Preparation method, pharmaceutical composition and application of CDK2 / 4 / 6 proteolysis targeting chimera

Abstract

The invention discloses a CDK2 / 4 / 6 proteolysis targeting chimera as well as a preparation method, a pharmaceutical composition and application thereof, and the chemical structure of the CDK2 / 4 / 6 proteolysis targeting chimera is shown as a formula I, the CDK2 / 4 / 6 proteolysis targeting chimera or the isomer, the pharmaceutically acceptable salt, the solvate, the prodrug, the deuterated compound or the polymorphic substance thereof can induce selective degradation of CDK2, CDK4 and CDK6 through ubiquitin-proteasome, so that the phosphorylation level of RB1 is reduced, release of a transcription factor E2F is further inhibited, the process of cells from a G1 phase to an S phase is blocked, and the targeting effect of the cells is improved. Compared with an inhibitor, the compound is more efficient and low in toxicity, and can be used for treating various cancers. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a CDK2 / 4 / 6 protein hydrolysis targeting chimera, a preparation method thereof, a pharmaceutical composition and uses thereof. Background Art

[0002] Cancer remains a major public health problem worldwide and the leading cause of global disease burden. Lung cancer is the most common cancer globally, followed by breast cancer, colorectal cancer, prostate cancer, and gastric cancer.

[0003] Cyclin-dependent kinases (CDKs) collaborate with their cyclin partners to regulate cell cycle progression and transcription, allowing cell cycle events to proceed in an orderly manner. Among them, CDK2, CDK4, and CDK6 are involved in the critical transition from G1 to S phase and play a key role in the occurrence and progression of various cancers. In terms of drug development targeting CDKs, the fastest progress is in the development of CDK4 / 6 inhibitors. Currently, selective CDK4 / 6 inhibitors such as Palbociclib (PD-0332991), Ribociclib (LEE011), and Abemaciclib (LY2835219) have been approved for the treatment of breast cancer. However, natural and acquired resistance to CDK4 / 6 inhibitors remain a difficult problem in clinical practice. Increased activity of the Cyclin-E-CDK2 complex is one of the reasons for CDK4 / 6i resistance. Studies have shown that the combination of CDK2 inhibitors and CDK4 / 6 inhibitors is effective against cell lines that are resistant to CDK4 / 6 inhibitors due to overexpression of Cyclin E. Therefore, a series of targeted inhibitors against CDK2 / 4 / 6 have been developed one after another.

[0004] Proteolysis-targeting chimeras (PROTACs) are a novel drug development technology characterized by high efficacy, selectivity, targeting undruggable targets, and the ability to overcome acquired drug resistance. PROTACs consist of a linker and ligands attached to each end of the linker: one ligand recruits and binds to the protein of interest (POI), while the other recruits and binds to the E3 ubiquitin ligase. PROTACs bind both the POI and the ligase simultaneously, inducing ubiquitination of the POI. The POI is then degraded by the ubiquitin–proteasome system (UPS), after which the PROTAC is released for the next round of POI degradation. PROTAC molecules offer the following advantages over traditional small molecule inhibitors: 1. They have the potential to target targets that are undruggable for small molecule inhibitors; 2. They can overcome mutation-induced drug resistance by eliminating pathologically relevant proteins; 3. Their catalytic mechanism of action and event-driven pharmacology enable high efficacy at low doses; and 4. They achieve high selectivity through protein-protein interactions between the target protein and the E3 ubiquitin ligase.

[0005] Recently, more and more studies have shown that CDK4 / 6 inhibitor resistance caused by CDK6 overexpression can be overcome by designing PROTAC molecules targeting CDK4 / 6. Therefore, based on the unique mechanism of action of PROTAC degraders themselves, the development of CDK2 / 4 / 6 PROTAC molecules has better application prospects than CDK2 / 4 / 6 inhibitors.

[0006] It should be noted that the information disclosed in the above background technology section is only used to enhance the understanding of the background of the present disclosure, and therefore may include information that does not constitute prior art known to ordinary technicians in the field. Summary of the Invention

[0007] The purpose of the present invention is to provide a CDK2 / 4 / 6 protein hydrolysis targeted chimera and its preparation method, pharmaceutical composition and use, which can induce the selective degradation of CDK2, CDK4 and CDK6 through ubiquitin-proteasome, thereby reducing the phosphorylation level of RB1, thereby inhibiting the release of transcription factor E2F, blocking the progression of cells from G1 phase to S phase, and reducing cell proliferation of cancer cell lines. Compared with inhibitors, it is more efficient and less toxic, and can be used to treat various cancers.

[0008] To achieve the above objectives, the present invention provides a CDK2 / 4 / 6 proteolysis targeting chimera or its isomers, pharmaceutically acceptable salts, solvates, prodrugs, deuterated compounds or polymorphs, wherein the chemical structure of the chimera is shown in Formula I:

[0009]

[0010] Wherein, R1 is selected from any one of the following structures: hydrogen, deuterium, cyano, halogen, amino, hydroxyl, nitro, CHF2, CF3, C(O)R a 、C(O)NHR a 、C(O)N(R a )2, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, 3-8 membered cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl, or at least one H in the amino, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C1-C6 alkoxy, 3-8 membered cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl is optionally substituted with Ra;

[0011] Among them, the R a is selected from deuterium, nitro, cyano, hydroxy, amino, trifluoromethyl, halogen, C1-C6 alkyl or C1-C6 deuterated alkyl, or at least one H in the C1-C6 alkyl and C1-C6 deuterated alkyl is optionally substituted with halogen;

[0012] R2 is selected from hydrogen, halogen, C1-C6 alkyl or C1-C6 deuterated alkyl, wherein at least one H in the C1-C6 alkyl is optionally substituted by halogen;

[0013] R3 is selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, 3-7-membered cycloalkyl, C3-C8 alkenyl, 5-8-membered cycloalkenyl, 6-10-membered aryl, 5-10-membered heteroaryl, or at least one H in the aryl, C1-C8 alkyl, C1-C8 alkoxy, 3-7-membered cycloalkyl, C3-C8 alkenyl, 5-8-membered cycloalkenyl, 6-10-membered aryl, 5-10-membered heteroaryl is optionally replaced by R a replace;

[0014] R4 is selected from any one of the following structures:

[0015]

[0016] wherein m and f are each independently an integer of 1-4, n is an integer of 0-14, W and Q are each independently selected from CH or N, and L is selected from O or S;

[0017] Y is selected from -S(O)2-, -SO-, -S(O)2NR e -, -S-, -CH2- or -C(O)-; wherein R eis selected from hydrogen, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl, or at least one H in the C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl is optionally replaced by R a replace;

[0018] R5 is selected from any one of the following structures:

[0019] wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N;

[0020] Linker is selected from any of the following structures:

[0021]

[0022] f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, n1 is an integer of 1-14, W2 and Q2 are each independently selected from CH or N, and L1 is selected from O or S;

[0023] The E3 ubiquitin ligase ligand is selected from any one of the following structures:

[0024]

[0025] In formula i, ring A is selected from any one of the following structures:

[0026]

[0027] where R b and R f are independently selected from any one of the following structures: hydrogen, alkyl, alkoxy, halogen, haloalkyl, haloalkoxy or cyano, or R b and R f Combined to form -C=O;

[0028] R c 、R d are independently selected from any one of the following structures: hydrogen, alkyl, alkoxy, halogen, haloalkyl, haloalkoxy or cyano;

[0029] G is selected from a single covalent bond, -O-, -C(=O)-, or -NH-;

[0030] The C ring is selected from the following structures: m2 is an integer from 1 to 4;

[0031] R h selected from hydrogen or alkyl;

[0032] In formula ii, W3 is selected from CH or N;

[0033] Z is selected from a single covalent bond connecting W and ring B, -CH2-, -NH-, -O- or -NHC(O)-, wherein the NH end of -NHC(O)- is connected to W;

[0034] X is selected from a single covalent bond connecting the linker and the E3 ubiquitin ligase, alkylene, -O-, -(O-alkylene)-, -(alkylene-O)-, -NH-, -N(alkyl)-, -C(O)NH-;

[0035] Ring B is selected from C6-C10 monocyclic aryl, C5-C10 monocyclic heteroaryl, C9-C10 fused bicyclic heteroaryl, or at least one H in the C6-C10 monocyclic aryl, C5-C10 monocyclic heteroaryl, C9-C10 fused bicyclic heteroaryl is optionally replaced by R a replace.

[0036] The terminology in this invention is explained as follows:

[0037] The term "deuterated" as used herein means that one or more hydrogen atoms of the group mentioned are replaced by deuterium atoms;

[0038] The term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine;

[0039] As used herein, the term "alkyl" refers to a saturated aliphatic hydrocarbon group having a specific number of carbon atoms, both branched and straight. For example, the definition of "C1-C6 alkyl" includes saturated aliphatic hydrocarbon groups having 1, 2, 3, 4, 5, or 6 carbon atoms in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0040] The term "alkylene" as used herein refers to a straight-chain or branched divalent saturated hydrocarbon group consisting of carbon and hydrogen;

[0041] The term "alkoxy" as used herein refers to a straight or branched chain alkoxy group having the structural formula alkyl-O-. Optionally, the alkyl portion of the alkoxy group may contain 1 to 10 carbon atoms. Representative examples of "alkoxy" include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, 2-pentoxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, 3-methylpentoxy, and the like. The term "C1-C6 alkoxy" refers to a straight or branched chain alkoxy group containing 1 to 6 carbon atoms;

[0042] The term "heteroaryl" as used herein refers to a 5- to 10-membered monocyclic, bicyclic, or polycyclic group containing at least one aromatic ring having 1 or more (e.g., 1 to 6, 1 to 5, 1 to 4, or 1 to 3) heteroatoms independently selected from oxygen, nitrogen, and sulfur. Bicyclic or polycyclic heteroaryl groups include bicyclic, tricyclic, or tetracyclic heteroaryl groups, one of which is an aromatic ring having one or more heteroatoms independently selected from O, S, and N, and the other rings may be saturated, partially unsaturated, or aromatic and may be carbocyclic or contain one or more heteroatoms independently selected from O, S, and N.

[0043] The term "aryl" as used herein refers to a monovalent aromatic hydrocarbon group containing 5 to 20 (e.g., 5 to 14, 5 to 15, 5 to 10, 5 to 9, 5 to 8, 6 to 15, or 6 to 10) carbon atoms and optionally containing one or more fused rings, such as phenyl, theanyl, or fluorenyl. In the present disclosure, optionally, the "aryl" is a substituted aryl;

[0044] The term "cycloalkyl" as used herein refers to a saturated or partially unsaturated (ie, having one or more double bonds, but not fully conjugated) monocyclic, bicyclic, tricyclic, or polycyclic hydrocarbon radical, which in some embodiments has 3 to 8 carbon atoms.

[0045] In some embodiments, polymorphic forms of the chimeras of the present invention or salts of the compounds of the present disclosure are also provided. The salts of the chimeras of the present invention can be pharmaceutically acceptable salts, including but not limited to hydrohalides (including hydrochlorides, hydrobromides), sulfates, citrates / citrate salts, maleates, methanesulfonates, lactates, lactobionates, L-tartrates, fumarates, L-malates, α-ketoglutarate, hippurates, D-glucuronates, D-gluconates, α-D-glucoheptonates, glycolates, mucates, L-ascorbic acid salts, orotates, picrates, glycinates, alanine salts, arginine salts, cinnamates, laurates, sebacates, benzenesulfonates, methanesulfonates, ethanesulfonates, edisylate, formates, acetates, 2, 2-dichloroacetate, pivalate, propionate, valerate, palmitate, triphenylacetate, 2-ethyl-succinate, iodate, nicotinate, L-pyroglutamate, L-proline, ferulate, 2-hydroxyethanesulfonate, nitrate, gentisate, cholate, salicylate, terephthalate, glutarate, adipate, stearate, oleate, undecylenate, dodecylsulfonate, phosphate, thiocyanate, dihydrogenphosphate, pyrophosphate, metaphosphate, oxalate, malonate, benzoate, mandelate, succinate, pyruvate, p-chlorobenzenesulfonate, 1,5-naphthalenedisulfonate, 3-hydroxy-2-naphthoate, 1-hydroxy-2-naphthoate, 2-naphthalenesulfonate, trifluoroacetate, glycolate or p-toluenesulfonate, etc.

[0046] In some embodiments, the chimera of the present invention can exist in the form of an unsolvated or solvated form, in pharmaceutically acceptable solvents such as water, ethanol, and the like.

[0047] In some embodiments, the compounds of the present disclosure can be prepared as prodrugs or prodrugs. Prodrugs can be converted into parent drugs in the body to exert their effects.

[0048] In some embodiments, isotopically labeled compounds of the present disclosure are also provided. Examples of isotopes include deuterium (D or 2 H).

[0049] Preferably, the R1 is selected from any one of the following structures: C(O)R a 、C(O)NHR a or C(O)N(R a )2; wherein, the R a Selected from C1-C6 alkyl; said R2 is selected from C1-C6 alkyl; said R3 is selected from 3-7 membered cycloalkyl; said R4 is selected from wherein m and f are each independently an integer of 1-4, W is selected from CH or N; Y is selected from -S(O)2-; and R5 is selected from any one of the following structures:

[0050] wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N;

[0051] The Linker is selected from any one of the following structures: wherein f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, and W2 and Q2 are each independently selected from CH or N.

[0052] Preferably, the chemical structure of the chimera is as shown in Formula II:

[0053]

[0054] The Linker is selected from any one of the following structures: f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, W2 and Q2 are each independently selected from CH or N;

[0055] The R5 is selected from any one of the following structures:

[0056] wherein f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, and W2 and Q2 are each independently selected from CH or N.

[0057] Preferably, the Linker is selected from any one of the following structures:

[0058]

[0059] wherein j is an integer from 0 to 14, W2 and Q2 are each independently selected from CH or N;

[0060] The R5 is selected from any one of the following structures:

[0061] wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N.

[0062] More preferably, in the Linker, j is an integer from 0 to 1, W2 and Q2 are each independently selected from CH or N; in the R5, f1 is 1, f3 is 2, m3 is 2, and W1 and Q1 are each independently selected from CH or N.

[0063] Preferably, the E3 ubiquitin ligase ligand is selected from any one of the following structures:

[0064]

[0065] Preferably, the chemical structure of the chimera is selected from any one of the following structures:

[0066]

[0067]

[0068] A second object of the present invention is to provide a method for preparing the CDK2 / 4 / 6 proteolysis-targeted chimera or its isomers, pharmaceutically acceptable salts, solvates, prodrugs, deuterated compounds or polymorphs, wherein the method for preparing the chimera comprises:

[0069]

[0070] A pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 having a chemical structure as shown in Formula III, an intermediate 2 containing an E3 ubiquitin ligase ligand and a linker structure, and DIEA are reacted in an organic solvent with or without the presence of HATU to obtain the chimera.

