PARP1 selective protein degradation agent and application thereof in tumor resistance

Abstract

The invention discloses a PARP1 selective protein degradation agent and an application of the PARP1 selective protein degradation agent in tumor resistance. The degradation agent has the following structural formula. Wherein the 3-ethyl-7-((4-(pyridine-3-yl) piperazine-1-yl) methyl) quinoxaline-2 (1H)-ketone part is a PARP1 target protein ligand, and L is a connecting unit; and B is an E3 ubiquitin ligase ligand. The PARP1 selective protein degradation agent prepared by the preparation method disclosed by the invention can be used for effectively degrading PARP1 protein, inhibiting cell proliferation and inducing tumor cell apoptosis. Meanwhile, when the compound is combined with chemotherapeutic drugs, the effect of enhancing the chemotherapeutic effect is achieved, almost no physiological toxicity exists, and the compound is expected to provide an ideal way for improving treatment of various diseases caused by PARP1 excessive activation. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to a PARP1 selective protein degrader and an application thereof in anti-tumor treatment. Background Art

[0002] In the process of life activities, the genome of organisms is susceptible to exogenous or endogenous factors, resulting in DNA damage, of which endogenous damage includes DNA replication errors, hydrolysis DNA damage, cell's own biochemical processes and excessive reactive oxygen (RSO) production, etc., and exogenous damage includes physical damage, chemical damage, etc. The types of DNA damage caused by these exogenous or endogenous damage factors can be specifically divided into five types: including base mutations, DNA cross-links, DNA single-strand breaks (SSBs), DNA double-strand breaks (DSBs) and base mismatches.

[0003] During the long evolutionary process, living organisms have developed repair mechanisms for different types of DNA damage to cope with nuclear DNA and mitochondrial DNA (mtDNA) damage and maintain appropriate chromosome structure and stability. These repair mechanisms specifically include direct repair, base excision repair (BER), nucleotide excision repair (NER), single strand annealing repair (SSA), mismatch repair (MMR), homologous recombination (HR), and non-homologous end joining (NHEJ). Among them, DNA single-strand breaks repair (SSBR) is a mechanism for dealing with single-strand breaks in the DNA double helix. SSBR mainly includes base excision repair (BER), mismatch repair (MMR) and nucleotide excision repair (NER). In the process of studying this repair mechanism, it was also found that if single-strand breaks in DNA cannot be repaired in time, it will usually lead to fatal double-strand breaks in DNA. The DNA double-strand breaks repair (DSBR) mechanism mainly includes homologous recombination (HR) and non-homologous end joining (NHEJ). These two types of DNA damage repair mechanisms are crucial to maintaining normal cell survival. Therefore, researchers have developed a series of methods to regulate these mechanisms for different mechanisms of DNA damage repair to treat diseases such as tumors, such as blocking the DNA repair pathway of tumor cells and intervening in proteins involved in DNA repair.

[0004] Among the many proteins involved in DNA repair, the Poly (ADP-ribose) polymerases (PARPs) family plays an important role in DNA repair. Among them, poly ADP ribose polymerase-1 (PARP1) is the most abundant and best characterized member of the poly ADP ribosylase (PARP) family. As a DNA-dependent ribozyme, PARP1 plays a key role in the signal transduction and repair of DNA damage. After DNA damage caused by the cell's own metabolic activities, chemical toxins or ionizing radiation, PARP1 is rapidly activated, and then recognizes and binds to DNA single-strand breaks (SSBs), where it catalyzes the cleavage of nicotinamide adenine dinucleotide (NAD +) is broken into ADP ribose and nicotinamide, and then the ADP ribosome is transferred to synthesize poly ADP nucleopolysaccharide chains on itself or several receptor proteins. These polymer chains with more negative charges and greater steric hindrance can separate PARP1 from DNA breaks by reducing the affinity between PARP1 and DNA, and then guide DNA repair factors to bind to DNA gaps to repair the damaged parts, which is a key process in the base excision repair pathway of DNA repair. PARP1 knockout cancer cells and animals show high sensitivity to radiation or cytotoxic agents. Initially, PARP inhibitors (PARPi) were developed as adjuvant components of standard chemotherapy and radiotherapy for tumor treatment. Subsequent studies have found that cancer cells carrying BRCA-1 or -2 mutations are very sensitive to PARPi, and the corresponding physiological process is also called "synthetic lethality." To date, six PARP inhibitors, including olaparib, rucaparib, niraparib, and talazoparib, have been approved for marketing. As the first oral potent PARP1 / 2 inhibitor, olaparib has been approved for clinical treatment of BRCA mutation-related tumor diseases.

