Compounds, degradation agents targeting rad51 protein, and uses thereof

Abstract

This invention discloses compounds, degraders targeting RAD51 protein, and their applications, relating to the field of drug synthesis technology. The compounds of this invention are obtained by covalently linking a CRBN derivative with a RI-1 (5) or RI-2 (11) via a flexible linker. The compounds provided by this invention utilize a RI-1 or RI-2 derivative with RAD51 inhibitory activity to covalently link a CRBN ligand with a flexible linker of adjustable length, successfully constructing a novel PROTAC molecule. This compound can simultaneously bind to the target protein and E3 ligase, forming a ternary complex, achieving efficient and selective degradation of the target protein. This invention provides a new solution for treating drug-resistant cancers.

Description

Technical Field

[0001] This invention relates to the field of drug synthesis technology, and in particular to compounds, degrading agents targeting RAD51 protein, and their applications. Background Technology

[0002] RAD51 is a core molecule in DNA damage homologous recombination repair, mediating the formation of RAD51-ssDNA nucleoprotein filaments and their invasion into the DNA double helix. It forms the basis for subsequent d-loop formation and various damage repair pathways, and is highly correlated with tumor formation, metastasis, and drug resistance (Cells 2023, 12: 1169−1189). Studies have shown that specifically reducing RAD51 expression can sensitize tumor cells to radiotherapy and chemotherapy, reduce tumor cell invasion and metastasis, and that combination therapy with RAD51 inhibitors and PARP inhibitors has potential clinical therapeutic value (Nature 2023, 619: 650–657).

[0003] To date, there are relatively few reports on small molecule inhibitors with pharmacological activity at the cellular level targeting RAD51. Most existing RAD51 inhibitors exhibit poor or moderate activity. These compounds can affect the enzyme activity of RAD51 to varying degrees, thereby inhibiting tumor cell growth or enhancing treatment sensitivity when combined with radiotherapy and chemotherapy (BBA-Rev. Cancer, 2021, 1876: 18859–18867.). Representative RAD51 inhibitors include DIDS, IBR2, Halenaquinone, RS-1, RI-1, RI-2, BO2, RI(dl)-1, and Cambridge7m (OncoTargets Ther. 2022, 15: 1509–1518.). There are also no reports on RAD51 degrading agents, either domestically or internationally. Furthermore, protein degradation-targeting chimeric (PROTACs) technology has emerged as a highly promising new drug development technology, capable of achieving specific and efficient degradation of disease-related proteins, thereby achieving therapeutic effects (Nat. Rev. Cancer. 2021, 21: 638–654.). Targeting the degradation of the RAD51 protein in cancer using PROTACs technology, combined with radiotherapy and chemotherapy, holds promise for enhancing chemosensitivity, overcoming tumor drug resistance, and becoming a feasible new strategy in cancer treatment (Chem. Soc. Rev. 2022, 51: 5214−5236.). Given the unique mechanism of action of PROTAC degraders, they possess significant advantages in overcoming cancer drug resistance, reducing side effects, and targeting untreatable targets. It is necessary to utilize PROTACs technology to design and synthesize new compounds targeting RAD51 to develop new drugs for cancer treatment. Summary of the Invention

[0004] The technical problem to be solved by this invention is to address the defects or deficiencies mentioned in the background art by providing compounds, degrading agents targeting RAD51 protein, and their applications. This invention proposes the following technical solutions: A compound obtained by covalently linking a CRBN derivative with a flexible chain linker of formula RI-1 (5) or RI-2 (11); .

[0005] Furthermore, the compound has the following general structural formula: ; In the formula, X1 is selected from bromine, chlorine, hydroxyl, methyl, trifluoromethyl, straight-chain alkyl or p-methoxyphenyl; X2 is selected from hydrogen or fluorine; X3 is selected from hydrogen or fluorine; Y is O or none; M is CH2 or O; R is selected from halogen, methyl, methoxy, trifluoromethyl, phenyl, hydroxyl, amino, benzyl, nitro, nitrile, 2 / 3-pyrrole, 2 / 3-thienyl, 2 / 3-pyridyl, 2 / 3-furanyl, N-pyrazolyl or pyridyl; The value of m ranges from 0 to 10; the value of n ranges from 0 to 10.

[0006] Furthermore, the compound is selected from at least one of the compounds shown in the following structural formulas: .

[0007] The present invention also provides the use of the said compound, or a pharmaceutically acceptable salt thereof, in the preparation of a drug that targets and degrades the DNA homologous recombinant protein RAD51.

[0008] The present invention also provides the use of the said compound, or a pharmaceutically acceptable salt thereof, in the preparation of antitumor and cancer drugs.

[0009] Furthermore, the cancers mentioned include liver cancer.

[0010] The present invention also provides a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier; wherein the active ingredient is the compound or a pharmaceutically acceptable salt thereof.

[0011] The present invention also provides a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier; wherein the active ingredient is: the compound or a pharmaceutically acceptable salt thereof, and one or more therapeutically active ingredients.

[0012] The present invention also provides a targeted degradation agent for the DNA homologous recombinant protein RAD51, comprising the compound thereon or a pharmaceutically acceptable salt thereof.

[0013] Further, the pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt; the inorganic acid salt includes at least one of hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, nitrate, phosphate, and acid phosphate; the organic acid salt includes at least one of formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, salicylate, picrate, glutamate, ascorbate, camphorate, and camphorsulfonate.

[0014] Compared with the prior art, the technical effects achieved by the present invention include: The compounds provided by this invention are novel PROTAC molecules successfully constructed by covalently linking RI-1 or RI-2 derivatives with RAD51 inhibitory activity to a CRBN ligand via a flexible linker of adjustable length. These compounds can simultaneously bind to the target protein and E3 ligase, forming a ternary complex that utilizes the body's natural ubiquitin-proteasome system to achieve efficient and selective degradation of the target protein. The compounds designed using targeted protein chemical knockdown technology contain both a target protein inhibitor and a CRBN ligand, effectively delaying or overcoming drug resistance in tumor cells caused by target protein mutations or overexpression, providing a new solution for treating drug-resistant cancers. The introduction of the flexible linker helps to regulate the flexible structure of the compound, maximizing its degradation activity.

[0015] The compounds provided by this invention lay a solid foundation for the subsequent development of innovative drugs to treat cancer or diseases related to the specific target protein RAD51. Detailed Implementation

[0016] Unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of this invention, definitions of some terms are provided below. When the definitions and interpretations of terms provided in this invention differ from the meanings commonly understood by those skilled in the art, the definitions and interpretations provided in this invention shall prevail.

[0017] As used herein, unless the context clearly specifies otherwise, singular forms of words such as “a,” “a kind,” and “the” include their corresponding plural indicators.

