) glycerol, 0.1 M β-mercaptoethanol) was added. After lysis, the samples were boiled at 100°C for 20 min. Equal volumes of sample were aspirated and subjected to SDS-PAGE electrophoresis. Following electrophoresis, proteins were transferred to nitrocellulose membranes using a GenScript Express transfer system. Bands were excised based on protein size, blocked with TBST containing 5% nonfat dry milk for 1 h, and incubated with primary antibodies overnight at 4°C. Excess primary antibody was washed off with TBST for 10 minutes three times, followed by incubation with secondary antibody at room temperature for 1 hour, followed by washing with TBST for 10 minutes three times. Finally, the bands were stained and photographed using a Bio-Rad chromatometer.

[0383] The results are shown in Figure 1, Figure 2B, Figure 3, Figure 4, Figure 7 (WSU-DLCL2 cells), and Figure 5A (WSU-DLCL2 cells) and B (U-2932 cells), and Figure 6 (GP2D cells).

[0384] CDK9 is primarily involved in transcriptional regulation. Heterodimers composed of CDK9 and cyclins (T1, T2a, T2b, and K) form the positive transcription elongation factor (P-TEFb). CDK9 has two isoforms (CDK9 42 and CDK9 55), with approximately 80% of CDK9 binding to cyclin T1. Figure 1A shows that compounds Y44, Y45, Y41, Y35, and Y33 can degrade both CDK9 isoforms, CDK9 55 and CDK9 42, at a concentration of 100 nM. They can also degrade cyclin T1 and downregulate levels of the CDK9 downstream protein Mcl-1. The CDK9 inhibitor SNS-032, however, has no protein degradation activity. The positive control, THAL-SNS-032, can degrade CDK9 but cannot degrade cyclin T1 or downregulate Mcl-1 levels. Figure 1B shows that at a concentration of 1 μM, compounds Y1-Y5 can effectively degrade CDK9 and cyclin T1 and downregulate Mcl-1 levels.

[0385] Figure 2B shows that at a concentration of 100 nM, compound Y35 can effectively induce the degradation of CDK9 and cyclin T1 and downregulate the level of CDK9 downstream protein Mcl-1; when cells are co-treated with Y35 and the autophagy inhibitor bafilomycin A1 (BafA1), the degradation effect of Y35 on CDK9 and cyclin T1 disappears.

[0386] Figure 3 shows that at a concentration of 100 nM, compounds Y44, Y82-Y86 can degrade CDK9, among which Y44, Y83 and Y84 can also degrade cyclin T1 while degrading CDK9, thereby downregulating the level of Mcl-1.

[0387] Figure 4A shows that the degrader Y53 can effectively degrade CDK2 compared with the CDK2 inhibitor SY-5609a.

[0388] Figure 4B shows that compound Y67 can degrade the levels of CDK7 and cyclin H at a concentration of 10 μM. Figure 4C shows that compound Y62 can degrade CDK2 and CDK6 and their corresponding cyclin A2 / E1 / D1 at a concentration of 10 μM. Figure 4D shows that compounds Y59-Y61 can degrade CDK2 / 4 / 6 and their corresponding cyclin A2 / E1 / D1 at a concentration of 10 μM.

[0389] Figure 5A shows that compounds Y47 and Y50 can degrade EZH2, EED, SUZ12, and EZH1, with greater efficacy than the positive control MS177 (CAS No. 2225938-86-1, a small molecule EZH2 degrader). The EZH2 inhibitor C24 showed no degradation. Figure 5B shows that at a concentration of 10 μM, Y50 and Y87-Y89 can effectively degrade EZH2, EED, SUZ12, and EZH1, with greater efficacy than the positive control MS177.

[0390] FIG6 shows that compound Y69 can degrade Ras at a concentration of 10 μM and downregulate the level of pERK.

[0391] Figure 7A shows that at a concentration of 100 nM, compounds Y83, Y95, and Y96 can effectively induce the degradation of multiple proteins including CDK2, cyclin A2, cyclin E1, CDK5, CDK6, cyclin D1, CDK7, CDK9, and cyclin T1. Figure 7B shows that at a concentration of 100 nM, compounds Y84, Y100-Y106 can effectively induce the degradation of multiple proteins including CDK2, cyclin A2, cyclin E1, CDK4, CDK5, CDK6, cyclin D1, CDK7, CDK9, and cyclin T1.