[0005] However, there are still multiple challenges in the use of PARPi that limit its therapeutic efficacy, including the safety of long-term use and the acquisition of drug resistance. In terms of drug safety, PARPi, as an anti-tumor drug, not only inhibits PARP enzyme activity, but also causes PARP trapping during DNA damage repair. The PARP trapping mechanism is that when the PARP protein binds to DNA, PARPi prevents the PARP protein from dissociating from the DNA break to form a stable PARP1-PARPi complex, which may block and destroy DNA replication. Therefore, it is more toxic to normal cells than unrepaired SSBs. Researchers are unable to evaluate the specific effects of these two pathways (PARP catalytic inhibition and trapping) on ​​PARP inhibitor-mediated cytotoxicity. At the same time, PARPi's inhibition of PARP2 may induce adverse reactions related to the blood system. In addition to the safety of medication, the drug resistance of tumors caused by long-term use of PARPi cannot be ignored. The development of resistance to PARP inhibitors is a complex process. One theory is that the secondary mutation of BRCA1 / 2 genes in cells restores their protein function and thus develops resistance to PARP inhibitors. Once resistance develops, the efficacy of PARPi will be significantly reduced. In order to achieve the goal of selectively blocking the function of PARP and reducing the occurrence of PARP capture, while improving the resistance of tumor cells to PARPi, PROTAC technology has become one of the strategies considered by researchers.

[0006] Treating diseases by degrading target pathogenic proteins is a new model in the current field of drug development. Proteolysis Targeting Chimera (PROTAC) technology is a representative technology currently used for targeted tumor therapy. PROTAC molecules are bifunctional small molecules containing two ligands, which are connected together by a connector and specifically bind to the target protein and E3 ubiquitin ligase respectively. These molecules can first drive the target protein to the E3 ubiquitin ligase to form a ternary complex, resulting in polyubiquitination of the target protein, and then proteasome-mediated degradation. The PROTAC strategy has made great progress in inducing degradation of target proteins in vitro and in vivo, such as BET, AR, ER, and CDK9, which is attributed to the discovery of several efficient small molecule ligands of E3 ubiquitin ligases, such as thalidomide and pomalidomide. Compared with traditional small molecule inhibitors, PROTAC compounds have their unique advantages. Using PROTAC technology, researchers have developed many PROTAC compounds that can efficiently degrade pathogenic proteins in vivo and in vitro. Currently, several anti-tumor drugs developed based on this technology have entered clinical research. The present invention uses PROTAC technology to design PARP1 degraders, in order to overcome the defects of small molecule PARP1 inhibitors and provide a new method for tumor targeted therapy. Summary of the invention

[0007] The purpose of the embodiments of the present application is to address the above-mentioned deficiencies in the prior art, provide a PARP1 selective protein degrader and its application in anti-tumor, and provide a compound that can not only effectively inhibit the proliferation activity of tumor cells but also selectively degrade PARP1 protein.

[0008] The purpose of the embodiments of the present application is to address the above-mentioned deficiencies in the prior art, provide a PARP1 selective protein degrader and its application in anti-tumor, and provide a compound that can not only effectively inhibit the proliferation activity of tumor cells but also selectively degrade PARP1 protein.

[0009] To achieve the above purpose, the technical solution adopted by the present invention to solve the technical problem is:

[0010] A compound having the structural formula:

[0011]

[0012] The 3-ethyl-7-((4-(pyridin-3-yl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one moiety is a PARP1 target protein ligand;

[0013] L is a linking unit, comprising a bond or chemical linking part covalently coupled in a compound molecule, including at least one or a combination of a linear diamine linking segment, a linear amino acid linking segment or a ring structure linking segment; or a compound formed by a linear amino acid of a certain length, a linear diamine, piperazine, piperidine, pyrrolidine and azetidine;

[0014] B is the CRBN E3 ubiquitin ligase ligand;

[0015] The general structural formula of the E3 ubiquitin ligase ligand is:

[0016]

[0017] wherein Z is one of -CH2- and -C(=O)-, and X and Y are each independently one of H or F.