[0071] A third object of the present invention is to provide a method for preparing a pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1, wherein the chemical structure of the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 is as shown in Formula IV or Formula V. The method for preparing the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 shown in Formula IV comprises:

[0072]

[0073] Step 1: Synthesis of 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0074] Under an inert atmosphere, 6-bromo-2-chloro-8-cyclopentyl-5-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one is reacted in N-methylpyrrolidone in the presence of di(tri-tert-butylphosphine)palladium at 45°C with tributyl(1-ethoxyvinyl)tin to obtain 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one;

[0075] Step 2: Synthesis of 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0076] 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one was dissolved in THF, and aqueous hydrochloric acid was added. The mixture was stirred at room temperature to react to obtain 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one;

[0077] Step 3: Synthesis of tert-butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate

[0078] 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one and 1-tert-butoxycarbonyl-4-aminopiperidine were reacted in DMF with stirring at 80° C. in the presence of DIEA to obtain tert-butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate;

[0079] Step 4: Synthesis of 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one

[0080] tert-Butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate and dioxane hydrochloride solution were stirred at room temperature to react to obtain 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one;

[0081] Step 5: Synthesis of pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula IV

[0082] 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one was reacted in DCM with the compound represented by formula d in an ice bath under the action of DIEA to obtain pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula IV;

[0083] The preparation method of the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 shown in Formula V comprises:

[0084]

[0085] Step 1: Synthesis of tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate

[0086] 6-Acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one was reacted in anhydrous DCM with butyl 4-chlorosulfonylpiperidinecarboxylate in the presence of DIEA, with stirring at room temperature under an ice bath to obtain tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate;

[0087] Step 2: Synthesis of pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula V

[0088] Tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate and HCl·dioxane solution were stirred and reacted at room temperature to obtain pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula V.

[0089] The fourth object of the present invention is to provide a pharmaceutical composition comprising the CDK2 / 4 / 6 proteolysis targeting chimera or its isomers, pharmaceutically acceptable salts, solvates, prodrugs, deuterated compounds or polymorphs.

[0090] In some embodiments, the pharmaceutical composition disclosed herein comprises as an active ingredient a chimera represented by Formula I of the present invention or a pharmaceutically acceptable salt, solvate, isotopically enriched analog, polymorph, prodrug, stereoisomer (including enantiomer) or mixture of stereoisomers thereof, and at least one pharmaceutically acceptable carrier.

[0091] In some embodiments, pharmaceutically acceptable carriers include, but are not limited to, fillers, stabilizers, dispersants, suspending agents, diluents, excipients, thickeners, colorants, solvents, or encapsulating materials. A carrier must be "acceptable" if it is compatible with the other ingredients of the formulation (including the compounds useful in the present invention) and not harmful to the patient. Some examples of materials that are pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerol sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; phosphate buffered saline surfactants; polyethylene oxide, polyvinyl pyrrolidone, polyacrylamide, poloxamer; and other nontoxic compatible substances used in pharmaceutical formulations.

[0092] The pharmaceutical composition of the present invention further includes at least one second therapeutic agent, such as an anticancer agent. The second therapeutic agent can be combined with the chimera of Formula I described herein to treat the diseases or conditions described herein. The second therapeutic agent includes, but is not limited to, a chemotherapeutic agent, an immunotherapeutic agent, a gene therapy agent, and the like.

[0093] The fifth object of the present invention is to provide the use of the CDK2 / 4 / 6 protein hydrolysis targeting chimera or its isomers, pharmaceutically acceptable salts, solvates, prodrugs, deuterated compounds or polymorphs, or the pharmaceutical composition in the preparation of drugs for preventing and / or treating cancer.

[0094] The pharmaceutical composition of the present invention comprising the chimera of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient can be prepared into a suitable dosage form according to an appropriate route of administration (including but not limited to nasal administration, inhalation administration, topical administration, oral administration, oral mucosal administration, rectal administration, pleural cavity administration, peritoneal administration, vaginal administration, intramuscular administration, subcutaneous administration, transdermal administration, epidural administration, intrathecal administration and intravenous administration), such as spray preparations, patches, tablets (e.g., conventional tablets, dispersible tablets, orally disintegrating tablets), capsules (e.g., soft capsules, hard capsules, enteric-coated capsules), dragees, lozenges, powders, granules, powder injections, suppositories or liquid preparations (e.g., suspensions, such as aqueous or oily suspensions; emulsions or syrups, or conventional injection forms, such as injectable solutions, such as sterile injectable solutions prepared according to methods known in the art using water, Ringer's solution or isotonic sodium chloride solution as a carrier or solvent) or lyophilized compositions. Those skilled in the art can also prepare the chimera of the present invention into conventional, dispersible, chewable, orally rapidly disintegrating or rapidly dissolving preparations, or sustained-release capsules or controlled-release capsules as needed.

[0095] The chimera of Formula I described herein, or a pharmaceutically acceptable salt, solvate, isotopically enriched analog, polymorph, prodrug, stereoisomer (including enantiomer), or mixture of stereoisomers thereof, can be used as a medicament. In particular, the chimera of Formula I described herein, or a pharmaceutically acceptable salt, solvate, isotopically enriched analog, polymorph, prodrug, stereoisomer (including enantiomer), or mixture of stereoisomers thereof, can be used to prepare a medicament for treating and / or preventing a disease or condition selected from the group consisting of tumors or cancers.

[0096] In some embodiments, the disease or condition includes, but is not limited to: myeloma, including multiple myeloma, plasma cell myeloma, smoldering myeloma, smoldering multiple myeloma; transplant-related cancers; neutropenia; leukemia, including chronic myeloid leukemia, B-cell chronic lymphocytic leukemia, anemia associated with leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), monocytic leukemia, myelomonocytic leukemia, acute lymphoblastic leukemia (ALL), acute T-lymphocytic leukemia, T-lymphocytic leukemia , chronic lymphocytic leukemia (CLL); lymphomas, including diffuse large B-cell lymphoma, non-Hodgkin's lymphoma, anaplastic lymphoma, anaplastic large cell lymphoma, immunoblastic T-cell lymphoma CD20-positive lymphoma, mantle cell lymphoma, follicular lymphoma (FL), Burkitts lymphoma, marginal zone lymphoma (MZL), primary lymphoma, B-cell lymphoma, relapsed B-cell non-Hodgkin's lymphoma, relapsed diffuse large B-cell lymphoma, relapsed mediastinal (thymic) large B-cell lymphoma, primary mediastinal (thymic) large B-cell lymphoma B-cell lymphoma, relapsed transformed non-Hodgkin's lymphoma, refractory B-cell non-Hodgkin's lymphoma, refractory diffuse large B-cell lymphoma, refractory primary mediastinal (thymic) large B-cell lymphoma, refractory transformed non-Hodgkin's lymphoma; thyroid cancer; melanoma; lung cancer, including lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung cancer and small cell lung cancer; inflammatory myofibroblastic tumor; colorectal cancer; intestinal cancer; glioma; astroblastoma; glioma; peripheral neuroepithelial tumor; ovarian cancer; bronchogenic carcinoma; prostate cancer; breast cancer, including triple-negative breast cancer, Patients with incidental breast cancer, ductal carcinoma, and Cowden disease; pancreatic cancer; central nervous system cancers; neuroblastoma; extramedullary plasmacytoma; plasmacytoma; gastric cancer; gastrointestinal stromal tumors; esophageal cancer; colorectal adenocarcinoma; esophageal squamous cell carcinoma; liver cancer; renal cell carcinoma; bladder cancer; endometrial cancer; head and neck cancer; brain cancer; oral cancer; sarcomas, including rhabdomyosarcoma, various adipose tumors, Ewing sarcoma / primitive neuroectodermal tumor (Ewing / PNETS), and leiomyosarcoma; urothelial carcinoma; basal cell carcinoma; oral squamous cell carcinoma; bile duct cancer; bone cancer; cervical cancer; and skin cancer.

[0097] Preferably, the disease or disorder is selected from non-small cell lung cancer and breast cancer.

[0098] The chimera of Formula I as the active ingredient is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount for the indication to be treated to the subject without causing severe toxic effects in the treated subject. The dosage of the active compound for all diseases or conditions mentioned in the present invention is, for example, about 25 mg / kg subject body weight / day.

[0099] The chimera of Formula I of the present invention or a pharmaceutically acceptable salt thereof can be conveniently administered in any suitable dosage form, and the specifications of suitable dosage forms include but are not limited to less than 1 mg, 1 mg to 3000 mg, 5 mg to 1000 mg, for example, 5 to 500 mg, 25 to 250 mg of active ingredient per unit dosage form.

[0100] A "therapeutically effective amount" of the chimeras of the present invention depends on a variety of factors, including the activity of the compound being used, the metabolic stability and duration of action of the compound, the age, sex, and weight of the patient, the patient's overall medical condition, the route and timing of administration, the rate of excretion, concomitant medications, and the progression of the disease or condition being treated. Those skilled in the art will be able to determine an appropriate dosage based on these and other factors. It will be understood that the choice of one or more active compounds and / or compositions and their dosages will depend on the individual's individual circumstances (generally, the optimal effect will be achieved for that individual's circumstances).

[0101] The patients or subjects for treatment mentioned above refer to animals, such as mammals, including but not limited to primates (such as humans), cows, sheep, goats, horses, dogs, cats, rabbits, guinea pigs, rats, mice, etc.

[0102] The CDK2 / 4 / 6 proteolysis-targeted chimera of the present invention, its preparation method, pharmaceutical composition and use have the following advantages:

[0103] The chimera of the present invention induces the selective degradation of CDK2, CDK4 and CDK6 through ubiquitin-proteasome, thereby reducing the phosphorylation level of RB1, thereby inhibiting the release of transcription factor E2F, blocking the progression of cells from G1 phase to S phase, reducing cell proliferation of cancer cell lines, and promoting cancer cell apoptosis. It has been well verified at the cell, organoid and animal model levels, is more efficient and less toxic than inhibitors, and can be used to treat a variety of cancers. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] Figure 1 The results show the effects of the PROTAC drug prepared in this embodiment of the present invention on the degradation ability of CDK2, 4, and 6 in A549 cells and the phosphorylation level of RB1.

[0105] Figure 2 The results show the effects of the PROTAC drug prepared in this embodiment of the present invention on the degradation ability of CDK2, 4, and 6 in NCI-H1975 cells and the phosphorylation level of RB1.

[0106] Figure 3 The results show the effects of the PROTAC drug prepared in this embodiment of the present invention on the degradation ability of CDK2, 4, and 6 in NCI-H1299 cells and the phosphorylation level of RB1.

[0107] Figure 4 These are the results of the effects of the compounds WYY-03-127, WYY-03-120, WYY-03-185 and WYY-03-186 of the present invention on the degradation ability of CDK2, 4 and 6 in A549 cells and on the phosphorylation level of RB1 over time.

[0108] Figure 5 The results show that the degradation ability of compounds WYY-03-127 and 3600 on CDK2, 4, and 6 in A549 cells and their effects on the phosphorylation level of RB1 change with concentration.

[0109] Figure 6 These are the results of the inhibitory effects of proteasome inhibitors MLN4924, Thalidomide, and MG132 on the ubiquitination and degradation ability of compound WYY-03-127.

[0110] Figure 7 These are the results of a clone formation experiment showing the inhibitory effect of compounds WYY-03-127 and WYY-03-120 on A549 cell proliferation.

[0111] Figure 8 The results show the effects of intraperitoneal injection of the compound on tumor volume (A) and mouse body weight (B) in the A549 tumor cell mouse tumor model. DETAILED DESCRIPTION

[0112] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0113] Throughout this disclosure, all features, such as values, amounts, amounts, and concentrations, specified in numerical ranges or percentage ranges are provided for simplicity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to encompass and specifically disclose all possible subranges and individual values ​​within those ranges (including integers and fractions).

[0114] The features described in this disclosure may be combined in any manner, and as long as there are no conflicts between the combinations of these features, all possible combinations should be considered within the scope of this specification. Each feature disclosed in this specification may be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the features disclosed are merely general examples of equivalent or similar features.

[0115] The technical solutions of the present invention are described in detail below through specific embodiments, but the protection scope of the present invention includes but is not limited to these.

[0116] The structures of the following compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). 1 H NMR was measured by Bruker AVANCE NEO 600 (600 MHz) and Bruker biospin AV400 (400 MHz). 13 C NMR was measured by Bruker AVANCE NEO 600 (600 MHz). 1 H NMR and 13 C NMR chemical shifts are in ppm, coupling constants are in Hz, and tetramethylsilane is the internal standard; 1 Relative representation of peak types in H NMR: s, singlet; d, doublet; t, triplet; m, multiplet.

[0117] The abbreviations involved in the following embodiments are as follows:

[0118] NMP: N-methylpyrrolidone; Pd(t-Bu3P)2: bis(tri-tert-butylphosphine)palladium; EA: ethyl acetate; THF: tetrahydrofuran; DMF: N,N-dimethylformamide; DIEA: N,N-diisopropylethylamine; dioxane: dioxane; MeOH: methanol; DCM: dichloromethane; Pd(dppf)Cl2: 1,1-bis(diphenylphosphine)diferronichloridopalladium; EtOH: ethanol; Pd(dppf)C l2·DCM: [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex; TBAB: tetrabutylammonium bromide; HOAc: acetic acid; Tol: toluene; BINAP: 1,1'-binaphthyl-2,2'-bisdiphenylphosphine; DMSO: dimethyl sulfoxide; Pd-PEPPSI-IPent(Cl): [1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl) dichloropalladium(II).

[0119] The solvents and reagents involved in the following examples are as follows:

[0120] The solvents used in the reaction, such as EtOH, DMSO, DMF, EtOH, MeOH, EA, and other reaction substrates and reagents, can be obtained from commercial sources. The water used in the reaction is preparative-grade deionized water.

[0121] It should be noted that if specific conditions are not specified in the examples, conventional conditions or conditions recommended by the manufacturer shall be followed. The starting materials or reagents used in each scheme can be purchased from commercial sources or prepared by methods known to those skilled in the art.

[0122] 1. Synthesis of Pyrido[2,3-d]pyrimidin-7(8H)-one Intermediates

[0123] Example 1: 6-Acetyl-2-((1-((4-(bromomethyl)phenyl)sulfonyl)piperidin-4-yl)amino)-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0124]

[0125] Step 1: Synthesis of 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0126] 6-Bromo-2-chloro-8-cyclopentyl-5-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one (20.0 g, 58.3 mmol, 1.0 eq) was weighed into a 1000 mL eggplant-shaped flask and dissolved in N-methylpyrrolidone (300 mL). Tributyl(1-ethoxyethylene)tin (25 mL, 75.8 mmol, 1.3 eq) was then added. Di(tri-tert-butylphosphine)palladium (1.49 g, 2.9 mmol, 0.05 eq) was then added under N2 protection. The mixture was heated to 45°C for 24 hours. After TLC and LCMS showed the reaction was complete, saturated potassium fluoride solution (600 mL) was added to quench the reaction and stirred overnight. The reaction solution was then extracted with EA (300 mL x 3). The organic phase was washed with saturated NaCl solution (300 mL), dried over anhydrous Na2SO4, and EA was removed by distillation under reduced pressure. A yellow oily liquid was obtained by column chromatography (5% to 9% EA in PE) with a yield of 78.4%. 1 H NMR(600MHz, DMSO-d6)δ9.08(s,1H),5.73(tt,J=9.7,7.7Hz,1H),4.52–4.50(m,1H),4.11(d,J=2. 4Hz,1H),3.85(q,J=7.0Hz,2H),2.44(s,3H),2.15–1.59(m,8H),1.25(t,J=7.0Hz,3H); LCMS(ESI)C 17 H 20 ClN3O2 + [M+H] + =334.23.