[0018] Unless the context clearly specifies otherwise, the term “or” is used to mean the terms “and / or” and is used interchangeably with them.

[0019] In the following examples, unless otherwise specified, all starting materials and reagents are commercially available or synthesized according to known methods, and the commercially available materials and reagents are used directly without further purification.

[0020] Example 1 The synthetic route for preparing compound 4a is as follows: .

[0021] The specific steps are as follows: 32 mg (0.20 mmol) of tert-butyl 3-oxopropionate and 72 mg (0.20 mmol) of compound 1 were dissolved in a mixed solvent DCM / MeOH (5:1, v / v). The solution was cooled to 5 °C, and a few drops of HOAc were added. After stirring the reaction solution for 10 minutes, Na(AcO)3BH (128 mg, 0.6 mmol) was added. The temperature was slowly restored to room temperature, and the reaction was stirred for 12 h. A saturated NaHCO3 solution was added to the reaction mixture, and after stirring for 5 minutes, the mixture was extracted with dichloromethane. The organic phases were combined, concentrated, and purified by column chromatography (DCM / MeOH = 50:1 to 25:1) to give yellow compound 2a, with a yield of 64 mg and a yield of 79%.

[0022] Compound 3 (34 mg, 0.1 mmol) and intermediate compound 2a (40 mg, 0.1 mmol) were dissolved in DMF (2 mL), and condensing agents HATU (76 mg, 0.1 mmol) and DIPEA (0.2 mL) were added. After stirring, the mixture was allowed to react overnight at room temperature for about 10 h. After the reaction was completed by TLC monitoring, 15 mL of dichloromethane was added for extraction, followed by washing with saturated brine. The organic phases were combined, concentrated, and passed through a silica gel column to give compound 4a in a yield of 60 mg (81%) as a bright yellow solid.

[0023] 3-(4-(2-aminoethyl)piperazin-1-yl)-4-chloro-1-(3,4-dichlorophenyl)- 1H-pyrrole-2,5-dione (2a). Yellowish solid (64 mg, 79% yield). 1HNMR (600MHz, DMSO-d6): δ 7.18 (d, J = 6.0 Hz, 1H), 6.74 (s, 1H), 6.52 (d, J = 6.0 Hz,1H), 3.81-3.92 (m, 4H), 3.49-3.54 (m, 4H), 2.89-3.04 (m, 4H). N-(2-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)amino)acetamide (4a). Yellowish solid (30 mg, 81%yield). 1 HNMR (600 MHz, DMSO-d6): δ 11.12 (brs, 1H), 7.79 (d, J = 8.0 Hz,1H), 7.69 (s, 1H), 7.61-7.64 (m, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.12 (d, J =8.0 Hz, 1H), 7.08-7.10 (m, 1H), 5.06-5.10 (m, 1H), 4.25-4.26 (m, 2H), 3.93-4.01 (m, 3H), 3.70-3.72 (m, 3H), 3.15-3.16 (m, 2H), 2.88-2.94 (m, 2H), 2.44-2.46 (m, 2H), 2.04-2.06 (m, 2H), 1.22-1.25 (m, 2H). 13 C NMR (150 MHz, DMSO-d6): δ 173.4, 170.6, 169.1, 167.8, 167.4, 164.9, 163.8, 145.9, 142.2, 136.8,131.9, 131.8, 131.6, 131.4, 130.8, 129.0, 127.6, 118.8, 111.6, 110.2, 94.6,53.6, 49.1, 48.0, 44.4, 42.6, 39.4, 31.6, 22.5. HRMS (EI): calcd. for[C 31 H 29 Cl3N7O7] (M+H)+, 716.1189; found, 716.1180. Example 2 Compound 4b was prepared, and its structural formula is as follows: .

[0024] Following the method described in Example 1, compound 1 was reacted with the corresponding aminoaldehyde derivative, followed by a reducing amination reaction and TFA deprotection to synthesize intermediate 2b; intermediate 2b was then reacted with compound 3 to obtain compound 4b, totaling 53 mg with a yield of 72%, as a yellow solid powder.

[0025] 3-(4-(3-aminopropyl)piperazin-1-yl)-4-chloro-1-(3,4-dichlorophenyl)- 1H-pyrrole-2,5-dione (2b). Yellowish solid (66 mg, 81% yield). 1 HNMR (600MHz, DMSO-d6): δ 7.16 (d, J = 6.0 Hz, 1H), 6.75 (s, 1H), 6.54 (d, J = 6.0 Hz,1H), 3.82-3.91 (m, 4H), 3.49-3.53 (m, 4H), 2.90-3.04 (m, 4H), 171-1.72 (m, 2H). N-(3-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)propyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)amino)acetamide (4b). Yellowish solid (53 mg, 72%yield). 1 HNMR (600 MHz, DMSO-d6): δ 11.11 (brs, 1H), 7.80 (d, J = 8.0 Hz,1H), 7.70 (s, 1H), 7.61-7.64 (m, 1H), 7.49 (d, J = 8.0 Hz, 1H), 7.12 (d, J =8.0 Hz, 1H), 7.08-7.12 (m, 1H), 5.06-5.10 (m, 1H), 4.24-4.25 (m, 2H), 3.94-4.01 (m, 3H), 3.71-3.74 (m, 3H), 3.15-3.16 (m, 2H), 2.90-2.93 (m, 2H), 2.42-2.45 (m, 2H), 2.04-2.06 (m, 2H), 1.72-1.74 (m, 2H), 1.22-1.25 (m, 2H). 13C NMR (150 MHz, DMSO-d6): δ 173.6, 170.5, 169.2, 167.8, 167.6, 165.0, 163.8, 146.0,142.1, 136.8, 131.9, 131.8, 131.6, 131.5, HRMS (EI):calcd. for [C 32 H 31 Cl3N7O7] (M+H)+, 730.1345; found, 730.1346. Example 3 Compound 4c was prepared, and its structural formula is as follows: .

[0026] Following the method described in Example 1, compound 1 was reacted with the corresponding amino aldehyde derivative, followed by a reducing amination reaction and TFA deprotection to synthesize intermediate 2c; intermediate 2c was then reacted with compound 3 to obtain compound 4c, totaling 50 mg with a yield of 68%, as a yellow solid powder.