[0018] The connecting unit L is one of the following structures:

[0019] -NHCH2(CH2)nCH2NH-; -NH(CH2 CH2O)n(CH2) m NH-;

[0020] -NHCH2(CH2)n CH2C(=O)NH-; -NH(CH2 CH2O)n(CH2) m C(=O)-;

[0021] -NH(CH2 CH2O)n(CH2) m C(=O)NH(CH2) m C(=O)NH-;

[0022]

[0023]

[0024] The n is 2 to 10; m, s, and t are each independently selected from 0, 1, 2, and 3; X1 and X2 are each independently selected from CH or N.

[0025] Furthermore, the linker unit L is composed of a straight-chain diamine; the E3 ubiquitin ligase ligand is the CRBN ligand thalidomide and its derivatives; the structural formula of the compound is:

[0026]

[0027]

[0028] Furthermore, the straight-chain diamine linking segment is a linking segment derived from aliphatic chain diamine or polyethylene glycol diamine.

[0029] Furthermore, the linear diamine linking segment is at least one of linking segments from butanediamine, pentanediamine, hexanediamine, heptanediamine, octanediamine, nonanediamine, decanediamine, undecanediamine, dodecanediamine or diamines containing oxygen atoms.

[0030] Furthermore, X and Y are either a H atom or a fluorine atom.

[0031] Furthermore, the linker unit L is composed of a straight-chain amino acid linker segment; the E3 ubiquitin ligase ligand is the CRBN ligand lenalidomide; and the structural formula of the compound is:

[0032]

[0033] Furthermore, the straight-chain amino acid linker is at least one of a linker selected from glycine, β-alanine, aminobutyric acid, aminovaleric acid, aminocaproic acid, aminoheptanoic acid, aminooctanoic acid, aminononanoic acid, aminodecanoic acid, aminoundecanoic acid, aminododecanoic acid or an amino acid containing polyethylene glycol.

[0034] Furthermore, the linking unit L is composed of a ring structure linking segment or a combination of a ring structure linking segment and a fatty chain linking segment; the E3 ubiquitin ligase ligand is the CRBN ligand thalidomide and its derivatives; the structural formula of the compound is:

[0035]

[0036] Furthermore, the ring structure is an aliphatic ring structure.

[0037] Furthermore, the aliphatic ring structure is at least one of piperazine, piperidine, pyrrolidine and azetidine.

[0038] Furthermore, the fatty chain is a straight-chain amino acid compound.

[0039] Furthermore, the straight-chain amino acid is one of aminocaprylic acid or aminodecanoic acid.

[0040] Furthermore, X and Y are either a H atom or a fluorine atom.

[0041] Specifically, the E3 is selected from the following structures:

[0042]

[0043] More specifically, the connecting unit L is selected from the following structures:

[0044]

[0045]

[0046] A drug for inhibiting tumor cell proliferation, comprising the above compound and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate, enantiomer.

[0047] A PARP1 protein degrader comprises the above compound and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate, enantiomer.

[0048] Further, the concentration of the compound in the degrading agent is greater than 1 μM.

[0049] An anti-tumor drug comprises the above compound and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate, enantiomer, and pharmaceutically acceptable auxiliary components.

[0050] Furthermore, the tumor is fallopian tube cancer, colorectal cancer, prostate cancer or esophageal cancer.

[0051] A combined anti-tumor drug comprises the above compound and a chemotherapy drug used in combination therewith.

[0052] Furthermore, the chemotherapy drug is cisplatin.

[0053] The beneficial effects of the present invention are:

[0054] The PARP1 selective protein degrader prepared by the present invention can effectively degrade PARP1 protein, inhibit cell proliferation, and induce apoptosis of tumor cells. At the same time, it has the effect of enhancing the efficacy of chemotherapy when used in combination with chemotherapy drugs, and has almost no physiological toxicity. The compound is expected to provide an ideal way to improve the treatment of various diseases caused by excessive activation of PARP1.