[0392] Test Example 3: Co-Immunoprecipitates Experiment

[0393] After treatment with the compounds, WSU-DLCL2 cells were lysed on ice for 1 hour with NP-40 (purchased from Beyotime, Catalog No.: P0013F) supplemented with phosphatase inhibitors (purchased from Roche, Catalog No.: 4906845001) and protease inhibitor cocktail (purchased from Roche, Catalog No.: 04693132001) and centrifuged at 12,000 g for 10 minutes at 4°C. The protein concentration in the supernatant was measured using a BCA protein assay kit (Thermo Scientific, 23225), and equal amounts of protein were incubated with primary antibodies (Normal Rabbit IgG (purchased from Cell Signaling Technology, Catalog No.: 2729), CDK9 rabbit mAb (purchased from Abclonal, Catalog No.: A11145) at 4°C overnight, followed by the addition of protein A / G magnetic beads (purchased from Thermo Scientific, Catalog No.: 88803) and incubation at 4°C for another 4 hours. The immunoprecipitate was washed three times with NP-40 and PBS, then boiled with SDS-PAGE loading buffer and subjected to immunoblotting.

[0394] The results are shown in Figure 2 A. At a concentration of 1 μM, Y2 and Y3 can increase the interaction between CDK9 and LC3B.

[0395] Figure 2 A and B together demonstrate the LC3B-dependent autophagy-lysosomal degradation pathway of this class of compounds.

Claims

1. A protein degradation agent represented by formula (1) or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, LCBM-Linker-POIL (1); in: LCBM represents a part that binds to LC3; it is a structure shown in formula (A), in: Y 1 and Y 2 Each independently selected from O or S; The Ar ring is selected from a C6-C10 aryl group or a 5-10 membered heteroaryl group; R 1 is n substituents on the Ar ring, n is an integer selected from 0-4; Each R 1 Each is independently selected from halogen, -CN, -OH, -NH2, -NO2, -COOH, unsubstituted or substituted C1-C20 alkyl, unsubstituted or substituted C1-C20 alkoxy, C1-C20 alkyl-NH-, (C1-C20 alkyl)(C1-C20 alkyl)N-, C1-C20 alkoxycarbonyl-NH-, unsubstituted or substituted C3-C16 cycloalkyl, unsubstituted or substituted 3-16 membered heterocyclyl, unsubstituted or substituted C6-C14 aryl, unsubstituted or substituted 5-15 membered heteroaryl, wherein the substitution means that one or more hydrogen in the defined group is replaced by one or more substituents selected from halogen, -NH2, -OH; R 2 Selected from hydrogen, unsubstituted or substituted C1-C20 alkyl, amino C1-C20 alkyl, (C1-C6 alkyl)(C1-C6 alkyl)N-C1-C16 alkyl, C1-C6 alkyl-NH-C1-C16 alkyl, unsubstituted or substituted C3-C16 cycloalkyl, unsubstituted or substituted C3-C16 cycloalkylC1-C6 alkyl, unsubstituted or substituted 3-16 membered heterocycloalkyl, unsubstituted or substituted 3-10 membered heterocycloalkyl C1-C6 alkyl, unsubstituted or substituted C6-C14 aryl, unsubstituted or substituted 5-15 membered heteroaryl, unsubstituted or substituted C6-C14 aryl C1-C6 alkyl, unsubstituted or substituted 5-15 membered heteroaryl C1-C6 alkyl, wherein the substitution means that one or more hydrogen atoms in the defined group are replaced by one or more substituents selected from halogen, amino, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, trifluoromethyl; wherein the heteroaryl and heterocyclic groups contain 1 to 4 heteroatoms selected from N, O and S; Indicates connecting to linker from here; Linker represents a covalently linked moiety; POIL represents the target protein ligand portion that binds to CDK or EZH2 or RAS.

2. The protein degradation agent according to claim 1 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: In formula (A), Each R 1 Each is independently selected from halogen, -CN, -OH, -NH2, -NO2, -COOH, unsubstituted or substituted C1-C10 alkyl, unsubstituted or substituted C1-C10 alkoxy, C1-C10 alkyl-NH-, (C1-C10 alkyl)(C1-C10 alkyl)N-, C1-C10 alkoxycarbonyl-NH-, unsubstituted or substituted C3-C10 cycloalkyl, unsubstituted or substituted 3-10 membered heterocyclyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, wherein the substitution means that one or more hydrogen in the defined group is replaced by one or more substituents selected from halogen, -NH2, -OH; and / or R 2 Selected from hydrogen, unsubstituted or substituted C1-C10 alkyl, amino C1-C10 alkyl, (C1-C6 alkyl)(C1-C6 alkyl)N-C1-C10 alkyl, C1-C6 alkyl-NH-C1-C10 alkyl, unsubstituted or substituted C3-C10 cycloalkyl, unsubstituted or substituted C3-C10 cycloalkylC1-C6 alkyl, unsubstituted or substituted 3-10 membered heterocycloalkyl, unsubstituted or substituted 3-10 membered heterocycloalkyl C1-C6 alkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, unsubstituted or substituted C6-C10 aryl C1-C6 alkyl, unsubstituted or substituted 5-10 membered heteroaryl C1-C6 alkyl, wherein the substitution means that one or more hydrogen in the defined group is replaced by one or more substituents selected from halogen, -NH2, -OH, C1-C3 alkyl, C1-C3 alkoxy, trifluoromethyl.