[0027] 3-(4-(4-aminobutyl)piperazin-1-yl)-4-chloro-1-(3,4-dichlorophenyl)- 1H-pyrrole-2,5-dione (2c). Yellowish solid (69 mg, 80% yield). 1 HNMR (600MHz, DMSO-d6): δ 7.14 (d, J = 6.0 Hz, 1H), 6.76 (s, 1H), 6.54 (d, J = 6.0 Hz,1H), 3.85-3.90 (m, 4H), 3.49-3.53 (m, 4H), 2.91-3.06 (m, 4H), 1.78-1.79 (m, 2H), 172-1.73 (m, 2H). N-(4-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)butyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)amino)acetamide (4c). Yellowish solid (50 mg, 68%yield). 1HNMR (600 MHz, DMSO-d6): δ 11.10 (brs, 1H), 7.80 (d, J = 8.0 Hz,1H), 7.71 (s, 1H), 7.63-7.65 (m, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.12 (d, J =8.0 Hz, 1H), 7.08-7.11 (m, 1H), 5.06-5.10 (m, 1H), 4.26-4.27 (m, 2H), 3.94-4.01 (m, 3H), 3.72-3.74 (m, 3H), 3.14-3.16 (m, 2H), 2.90-2.93 (m, 2H), 2.43-2.45 (m, 2H), 2.05-2.06 (m, 2H), 1.72-1.73 (m, 2H), 1.51-1.52 (m, 2H), 1.21-1.26 (m, 2H). 13 C NMR (150 MHz, DMSO-d6): δ 174.1, 170.5, 169.2, 168.0, 167.6,164.9, 163.8, 146.0, 142.2, 136.8, 132.0, 131.7, 131.6, 131.4, HRMS (EI): calcd. for [C 33 H 33 Cl3N7O7] (M+H)+, 744.1502; found, 744.1506. Example 4 Compound 4d was prepared, and its structural formula is as follows: .

[0028] Following the method described in Example 1, compound 1 was reacted with the corresponding aminoaldehyde derivative, followed by a reducing amination reaction and TFA deprotection to synthesize intermediate 2d; intermediate 2d was then reacted with compound 3 to obtain compound 4d, totaling 48 mg, with a yield of 63%, as a yellow solid powder.

[0029] 3-(4-(5-aminopentyl)piperazin-1-yl)-4-chloro-1-(3,4-dichlorophenyl)- 1H-pyrrole-2,5-dione (2d). Yellowish solid (64 mg, 72% yield). 1 HNMR (600MHz, DMSO-d6): δ 7.16 (d, J = 6.0 Hz, 1H), 6.77 (s, 1H), 6.54 (d, J = 6.0 Hz,1H), 3.84-3.89 (m, 4H), 3.49-3.52 (m, 4H), 2.92-3.05 (m, 4H), 1.78-1.80 (m,2H), 172-1.74 (m, 2H), 1.35-1.36 (m, 2H). N-(5-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)pentyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)amino)acetamide (4d). Yellowish solid (48 mg, 63%yield). 1 HNMR (600 MHz, DMSO-d6): δ 11.11 (brs, 1H), 7.80 (d, J = 8.0 Hz,1H), 7.72 (s, 1H), 7.62-7.64 (m, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.16 (d, J =8.0 Hz, 1H), 7.08-7.11 (m, 1H), 5.09-5.12 (m, 1H), 4.26-4.27 (m, 2H), 3.93-4.02 (m, 3H), 3.72-3.74 (m, 3H), 3.15-3.18 (m, 2H), 2.90-2.94 (m, 2H), 2.43-2.46 (m, 2H), 2.05-2.08 (m, 2H), 1.72-1.74 (m, 2H), 1.51-1.53 (m, 2H), 1.27-1.30 (m, 2H), 1.22-1.25 (m, 2H). 13C NMR (150 MHz, DMSO-d6): δ 174.9, 170.6,169.2, 168.1, 167.7, 164.9, 163.9, 146.2, 142.2, 136.9, 132.1, 131.9, 131.6,131.3, HRMS (EI): calcd. for[C 34 H 35 C l3 N7O7] (M+H)+, 758.1658; found, 758.1660. Example 5 Compound 4e was prepared, and its structural formula is as follows: .

[0030] Following the method described in Example 1, compound 1 was reacted with the corresponding aminoaldehyde derivative, followed by a reducing amination reaction, and TFA deprotection to synthesize intermediate 2e; intermediate 2e was then reacted with compound 3 to obtain compound 4e, totaling 56 mg with a yield of 73%, as a yellow solid powder.

[0031] 3-(4-(2-(2-aminoethoxy)ethyl)piperazin-1-yl)-4-chloro-1-(3,4- dichlorophenyl)-1H-pyrrole-2,5-dione (2e). Yellowish solid (72 mg, 81%yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.18 (d, J = 6.0 Hz, 1H), 6.78 (s, 1H), 6.56 (d, J = 6.0 Hz, 1H), 3.84-3.89 (m, 4H), 3.49-3.52 (m, 4H), 3.35-3.36 (m,2H), 2.92-3.05 (m, 4H), 2.78-2.80 (m, 2H). N-(2-(2-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin -4-yl)amino)acetamide (4e). Yellowish solid (56 mg, 73%yield). 1HNMR (600 MHz, DMSO-d6): δ 11.10 (brs, 1H), 7.80 (d, J = 8.0 Hz,1H), 7.74 (s, 1H), 7.63-7.64 (m, 1H), 7.56 (d, J = 8.0 Hz, 1H), 7.16 (d, J =8.0 Hz, 1H), 7.08-7.10 (m, 1H), 5.09-5.12 (m, 1H), 4.24-4.26 (m, 2H), 3.93-4.02 (m, 3H), 3.72-3.74 (m, 3H), 3.15-3.20 (m, 2H), 3.05-3.08 (m, 2H), 2.90-2.96 (m, 2H), 2.72-2.74 (m, 2H), 2.42-2.46 (m, 2H), 2.22-2.28 (m, 2H), 1.29-1.35 (m, 2H). 13 C NMR (150 MHz, DMSO-d6): δ 175.0, 170.8, 169.2, 168.4, 167.8,164.9, 163.9, 146.6, 142.3, 136.9, 132.4, 131.9, 131.4, 131.3, HRMS (EI): calcd. for [C 33 H 33 C l3 N7O8] (M+H)+, 760.1451; found,760.1454. Example 6 The synthetic route for preparing compound 6a is as follows: Compound 5 (33 mg, 0.1 mmol) and intermediate compound 2a (40 mg, 0.1 mmol) were dissolved in DMF (2 mL), and condensing agents HATU (76 mg, 0.2 mmol) and DIPEA (0.2 mL) were added. After stirring, the mixture was allowed to react overnight at room temperature for about 10 h. After the reaction was completed by TLC monitoring, 15 mL of dichloromethane was added for extraction, followed by washing with saturated brine. The organic phases were combined, concentrated, and passed through a silica gel column to give compound 6a in a yield of 50 mg (69%) as a bright yellow solid.