[0127] Step 2: Synthesis of 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0128] The compound 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one (15.25 g, 45.7 mmol, 1.0 eq) was weighed and dissolved in a 500 mL eggplant flask with THF (150 mL). Then, 1 mol / L aqueous hydrochloric acid solution (150 mL) was added and stirred at room temperature for 1 hour. After TLC and LCMS monitoring showed that the reaction was complete, the reaction was quenched with saturated NaHCO3 solution, and the reaction solution was extracted with EA (200 mL × 3). The organic phase was washed with saturated NaCl solution (400 mL) and dried over anhydrous Na2SO4. The EA was removed by distillation under reduced pressure and the target product was purified by column chromatography to obtain a light yellow solid with a yield of 54.7%. 1 HNMR(400MHz,DMSO-d6)δ9.23(s,1H),5.84–5.75(m,1H),2.50(s,3H),2.41(s,3H),2.10–1.52(m,8H); LCMS(ESI)C 15 H 16 ClN3O2 + [M+H] + =306.14.

[0129] Step 3: Synthesis of tert-butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate

[0130] The compounds 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one (3.00 g, 9.8 mmol, 1.0 eq) and 1-tert-butoxycarbonyl-4-aminopiperidine (3.94 g, 19.7 mmol, 2.0 eq) were weighed separately and placed in a 250 mL eggplant flask. After dissolution with 60 mL of DMF, DIEA (5.1 mL, 29.5 mmol, 3.0 eq) was added and stirred in an 80°C oil bath for 4 hours. After TLC and LCMS showed the reaction was complete, water (160 mL) was added to quench the reaction. The reaction solution was extracted with EA (300 mL × 3). The organic phase was washed with saturated NaCl solution (300 mL) and dried over anhydrous Na2SO4. The EA was removed by distillation under reduced pressure and the product was isolated by column chromatography to obtain a white powder with a yield of 84.5%. 1H NMR(600MHz,DMSO-d6)δ8.80(d,J=27.4Hz,1H),5.95–5.69(m,1H),4.09–3.99(m,1H),3.98–3.89(m,2H),2.96–2.75(m,2H),2.40(s,3H),2 .25(s,3H),2.20–2.12(m,1H),2.00–1.90(m,2H),1.89–1.81(m,2H),1.79–1.71(m,2H),1.68–1.53(m,2H),1.46–1.33(m,12H); LCMS(ESI)C 25 H 35 N5O4 + [M+H] + =470.28.

[0131] Step 4: Synthesis of 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one

[0132] Compound 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (2.58 g, 5.49 mmol, 1 eq) was weighed into a 50 mL eggplant-shaped flask, and dioxane hydrochloride solution (20 mL, 4 mol / L) was added. The mixture was stirred at room temperature for 3 hours. After LCMS showed that the reaction was complete, the solvent was removed under reduced pressure, and MeOH was added and distilled under reduced pressure three times to remove residual hydrochloric acid. The mixture was dried in vacuo to obtain a white solid powder with a yield of 95.3%. 1 H NMR(400MHz,DMSO-d6)δ8.83(s,1H),5.92(s,1H),3.56–3.54(m,1H),2.99-3 .16(m,2H),2.40–2.21(m,5H),2.26(s,3H),2.21–1.49(m,12H); LCMS(ESI)C 20 H 27 N5O2 + [M+H] + =370.51.

[0133] Step 5: 6-Acetyl-2-((1-((4-(bromomethyl)phenyl)sulfonyl)piperidin-4-yl)amino)-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0134] The compound 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (500 mg, 1.23 mmol, 1.0 eq) was weighed into a 25 mL eggplant flask, DCM (10 mL) was added, and then DIEA (1.1 mL, 6.15 mmol, 5.0 eq) was added. Then, 4-bromomethylbenzenesulfonyl chloride (332 mg, 1.23 mmol, 1.0 eq) was added in batches in an ice bath. The reaction was carried out in an ice bath for 30 min. After the completion of the reaction monitored by TLC and LCMS, the solvent was distilled off under reduced pressure and dried in vacuo to give 632 mg of a crude product as a white powder, which was the target compound. 1 H NMR(400MHz, DMSO-d6)δ8.76(s,1H),7.78(d,J=8.2Hz,2H),7.72(d,J=7.7Hz,2H),5.92–5.60(m,1H),4.88(s,2H),3.9 0–3.64(m,1H),3.64–3.48(m,2H),2.48–2.41(m,2H),2.38(s,3H),2.28–2.08(m,5H),1.97–1.44(m,10H); LCMS(ESI)C 27 H 32 BrN5O4S + [M+H] + =602.08.

[0135] Example 2: 6-Acetyl-8-cyclopentyl-5-methyl-2-((1-(piperidin-4-ylsulfonyl)piperidin-4-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one hydrochloride

[0136]

[0137]

[0138] Step 1: Synthesis of tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate

[0139] 6-Acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (1.00 g, 2.46 mmol, 1.0 eq) was weighed into an eggplant-shaped flask. Ultra-dry DCM (16 mL) was added, followed by DIEA (2.2 mL, 12.30 mmol, 5.0 eq). Butyl 4-chlorosulfonylpiperidinecarboxylate (698 mg, 2.46 mmol, 1.0 eq) was then added portionwise under ice-cooling. The mixture was then stirred at room temperature for 3 hours. After completion of the reaction, the mixture was dried and purified by column chromatography to obtain a white powder with a yield of 84.2%. 1 HNMR(600MHz,Chloroform-d)δ8.62(s,1H),5.89–5.73(m,1H),5.31–5.07(m,1H),4.38–3.94(m,3H),3.92–2.96(m ,6H),2.78–2.60(m,2H),2.52(s,3H),2.32(s,5H),2.17–1.77(m,8H),1.69–1.58(m,4H),1.46(s,9H); LCMS(ESI)C 30 H 44 N6O6S + [M+H] + =617.16.

[0140] Step 2: Synthesis of 6-acetyl-8-cyclopentyl-5-methyl-2-((1-(piperidin-4-ylsulfonyl)piperidin-4-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one hydrochloride

[0141] The compound tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate (840 mg, 1.36 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HCl·dioxane solution (2 mL) was added. The mixture was stirred at room temperature for 3 hours. After LCMS showed that the reaction was complete, the mixture was spin-dried and freeze-dried to give a white solid powder, which was the target compound. 1HNMR(600MHz,DMSO-d6)δ8.84(s,1H),6.13–6.02(m,1H),5.83(tt,J=10.6,2.7Hz,1H),3.68(dt,J=13.0,4.1Hz,2H),3.54(tt,J=10.6,3.3Hz ,1H),3.34–3.22(m,2H),3.14–3.07(m,2H),2.95–2.85(m,2H),2.41(s,3H),2.26(s,5H),2.14–2.08(m,2H),2.00–1.53(m,12H); LCMS(ESI)C 25 H 36 N6O4S + [M+H] + =517.57.

[0142] 2. Synthesis of E3 Ubiquitin Ligase Ligands

[0143] Example 3: 3-(6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione hydrochloride

[0144]

[0145]

[0146] Step 1: Synthesis of tert-butyl 4-(2',6'-bis(benzyloxy)-[3,3'-bipyridyl]-6-yl)piperazine-1-carboxylate

[0147] 6-(4-Boc-1-piperazinyl)pyridine-3-boronic acid pinacol ester (500 mg, 1.28 mmol, 1.2 eq), 2,6-bis(benzyloxy)-3-bromopyridine (395 mg, 1.07 mol, 1.0 eq), and KCO (295 mg, 2.14 mmol, 1.0 eq) were weighed into a 25 mL eggplant flask. Ultra-dry dioxane (10 mL) and water (1.5 mL) were added. After expelling N2 three times, the catalyst Pd(dppf)Cl2 (39 mg, 54 μmol, 0.05 eq) was added. The N2 was again expelled three times, and the mixture was heated under reflux in a 95°C oil bath overnight. After LCMS monitoring indicated sufficient reaction, the reaction solution was cooled to room temperature and filtered. The filter cake was washed thoroughly with EA, and the filtrate was spin-dried. Purification by column chromatography afforded a white solid in a 78.6% yield. 1H NMR (400MHz, DMSO-d6) δ8.32–8.30(m,1H),7.77–7.71(m,2H),7.45–7.30(m,10H),6.86(d,J=8.8Hz,1H),6. 53(d,J=8.0Hz,1H),5.38(d,J=10.9Hz,4H),3.51–3.46(m,4H),3.43–3.38(m,4H),1.42(s,9H); LCMS(ESI)C 33 H 36 N4O4 + [M+H] + =553.39.

[0148] Step 2: Synthesis of tert-butyl 4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazine-1-carboxylate

[0149] The compound tert-butyl 4-(2',6'-bis(benzyloxy)-[3,3'-bipyridyl]-6-yl)piperazine-1-carboxylate (556 mg, 1.01 mmol, 1.0 eq) was weighed into a 50 mL eggplant-shaped flask, and EtOH (15 mL) and EA (15 mL) were added, followed by 10% palladium carbon (55% in water, 500 mg). H2 was exchanged three times, and the reaction solution was stirred at room temperature for 16 hours. After TLC and LCMS showed that the reaction was sufficient, the reaction solution was filtered, the filter cake was rinsed with EA, the filtrate was evaporated under reduced pressure, and purified by column chromatography to obtain a light pink solid with a yield of 63.5%. 1 H NMR (400MHz, DMSO-d6) δ10.82(s,1H),7.97(d,J=2.4Hz,1H),7.42(dd,J=8.8,2.5Hz,1H),6.82(d,J=8.7Hz,1H),3.75(dd,J=12.2,4.9Hz,1H) ,3.48–3.43(m,4H),3.42–3.37(m,4H),2.73–2.62(m,1H),2.55–2.51(m,1H),2.23–2.13(m,1H),2.00–1.93(m,1H),1.42(s,9H); LCMS(ESI)C 19 H 26 N4O4 + [M+H] + =375.19.

[0150] Step 3: Synthesis of 3-(6-(piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione hydrochloride

[0151] The synthesis of compound 4-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (240 mg, 641 mmol, 1.0 eq) was weighed into a 25 mL eggplant-shaped flask, and dioxane hydrochloride solution (3 mL, 4 mol / L) and DCM (4 mL) were added. The mixture was stirred at room temperature for 2 hours. After LCMS showed that the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, MeOH was added and dried three times to remove excess hydrochloric acid, and the crude product was dried in vacuo to obtain the target compound. 1 H NMR (400MHz, DMSO-d6) δ10.93(s,1H),7.99(d,J=2.2Hz,1H),7.88(dd,J=9.3,2.3Hz,1H),7.30(d,J=9.2Hz,1H),3.96–3.93(m,4H ),3.57–3.55(m,1H),3.24–3.21(m,4H),2.73–2.64(m,1H),2.60–2.55(m,1H),2.33–2.22(m,1H),2.00–1.92(m,1H); LCMS(ESI)C 14 H 18 N4O2 + [M+H] + =275.35.

[0152] Example 4: 3-(3-(piperazin-1-yl)phenyl)piperidine-2,6-dione hydrochloride

[0153]

[0154] Step 1: tert-Butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)piperazine-1-carboxylate

[0155] 1-BOC-4-(3-bromophenyl)piperazine (500 mg, 1.47 mmol, 1.0 eq), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (673 mg, 1.61 mmol, 1.1 eq), and KCO (672 mg, 161 mmol, 1.1 eq) were weighed into a 25 mL eggplant flask. Water (2 mL) and ultra-dry dioxane (8 mL) were added. The system was purged with N2 three times, and Pd(dppf)Cl2·DCM (122 mg, 0.15 mmol, 0.1 eq) was added. The N2 was then purged again three times, and the system was stirred at 110°C overnight. After TLC and LCMS showed that the reaction was completed, the product was quenched with water (12 mL) and extracted with EA (8 mL × 3). The organic phase was washed with saturated NaCl solution (10 mL × 2), dried over anhydrous Na2SO4, and then spin-dried. The product was purified by column chromatography to obtain a white solid with a yield of 72.7%. 1 H NMR(600MHz,DMSO-d6)δ7.75(d,J=8.1Hz,1H),7.48–7.29(m,10H),7.25–7.21(m,1H),7.12–7.10(m,1H),6.96(dd,J=7.6,1.5Hz,1H),6.8 6(dd,J=8.2,2.5Hz,1H),6.55(d,J=8.0Hz,1H),5.40(s,2H),5.37(s,2H),3.41–3.38(m,4H),3.02–2.92(m,4H),1.43(s,9H); LCMS(ESI)C 34 H 37 N3O4 + [M+H] + =552.15.

[0156] Step 2: tert-Butyl 4-(3-(2,6-dioxopiperidin-3-yl)phenyl)piperazine-1-carboxylate

[0157] The compound tert-butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)phenyl)piperazine-1-carboxylate (590 mg, 1.07 mmol, 1.0 eq) was weighed and placed in a 25 mL eggplant-shaped flask. EA (5 mL), EtOH (5 mL), and ultra-dry THF (5 mL) were added respectively, and then 10% Pd / C (200 mg) was added to replace H2 three times. The mixture was stirred at room temperature for 16 hours. After TLC and LCMS showed that the reaction was complete, the reaction solution was filtered and the filter cake was washed with EA. The filtrate was distilled under reduced pressure to obtain about 400 mg of a crude white solid. 1H NMR(600MHz,DMSO-d6)δ10.79(s,1H),7.19–7.15(m,1H),6.85–6.81(m,2H),6.66–6.64(m,1H),3.81–3.71(m,1H),3.48–3.41( LCMS(ESI)C 20 H 27 N3O4 + [M+H] + =374.24.

[0158] Step 3: 3-(3-(Piperazin-1-yl)phenyl)piperidine-2,6-dione hydrochloride

[0159] The compound tert-butyl 4-(3-(2,6-dioxopiperidin-3-yl)phenyl)piperazine-1-carboxylate (400 mg) was weighed into a 25 mL eggplant flask, DCM (4 mL) was added, and then HCl·dioxane solution (4 mL) was added. The reaction was stirred at room temperature for 2 hours. After LCMS showed that the reaction was complete, the solvent was removed by distillation under reduced pressure, an appropriate amount of MeOH was added and the mixture was dried by spin drying. This was repeated three times and then vacuum dried to give the crude product as a white powder, thereby obtaining the target compound. 1 H NMR (400MHz, DMSO-d6) δ10.82(s,1H),7.25–7.17(m,1H),6.97–6.84(m,2H),6.72(d,J=7.5Hz,1H),3.40–3.31(m ,4H),3.24–3.16(m,4H),2.72–2.59(m,1H),2.50–2.43(m,1H),2.27–2.13(m,1H),2.07–1.96(m,1H); LCMS(ESI)C 15 H 19 N3O2 + [M+H] + =274.13.

[0160] Example 5: 1-(7-Fluoro-1-methyl-6-(piperidin-4-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione hydrochloride

[0161]

[0162] Step 1: Synthesis of 6-bromo-5-fluoro-1-methyl-1H-indole-3-amine

[0163] NaHCO₃ (7.70 g, 91.6 mmol, 4.0 eq) was weighed and placed in a 25 mL eggplant-shaped flask. Water (4 mL) was added, followed by methylhydrazine sulfate (9.90 g, 68.7 mmol, 3.0 eq). The mixture was stirred for half an hour to liberate the methylhydrazine. EtOH (2 mL) was then added to precipitate the salts in the system. The mixture was filtered, and the filtrate was transferred to a 100 mL pressure flask. 4-Bromo-2,5-difluorobenzonitrile (5.00 g, 22.9 mmol, 1.0 eq) dissolved in EtOH (35 mL) was then added to the pressure flask. The reaction system was stirred at 80°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature, water (60 mL) was added, and the mixture was filtered. The filter cake was washed with an appropriate amount of water. The residue was collected, spin-dried, and vacuum-dried to obtain the crude product as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.83 (d, J = 5.6 Hz, 1H), 7.64 (d, J = 8.9 Hz, 1H), 5.53 (s, 2H), 3.73 (s, 3H); LCMS (ESI) C8H7BrFN3 + [M+H] + =244.16.