[0055] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 The results of the test on the degradation of intracellular PARP1 / 2 protein by compounds P3 and P4;

[0057] Figure 2 The degradation effect of compounds P3 and P4 on cellular PARP1 protein and their half-maximal degradation concentration DC 50 Test results;

[0058] Figure 3 IC is the half inhibition rate of compounds P3 and P4 on tumor cell line MDA-MB-436 cells 50 Test results;

[0059] Figure 4The results are the half inhibition rate test results of compounds P3 and P4 combined with cisplatin on the tumor cell line MDA-MB-436 cells. DETAILED DESCRIPTION

[0060] Here, exemplary embodiments are described in detail, and examples thereof are shown in the accompanying drawings. When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The implementations described in the following exemplary embodiments do not represent all implementations consistent with the present application.

[0061] The terms used in this application are for the purpose of describing specific embodiments only and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application and the appended claims are also intended to include plural forms unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0062] It should be understood that although the terms first, second, third, etc. may be used in the present application to describe various information, these information should not be limited to these terms. These terms are only used to distinguish the same type of information from each other. For example, without departing from the scope of the present application, the first information may also be referred to as the second information, and similarly, the second information may also be referred to as the first information. Depending on the context, the word "if" as used herein may be interpreted as "at the time of" or "when" or "in response to determining".

[0063] Example 1 Preparation of PARP1 small molecule ligand

[0064] Its synthetic route is:

[0065]

[0066]

[0067] 1 (19.9 g, 100 mmol) and 2 (15.3 g, 110 mmol) were added to a 500 mL round-bottom flask, dissolved with 200 mL THF, and the reaction solution was cooled to 0 ° C. NaHCO3 (9.24 g, 110 mmol) was added and reacted at room temperature for 12 h. TLC monitored the complete reaction of the raw materials, and 300 mL of water was added to the reaction solution. The reaction solution was then filtered, the filter cake was washed with a small amount of EA, and the filtrate was separated. The organic phase was retained, and the aqueous phase was extracted with ethyl acetate (3×20 mL). The organic phases were combined and the solvent was evaporated under reduced pressure to obtain the intermediate crude product 3 (26.8 g, 92% yield, yellow solid). 1H NMR (400MHz, CDCl3) δ8.90(d,J=1.6Hz,1H),8.68(d,J=6.8Hz,1H),8.05(dd,J=9.2,1.2Hz,1H),6.75(d,J=9.2Hz,1H),4.28( dd,J=13.2,6.4Hz,1H),3.90(s,3H),3.80(s,3H),2.03(ddd,J=21.6,14.4,7.2Hz,2H),1.06(t,J=7.6Hz,3H); HRMS(ESI):m / z calcd for C 13 H 16 N2O6[M+H] + :297.1008,found:297.1011.

[0068] In a 1000mL three-necked flask, add 3 (29.6g, 100mmol), Fe (44.6g, 800mmol), NH4Cl (42.8g, 800mmol) and dissolve with 500mL MeOH, then place the reaction under a mechanical stirring device. React at 85°C for 12h. After TLC monitoring, the reaction of the raw materials is complete, the methanol is evaporated under reduced pressure, and it is redissolved with 200mL DMF and heated to 90°C. The reaction solution is filtered while hot, the filter cake is washed with a small amount of hot DMF solution, poured into sufficient ice water while hot, and the filter cake is retained by suction filtration. After drying, the intermediate crude product 4 (16.4g, 70% yield, white solid) is obtained. 1 H NMR (400MHz, DMSO-d6) δ7.82–7.58(m,1H),7.39–7.25(m,2H),4.83(s,2H),2.91–2.71(m,3H),1.32–1.11(m,5H); HRMS(ESI):m / z calcd forC 12 H 14 N2O3[M+H] + :235.1077,found:235.1078.