[0032] N-(2-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl) piperazin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)oxy) acetamide (6a). Yellowish solid (50 mg, 69% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.16 (brs, 1H), 7.79-7.82 (m, 2H), 7.70 (d, J =8.0 Hz), 1H), 7.51-7.54 (m, 1H), 7.44 (d, J = 8.0 Hz, 2H), 5.26-5.27 (m, 2H),5.10-5.14 (m, 1H), 3.94-4.06 (m, 4H), 3.62-3.78 (m, 3H), 3,11-3.12 (m, 2H),2.88-2.91 (m, 2H), 2.42-2.46 (m, 2H), 2.03-2.09 (m, 1H), 1.25-1.27 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 173.4, 170.4, 167.9, 165.7, 165.4, 164.8, 164.0,156.1, 142.6, 137.4, 133.6, 131.9, 131.4, 131.3, 130.6, 128.7, 127.4, 120.6,116.8, 115.6, 94.6, 66.5, 56.7, 49.8, 48.0, 44.9, 42.0, 40.7, 31.8, 22.9.HRMS (EI): calcd. for [C 31 H 28 [Cl3N6O8] (M+H +, 717.1029; found, 717.1030. Example 7 Compound 6b was prepared, and its structural formula is as follows: .

[0033] Following the method described in Example 6, intermediate 2b was reacted with compound 5 to obtain compound 6b, totaling 52 mg, with a yield of 71%, as a yellow solid powder.

[0034] N-(3-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)propyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)oxy)acetamide (6b). Yellowish solid (52 mg, 71% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.14 (brs, 1H), 7.80-7.82 (m, 2H), 7.72 (d, J =8.0 Hz), 1H), 7.51-7.53 (m, 1H), 7.46 (d, J = 8.0 Hz, 2H), 5.26-5.28 (m, 2H),5.10-5.16 (m, 1H), 3.94-4.05 (m, 4H), 3.64-3.79 (m, 3H), 3,11-3.12 (m, 2H),2.89-2.91 (m, 2H), 2.41-2.46 (m, 2H), 2.04-2.08 (m, 1H), 1.71-1.73 (m, 2H), 1.25-1.28 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 173.6, 170.7, 167.9, 165.8,165.4, 164.9, 164.0, 156.6, 142.7, 137.8, 133.6, 131.9, 131.6, 131.3, 130.9,128.7, 128.4, 120.6, 116.9, 115.6, 94.6, 67.5, 568, 49.8, 49.0, 44.9, 42.0,40.8, 31.8, 31.2, 23.6. HRMS (EI): calcd. for [C 32 H 30 C l3[N6O8] (M+H) + , 731.1185;found, 731.1186. Example 8 Compound 6c was prepared, and its structural formula is as follows: .

[0035] Following the method described in Example 6, intermediate 2c was reacted with compound 5 to obtain compound 6c, totaling 54 mg, with a yield of 72%, as a yellow solid powder.

[0036] N-(4-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)butyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)oxy)acetamide (6c). Yellowish solid (54 mg, 72% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.15 (brs, 1H), 7.80-7.83 (m, 2H), 7.72 (d, J =8.0 Hz), 1H), 7.51-7.54 (m, 1H), 7.47 (d, J = 8.0 Hz, 2H), 5.27-5.29 (m, 2H),5.10-5.17 (m, 1H), 3.95-4.04 (m, 4H), 3.63-3.79 (m, 3H), 3,10-3.12 (m, 2H),2.90-2.91 (m, 2H), 2.42-2.47 (m, 2H), 2.04-2.09 (m, 1H), 1.71-1.74 (m, 2H),1.51-1.53 ​​(m, 2H), 1.26-1.28 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 173.6,170.8, 167.9, 165.9, 165.4, 165.0, 164.9, 156.8, 142.7, 138.6, 133.6, 132.0,131.6, 131.4, 130.9, 128.8, 128.1, 120.6, 117.0, 115.6, 94.8, 67.5, 568,49.8, 49.0, 46.6, 42.0, 40.9, 31.8, 31.6, 27.6, 23.9. HRMS (EI): calcd. for[C33 H 32 C l3 [N6O8] (M+H) + , 745.1342; found, 745.1344. Example 9 Compound 6d was prepared, and its structural formula is as follows: .

[0037] Following the method described in Example 6, intermediate 2d was reacted with compound 5 to obtain compound 6d, totaling 52 mg, with a yield of 68%, as a yellow solid powder.

[0038] N-(5-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)pentyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)oxy)acetamide (6d). Yellowish solid (52 mg, 68% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.14 (brs, 1H), 7.80-7.84 (m, 2H), 7.74 (d, J =8.0 Hz), 1H), 7.50-7.53 (m, 1H), 7.48 (d, J = 8.0 Hz, 2H), 5.27-5.29 (m, 2H),5.10-5.16 (m, 1H), 3.98-4.03 (m, 4H), 3.64-3.76 (m, 3H), 3,10-3.12 (m, 2H),2.88-2.90 (m, 2H), 2.42-2.46 (m, 2H), 2.04-2.08 (m, 1H), 1.72-1.74 (m, 2H),1.51-1.54 (m, 2H), 1.34-1.36 (m, 2H), 1.26-1.28 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.1, 171.4, 168.4, 166.4, 165.4, 165.1, 164.9, 156.9, 142.7,138.6, 134.2, 132.0, 131.9, 131.4, 130.9, 128.9, HRMS (EI): calcd. for [C 34 H 34 C l3 [N6O8] (M+H) + , 759.1498; found, 759.1498. Example 10 Compound 6e was prepared, and its structural formula is as follows: .

[0039] Following the method described in Example 6, intermediate 2e was reacted with compound 5 to obtain compound 6e, totaling 48 mg, with a yield of 64%, as a yellow solid powder.