[0164] Step 2: Synthesis of 3-((6-bromo-5-fluoro-1-methyl-1H-indazol-3-yl)amino)propanoic acid

[0165] The compound 6-bromo-5-fluoro-1-methyl-1H-indole-3-amine (2.50 g, 10.2 mmol, 1.0 eq) was weighed and placed in a 100 mL eggplant-shaped flask. Freshly prepared 2M dilute hydrochloric acid solution (25 mL) was added, followed by tetrabutylammonium bromide (330 mg, 1.0 mmol, 0.1 eq) and acrylic acid (1.0 mL, 15.3 mmol, 1.5 eq). The reaction solution was heated to 100°C and stirred overnight. Afterwards, the reaction solution was cooled to room temperature and the pH was adjusted to 8 with saturated NaHCO₃ solution and then to 5 with acetic acid. The solution was then extracted with EA (20 mL x 3). The organic phase was washed with saturated NaCl solution (50 mL x 1) and dried over anhydrous Na₂SO₄. After spin drying, 2.7 g of the crude brown oil was obtained. 1 H NMR (400MHz, DMSO-d6) δ7.82(d,J=5.6Hz,1H),7.65(d,J=9.0Hz,1H),6.23(s,1H),3.75(s,3H),3.47–3.43(m,2H),2.61–2.56(m,2H); LCMS(ESI)C 11 H 11 BrFN3O2 + [M+H] + =316.11.

[0166] Step 3: Synthesis of 1-(6-bromo-7-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0167] The compound 3-((6-bromo-5-fluoro-1-methyl-1H-indazol-3-yl)amino)propionic acid (2.70 g, 8.5 mmol, 1.0 eq) was weighed and placed in a 100 mL eggplant-shaped flask, AcOH (25 mL) was added to form a suspension, and sodium isocyanate (1.11 g, 17.0 mmol, 2.0 eq) was added. The reaction solution was then heated to 60° C. and stirred for 16 hours. Concentrated hydrochloric acid (25 mL) was added and stirred at 60° C. for 3 hours. The reaction solution was then cooled to room temperature, diluted with an appropriate amount of water, filtered, and the filter cake was washed with water. The filter residue was collected, spin-dried, and vacuum-dried to obtain a white solid crude product. 1 HNMR(400MHz, DMSO-d6)δ8.16(d,J=5.6Hz,1H),7.61(d,J=9.1Hz,1H),3.99(s,3H),3.95–3.88(m,2H),2.78–2.73(m,2H); LCMS(ESI)C 12 H 10 BrFN4O2 + [M+H] + =341.24.

[0168] Step 4: Synthesis of tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-7-fluoro-1-methyl-1H-indazol-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate

[0169] 1-(6-Bromo-7-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (1.50 g, 4.4 mmol, 1.0 eq), Na2CO3 (2.19 g, 13.2 mmol, 3.0 eq), and N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (1.36 g, 4.4 mmol, 1.0 eq) were weighed into a 100 mL eggplant flask. Ultra-dry dioxane (30 mL) and water (5 mL) were added. Under a N2 atmosphere, Pd(dppf)Cl2·DCM (359 mg, 440 μmol, 0.1 eq) was added. The N2 atmosphere was then purged three times, and the mixture was heated to 80°C and stirred overnight. After LCMS and TLC showed that the reaction was complete, the reaction solution was filtered and the filter cake was washed with an appropriate amount of EA. The filtrate was collected and concentrated, washed with water (20 mL × 1) and saturated NaCl solution (30 mL × 1), and then dried over anhydrous Na2SO4. After spin-drying, it was purified by column chromatography and dried in vacuo to obtain a white powder with a yield of 48%.1 H NMR (400MHz, DMSO-d6) δ10.60(s,1H),7.60(d,J=6.1Hz,1H),7.42(d,J=11.4Hz,1H),6.05(s,1H),4.03(t,J =4.5Hz,2H),4.00(s,3H),3.93–3.88(m,2H),3.61–3.53(m,2H),2.79–2.72(m,2H),1.44(s,9H); LCMS(ESI)C 22 H 26 FN5O4 + [M+H] + =444.16.

[0170] Step 5: Synthesis of tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-7-fluoro-1-methyl-1H-indazol-6-yl)piperidine-1-carboxylate

[0171] The compound tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-7-fluoro-1-methyl-1H-indazol-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate (483 mg, 1.09 mmol, 1.0 eq) was weighed and placed in a 100 mL eggplant flask. MeOH (20 mL) and EA (30 mL) were added, followed by 10% Pd / C (552 mg). H2 was exchanged three times, and the reaction was stirred at room temperature for 16 hours. After TLC and LCMS showed the reaction was complete, the reaction solution was filtered, the filter cake was washed with EA, and the filtrate was collected and dried to give 280 mg of crude product. 1 H NMR (600MHz, DMSO-d6) δ7.59(d,J=5.9Hz,1H),7.37(d,J=10.8Hz,1H),3.99(s,3H),3.92–3. 86(m,2H),3.10–2.97(m,1H),2.78–2.70(m,2H),1.84–1.58(m,4H),1.42(s,9H); LCMS(ESI)C 22 H 28 FN5O4 + [M+H] + =446.13.

[0172] Step 6: Synthesis of 1-(7-fluoro-1-methyl-6-(piperidin-4-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione hydrochloride

[0173] tert-Butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-7-fluoro-1-methyl-1H-indazol-6-yl)piperidine-1-carboxylate (280 mg, 629 μmol, 1.0 eq) was placed in a 25 mL eggplant flask. DCM (4 mL) and HCl·dioxane solution (1 mL) were added and stirred at room temperature for 3 hours. LCMS indicated the reaction was complete. The solvent was then removed by spin drying, and the mixture was slurried with methyl tert-butyl ether, filtered, spin dried, and dried under vacuum to obtain 228 mg of a crude white solid powder, the target compound. 1 H NMR (400MHz, DMSO-d6) δ10.60(s,1H),7.50(d,J=5.9Hz,1H),7.42(d,J=11.0Hz,1H),4.01(s,3H),3.94 –3.86(m,2H),3.25–3.14(m,1H),3.12–2.99(m,2H),2.79–2.71(m,2H),2.09–1.95(m,4H); LCMS(ESI)C 17 H 20 FN5O2 + [M+H] + =346.18.

[0174] Example 6: 1-(2-methoxy-5-(piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione hydrochloride

[0175]

[0176] Step 1: Synthesis of 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid

[0177] 3-Amino-4-methoxybenzoic acid (4.50 g, 27 mmol, 1.0 eq) was weighed and placed in a 250 mL eggplant-shaped flask. Toluene (50 mL) was added, followed by acrylic acid (2.78 mL, 41 mmol, 1.5 eq). The mixture was stirred at 100°C for 24 hours. LCMS indicated complete consumption of the substrate. The reaction solution was cooled to room temperature, and urea (8.10 g, 135 mmol, 5.0 eq) and AcOH (50 mL) were added. The mixture was stirred at room temperature for 24 hours. After TLC indicated completion of the reaction, the solvent was removed by spin drying and the mixture was purified by column chromatography to obtain a brown solid in a yield of 15.3%. 1HNMR (400MHz, DMSO-d6) δ10.37(s,1H),9.95(s,1H),7.91(dd,J=8.6,2.2Hz,1H),7.82(d,J=2. 2Hz,1H),7.20(d,J=8.8Hz,1H),3.87(s,3H),3.61–3.57(m,2H),2.46–2.40(m,2H); LCMS(ESI)C 12 H 12 N2O5 + [M+H] + =265.25.

[0178] Step 2: Synthesis of tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)piperazine-1-carboxylate

[0179] 3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid (250 mg, 946 μmol, 1.0 eq), HATU (360 mg, 946 μmol, 1.0 eq), and N-BOC-piperazine (211 mg, 1.14 mmol, 1.2 eq) were weighed and placed in a 50 mL eggplant flask. DMF (10 mL) and DIEA (494 μL, 2.84 mmol, 3.0 eq) were added and stirred at room temperature for 4 hours. LCMS and TLC indicated the reaction was complete. The mixture was quenched with water (30 mL) and extracted with EA (10 mL x 3). The organic phase was washed with saturated NaCl solution (20 mL x 2), dried over anhydrous NaSO, and then spin-dried to obtain 159 mg of the crude product. 1 H NMR (400MHz, DMSO-d6) δ10.35(s,1H),7.41(dd,J=8.5,2.2Hz,1H),7.37(d,J=2.2Hz,1H),7.17(d,J=8.5 Hz,1H),3.85(s,3H),3.64–3.42(m,8H),2.91–2.85(m,2H),2.73–2.65(m,2H),1.41(s,9H); LCMS(ESI)C 21 H 28 N4O6 + [M+H] + =433.09.

[0180] Step 3: Synthesis of 1-(2-methoxy-5-(piperazine-1-carbonyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione hydrochloride

[0181] The compound tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)piperazine-1-carboxylate (159 mg, 368 μmol, 1.0 eq) was weighed into an eggplant-shaped flask, HCl·dioxane solution was added, and the mixture was stirred at room temperature for 3 hours. After LCMS showed that the reaction was complete, the solvent was dried and vacuum dried to give 126 mg of a crude product as a white solid, which was the target compound. 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),7.46(dd,J=8.5,2.2Hz,1H),7.43(d,J=2.1Hz,1H),7.18(d,J=8.6Hz,1H),3.85 (s,3H),3.77–3.67(m,4H),3.59–3.57(m,2H),3.25–3.20(m,1H),2.94–2.85(m,3H),2.72–2.64(m,2H).; LCMS(ESI)C 16 H 20 N4O4 + [M+H] + =333.27.

[0182] Example 7: N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(piperazin-1-yl)benzamide hydrochloride

[0183]

[0184] Step 1: Synthesis of tert-butyl 4-(3-fluoro-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate

[0185] 4-Bromo-2-fluoroacetophenone (500 mg, 2.15 mmol, 1.0 eq), N-BOC-piperazine (602 mg, 3.23 mmol, 1.5 eq), cesium carbonate (736 mg, 2.26 mmol, 1.05 eq), and BINAP (133 mg, 220 μmol, 0.1 eq) were placed in a 25 mL eggplant flask and toluene (10 mL) was added. Under a nitrogen atmosphere, palladium acetate (24 mg, 110 μmol, 0.05 eq) was added, the nitrogen was replaced three times, and the mixture was heated at 60°C for 20 hours. After LCMS and TLC showed complete consumption of the substrate, the reaction solution was filtered and concentrated, and purified by column chromatography to obtain a white solid in a 90.9% yield. 1 H NMR (400MHz, DMSO-d6) δ7.71 (dd, J=7.9, 0.5Hz, 1H), 6.82–6.72 (m, 2H), 3.76 (s, 3H), 3.45–3.40 (m, 4H), 1.41 (s, 9H); LCMS (ESI) C17 H 23 FN2O4 + [M+H] + =339.27.

[0186] Step 2: Synthesis of 4-(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-2-fluorobenzoic acid

[0187] The compound tert-butyl 4-(3-fluoro-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (630 mg, 1.86 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and MeOH (6 mL) was added followed by 2M NaOH solution (6 mL). The mixture was heated with stirring at 50°C for 5 hours. After LCMS showed that the reaction was complete, an appropriate amount of 1M dilute hydrochloric acid solution was added to adjust the pH to 5. The solid in the mixed system was filtered, the filter cake was washed with water, the filter residue was collected, and the residue was spin-dried to obtain 550 mg of a crude product as a white solid powder. 1 H NMR (400MHz, DMSO-d6) δ7.69 (dd, J=7.9, 0.5Hz, 1H), 6.85–6.64 (m, 2H), 3.48–3.39 (m, 8H), 1.41 (s, 9H); LCMS (ESI) C 16 H 21 FN2O4 + [M+H] + =324.19.

[0188] Step 3: Synthesis of tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)piperazine-1-carboxylate

[0189] The compound 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-fluorobenzoic acid (550 mg, 1.70 mmol, 1.0 eq) and HATU (646 mg, 1.70, 1.0 eq) were weighed into an eggplant-shaped flask, DMF (6 mL) was added and stirred at room temperature for 5 minutes, and then 3-aminopiperidine-2,6-dione hydrochloride (280 mg, 1.70 mmol, 1.0 eq) and DIEA (841 μL, 5.10 mmol, 3.0 eq) were added and stirred at room temperature for 3 hours. After LCMS showed that the reaction was complete, an appropriate amount of water was added to the reaction solution, filtered, the filter cake was washed with water, the filter residue was collected, and the crude product 700 mg was obtained after spin drying as a white solid. 1H NMR (400MHz, DMSO-d6) δ10.86(s,1H),8.10(dd,J=8.0,0.5Hz,1H),7.64(dd,J=8.0,1.3Hz,1H),6.88–6.76(m,2H),4.73(ddd, J=12.6,7.9,5.7Hz,1H),3.47–3.42(m,4H),3.32–3.28(m,4H),2.82–2.52(m,2H),2.17–1.98(m,2H),1.43(s,9H); LCMS(ESI)C 21 H 27 FN4O5 + [M+H] + =435.16.

[0190] Step 4: Synthesis of N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(piperazin-1-yl)benzamide hydrochloride

[0191] tert-Butyl 4-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)piperazine-1-carboxylate (700 mg, 1.61 mmol, 1.0 eq) was weighed into an eggplant-shaped flask. DCM (5 mL) and HCl·dioxane solution (5 mL) were added and stirred at room temperature for 3 hours. LCMS showed complete consumption of the substrate. The solvent was evaporated and the crude product was dried under vacuum to obtain a white powder, which was the target compound. 1 HNMR (400MHz, DMSO-d6) δ10.85(s,1H),9.29(s,2H),8.15(dd,J=8.0,0.5Hz,1H),7.66(dd,J=8.0,1.3Hz,1H),6.87(dd,J=1.3,0.5Hz,1H),4.73(dd, J=7.8,5.4Hz,1H),3.58–3.51(m,4H),3.22–3.13(m,4H),2.77(ddd,J=17. 2,13.3,5.6Hz,1H),2.53(d,J=3.9Hz,1H),2.18–1.96(m,2H); LCMS(ESI)C 16 H 19 FN4O3 + [M+H] + =335.15.