3. The protein degradation agent according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: The structure represented by formula (A) is selected from the structure represented by the following formula (A-1): Wherein, the Ar ring is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl; R 1 is n substituents on the Ar ring, n is selected from an integer of 0-2, preferably, n is 0 or 1; Each R 1 Each is independently selected from halogen, -NO2, -COOH, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, wherein the substitution means that one or more hydrogens in the defined group are replaced by one or more substituents selected from halogen, -NH2, -OH; preferably, R 1 Selected from halogen, -NO2, methyl, methoxy; R 2 is selected from hydrogen, unsubstituted or substituted C1-C10 alkyl, unsubstituted or substituted C3-C10 cycloalkyl C1-C6 alkyl, unsubstituted or substituted 3-10 membered heterocyclyl C1-C6 alkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 5-10 membered heteroaryl, unsubstituted or substituted C6-C10 aryl C1-C6 alkyl, unsubstituted or substituted 5-10 membered heteroaryl C1-C6 alkyl, wherein the substitution means that one or more hydrogens in the defined group are replaced by one or more substituents selected from halogen, amino, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, trifluoromethyl; preferably, R 2 Selected from hydrogen, ethyl, t-pentyl, cyclohexylmethyl, N-methylpiperidinylmethyl, N,N-dimethylaminoethyl, phenyl, benzyl, benzyl substituted by methoxy or trifluoromethyl, phenethyl, pyridyl, N-methylpyrazolyl.

4. The protein degradation agent according to any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: The structure shown in formula (A) is selected from the following structures:

5. The protein degradation agent or pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof according to claim 1, characterized in that: POIL represents a target protein ligand portion that binds to CDK, wherein the CDK includes CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, CDK14, CDK15, CDK16, CDK17, CDK18, CDK19, CDK20, CDK21, preferably CDK9, CDK7, CDK2, CDK4, CDK6; or The POIL part is a CDK probe, a CDK inhibitor, an EZH2 probe, an EZH2 inhibitor, a RAS probe, a RAS inhibitor, including the following compounds:

6. The protein degradation agent according to claim 1 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: Linker is a chemical bond or a linear or branched alkylene or saturated cyclic alkyl structure containing 1-50, preferably 2-16 carbon atoms, wherein one or more carbon atoms, especially 1-6, more especially 1 or 2 carbon atoms are optionally replaced by heteroatoms selected from O, S, NR a , PR a , preferably O, S or NR a , more preferably O or NR a , especially O, where R a is H or C1-C3 alkyl; or, one or more carbon atoms, particularly 1-6, more particularly 1 or 2 carbon atoms, are optionally replaced by -C(=O)-, -C(=S)-, -S(=O)-, -SO2-, or a 3- to 6-membered ring having 0 to 4 heteroatoms selected from O, S, N, P; Preferably, Linker is selected from: -(CH2) n -、-(CH2CH2O) m -(CH2) n -、-CO-(CH2) n -、 wherein each n is independently an integer selected from 1-20, preferably an integer from 1-16, and more preferably an integer from 1-8; Each m is independently an integer of 1-10, preferably an integer of 1-6, more preferably an integer of 1-2; p is an integer of 0-10, preferably an integer of 0-6, more preferably an integer of 0-2; Further preferably, Linker is selected from the following structures:

7. The protein degradation agent according to any one of claims 1 to 6 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: The protein degradation agent represented by formula (1) is selected from the following structures: Wherein, n is an integer of 1-16, preferably an integer of 1-8; m is an integer of 1-6, preferably 1 or 2; R 1 , R 2 The definition as described in any one of claims 1 to 4.

8. The protein degradation agent according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: The protein degradation agent is selected from the following structures:

9. A pharmaceutical composition comprising a therapeutically effective amount of a protein degrader selected from any one of claims 1 to 8, or one or more of its pharmaceutically acceptable salts, stereoisomers, solvates, polymorphs, tautomers, isotopic compounds, metabolites or prodrugs, and an optional pharmaceutically acceptable carrier.

10. The protein degrader according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, or the pharmaceutical composition according to claim 8 for one or more of the following uses: a. Use in the preparation of a product for degrading one or more proteins in CDK9, cyclin T1, CDK9-cyclin T1 complex, EZH2, EED, SUZ12, EZH1, CDK2, CDK4, CDK5, CDK6, CDK7, cyclin H, cyclin A2, cyclin E1, cyclin D1 or RAS; b. Use in the preparation of a medicament for treating, preventing and / or ameliorating a disease or condition related to CDK or EZH2 or RAS.

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