[0040] N-(2-(2-(4-(4-chloro-1-(3,4-dichlorophenyl)-2,5-dioxo-2,5-dihydro-1H- pyrrol-3-yl)piperazin-1-yl)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin -4-yl)oxy)acetamide (6e). Yellowish solid (48 mg, 64% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.16 (brs, 1H), 7.82-7.86 (m, 2H), 7.76 (d, J =8.0 Hz), 1H), 7.50-7.53 (m, 1H), 7.49 (d, J = 8.0 Hz, 2H), 5.28-5.29 (m, 2H),5.10-5.16 (m, 1H), 3.98-4.02 (m, 4H), 3.68-3.77 (m, 3H), 3,11-3.14 (m, 2H),2.96-2.98 (m, 2H), 3.41-2.47 (m, 2H), 2.05-2.08 (m, 2H), 1.74-1.78 (m, 2H),2.51-2.56 (m, 2H), 3.24-3.36 (m, 2H). 13C NMR (150 MHz, DMSO- d 6 ): δ 174.3,171.0, 168.5, 166.4, 165.6, 165.1, 164.9, 157.0, 142.7, 138.8, 134.2, 132.1,131.9, 131.6, 130.9, 128.9, 128.7, 120.6, 118.0, 116.4, 94.9, 68.6, 56.8,50.9, 49.0, 46.8, 42.6, 32.8, 31.6, 29.6. HRMS (EI): calcd. for [C 33 H 32 C l3 [N6O9](M+H) + , 761.1291; found, 761.1294. Example 11 The synthetic route for preparing compound 9a is as follows: The specific steps are as follows: (1) 32 mg (0.20 mmol) of tert-butyl 3-oxopropionate and 86 mg (0.20 mmol) of compound 7 were weighed and dissolved in a mixed solvent DCM / MeOH (5:1, v / v). The solution was cooled to 5 °C, and a few drops of HOAc were added. After stirring the reaction solution for 10 minutes, Na(AcO)3BH (128 mg, 0.6 mmol) was added. The temperature was slowly restored to room temperature, and the reaction was stirred for 12 h. A saturated NaHCO3 solution was added to the reaction mixture, and after stirring for 5 minutes, the mixture was extracted with dichloromethane. The organic phases were combined, concentrated, and purified by column chromatography (DCM / MeOH = 50:1 to 25:1) to give yellow compound 8a, with a yield of 70 mg and a yield of 74%.

[0041] (2) Compound 3 (34 mg, 0.1 mmol) and intermediate compound 8a (48 mg, 0.1 mmol) were dissolved in DMF (2 mL), and condensing agents HATU (76 mg, 0.1 mmol) and DIPEA (0.2 mL) were added. After stirring, the mixture was allowed to react overnight at room temperature for about 10 h. After the reaction was completed by TLC monitoring, 15 mL of dichloromethane was added for extraction, followed by washing with saturated brine. The organic phases were combined and concentrated, and then passed through a silica gel column to obtain compound 9a, with a yield of 49 mg and a yield of 62%, as a bright yellow solid.

[0042] 3-(4-(2-aminoethyl)piperazin-1-yl)-1-(3,4-dichlorophenyl)-4-(4- methoxyphenyl) -1H-pyrrole-2,5-dione (8a). Yellowish solid (70 mg, 74%yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.42-7.43 (m, 2H), 7.17 (d, J = 6.0 Hz,1H), 6.81-6.83 (m, 2H), 6.76 (s, 1H), 6.54 (d, J = 6.0 Hz, 1H), 3.81-3.91 (m,4H), 3.50-3.54 (m, 4H), 2.90-3.03 (m, 4H). N-(2-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)- 1,3-dioxo-isoindolin-4-yl)amino)acetamide (9a). Yellowish solid (49 mg, 62%yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.12 (brs, 1H), 7.80 (d, J = 8.0 Hz, 1H),7.76 (d, J = 8.0 Hz, 2H), 7.69 (s, 1H), 7.64-7.68 (m, 1H), 7.58 (d, J = 8. 0Hz, 2H), 7.45 (d, J = 8.0 Hz, 1H), 7.12 (d, J = 8.0 Hz, 1H), 7.09-7.11 (m,1H), 5.08-5.10 (m, 1H), 4.26-4.28 (m, 2H), 3.94-4.03 (m, 3H), 3.70-3.74 (m,3H), 3.65 (s, 3H), 3.12-3.14 (m, 2H), 2.88-2.95 (m, 2H), 2.42-2.45 (m, 2H),2.04-2.06 (m, 2H), 1.24-1.26 (m, 2H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 173.8,170.8, 169.6, 168.0, 167.6, 165.8, 164.9, 154.7, 145.9, 144.2, 137.4, 133.5,131.8, 131.7, 131.4, 130.6, 129.6, HRMS (EI):calcd. for [C 38 H 36 C l2 [N7O8] (M+H) + , 788.1997; found, 788.1998. Example 12 Compound 9b was prepared, and its structural formula is as follows: .

[0043] Following the method described in Example 11, intermediate 8b was reacted with compound 3 to obtain compound 9b, totaling 58 mg, with a yield of 72%, as a yellow solid powder.

[0044] 3-(4-(3-aminopropyl)piperazin-1-yl)-1-(3,4-dichlorophenyl)-4-(4- methoxy phenyl)-1H-pyrrole-2,5-dione (8b). Yellowish solid (79 mg, 81%yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.42-7.45 (m, 2H), 7.18 (d, J = 6.0 Hz,1H), 6.81-6.84 (m, 2H), 6.78 (s, 1H), 6.56 (d, J = 6.0 Hz, 1H), 3.82-3.91 (m,4H), 3.49-3.53 (m, 4H), 2.90-3.06 (m, 4H), 1.72-1.74 (m, 2H). N-(3-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)propyl)-2-((2-(2,6-dioxopiperidin-3- yl)-1,3-dioxoisoindolin-4-yl)amino)acetamide (9b). Yellowish solid (58 mg,72% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.11 (brs, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.70 (s, 1H), 7.63-7.69 (m, 1H), 7.58 (d, J =8. 0 Hz, 2H), 7.44 (d, J = 8.0 Hz, 1H), 7.18 (d, J = 8.0 Hz, 1H), 7.09-7.13(m, 1H), 5.08-5.09 (m, 1H), 4.26-4.29 (m, 2H), 3.95-4.03 (m, 3H), 3.71-3.76(m, 3H), 3.66 (s, 3H), 3.13-3.16 (m, 2H), 2.88-2.98 (m, 2H), 2.42-2.45 (m,2H), 2.04-2.08 (m, 2H), 1.72-1.74 (m, 2H), 1.23-1.27 (m, 2H). 13 C NMR (150MHz, DMSO- d 6 ): δ 173.9, 170.8, 170.6, 168.6, 167.4, 165.8, 164.4, 154.8,145.9, 144.2, 138.2, 133.5, 132.6, 131.7, 131.6, 130.6, 129.8, 129.5, 128.8,128.3, 118.7, 114.6, 111.5, 110.8, 94.6, 56.2, 53.8, 49.5, 48.4, 44.5, 42.5,39.6, 31.9, 30.6, 22.9. HRMS (EI): calcd. for [C 39 H 38 [Cl2N7O8] (M+H + , 802.2153;found, 802.2156. Example 13 Compound 9c was prepared, and its structural formula is as follows: .

[0045] Following the method described in Example 11, intermediate 8c was reacted with compound 3 to obtain compound 9c, totaling 53 mg, with a yield of 65%, as a yellow solid powder.