[0192] Example 8: 3-((3-Fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione hydrochloride

[0193]

[0194]

[0195] Step 1: Synthesis of tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate

[0196] tert-Butyl (1-(4-amino-2-fluorophenyl)piperidin-4-yl)carbamate (500 mg, 1.69 mmol, 1.0 eq) and 3-bromopiperidine-2,6-dione (427 mg, 5.08 mmol, 1.0 eq) were weighed into an eggplant-shaped flask and dissolved in DMF (8 mL). NaHCO₃ (1.42 g, 16.9 mmol, 10.0 eq) was then added and stirred at 80°C for 12 hours. After the reaction, the mixture was cooled to room temperature, water (25 mL) was added, and the mixture was extracted with EA (12 mL × 3). The organic phase was washed with saturated NaCl solution and dried over anhydrous Na₂SO₄. The mixture was then spin-dried and purified by column chromatography to obtain a green solid with a yield of 88.9%. 1 H NMR (400MHz, DMSO-d6) δ10.77(s,1H),6.83(dd,J=9.9,8.7Hz,1H),6.51(dd,J=14.9,2.6Hz,1H),6.41(dd,J=8.7,2.6Hz,1H),4.26(dd,J=7.7,4.8H z,1H),3.45–3.39(m,4H),2.81–2.75(m,4H),2.74–2.67(m,1H),2.60–2. 52(m,1H),2.16–2.02(m,1H),1.93–1.78(m,1H),1.41(s,9H); LCMS(ESI)C 20 H 27 FN4O4 + [M+H] + =407.31.

[0197] Step 2: Synthesis of 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione hydrochloride. Compound 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylic acid tert-butyl ester (610 mg, 1.56 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HCl·dioxane solution (5 mL) and DCM (5 mL) were added. The mixture was stirred at room temperature for 3 hours. After the reaction was completed as monitored by LCMS, the solvent was dried and vacuum-dried to obtain a pale green powder crude product, thereby obtaining the target compound. 1HNMR(400MHz, DMSO-d6)δ10.83(s,1H),6.98–6.89(m,1H),6.60(dd,J=14.9,2.6Hz,1H),6.51(dd,J=8.7,2.6Hz,1H),4.35–4.29(m, 1H),3.24–3.18(m,4H),3.16–3.09(m,4H),2.79–2.68(m,1H),2.62–2.54(m,1H),2.12–2.02(m,1H),1.94–1.81(m,1H); LCMS(ESI)C 15 H 19 FN4O2 + [M+H] + =307.28.

[0198] Example 9: 3-(1-oxo-5-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0199]

[0200]

[0201] Step 1: Synthesis of tert-butyl 4-(3-cyano-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate

[0202] 2-Cyanomethyl-4-fluoro-benzoic acid methyl ester (5.00 g, 27.91 mmol, 1.0 eq) and tert-butyl piperazine-1-carboxylate (6.20 g, 33.49 mmol, 1.2 eq) were weighed into a 250 mL eggplant-shaped flask, DMSO (100 mL) was added to dissolve the mixture, and then DIEA (15 mL, 84 mmol, 3.0 eq) was added. The reaction was stirred at 120° C. overnight. After LCMS and TLC monitoring results showed that the reaction was completed, water (100 mL) was added to quench the reaction, and the mixture was extracted with EA (100 mL×3). The organic phase was washed with saturated NaCl solution (200 mL×2), dried over anhydrous Na2SO4, and then spin-dried. The product was purified by column chromatography with a yield of 80.5%. 1 H NMR(400MHz, DMSO-d6)δ7.90(d,J=9.0Hz,1H),7.41(d,J=2.7Hz,1H),7.21(dd, J=9.1,2.7Hz,1H),3.82(s,3H),3.42(d,J=2.8Hz,8H),1.41(s,9H); LCMS(ESI)C 18 H 23 N3O4 + [M+H] + =346.07.

[0203] Step 2: Synthesis of tert-butyl 4-(3-formyl-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate

[0204] tert-Butyl 4-(3-cyano-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (7.00 g, 20.27 mmol, 1.0 eq) was weighed and added to an eggplant-shaped flask, followed by water (35 mL), pyridine (27 mL), and AcOH (3 mL). The system was purged with N2 three times before and after the addition of Raney nickel (20 mL). The reaction was allowed to proceed at 50°C overnight. LCMS indicated completion of the reaction. EA (50 mL) was added to dilute the reaction solution, which was then filtered and dried to afford a crude product (approximately 7.1 g). LCMS (ESI) indicated C 18 H 24 N2O5 + [M+H] + =349.18.

[0205] Step 3: Synthesis of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperazine-1-carboxylate

[0206] tert-Butyl 4-(3-formyl-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (7.1 g, 20.38 mmol, 1.0 eq) and 3-amino-2,6-piperidinedione hydrochloride (5.0 g, 30.57 mmol, 1.5 eq) were weighed into an eggplant-shaped flask and dissolved in DCM (140 mL). DIEA (7.1 mL, 40.76 mmol, 2.0 eq) and AcOH (3 mL) were then added. After stirring at room temperature for 2 hours, sodium cyanoborohydride (1.9 g, 30.57 mmol, 1.5 eq) was added and the reaction was allowed to proceed overnight at room temperature. LCMS indicated the reaction was complete. The mixture was quenched by the addition of saturated NaHCO₃ solution (150 mL) and extracted with DCM (40 mL x 3). The organic phase was washed with an appropriate amount of water, then with saturated NaCl solution, dried over anhydrous Na₂SO₄, and column chromatography to obtain a white solid in a yield of 17.2%. 1 H NMR (400MHz, DMSO-d6) δ10.97(s,1H),7.55(d,J=8.4Hz,1H),7.08(d,J=8.9Hz,2H),5.06(dd,J=13.3,5.1Hz,1H),4.40–4.15(m ,2H),3.54–3.45(m,4H),3.33–3.24(m,4H),3.01–2.54(m,2H),2.46–2.30(m,1H),2.01–1.92(m,1H),1.44(s,9H); LCMS(ESI)C 22 H 28N4O5 + [M+H] + =429.10;

[0207] Step 4: Synthesis of 3-(1-oxo-5-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0208] The compound tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperazine-1-carboxylate (1.40 g, 3.84 mmol, 1.0 eq) was weighed and added to an eggplant-shaped flask, and HCl·dioxane solution (15 mL) was added. After stirring at room temperature for 3 hours, LCMS showed that the reaction was complete. The product was spin-dried and freeze-dried to give a pale green solid crude product, which was the target compound, about 1.21 g. 1 H NMR (600MHz, DMSO-d6) δ10.95(s,1H),7.57(d,J=8.4Hz,1H),7.15(d,J=2.2Hz,1H),7.12(dd,J=8.6,2.3Hz,1H),5.05(dd,J=13.3,5.1 Hz,1H),4.40–4.19(m,2H),3.55–3.52(m,4H),3.22–3.18(m,4H),2.94–2.59(m,2H),2.42–2.32(m,1H),1.99–1.92(m,1H); LCMS(ESI)C 17 H 20 N4O3 + [M+H] + =329.21.

[0209] Example 10: 2-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)isoindoline-1,3-dione hydrochloride

[0210]

[0211] Step 1: Synthesis of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine-1-carboxylate

[0212] 2-(2,6-Dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (1.00 g, 3.62 mmol, 1.0 eq) and tert-butyl piperazine-1-carboxylate (670 mg, 3.62 mmol, 1.0 eq) were weighed into an eggplant-shaped flask. DMSO (16 mL) was added, followed by DIEA (1.8 mL, 10.86 mmol, 3.0 eq). The reaction was stirred at 90°C for 4 hours. After LCMS showed the reaction was complete, water (40 mL) was added to quench the reaction. The mixture was extracted with EA (20 mL x 3). The organic phase was then washed with saturated NaCl solution, dried over anhydrous Na2SO4, and dried by spin drying. The product was purified by column chromatography to obtain a yellow solid in a 63.8% yield. 1 H NMR(400MHz,DMSO-d6)δ11.11(s,1H),7.73–7.69(m,1H),7.41–7.32(m,2H),5.15–5.07(m,1H),3.50(s,4 LCMS(ESI)C 22 H 26 N4O6 + [M+H] + =443.33.

[0213] Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)isoindoline-1,3-dione hydrochloride

[0214] The compound tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine-1-carboxylate (1.02 g, 2.30 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HCl·dioxane solution (10 mL) was added. The mixture was reacted for 2 hours, dried by spin drying, and vacuum dried to obtain an orange powder to obtain the target compound. 1 HNMR(400MHz,DMSO-d6)δ11.13(s,1H),7.79–7.72(m,1H),7.48–7.39(m,2H),5.14–5.08(m,1H),3.53 –3.47(m,4H),3.30–3.23(m,4H),2.94–2.79(m,1H),2.64–2.52(m,2H),2.07–1.98(m,1H); LCMS(ESI)C 17 H 18 N4O4 + [M+H] + =344.29.

[0215] Example 11: 2-(1-(4-((2,6-dioxopiperidin-3-yl)amino)phenyl)-4-hydroxypiperidin-4-yl)acetic acid

[0216]

[0217] Step 1: Synthesis of tert-butyl 2-(4-hydroxy-1-(4-nitrophenyl)piperidin-4-yl)acetate

[0218] 2-(4-Hydroxypiperidin-4-yl)AcOH tert-butyl ester (hydrochloride) (1.88 g, 8.73 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and DMF (30 mL) was added. p-Fluoronitrobenzene (1.0 mL, 9.60 mmol, 1.1 eq) and DIEA (4.4 mL, 26.20 mmol, 3.0 eq) were then added. The reaction was stirred at 100°C overnight. After LCMS showed completion of the reaction, the product was quenched with water (90 mL) and extracted with EA (30 mL × 3). The organic phase was then washed with saturated NaCl solution (50 mL × 2), dried over anhydrous Na2SO4, and purified by column chromatography to give a yellow solid product in an 88.8% yield. 1 H NMR(400MHz,DMSO-d6)δ8.02(d,J=9.1Hz,2H),7.01(d,J=9.1Hz,2H),4.75(s,1H) ,3.80(d,J=13.3Hz,2H),2.34(s,2H),1.72–1.59(m,4H),1.34(s,9H); LCMS(ESI)C 17 H 24 N2O5 + [M+H] + =337.44.

[0219] Step 2: Synthesis of tert-butyl 2-(1-(4-aminophenyl)-4-hydroxypiperidin-4-yl)acetate

[0220] 2-(4-hydroxy-1-(4-nitrophenyl)piperidin-4-yl)acetic acid tert-butyl ester (2.61 g, 7.76 mmol, 1.0 eq) was weighed into an eggplant-shaped flask and dissolved in EA (25 mL). 10% Pd / C (200 mg) was then added, and the H2 solution was exchanged three times. The mixture was stirred at room temperature overnight. LCMS indicated the reaction was complete. The reaction mixture was diluted with EA, filtered, and dried to obtain a pale pink crude solid (approximately 2.18 g). 1HNMR(400MHz,DMSO-d6)δ6.71–6.63(m,2H),6.50–6.42(m,2H),4.53(s,2H),4.45(s,1H),3. 06–2.97(m,2H),2.89–2.78(m,2H),2.31(s,2H),1.77–1.53(m,4H),1.39(s,9H); LCMS(ESI)C 17 H 26 N2O3 + [M+H] + =307.10.

[0221] Step 3: Synthesis of tert-butyl 2-(1-(4-((2,6-dioxopiperidin-3-yl)amino)phenyl)-4-hydroxypiperidin-4-yl)acetate

[0222] The compound tert-butyl 2-(1-(4-aminophenyl)-4-hydroxypiperidin-4-yl)acetate (2.00 g, 6.53 mmol, 1.0 eq), 3-bromopiperidine-2,6-dione (1.88 g, 9.79 mmol, 1.5 eq), and NaHCO3 solid (1.65 g, 19.58 mmol, 3.0 eq) were weighed into an eggplant-shaped flask, and DMF (20 mL) was added to dissolve it. The mixture was stirred at 65 ° C for 16 hours. After the reaction was completed, it was quenched with water (60 mL), extracted with EA (35 mL × 3), and the organic phase was washed with saturated NaCl solution (60 mL × 2). Then, it was dried over anhydrous Na2SO4, spin-dried, and column chromatography gave a black solid product with a yield of 72.1%. 1 H NMR (400MHz, DMSO-d6) δ10.77(s,1H),6.74(d,J=8.5Hz,2H),6.58(d,J=8.4Hz,2H),4.46(s,1H),4.23–4.12(m,1H),3.11–3. 01(m,2H),2.90–2.85(m,2H),2.60–2.52(m,1H),2.32(s,2H),2.13–2.04(m,1H),1.93–1.53(m,6H),1.39(s,9H); LCMS(ESI)C 22 H 31 N3O5 + [M+H] + =418.18.

[0223] Step 4: Synthesis of 2-(1-(4-((2,6-dioxopiperidin-3-yl)amino)phenyl)-4-hydroxypiperidin-4-yl)acetic acid

[0224] The compound tert-butyl 2-(1-(4-((2,6-dioxopiperidin-3-yl)amino)phenyl)-4-hydroxypiperidin-4-yl)acetate (1.76 g, 5.74 mmol, 1.0 eq) was weighed and dissolved in 1,4-dioxane (4 mL). Then, HCl·dioxane solution (4 mL) was added and reacted at room temperature overnight. After the reaction was completed, the mixture was spin-dried and freeze-dried to obtain a dark green solid, which was the target compound. 1 H NMR(400MHz, DMSO-d6)δ6.76(d,J=2.1Hz,2H),6.75(d,J=2.0Hz,2H),3.61(s,1H),3.34–3.30(m,2H),2.89–2.8 7(m,2H),2.73–2.71(m,2H),2.58–2.54(m,1H),2.32(s,2H),2.08–2.04(m,1H),1.93–1.87(m,4H); LCMS(ESI)C 18 H 23 N3O5 + [M+H] + =362.09.

[0225] Example 12: 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid

[0226]

[0227]

[0228] Step 1: Synthesis of tert-butyl 1-(5-bromopyridin-2-yl)piperidine-4-carboxylate

[0229] 2-Fluoro-5-bromopyridine (1.7 mL, 17.05 mmol, 1.0 eq) and tert-butyl piperidine-4-carboxylate hydrochloride (5.68 g, 25.57 mmol, 1.5 eq) were weighed into an eggplant-shaped flask and dissolved in DMSO (40 mL). DIEA (12.1 mL, 68.18 mmol, 4.0 eq) was then added and stirred at 90°C for 24 hours. After completion of the reaction, the mixture was quenched with water (120 mL) and extracted with EA (50 mL × 3). The organic phase was washed with saturated NaCl solution (100 mL × 2), dried over anhydrous Na2SO4, and spin-dried. A white solid was obtained by column chromatography in a yield of 54.3%. 1H NMR (600MHz, DMSO-d6) δ8.14(dd,J=2.7,0.7Hz,1H),7.64(dd,J=9.1,2.6Hz,1H),6.83(dd,J=9.2,0.7Hz,1H),4.15–4.08 (m,2H),2.98–2.87(m,2H),2.47(tt,J=11.1,4.0Hz,1H),1.85–1.77(m,2H),1.50–1.43(m,2H),1.39(s,9H); LCMS(ESI)C 15 H 21 BrN2O2 + [M+H] + =341.18.