[0046] 3-(4-(4-aminobutyl)piperazin-1-yl)-1-(3,4-dichlorophenyl)-4-(4- methoxyphenyl)-1H-pyrrole-2,5-dione (8c). Yellowish solid (74 mg, 74% yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.42-7.46 (m, 2H), 7.20 (d, J = 6.0 Hz, 1H),6.81-6.84 (m, 2H), 6.78 (s, 1H), 6.57 (d, J = 6.0 Hz, 1H), 3.82-3.90 (m, 4H),3.49-3.51 (m, 4H), 2.90-3.06 (m, 4H), 1.71-1.73 (m, 2H), 1.51-1.52 (m, 2H). N-(4-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)butyl)-2-((2-(2,6-dioxopiperidin-3-yl)- 1,3-dioxoisoindolin-4-yl)amino)acetamide (9c). Yellowish solid (53 mg, 65%yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.12 (brs, 1H), 7.79 (d, J = 8.0 Hz, 1H),7.78 (d, J = 8.0 Hz, 2H), 7.70 (s, 1H), 7.64-7.69 (m, 1H), 7.57 (d, J = 8. 0Hz, 2H), 7.46 (d, J = 8.0 Hz, 1H), 7.18 (d, J = 8.0 Hz, 1H), 7.10-7.14 (m,1H), 5.08-5.10 (m, 1H), 4.25-4.28 (m, 2H), 3.95-4.04 (m, 3H), 3.71-3.76 (m,3H), 3.68 (s, 3H), 3.12-3.14 (m, 2H), 2.88-2.95 (m, 2H), 2.42-2.46 (m, 2H),2.04-2.08 (m, 2H), 1.71-1.73 (m, 2H), 1.52-1.53 (m, 2H), 1.24-1.27 (m, 2H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.0, 170.8, 170.7, 168.6, 167.6, 165.8, 164.4,155.6, 145.9, 144.3, 138.2, 133.8, 132.6, 131.7, 131.5, 130.6, 130.0, 129.5,128.8, 128.4, 118.7, 114.8, 111.5, 110.9, 94.6, 56.4, 53.3, 49.5, 48.7, 44.5,42.5, 39.8, 31.9, 30.9, 25.8, 22.9. HRMS (EI): calcd. for [C 40 H 40 C l2 [N7O8] (M+H) + , 816.2310; found, 816.2314. Example 14 Compound 9d was prepared, and its structural formula is as follows: .

[0047] Following the method described in Example 11, intermediate 8d was reacted with compound 3 to obtain compound 9d, totaling 52 mg, with a yield of 62%, as a yellow solid powder.

[0048] 3-(4-(5-aminopentyl)piperazin-1-yl)-1-(3,4-dichlorophenyl)-4-(4- methoxyphenyl)-1H-pyrrole-2,5-dione (8d). Yellowish solid (80 mg, 78% yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.43-7.46 (m, 2H), 7.18 (d, J = 6.0 Hz, 1H), 6.82-6.84 (m, 2H), 6.79 (s, 1H), 6.60 (d, J = 6.0 Hz, 1H), 3.82-3.89 (m, 4H), 3.47-3.51 (m, 4H), 2.92-3.05 (m, 4H), 1.72-1.73 (m, 2H), 1.50-1.51 (m, 2H), 1.28-1.29 (m, 2H). N-(5-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)pentyl)-2-((2-(2,6-dioxopiperidin-3- yl)-1,3-dioxoisoindolin-4-yl)amino)acetamide (9d). Yellowish solid (52 mg, 62% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.13 (brs, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.70 (s, 1H), 7.66-7.70 (m, 1H), 7.56 (d, J =8. 0 Hz, 2H), 7.46 (d, J = 8.0 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.10-7.14(m, 1H), 5.09-5.10 (m, 1H), 4.26-4.28 (m, 2H), 3.94-4.06 (m, 3H), 3.72-3.76(m, 3H), 3.65 (s, 3H), 3.12-3.14 (m, 2H), 2.88-2.96 (m, 2H), 2.42-2.46 (m,2H), 2.05-2.10 (m, 2H), 1.71-1.74 (m, 2H), 1.52-1.53 ​​(m, 2H), 1.35-1.36 (m,2H), 1.24-1.26 (m,2H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.1, 170.9, 170.7, 168.4, 167.6, 166.7, 164.4, 155.8, 145.9, 144.3, 138.7, 133.6, 132.6, 131.7, 131.4, 130.6, 130.0, 129.8, 128.6, 128.4, 118.7, 114.8, 111.6, 110.9, 94.6, 56.9, 53.3, 49.5, 48.8, 44.5, 43.4, 39.7, 31.9, 31.6, 25.9, 24.7, 22.9. HRMS(EI): calcd. for [C 41 H 42 C l2 [N7O8] (M+H) + , 830.2466; found, 830.2468. Example 15 Compound 9e was prepared, and its structural formula is as follows: .

[0049] Following the method described in Example 11, intermediate 8e was reacted with compound 3 to obtain compound 9e, totaling 43 mg, with a yield of 52%, as a yellow solid powder.

[0050] 3-(4-(2-(2-aminoethoxy)ethyl)piperazin-1-yl)-1-(3,4-dichlorophenyl)- 4-(4-methoxyphenyl)-1H-pyrrole-2,5-dione (8e). Yellowish solid (74 mg, 71%yield). 1 HNMR (600 MHz, DMSO-d6): δ 7.44-7.46 (m, 2H), 7.19 (d, J = 6.0 Hz, 1H), 6.82-6.84 (m, 2H), 6.78 (s, 1H), 6.59 (d, J = 6.0 Hz, 1H), 3.82-3.89 (m,4H), 3.47-3.51 (m, 4H), 3.72-3.74 (m, 2H), 3.50-3.56 (m, 2H), 2.92-3.05 (m,4H). N-(2-(2-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)ethoxy)ethyl)-2-((2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamide (9e). Yellowishsolid (43 mg, 52% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.12 (brs, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.73 (s, 1H), 7.66-7.70 (m,1H), 7.58 (d, J = 8. 0 Hz, 2H), 7.47 (d, J = 8.0 Hz, 1H), 7.18 (d, J = 8.0Hz, 1H), 7.10-7.16 (m, 1H), 5.09-5.10 (m, 1H), 4.27-4.28 (m, 2H), 3.95-4.08(m, 3H), 3.88-3.96 (m, 2H), 3.73-3.76 (m, 3H), 3.71-3.73 (m, 2H), 3.65 (s,3H), 3.45-3.48 (m, 2H), 3.12-3.14 (m, 2H), 3.05-3.09 (m, 2H), 2.52-2.53 (m,2H), 2.25-2.26 (m, 2H). 13 C NMR (150 MHz, DMSO- d6 ): δ 174.4, 170.9, 170.6,168.6, 167.6, 166.8, 164.4, 155.6, 145.9, 144.4, 138.7, 133.7, 132.6, 131.7,131.4, 130.9, 130.6, 129.8, 128.8, 128.2, 118.6, 114.8, 113.6, 110.9, 94.7,59.8, 57.0, 54.2, 53.8, 53.3, 51.7, 51.6, 49.8, 48.8, 45.9, 29.7. HRMS (EI):calcd. for [C 40 H 40 [Cl2N7O9] (M+H + , 832.2259; found, 832.2260. Example 16 The synthetic route for preparing compound 6a is as follows: Compound 5 (33 mg, 0.1 mmol) and intermediate compound 8a (48 mg, 0.1 mmol) were dissolved in DMF (2 mL), and condensing agents HATU (76 mg, 0.2 mmol) and DIPEA (0.2 mL) were added. After stirring, the mixture was allowed to react overnight at room temperature for about 10 h. After the reaction was completed by TLC monitoring, 15 mL of dichloromethane was added for extraction, followed by washing with saturated brine. The organic phases were combined, concentrated, and passed through a silica gel column to give compound 10a in a yield of 52 mg (66%) as a bright yellow solid.

[0051] N-(2-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)- 1,3-dioxoisoindolin-4-yl)oxy)acetamide (10a). Yellowish solid (52 mg, 66%yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.14 (brs, 1H), 7.79-7.80 (m, 2H), 7.74-7.76 (d, J = 8.0 Hz, 2H), 7.70 (d, J = 8.0 Hz), 1H), 7.63-7.65 (d, J = 8.0Hz, 2H), 7.50-7.53 (m, 1H), 7.46 (d, J = 8.0 Hz, 2H), 5.26-5.28 (m, 2H), 5.10-5.13 (m, 1H), 3.96-4.07 (m, 4H), 3.60-3.78 (m, 3H), 3.56 (s, 3H), 2.87-2.90 (m, 2H), 3.12-3.13 (m, 2H), 2.12-2.14 (m, 2H), 2.01-2.09 (m, 1H), 1.24-1.28 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.4, 171.3, 168.8, 164.3, 165.8,164.8, 163.9, 159.7, 156.6, 142.9, 138.7, 132.9, 132.6, 131.9, 131.8, 131.0,130.8, HRMS (EI): calcd. for[C 38 H 35 C l2 [N6O9] (M+H) + , 789.1837; found, 789.1838. Example 17 Compound 10b was prepared, and its structural formula is as follows: .

[0052] Following the method described in Example 16, intermediate 8b was reacted with compound 5 to obtain compound 10b, totaling 54 mg, with a yield of 67%, as a yellow solid powder.

[0053] N-(3-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)propyl)-2-((2-(2,6-dioxopiperidin-3- yl)-1,3-dioxoiso indolin-4-yl)oxy)acetamide (10b). Yellowish solid (54 mg, 67% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.12 (brs, 1H), 7.79-7.80 (m, 2H), 7.74-7.77 (d, J = 8.0 Hz, 2H), 7.70 (d, J = 8.0 Hz), 1H), 7.64-7.66 (d, J =8.0 Hz, 2H), 7.50-7.53 (m, 1H), 7.48 (d, J = 8.0 Hz, 2H), 5.26-5.28 (m, 2H), 5.10-5.14 (m, 1H), 3.96-4.08 (m, 4H), 3.60-3.74 (m, 3H), 3.55 (s, 3H), 2.87-2.90 (m, 2H), 3.12-3.14 (m, 2H), 2.12-2.16 (m, 2H), 2.01-2.09 (m, 1H), 1.72-1.74 (m, 2H), 1.25-1.27 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.6, 171.5,168.8, 164.4, 165.8, 164.9, 164.1, 159.7, 156.8, 142.9, 138.9, 132.9, 132.8,132.7, 131.9, 131.6, 130.8, 130.4, 128.0, 127.3, 119.9, 117.6, 116.2, 115.8,95.1, 67.8, 58.4, 53.4, 49.6, 48.5, 44.7, 42.4, 39.5, 31.6, 30.8, 22.8. HRMS(EI): calcd. for [C 39 H 37 C l2 [N6O9] (M+H) + , 803.1994; found, 803.1996. Example 18 Compound 10c was prepared, and its structural formula is as follows: Following the method described in Example 16, intermediate 8c was reacted with compound 5 to obtain compound 10c, totaling 51 mg, with a yield of 63%, as a yellow solid powder.

[0054] N-(4-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)butyl)-2-((2-(2,6-dioxopiperidin-3-yl)- 1,3-dioxoisoindolin-4-yl)oxy)acetamide (10c). Yellowish solid (51 mg, 63%yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.12 (brs, 1H), 7.79-7.80 (m, 2H), 7.74-7.78 (d, J = 8.0 Hz, 2H), 7.70 (d, J = 8.0 Hz), 1H), 7.63-7.64 (d, J = 8.0Hz, 2H), 7.50-7.53 (m, 1H), 7.46 (d, J = 8.0 Hz, 2H), 5.26-5.28 (m, 2H), 5.09-5.12 (m, 1H), 3.96-4.08 (m, 4H), 3.60-3.73 (m, 3H), 3.56 (s, 3H), 2.88-2.91 (m, 2H), 3.12-3.14 (m, 2H), 2.13-2.15 (m, 2H), 2.01-2.09 (m, 1H), 1.72-1.75 (m, 2H), 1.51-1.52 (m, 2H),1.26-1.28 (m, 1H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.8, 171.5, 168.9, 168.4, 165.6, 164.9, 164.3, 158.6, 156.8, 143.0,138.9, 132.9, 132.8, 132.6, 131.9, 131.4, 130.9, 130.4, 127.9, 127.3, 119.9,118.0, 116.2, 115.8, 95.2, 67.8, 58.6, 53.5, 49.6, 48.8, 44.6 42.4, 39.4,31.1, 30.8, 27.6, 22.4. HRMS (EI): calcd. for [C 40 H 39 C l2[N6O9] (M+H) + , 817.2150;found, 817.2154. Example 19 Compound 10d was prepared, and its structural formula is as follows: .

[0055] Following the method described in Example 16, intermediate 8d was reacted with compound 5 to obtain compound 10d, totaling 55 mg, with a yield of 66%, as a yellow solid powder.