[0230] Step 2: Synthesis of tert-butyl 1-(2',6'-bis(benzyloxy)-[3,3'-bipyridyl]-6-yl)piperidine-4-carboxylate

[0231] The compounds tert-butyl 1-(5-bromopyridin-2-yl)piperidine-4-carboxylate (3.16 g, 9.26 mmol, 1.0 eq), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (4.25 g, 10.19 mmol, 1.1 eq), and Cs2CO3 (9.05 g, 27.18 mmol, 3.0 eq) were weighed into an eggplant-shaped flask, and ultra-dry 1,4-dioxane (50 mL) and water (10 mL) were added. After N2 was replaced three times, Pd(dppf)Cl2·DCM (113 mg, 0.14 mmol, 0.015 eq) was added, and N2 was replaced three more times. The system was stirred at 110°C for 16 hours. After the reaction was completed, the reaction solution was diluted with an appropriate amount of EA, filtered, spin-dried, and subjected to column chromatography to obtain a yellow solid with a yield of 33.0%. 1 H NMR (600MHz, DMSO-d6) δ8.28(d,J=2.5Hz,1H),7.74–7.69(m,2H),7.45–7.27(m,10H),6.85(d,J=8.9Hz,1H),6.53(d,J=8.1Hz,1H),5.39(s ,2H),5.36(s,2H),4.21–4.16(m,2H),2.96–2.91(m,2H),2.49–2.45(m,1H),1.86–1.79(m,2H),1.53–1.47(m,2H),1.40(s,9H); LCMS(ESI)C 34 H 37 N3O4 + [M+H] + =552.46.

[0232] Step 3: Synthesis of tert-butyl 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylate

[0233] Weigh tert-butyl 1-(2',6'-bis(benzyloxy)-[3,3'-bipyridyl]-6-yl)piperidine-4-carboxylate (1.69 g, 3.06 mmol, 1.0 eq) into an eggplant-shaped flask. Add EA (5 mL), EtOH (5 mL), and THF (10 mL). Then, add 10% Pd / C (1.5 g). Replace H2 three times and stir at room temperature overnight. After the reaction is complete, filter the mixture, wash the filter cake with a mixture of DCM and MeOH, and collect the filtrate to dryness to obtain a crude brown oil. 1 H NMR (600MHz, DMSO-d6) δ7.94(d,J=2.5Hz,1H),7.37(dd,J=8.8,2.5Hz,1H),6.80(d,J=8.9Hz,1H),4.22–4.07(m,2H),3.75–3.71(m,1H),2 .93–2.87(m,2H),2.71–2.60(m,1H),2.21–2.10(m,1H),2.00–1.94(m,1H),1.84–1.79(m,2H),1.51–1.45(m,2H),1.39(s,9H); LCMS(ESI)C 34 H 37 N3O4 + [M+H] + =552.46.

[0234] Step 4: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid

[0235] Weigh the compound tert-butyl 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylate (865 mg, 2.73 mmol, 1.0 eq), add HCl·dioxane solution (10 mL), and stir at room temperature overnight. After the reaction is completed, spin dry and vacuum dry to obtain a black solid, which is the target compound. 1H NMR (400MHz, DMSO-d6) δ10.91(d,J=2.6Hz,1H),7.90(d,J=2.7Hz,1H),7.77(dd,J=9.2,2.3Hz,1H),7.26(d,J=9.7Hz,1H),4.13–4.07( LCMS(ESI)C 16 H 19 N3O4 + [M+H] + =318.12.

[0236] Example 13: 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid

[0237]

[0238] Step 1: Synthesis of tert-butyl 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylate

[0239] 2-(2,6-Dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione (1.00 g, 3.62 mmol, 1.0 eq) and tert-butyl piperidine-4-carboxylate hydrochloride (872 mg, 4.71 mmol, 1.3 eq) were weighed into an eggplant-shaped flask, DMSO (10 mL) was added, and then DIEA (1.8 mL, 10.86 mmol, 3.0 eq) was added. The mixture was stirred at 90 ° C for 12 hours. After the reaction was completed, water (30 mL) was added to quench the reaction. The reaction solution was extracted with EA (15 mL × 3), and the organic phase was washed with saturated NaCl solution, dried over anhydrous Na2SO4, and then column chromatography was performed to obtain a yellow solid with a yield of 92.7%. 1H NMR (600MHz, DMSO-d6) δ11.07(s,1H),7.66(d,J=8.5Hz,1H),7.32(d,J=2.4Hz,1 H),7.24(dd,J=8.6,2.4Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),4.00–3.90(m,2H) ,3.11–3.02(m,2H),2.90–2.80(m,1H),2.62–2.58(m,1H),2.58–2.52(m,2H),2. 04–1.97(m,1H),1.90–1.83(m,2H),1.62–1.51(m,2H),1.40(s,9H); LCMS(ESI)C 23 H 27 N3O6 + [M+H] + =442.45.

[0240] Step 2: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid

[0241] The compound tert-butyl 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylate (1.48 g, 3.35 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HCl·dioxane solution (15 mL) was added. The mixture was stirred at room temperature overnight. After the reaction was completed, the crude product was dried to obtain the target compound. 1 HNMR (400MHz, DMSO-d6) δ11.08(s,1H),7.66(d,J=8.5Hz,1H),7.33(d,J=2.3Hz,1H),7.24(dd,J=8.6,2.4Hz,1H),5.09–5.03(m, 1H),3.12–3.05(m,2H),2.94–2.81(m,2H),2.56(dq,J=10.5,5.1,4.5Hz,2H),2.06–1.86(m,4H),1.65–1.54(m,2H); LCMS(ESI)C 19 H 19 N3O6 + [M+H] + =386.11.

[0242] Example 14: 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)-4-hydroxypiperidin-4-yl)acetic acid

[0243]

[0244] Step 1: Synthesis of tert-butyl 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)-4-hydroxypiperidin-4-yl)acetate

[0245] The compounds 1-(6-bromo-7-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (100 mg, 293 μmol, 1.0 eq), tert-butyl 2-(4-hydroxypiperidin-4-yl)acetate (130 mg, 602 μmol, 2.0 eq), and Cs2CO3 (244 mg, 749 μmol, 2.5 eq) were weighed into a pressure bottle, and ultra-dry 1,4-dioxane (3 mL) was added. Under a N2 atmosphere, Pd-PEPPSI-IPent(Cl) (14 mg, 15 μmol, 0.05 eq) was added, and the mixture was stirred at 110°C for 16 hours. After the reaction, the mixture was quenched with water (10 mL), extracted with EA (5 mL × 3), and the organic phase was washed with saturated NaCl solution (100 mL × 2), dried over anhydrous Na2SO4, and spin-dried. The product was obtained as a yellow solid by preparative thin-layer chromatography with a yield of 28.8%. 1 H NMR (600MHz, DMSO-d6) δ10.52(s,1H),7.32(d,J=12.8Hz,1H),7.10(d,J=7.1Hz,1H),4.59(s,1H),3.93(s,3H),3 .90–3.86(m,2H),3.17–3.01(m,4H),2.76–2.71(m,2H),2.38(s,2H),1.88–1.68(m,4H),1.42(s,9H); LCMS(ESI)C 23 H 30 FN5O5 + [M+H] + =476.29.

[0246] Step 2: Synthesis of 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)-4-hydroxypiperidin-4-yl)acetic acid

[0247] The compound tert-butyl 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)-4-hydroxypiperidin-4-yl)acetate (40 mg, 190 μmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HCl·dioxane solution (2 mL) was added. The mixture was stirred at room temperature overnight. After the reaction was completed, the crude product was dried to obtain the target compound.1 HNMR (400MHz, DMSO-d6) δ10.53(s,1H),7.33(d,J=12.8Hz,1H),7.12(d,J=7.0Hz,1H),3.94(s,3H),3.9 0–3.86(m,2H),3.18–2.99(m,4H),2.76–2.71(m,2H),2.45–2.39(m,2H),1.91–1.70(m,4H); LCMS(ESI)C 19 H 22 FN5O5 + [M+H] + =420.04.

[0248] Example 15: 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)piperidin-4-yl)acetic acid

[0249]

[0250] Step 1: Synthesis of tert-butyl 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)piperidin-4-yl)acetate

[0251] The compound 1-(6-bromo-7-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (100 mg, 293 μmol, 1.0 eq), 4-piperidinyl AcOH tert-butyl ester (120 mg, 602 μmol, 2.0 eq), and Cs2CO3 (244 mg, 749 μmol, 2.5 eq) were weighed into a sealed tube. Then, ultra-dry 1,4-dioxane (5 mL) was added, and the N2 atmosphere was evacuated three times. The catalyst Pd-PEPPSI-IPent(Cl) (14 mg, 15 μmol, 0.05 eq) was added, and the N2 atmosphere was evacuated three more times. The mixture was stirred at 110°C for 16 hours. After the reaction, the mixture was cooled to room temperature and quenched with water (10 mL). The mixture was extracted with EA (5 mL × 3). The organic phase was washed with saturated NaCl solution (100 mL × 2), dried over anhydrous Na2SO4, and spin-dried. The product was purified by preparative thin-layer chromatography to obtain a yellow solid with a yield of 34.4%. 1H NMR (600MHz, DMSO-d6) δ10.52(s,1H),7.33(d,J=12.7Hz,1H),7.09(d,J=7.1Hz,1H),3.93(s,3H),3.91–3.87 (m,2H),3.43–3.39(m,2H),2.75–2.65(m,4H),2.23–2.17(m,2H),1.86–1.74(m,4H),1.42(s,9H); LCMS(ESI)C 23 H 30 FN5O4 + [M+H] + =460.22.

[0252] Step 2: Synthesis of 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)piperidin-4-yl)acetic acid

[0253] The compound tert-butyl 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)piperidin-4-yl)acetate (45 mg, 95 μmol, 1.0 eq) was weighed into a 25 mL eggplant-shaped flask, and HCl·dioxane solution (2 mL) was added. The mixture was stirred at room temperature overnight. After LCMS showed that the reaction was complete, the crude product was dried to obtain the target compound. 1 HNMR(400MHz,DMSO-d6)δ10.53(s,1H),7.34(d,J=12.6Hz,1H),7.12(dd,J=7.1,2.4Hz,1H),3.93(s,3H),3 .91–3.85(m,2H),3.43–3.40(m,2H),2.75–2.67(m,4H),2.36–2.30(m,2H),1.91–1.72(m,4H); LCMS(ESI)C 19 H 22 FN5O4 + [M+H] + =404.07.

[0254] Example 16: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione

[0255]

[0256] Step 1: Synthesis of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate

[0257] 2-(2,6-Dioxo-piperidin-3-yl)-5-fluoro-isoindole-1,3-dione (1.00 g, 3.62 mmol, 1.0 eq) and tert-butyl piperazine-1-carboxylate (670 mg, 3.62 mmol, 1.0 eq) were weighed into an eggplant flask. DMSO (16 mL) was added, followed by DIEA (1.8 mL, 10.86 mmol, 3.0 eq). The reaction was stirred at 90°C for 4 hours. After LCMS indicated completion of the reaction, the mixture was quenched with water (40 mL) and extracted with EA (20 mL x 3). The organic phase was washed with saturated NaCl solution, dried over anhydrous Na2SO4, and then spin-dried. The product was purified by column chromatography to obtain a yellow solid in a 72.4% yield. HRMS (ESI): m / z calculated for C 22 H 26 N4O6[M+H] + 442.1852, found 442.1847.

[0258] Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione hydrochloride

[0259] Weigh tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate (1.02 g, 2.30 mmol, 1.0 eq) into an eggplant-shaped flask and add HCl·dioxane solution (10 mL). React for 2 hours, spin dry, and vacuum dry to obtain an orange powder, which is the target compound. HRMS (ESI): m / z calculated for C 17 H 18 N4O4[M+H] + 342.1328, found 342.1326.

[0260] Example 17: Synthesis of 2-fluoro-N-(1-methyl-2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)benzamide hydrochloride

[0261]

[0262] Step 1: Synthesis of tert-butyl (2,6-dioxopiperidin-3-yl)carbamate

[0263] 3-N-tert-Butyloxycarbonylamino-2,6-dioxopiperidine (1 g, 4.38 mmol, 1.0 eq) and K2CO3 (1.66 g, 13.14 mmol, 3.0 eq) were weighed and placed in an eggplant-shaped flask. DMF (15 mL) was added to dissolve the mixture. Iodomethane (818 μL, 13.14 mmol, 3.0 eq) was slowly added dropwise in an ice bath. The reaction system was then heated to reflux at 50°C for 3 hours. After LCMS and TLC showed the reaction was complete, water (30 mL) was added to quench the reaction. The mixture was then extracted with EA (10 mL × 3) and DCM (10 mL × 3), dried over anhydrous Na2SO4, and spin-dried to obtain 1.01 g of the crude product.

[0264] Step 2: Synthesis of 3-amino-1-methyl-2,6-piperidinedione hydrochloride

[0265] The compound (2,6-dioxopiperidin-3-yl)carbamic acid tert-butyl ester (1.01 g, 4.16 mmol, 1.0 eq) was weighed into an eggplant-shaped flask, and DCM (10 mL) was added to dissolve it. Then, HCl·dioxane (4 M, 10 mL) was added. After LCMS showed that the reaction was complete, the solution was spin-dried and dried in vacuo to obtain the crude product as a yellow solid.

[0266] Step 3: Synthesis of tert-butyl 4-(3-fluoro-4-((1-methyl-2,6-dioxopiperidin-3-yl)carbamoyl)phenyl)piperazine-1-carboxylate

[0267] 4-(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-2-fluorobenzoic acid (200 mg, 617 μmol, 1.0 eq) was weighed into an eggplant-shaped flask, and HATU (235 mg, 617 μmol, 1.0 eq) was added. 3-Amino-1-methyl-2,6-piperidinedione hydrochloride (146 mg, 740 μmol, 1.2 eq) was then added. After dissolution in DMF (10 mL), DIEA (322 μL, 1.85 mmol, 3.0 eq) was added and stirred at room temperature for 1 hour. After completion of the reaction as indicated by LCMS and TLC, water (20 mL) was added for quenching. The mixture was extracted with EA (5 mL x 3), dried over anhydrous NaSO, and purified by column chromatography to afford a pale purple solid.

[0268] Step 4: Synthesis of 2-fluoro-N-(1-methyl-2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)benzamide hydrochloride

[0269] The compound tert-butyl 4-(3-fluoro-4-((1-methyl-2,6-dioxopiperidin-3-yl)carbamoyl)phenyl)piperazine-1-carboxylate (200 mg, 446 μmol, 1.0 eq) was weighed into an eggplant flask, DCM (2 mL) was added, and HCl·dioxane (4 M, 2 mL) was added. The reaction was stirred at room temperature for 1 hour. After LCMS showed that the reaction was complete, the reaction solution was spin-dried and freeze-dried to give the crude product as a black solid.

[0270] III. Synthesis Methods of Various CDK2 / 4 / 6 Protein Hydrolysis Targeting Chimeras in the Present Invention

[0271] The compounds of Formula I of the present invention and / or their pharmaceutically acceptable salts can be synthesized using commercially available raw materials by synthetic techniques known in the art. The synthetic methods described below illustrate the preparation methods of most compounds. Salts, racemates, enantiomers, phosphates, sulfates, hydrochlorides, and prodrug forms of the compounds of Formula I of the present invention can be prepared by those skilled in the art according to conventional techniques in the art.

[0272] Synthesis method A: Weigh the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate (1.0 eq) and the E3 ubiquitin ligase ligand (1.5 eq) into an eggplant-shaped flask, add DMF, then add DIEA (5.0 eq), stir at room temperature overnight. After LCMS shows that the reaction is complete, add water to quench the reaction, extract with EA three times, wash the organic phase twice with saturated NaCl solution, dry with anhydrous Na2SO4, spin dry, purify by preparative thin layer chromatography, and freeze-dry to obtain the final product.

[0273] Synthesis method B: Weigh the compound pyrido[2,3-d]pyrimidin-7(8H)-one intermediate (1.0 eq), HATU (1.0 eq) and E3 ubiquitin ligase ligand (1.0 eq) into an eggplant-shaped flask, add DMF, then add DIEA (5.0 eq), stir at room temperature for 1 hour. After the reaction is completed, quench the reaction solution with water, extract three times with EA, and then wash the organic phase twice with saturated NaCl solution, dry over anhydrous Na2SO4, spin dry, purify by preparative thin layer chromatography, and then vacuum dry to obtain the final product.

[0274] Table 1 Substrates and synthesis methods of various CDK2 / 4 / 6 proteolysis targeting chimeras in the present invention

[0275]

[0276]

[0277]

[0278] Example 18: 3-(6-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)pyridin-3-yl)piperidine-2,6-dione (WYY-03-120)

[0279] Characterization results of WYY-03-120: 1 H NMR(600MHz,Chloroform-d)δ10.07(s,1H),8.61(s,1H),8.03(s,1H),7.72(d,J=7.8Hz,2H),7.54(d,J=8.0Hz,2H),7.31(d,J=8.8Hz,1H), 6.64(d,J=8.9Hz,1H),5.80–5.62(m,1H),3.92–3.48(m,10H),2.76–2 .42(m,11H),2.34–2.14(m,7H),2.11–2.03(m,2H),1.89–1.49(m,8H); 13 C NMR(151MHz,Chloroform-d)δ202.97,174.03,173.34,161.73,160.27,158.93,157.28,155.86,147.57,143.89,137.3 1,129.63,127.86,121.89,107.18,62.42,53.23,47.67,45.10,32.12,30.49,27.93,26.37,25.41,14.03; LCMS(ESI)C 41 H 49 N9O6S + [M+H] + =796.35.

[0280] Example 19: 3-(3-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (WYY-03-121)

[0281] Characterization results of WYY-03-121: 1H NMR(600MHz,Chloroform-d)δ9.85(s,1H),8.63(s,1H),7.72(d,J=7.9Hz,2H),7.5 4(d,J=7.9Hz,2H),7.25–7.18(m,1H),6.84(dd,J=8.3,2.4Hz,1H),6.76–6.72(m,1 H),6.67(d,J=7.5Hz,1H),5.79–5.67(m,1H),3.90–3.58(m,6H),3.26–3.14(m,4H) ,2.72–2.45(m,11H),2.33–2.16(m,7H),2.08(d,J=13.7Hz,2H),1.92–1.52(m,8H); 13 C NMR(151MHz,Chloroform-d)δ202.87,173.96,173.38,162.63,161.68,160 .19,157.23,155.78,151.65,143.91,142.63,138.20,129.67,129.53,127 .77,119.17,116.01,115.27,106.23,62.25,53.35,53.12,49.00,48.14,4 7.63,44.96,36.54,31.52,30.92,27.85,26.51,25.33,13.96;LCMS(ESI)C 42 H 50 N8O6S + [M+H] + =795.24.

[0282] Example 20: 1-(6-(1-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperidin-4-yl)-5-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (WYY-03-122)

[0283] Characterization results of WYY-03-122: 1H NMR (600MHz, DMSO-d6) δ10.33(d,J=5.3Hz,1H),8.75(d,J=4.3Hz,1H),7.72(d ,J=7.9Hz,2H),7.61(d,J=7.4Hz,2H),7.41–7.37(m,1H),7.35(d,J=2.1Hz,1H) ,5.94–5.54(m,1H),3.84(s,3H),3.76–3.44(m,11H),2.73–2.64(m,2H),2.40( d,J=25.0Hz,8H),2.23(d,J=6.5Hz,5H),1.99–1.87(m,3H),1.76–1.39(m,8H); 13 CNMR(151MHz,DMSO-d6)δ202.43,170.62,160.95,160.27,158.46,154.78 ,153.64,151.80,142.70,141.43,137.98,137.73,131.58,129.37,127.5 6,115.59,109.40,106.31,104.77,53.61,52.33,51.98,47.16,45.15,43 .94,35.58,31.26,30.93,30.29,27.20,25.45,24.58,13.53; LCMS(ESI)C 44 H 51 FN 10 O6S + [M+H] + =867.25.

[0284] Example 21: 1-(5-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazine-1-carbonyl)-2-methoxyphenyl)dihydropyrimidine-2,4(1H,3H)-dione (WYY-03-123)

[0285] Characterization results of WYY-03-123: 1H NMR (600MHz, DMSO-d6) δ10.33(s,1H),8.75(d,J=4.3Hz,1H),8.03(d,J=7.7Hz,1H), 7.72(d,J=7.9Hz,2H),7.61(d,J=7.4Hz,2H),7.39(dd,J=8.4,2.2Hz,1H),7.35(d,J= 2.1Hz,1H),5.94–5.54(m,1H),3.84(s,3H),3.75–3.43(m,11H),2.73–2.64(m,2H), 2.40(d,J=25.0Hz,8H),2.23(d,J=6.5Hz,5H),1.99–1.87(m,3H),1.76–1.39(m,8H); 13 C NMR(151MHz,DMSO-d6)δ170.85,168.17,161.00,160.30,158.52,155.85,152.14,142.75,133.72,129.54,128.70,128.16,127.87–127 .35(m),112.09,104.81,61.06,56.00,52.36,47.17,45.16,44.32,31.30,31.10,30.60,30.30,27.66,27.22,25.47,24.58; LCMS(ESI)C 43 H 51 N9O8S + [M+H] + =854.22.

[0286] Example 22: 4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (WYY-03-127)

[0287] Characterization results of WYY-03-127: 1H NMR(600MHz,DMSO-d6)δ10.85(s,1H),8.74(d,J=7.0Hz,1H),8.06–8.02(m,1H),7.7 3(d,J=7.9Hz,2H),7.65–7.59(m,3H),6.81(d,J=9.1Hz,1H),6.76(d,J=15.5Hz,1H), 5.90–5.61(m,1H),4.76–4.70(m,1H),3.90–3.55(m,6H),3.31(d,J=5.3Hz,4H),2.92 –2.70(m,2H),2.38(s,6H),2.27–2.05(m,7H),2.04–1.89(m,4H),1.75–1.42(m,8H); 13 C NMR(151MHz,DMSO-d6)δ202.51,173.04,172.30,163.04,162.18,161.00,160.55,160.30,154.83,154.34,142.75,133.75,131.66,129.52,12 7.58,109.80,104.82,100.77,100.59,61.11,52.27(d,J=39.3Hz),49. 78,47.20,46.78,45.16,31.31,31.01,27.25,24.61,24.16; LCMS(ESI)C 43 H 50 FN9O7S + [M+H] + =856.54.

[0288] Example 23: 3-((4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (WYY-03-128)

[0289] Characterization results of WYY-03-128: 1H NMR(600MHz,Chloroform-d)δ9.92(s,1H),8.61(s,1H),7.72(d,J=7.9Hz,2H),7.55(d,J=7.8 Hz,2H),6.89–6.79(m,1H),6.46–6.33(m,2H),5.81–5.64(m,1H),4.65(d,J=4.1Hz,1H),4.04 –3.95(m,1H),3.90–3.79(m,1H),3.72–3.61(m,3H),3.05–2.96(m,3H),2.84–2.72(m,2H),2. 67–2.56(m,5H),2.52–2.43(m,4H),2.34–2.04(m,8H),2.01–1.94(m,1H),1.91–1.49(m,9H); 13 C NMR(151MHz,Chloroform-d)δ202.99,173.08,172.37,161.74,160.24,157.23,156.16,155.90,142.96,132.06,129.79,129.59,127. 85,120.68,109.27,102.58,102.41,62.39,54.92,53.44,52.73,51.25,44.99,31.66,31.24,27.94,25.75,25.41,14.04; LCMS(ESI)C 42 H 50 FN9O6S + [M+H] + =828.43.

[0290] Example 24: 3-(5-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (WYY-03-129)

[0291] Characterization results of WYY-03-129: 1H NMR(600MHz,Chloroform-d)δ10.10(s,1H),8.59(s,1H),7.71(dd,J=12.3,8.3Hz,3H),7 .54(d,J=8.0Hz,2H),6.96(dd,J=8.8,2.1Hz,1H),6.87(s,1H),5.81–5.63(m,1H),5.22–5 .08(m,1H),4.41–4.21(m,2H),3.90–3.58(m,5H),3.37–3.27(m,4H),2.87–2.74(m,2H), 2.65–2.55(m,5H),2.47(s,3H),2.34–2.22(m,6H),2.19–2.01(m,4H),1.91–1.49(m,8H); 13 C NMR(151MHz,Chloroform-d)δ202.97,172.52,170.79,169.66,161.73,160.23, 157.23,155.86,154.42,143.82(d,J=6.8Hz),142.65,135.04,129.62,127.89, 125.07,122.06,115.61,108.49,106.18,62.30,53.43,52.97,51.83,48.40,47 .17,45.01,31.70(d,J=15.4Hz),31.09,27.93,25.40,23.59,14.03; LCMS(ESI)C 44 H 51 N9O7S + [M+H] + =850.20.

[0292] Example 25: 4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)benzyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (WYY-03-133)

[0293] Characterization results of WYY-03-133: 1H NMR(600MHz,DMSO-d6)δ11.08(s,1H),8.73(d,J=9.6Hz,1H),7.80–7.71(m,2H),7.70 –7.58(m,3H),7.34(d,J=7.1Hz,1H),7.31(d,J=8.3Hz,1H),5.94–5.52(m,1H),5.13– 5.00(m,1H),3.87–3.56(m,5H),3.35–3.14(m,4H),2.91–2.82(m,1H),2.66–2.54(m, 5H),2.46–2.30(m,5H),2.22(d,J=8.4Hz,5H),2.07–1.86(m,4H),1.76–1.40(m,8H); 13 C NMR(151MHz,DMSO-d6)δ202.49,172.82,170.00,167.06,166.34,161.02,1 60.28,158.50,154.80,149.61,142.78,135.91,133.67,129.61,127.60,12 3.73,116.57,114.93,105.25,104.84,61.25,52.50,52.03,50.44,48.84, 47.22,45.25,31.29,30.98,30.34,27.21,25.48,24.54,22.09; LCMS(ESI)C 44 H 49 N9O8S + [M+H] + =864.24.

[0294] Example 26: WWZ-01-102

[0295] Characterization results of WWZ-01-102: HRMS (ESI): m / z calcd for C 44 H 49 N9O8S[M+H] + 863.3425,found 863.3423.

[0296] Example 27: WYY-04-065

[0297] Characterization results of WYY-04-065: HRMS (ESI): m / z calcd for C 44 H 52 FN9O7S[M+H] + 870.3772, found 870.3766;1 H NMR(600MHz,Chloroform-d)δ8.56(s,1H),7.95–7.90(m,1H),7.75–7.70(m,2H),7.56–7.52( m,2H),7.52–7.48(m,1H),6.74–6.62(m,1H),6.55–6.44(m,1H),4.76–4.67(m,1H),3.91–3.83 (m,1H),3.63(s,3H),3.35–3.30(m,4H),3.17(s,3H),2.94–2.84(m,2H),2.82–2.76(m,1H),2. 66–2.61(m,1H),2.60–2.56(m,4H),2.52–2.42(m,4H),2.34–2.06(m,8H),1.93–1.53(m,10H). 13 C NMR(151MHz,Chloroform-d)δ202.93,172.21,171.62,163.74,163.33,161. 70,160.44,157.47,155.75,155.10,155.03,143.75,142.39,135.11,132.91 ,132.88,129.52,128.03,127.88,110.32,109.44,100.84,100.65,62.22,52.71,52.01,47.36,38.68,36.57,31.77,31.61,31.51,27.30,24.59,13.98.

[0298] Example 28: 3-((4-(4-(2-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidin-1-yl)-2-oxoethyl)-4-hydroxypiperidin-1-yl)phenyl)amino)piperidine-2,6-dione (WYY-03-182)

[0299] Characterization results of WYY-03-182: 1H NMR(600MHz,Chloroform-d)δ8.63(s,1H),7.16–6.81(m,2H),6.75–6.39(m,2H),5.86–5.71(m,1H),5.19–4.85(m,1H),4 .64(d,J=79.1Hz,1H),4.21–3.58(m,6H),3.30–2.90(m,8H),2.56–2.39(m,6H),2.38–2.04(m,10H),2.02–1.48(m,16H); 13 C NMR(151MHz,Chloroform-d)δ203.25,173.41,170.86,161.77,160.38,157.74,155.94,142.57,119.84,114.98 ,67.88,59.16,53.53,47.97,44.71,42.44,40.42,36.85,31.72,29.83,28.13,25.71,14.09,1.16; LCMS(ESI)C 43 H 57 N9O8S + [M+H] + =860.25.

[0300] Example 29: 3-(6-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidin-1-yl)piperidin-1-yl)pyridin-3-yl)piperidine-2,6-dione (WYY-03-183)

[0301] Characterization results of WYY-03-183: 1H NMR(600MHz,Chloroform-d)δ9.96(s,1H),8.65(s,1H),8.01(d,J=2.5Hz,1H),7.32(dd,J=8.9,2.5Hz,1 H),6.68(d,J=8.9Hz,1H),5.83–5.76(m,1H),4.76–4.70(m,1H),4.35–4.25(m,2H),4.13–4.00(m,2H),3. 82–3.71(m,2H),3.67–3.61(m,1H),3.18–3.06(m,4H),2.95–2.86(m,4H),2.76–2.69(m,2H),2.68–2.62 (m,1H),2.60–2.53(m,1H),2.50–2.49(m,3H),2.38–2.27(m,6H),2.26–1.93(m,11H),1.72–1.57(m,7H); 13 C NMR(151MHz,Chloroform-d)δ203.02,174.08,173.35,173.18,161.75,160.29,158.54,157.26,155.96,147.20,142. 64,137.54,121.61,107.58,59.48,44.95,40.83,38.74,31.86,31.39,29.64,28.13,26.52,25.63,22.81; LCMS(ESI)C 41 H 53 N9O7S + [M+H] + =816.30.

[0302] Example 30: 5-(4-(4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidin-1-formyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (WYY-03-184)

[0303] Characterization results of WYY-03-184: 1H NMR(600MHz,Chloroform-d)δ10.49(s,1H),8.65(d,J=8.9Hz,1H),7.65(d,J=8.4Hz,1H),7.2 7–7.22(m,1H),7.05(d,J=8.8Hz,1H),5.87–5.74(m,1H),4.99–4.90(m,1H),4.77–4.66(m,1H ),4.14–4.02(m,2H),3.88–3.64(m,3H),3.20–3.10(m,4H),2.95–2.90(m,2H),2.85–2.74(m, 4H),2.50(s,5H),2.38–2.16(m,9H),2.14–2.05(m,5H),2.01–1.96(m,3H),1.75–1.59(m,7H); 13 C NMR(151MHz,Chloroform-d)δ203.07,172.66,169.62,168.14,167.43,161.74,157.30,155.92,155.35,134.43,129.51,125.52,119 .02,118.12,108.73,59.22,55.40,53.54,49.20,47.38,44.42,40.85,40.48,37.90,31.68,28.06,27.09,25.67,22.92; LCMS(ESI)C 44 H 53 N9O9S + [M+H] + =884.56.