[0056] N-(5-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)pentyl)-2-((2-(2,6-dioxopiperidin-3- yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (10d). Yellowish solid (55 mg, 66%yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.11 (brs, 1H), 7.79-7.80 (m, 2H), 7.74-7.76 (d, J = 8.0 Hz, 2H), 7.69 (d, J = 8.0 Hz), 1H), 7.64-7.66 (d, J = 8.0Hz, 2H), 7.50-7.53 (m, 1H), 7.48 (d, J = 8.0 Hz, 2H), 5.27-5.28 (m, 2H), 5.09-5.11 (m, 1H), 3.98-4.08 (m, 4H), 3.58-3.71 (m, 3H), 3.56 (s, 3H), 2.87-2.92 (m, 2H), 3.12-3.16 (m, 2H), 2.14-2.16 (m, 2H), 2.01-2.06 (m, 1H), 1.71-1.74 (m, 2H), 1.50-1.51 (m, 2H), 1.34-1.36 (m, 2H), 1.24-1.26 (m, 1H). 13 C NMR (150 MHz, DMSO-) d 6 ): δ 174.8, 171.7, 168.9, 165.8, 164.9, 164.8, 164.4, 158.6, 157.4, 143.0, 139.0, 132.9, 132.8, 132.7, 131.9, 131.6, 130.9, 130.6, 127.9, 127.8, 119.9, 119.4, 116.2, 115.3, 95.2, 68.2, 58.6, 53.6, 49.6, 48.9, 44.7, 42.4, 39.4, 31.9, 30.8, 27.7, 24.6, 22.7. HRMS (EI): calcd. for [C 41 H 41 C l2 [N6O9](M+H) + , 831.2307; found, 831.2308. Example 20 Compound 10e was prepared, and its structural formula is as follows: .

[0057] Following the method described in Example 16, intermediate 8e was reacted with compound 5 to obtain compound 10e, totaling 48 mg, with a yield of 58%, as a yellow solid powder.

[0058] N-(2-(2-(4-(1-(3,4-dichlorophenyl)-4-(4-methoxyphenyl)-2,5-dioxo-2,5- dihydro-1H-pyrrol-3-yl)piperazin-1-yl)ethoxy)ethyl)-2-((2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (10e). Yellowishsolid (48 mg, 58% yield). 1 HNMR (600 MHz, DMSO- d 6 ): δ 11.13 (brs, 1H), 7.81(d, J = 8.0 Hz, 1H), 7.78 (d, J = 8.0 Hz, 2H), 7.75 (s, 1H), 7.65-7.69 (m,1H), 7.57 (d, J = 8. 0 Hz, 2H), 7.47 (d, J = 8.0 Hz, 1H), 7.20 (d, J = 8.0Hz, 1H), 7.10-7.15 (m, 1H), 5.09-5.10 (m, 1H), 4.26-4.28 (m, 2H), 3.96-4.10(m, 3H), 3.87-3.95 (m, 2H), 3.74-3.76 (m, 3H), 3.72-3.73 (m, 2H), 3.66 (s,3H), 3.45-3.47 (m, 2H), 3.12-3.15 (m, 2H), 3.08-3.10 (m, 2H), 2.51-2.52 (m,2H), 2.24-2.27 (m, 2H). 13 C NMR (150 MHz, DMSO- d 6 ): δ 174.6, 170.6, 170.4,168.2, 167.6, 166.4, 164.1, 155.8, 146.0, 144.7, 138.8, 133.8, 132.4, 131.6,131.3, 130.9, 130.8, 129.8, 128.6, 128.2, 118.6, 115.0, 114.1, 110.9, 94.9,59.6, 57.0, 54.8, 53.7, 53.4, 51.9, 51.6, 49.7, 48.6, 46.0, 29.6. HRMS (EI):calcd. for [C 40 H 39 C l2 N6O 10 (M+H) + , 833.2099; found, 833.2099. Experiment to investigate the degradation efficiency of compounds on RAD51 protein SK-HEP-1 cells were seeded in 96-well black plates, with 6 × 10⁶ cells per well. 4After 12 h, add the appropriate dose of compound or DMSO (control wells) to each well. Incubate in an incubator for 36 h, then add 75 μL of 8% formaldehyde-TBS solution to each well and fix for 20 min at room temperature. Remove the solution from the wells, wash twice with 200 μL / well TBST, add 50 μL / well 0.1% Triton X PBS solution, and incubate for 15 min. Remove the solution from the 96-well plate, wash twice with 150 μL / well TBST. Then add 100 μL / well 1% H2O2 / TBS solution and incubate for 20 min at room temperature. Remove the solution and wash three times with an appropriate amount of TBST. Add 50 μL of blocking buffer to each well and incubate overnight at 4°C. Remove the blocking buffer, add 100 μL of blocking buffer containing 1:2000 diluted RAD51 antibody to each well (control group added blocking buffer containing DMSO), and incubate for 2 h at room temperature. The RAD51 primary antibody was recovered and washed twice with an appropriate amount of TBST. 150 μL of HRP-labeled rabbit antibody was added to each well, and the mixture was incubated at room temperature for 1 h. The secondary antibody was removed, and the mixture was washed three times with TBST. 100 μL of pre-prepared TMB substrate solution was added to each well, and the mixture was incubated at room temperature in the dark for 20 min. 10 μL of stop solution containing 1 N HCl was added to each well, and the mixture was shaken for 5 min. The absorbance values ​​at 450 and 570 nm were measured and recorded on a densitometer, and the relative content of RAD51 protein was calculated. The results are shown in Table 1.

[0059] Table 1. Degradation efficiency of various compounds on RAD51 protein at high and low concentrations As can be seen from the results in Table 1, the compounds provided by this invention exhibit varying degrees of degradation efficiency of the target protein RAD51 at different concentrations (300 nM and 3 μM), and generally possess target protein degradation activity. The activity is concentration-dependent, indicating that the compounds provided by this invention have good development potential.

[0060] In summary, the compounds provided in the embodiments of the present invention can significantly and selectively degrade RAD51 protein in tumor cells, and can be subsequently developed into innovative drugs for the treatment of cancer or diseases related to specific target protein RAD51.

[0061] In the above embodiments, the descriptions of each embodiment have different focuses. For parts that are not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0062] The above description describes specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in the present invention, and these modifications or substitutions should all be covered within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A compound, characterized in that, Selected from at least one of the compounds shown in the following structural formulas: 。 2. The use of the compound of claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of a drug targeting the degradation of the DNA homologous recombinant protein RAD51.

3. The use of the compound of claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of an antitumor drug.

4. The application as described in claim 3, characterized in that, The tumor includes cancer, and the cancer is selected from liver cancer.

5. A pharmaceutical composition, characterized in that, Includes the active ingredient and a pharmaceutically acceptable carrier; the active ingredient is: The compound of claim 1 or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition, characterized in that, Includes the active ingredient and a pharmaceutically acceptable carrier; the active ingredient is: The compound of claim 1 or a pharmaceutically acceptable salt thereof, and one or more therapeutically active ingredients.

7. A targeted degradation agent for the DNA homologous recombinant protein RAD51, characterized in that, Includes the compound of claim 1 or a pharmaceutically acceptable salt thereof.

Images