[0304] Example 31: 1-6-(4-(2-(4-(4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidin-1-yl)-2-oxoethyl)-4-hydroxypiperidin-1-yl)-5-fluoro-1-methyl-1H-indazole-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (WYY-03-185)

[0305] Characterization results of WYY-03-185: 1H NMR(600MHz,Chloroform-d)δ8.71(d,J=11.2Hz,1H),7.36–7.28(m,1H),6.8 3(d,J=6.8Hz,1H),5.91–5.76(m,1H),4.17–4.06(m,3H),4.01–3.92(m,3H), 3.86–3.65(m,2H),3.33–2.99(m,8H),2.93–2.83(m,2H),2.61–2.45(m,6H), 2.44–2.28(m,6H),2.18–2.08(m,3H),2.05–1.90(m,5H),1.88–1.54(m,11H); 13 C NMR(151MHz,Chloroform-d)δ203.02,170.72,161.77,160.27,157.24,155.99,152.26,151.61,142.88,141.11,138.79, 111.20,106.74,97.94,67.98,58.84,53.56,47.18,44.53,42.67,37.43,35.78,31.68,31.32,28.07,26.69; LCMS(ESI)C 44 H 56 FN 11 O8S + [M+H] + =918.26.

[0306] Example 32: 1-6-(4-(2-(4-(4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidin-1-yl)-2-oxoethyl)piperidin-1-yl)-5-fluoro-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (WYY-03-186)

[0307] Characterization results of WYY-03-186: 1H NMR(600MHz,Chloroform-d)δ8.66(d,J=14.5Hz,1H),7.32(d,J=12.2Hz,1H),6.78–6.53(m,1H),5.87–5.75(m,1H),4.13–3.92(m,5H),3.89–3.4 7(m,4H),3.31–2.91(m,6H),2.89–2.68(m,3H),2.63–2.49(m,4H),2.42 –2.20(m,8H),2.18–2.10(m,3H),2.04–1.88(m,5H),1.86–1.47(m,11H); 13 C NMR(151MHz,Chloroform-d)δ169.11,160.62,156.15,154.79,151.99,141.81,137.61,105.82,105.66,58. 09,52.39,50.55,43.46,39.49,34.68,31.69,31.41,30.55,30.18,28.68,26.92,25.66,24.68; LCMS(ESI)C 44 H 56 FN 11 O7S + [M+H] + =920.29.

[0308] 4. Biological activity detection experiment

[0309] Example 33: WB detection of the effect of PROTAC drugs on RB1 signaling

[0310] A549, NCI-H1299 and NCI-H1975 cells were seeded in 6-well plates (2.5×10 5 After the cells adhered, 0.5 μM of PROTAC drugs WYY-03-120, 121, 122, 123, 127, 128, 129, 133, 182, 183, 184, 185, and 186 were added. DMSO was used as a control. After 36 h of drug treatment, total cell protein was extracted for WB detection.

[0311] like Figures 1 to 3 As shown ( Figure 1102 is WWZ-01-102), which are the effects of the PROTAC drugs prepared in the examples of the present invention on the degradation ability of CDK2, 4, and 6 in A549, NCI-H1975, and NCI-H1299 cells, respectively, and on the phosphorylation level of RB1. It can be seen that the PROTAC drugs prepared in the examples of the present invention can target and degrade specific proteins (CDK2, 4, and 6). The bands at the positions of CDK2, 4, and 6, p-RB1 S807 / S811 (phosphorylation of RB1 protein at serine 807 / serine 811) and p-RB1-780 (phosphorylation of RB1 protein at serine 780) are somewhat weakened or even absent, especially WYY-03-120, 127, 185, and 186.

[0312] In addition, this experiment also studied the effects of PROTAC drugs on the degradation ability of CDK2, 4, and 6 and the phosphorylation level of RB1 over time (0, 5, 8, 12, and 24 hours) or drug concentration (100 nM, 500 nM, and 1 μM). The results are shown in Figure 4 and Figure 5 .like Figure 4 As shown in the figure, the degradation ability of the compounds WYY-03-127, WYY-03-120, WYY-03-185 and WYY-03-186 of the present invention on CDK2, 4, and 6 and the effect on the phosphorylation level of RB1 in A549 cells changes over time (the compound concentration is fixed at 0.5 μM). As time goes by, the degradation of CDK2, 4, and 6 is significant, and the phosphorylation level of RB1 decreases. Figure 5 As shown, the degradation ability of compounds WYY-03-127 and 3600 on CDK2, 4, and 6 in A549 cells and the effect on the phosphorylation level of RB1 vary with concentration (the positive control is PF-06873600, i.e., 3600 in the figure). As the concentration increases, the phosphorylation level of RB1 is significantly reduced compared with the positive control.

[0313]

[0314] A549 cells were seeded in 6-well plates. After the cells attached to the wall, one group was pretreated with 0.5 μM protease inhibitor MLN4924 for 2 h, and then WYY-03-127 (0.5 μM) was added. The other three groups were treated with DMSO, WYY-04-065 (0.5 μM), and WYY-03-127 (0.5 μM) for 24 h. Total cell protein was extracted for WB detection. Figure 6 Shown is the inhibitory effect of the proteasome inhibitor MLN4924 (0.5 μM) on the ubiquitination degradation ability of compound WYY-03-127 (WYY-04-065 is a negative control, i.e., 065 in the figure).

[0315]

[0316] Example 34: Clone formation experiment

[0317] A549 cells were seeded in 6-well plates at a density of 3000 cells / well. After the cells adhered, 0.5 μM PROTAC drugs (WYY-03-127 and WYY-03-120, PF-06873600 and Palbocilib as positive controls, and WYY-04-065 as a negative control) were added, along with DMSO as a control. The cells were cultured in a 37°C cell culture incubator for 6-9 days. After removing the culture medium, the cells were fixed with 4% formaldehyde and then stained with 0.1% crystal violet for 20 minutes. The cells were rinsed with water and allowed to dry naturally.

[0318] like Figure 7 As shown in FIG, the results of the clone formation experiment of the inhibitory effect of compounds WYY-03-127 and WYY-03-120 on the proliferation of A549 cells are shown. It can be seen that compared with the positive control, the compounds WYY-03-127 and WYY-03-120 of the present invention inhibit A549 cells more significantly, especially compound WYY-03-127.

[0319] Example 35: In vivo experiments in mice

[0320] 6-7 week old male BALB / c nude mice were purchased and raised under SPF conditions and acclimatized for 1 week before use. In the tumor formation experiment, the nude mice were divided into 3 groups, 5 in each group, and A549 cells (1.5×10 6 cells / mouse) were injected subcutaneously into the right axilla of BALB / c nude mice. Once tumors (average of about 50 mm) were detected, 3 ), the administration was started. Compound WYY-03-127 and PF-06873600 (positive control) powders were dissolved in 5% DMSO + 30% PEG300 + 5% Tween-80 + 60% saline. WYY-03-127 (25 mg / kg) and PF-06873600 (25 mg / kg) were intraperitoneally injected into mice, and the control group was given an equal amount of solvent (5% DMSO + 30% PEG300 + 5% Tween-80 + 60% saline). The drug was administered once every other day, and the size of the tumor was continuously measured with a vernier caliper. By V = 0.5 × L (length) × W (width) 2 The tumor volume was calculated and the mice were weighed.

[0321] like Figure 8As shown, the effects of intraperitoneal injection of the compound on tumor volume and mouse body weight in an A549 tumor cell mouse tumor model (PF-3600 in the figure is PF-06873600; the blank control is the solvent). It can be seen that compared with the positive control, the compound WYY-03-127 of the present invention can significantly reduce tumor volume and has little effect on mouse body weight.

[0322] Although the present invention has been described in detail through the above preferred embodiments, it should be understood that the above description is not intended to limit the present invention. After reading the above description, various modifications and substitutions of the present invention will become apparent to those skilled in the art. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. A CDK2 / 4 / 6 proteolysis targeting chimera or an isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph thereof, characterized in that: The chemical structure of the chimera is shown in Formula I: Wherein, R1 is selected from any one of the following structures: hydrogen, deuterium, cyano, halogen, amino, hydroxyl, nitro, CHF2, CF3, C(O)R a 、C(O)NHR a 、C(O)N(R a )2, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, 3-8 membered cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl, or at least one H in the amino, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C1-C6 alkoxy, 3-8 membered cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl is optionally substituted with Ra; Among them, the R a is selected from deuterium, nitro, cyano, hydroxy, amino, trifluoromethyl, halogen, C1-C6 alkyl or C1-C6 deuterated alkyl, or at least one H in the C1-C6 alkyl and C1-C6 deuterated alkyl is optionally substituted with halogen; R2 is selected from hydrogen, halogen, C1-C6 alkyl or C1-C6 deuterated alkyl, wherein at least one H in the C1-C6 alkyl is optionally substituted by halogen; R3 is selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, 3-7-membered cycloalkyl, C3-C8 alkenyl, 5-8-membered cycloalkenyl, 6-10-membered aryl, 5-10-membered heteroaryl, or at least one H in the aryl, C1-C8 alkyl, C1-C8 alkoxy, 3-7-membered cycloalkyl, C3-C8 alkenyl, 5-8-membered cycloalkenyl, 6-10-membered aryl, 5-10-membered heteroaryl is optionally replaced by R a replace; R4 is selected from any one of the following structures: wherein m and f are each independently an integer of 1-4, n is an integer of 0-14, W and Q are each independently selected from CH or N, and L is selected from O or S; Y is selected from -S(O)2-, -SO-, -S(O)2NR e -, -S-, -CH2- or -C(O)-; wherein R e is selected from hydrogen, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl, or at least one H in the C1-C6 alkyl, C1-C6 deuterated alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C6-C10 aryl or 5-10 membered heteroaryl is optionally replaced by R a replace; R5 is selected from any one of the following structures: wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N; Linker is selected from any of the following structures: f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, n1 is an integer of 1-14, W2 and Q2 are each independently selected from CH or N, and L1 is selected from O or S; The E3 ubiquitin ligase ligand is selected from any one of the following structures: In formula i, ring A is selected from any one of the following structures: where R b and R f are independently selected from any one of the following structures: hydrogen, alkyl, alkoxy, halogen, haloalkyl, haloalkoxy or cyano, or R b and R f Combined to form -C=O; R c 、R d are independently selected from any one of the following structures: hydrogen, alkyl, alkoxy, halogen, haloalkyl, haloalkoxy or cyano; G is selected from a single covalent bond, -O-, -C(=O)-, or -NH-; The C ring is selected from the following structures: m2 is an integer from 1 to 4; R h selected from hydrogen or alkyl; In formula ii, W3 is selected from CH or N; Z is selected from a single covalent bond connecting W and ring B, -CH2-, -NH-, -O- or -NHC(O)-, wherein the NH end of -NHC(O)- is connected to W; X is selected from a single covalent bond connecting the linker and the E3 ubiquitin ligase, alkylene, -O-, -(O-alkylene)-, -(alkylene-O)-, -NH-, -N(alkyl)-, -C(O)NH-; Ring B is selected from C6-C10 monocyclic aryl, C5-C10 monocyclic heteroaryl, C9-C10 fused bicyclic heteroaryl, or at least one H in the C6-C10 monocyclic aryl, C5-C10 monocyclic heteroaryl, C9-C10 fused bicyclic heteroaryl is optionally replaced by R a replace.

2. The CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to claim 1, characterized in that: The R1 is selected from any one of the following structures: C(O)R a 、C(O)NHR a or C(O)N(R a )2; wherein, the R a Selected from C1-C6 alkyl; Said R2 is selected from C1-C6 alkyl; Said R3 is selected from 3-7 membered cycloalkyl; The R4 is selected from wherein m and f are each independently an integer of 1-4, and W is selected from CH or N; Said Y is selected from -S(O)2-; R5 is selected from any one of the following structures: wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N; The Linker is selected from any one of the following structures: wherein f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, and W2 and Q2 are each independently selected from CH or N.

3. The CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to claim 1, characterized in that: The chemical structure of the chimera is shown in Formula II: The Linker is selected from any one of the following structures: f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, W2 and Q2 are each independently selected from CH or N; The R5 is selected from any one of the following structures: wherein f2 and m1 are each independently an integer of 1-4, j is an integer of 0-14, and W1 and Q1 are each independently CH or N.

4. The CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to claim 3, characterized in that: The Linker is selected from any one of the following structures: wherein j is an integer from 0 to 14, W2 and Q2 are each independently selected from CH or N; The R5 is selected from any one of the following structures: wherein f1, f3, and m3 are each independently an integer of 1 to 4, and W1 and Q1 are each independently selected from CH or N.

5. The CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to any one of claims 1 to 4, characterized in that: The E3 ubiquitin ligase ligand is selected from any one of the following structures:

6. The CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to claim 1, characterized in that: The chemical structure of the chimera is selected from any one of the following structures:

7. The method for preparing the CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to any one of claims 1 to 6, wherein: The method for preparing the chimera comprises: A pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 having a chemical structure as shown in Formula III, an intermediate 2 containing an E3 ubiquitin ligase ligand and a linker structure, and DIEA are reacted in an organic solvent with or without the presence of HATU to obtain the chimera.

8. A method for preparing a pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1, characterized in that: The chemical structure of the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 is shown in Formula IV or Formula V. The preparation method of the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 shown in Formula IV comprises: Step 1: Synthesis of 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one Under an inert atmosphere, 6-bromo-2-chloro-8-cyclopentyl-5-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one is reacted in N-methylpyrrolidone in the presence of di(tri-tert-butylphosphine)palladium at 45°C with tributyl(1-ethoxyvinyl)tin to obtain 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one; Step 2: Synthesis of 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one: 2-chloro-8-cyclopentyl-6-(1-ethoxyvinyl)-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one was dissolved in THF, and aqueous hydrochloric acid was added. The mixture was stirred at room temperature to obtain 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one. Step 3: Synthesis of tert-butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate 6-acetyl-2-chloro-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one and 1-tert-butoxycarbonyl-4-aminopiperidine were reacted in DMF with stirring at 80° C. in the presence of DIEA to obtain tert-butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate; Step 4: Synthesis of 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one tert-Butyl 4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate and dioxane hydrochloride solution were stirred at room temperature to react to obtain 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one; Step 5: Synthesis of pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula IV 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidin-7(8H)-one was reacted in DCM with the compound represented by formula d in an ice bath under the action of DIEA to obtain pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula IV; The preparation method of the pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 shown in Formula V comprises: Step 1: Synthesis of tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate 6-acetyl-8-cyclopentyl-5-methyl-2-(piperidin-4-ylamino)pyrido[2,3-d]pyrimidine-7(8H)- The ketone was reacted with butyl 4-chlorosulfonylpiperidinecarboxylate in anhydrous DCM in the presence of DIEA under ice-cooling and stirring at room temperature to obtain tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate; Step 2: Synthesis of pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula V Tert-butyl 4-((4-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-yl)sulfonyl)piperidine-1-carboxylate and HCl·dioxane solution were stirred and reacted at room temperature to obtain pyrido[2,3-d]pyrimidin-7(8H)-one intermediate 1 represented by formula V.

9. A pharmaceutical composition, characterized in that The method comprises the CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph as claimed in any one of claims 1 to 6.

10. Use of the CDK2 / 4 / 6 proteolysis targeting chimera or its isomer, pharmaceutically acceptable salt, solvate, prodrug, deuterated compound or polymorph according to any one of claims 1 to 6, or the pharmaceutical composition according to claim 7, in the preparation of a medicament for preventing and / or treating cancer.

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