[0069] 4 (23.4 g, 100 mmol) was added to a 250 mL round-bottom flask, dissolved with 100 mL 1,4-dioxane, and the reaction solution was cooled to 10 ° C. DDQ (24.9 g, 110 mmol) was slowly added in batches. After completion, the reaction was transferred to room temperature for 0.5 h. TLC monitored the complete reaction of the raw materials. Saturated NaHCO3 aqueous solution was added to the reaction solution to adjust the pH to 7-8. The reaction solution was then filtered, the filter cake was washed with a small amount of water, the filter cake was retained, and dried to obtain the intermediate crude product 5 (11.6 g, 50% yield, white solid). 1H NMR (400MHz, DMSO-d6) δ12.39 (s, 1H), 7.78 (d, J = 11.2Hz, 3H), 3.88 (s, 3H), 2.81 (s, 2H), 1.21 (s, 3H); HRMS (ESI): m / z calcd for C 12 H 12 N2O3[M+H] + :233.0848,found:233.0850.

[0070] Add 5 (23.2 g, 100 mmol) to a 250 mL round-bottom flask, dissolve it with 150 mL THF, cool the reaction solution to 0 ° C, slowly add LiAlH4 (6.0 g, 150 mmol) in batches, and transfer to room temperature for 0.5 h after completion. TLC monitors the complete reaction of the raw materials. Add a small amount of aqueous solution to the reaction solution to quench LiAlH4. Then filter the reaction solution, wash the filter cake with ethyl acetate, retain the filtrate, and evaporate the solvent under reduced pressure to obtain the intermediate crude product 6 (9.18 g, 45% yield, white solid). 1 H NMR(500MHz,DMSO-d6)δ12.30(s,1H),7.66(d,J=8.0Hz,1H),7.27(s,1H),7.18(dd,J=8.0,1.2Hz,1H),5 .40(t,J=5.5Hz,1H),4.59(d,J=5.5Hz,2H),2.79(q,J=7.0Hz,2H),1.22(t,J=7.0Hz,3H); HRMS(ESI):m / z calcd for C 11 H 12 N2O2[M+H] + :205.0899,found:205.0896.

[0071] 6 (2.0 g, 10 mmol) was added to a 100 mL round-bottom flask, dissolved with 20 mL HBr / AcOH, and the reaction solution was heated to 80°C for 2 h. After completion, ice water was added, and a large amount of solid precipitated in the reaction solution. The reaction solution was filtered, the filter cake was washed with water, and the filter cake was retained and dried to obtain the intermediate crude product 7 (2.1 g, 80% yield, white solid). 1H NMR (400MHz, DMSO-d6) δ8.34(t,J=6.4Hz,1H),7.88(d,J=8.8Hz,1H),7.69(t,J=9.6Hz,1H),7.35(dt ,J=13.6,6.8Hz,1H),3.63(s,2H),2.81(q,J=7.2Hz,2H),1.23(t,J=7.2Hz,3H); HRMS(ESI):m / zcalcd for C 11 H 11 BrN2O[M+H] + :267.0128,found:267.0126.

[0072] 8 (15.5 g, 100 mmol) and 9 (20.4 g, 110 mmol) were added to a 500 mL round-bottom flask, dissolved with 100 mL THF, and the reaction solution was heated to 60 ° C. K2CO3 (15.2 g, 110 mmol) was added and reacted at room temperature for 12 h. TLC monitored the complete reaction of the raw materials, and 100 mL of water was added to the reaction solution. The reaction solution was then extracted with ethyl acetate (3×100 mL), the organic phases were combined and dried, and purified by silica gel column (PE:EA=2:1), and then 30% TFA / DCM was added thereto, reacted for 0.5 h, and the solvent was evaporated under reduced pressure to obtain product 10 (18.1 g, 82% yield, white solid). 1 H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 7.60 (m, 1H), 7.32 (m, 1H), 7.06 (t, J = 7.2Hz, 1H), 2.07 (s, 3H), 1.40 (m, 8H); HRMS (ESI): m / z calcd forC 11 H 15 N3O2[M+H] + :222.1164,found:222.1164.

[0073] 7 (2.66 g, 10 mmol), 10 (2.21 g, 10 mmol), and Et3N (1.5 g, 11 mmol) were added to a 100 mL round-bottom flask, and dissolved with 20 mL CH3CN. The reaction solution was heated to 60 °C for 2 h. The reaction was complete by TLC detection. After cooling, a large amount of solid was observed to precipitate in the reaction solution. The reaction solution was filtered, the filter cake was washed with water, and the filter cake was retained and dried to obtain compound 11 (36.0 g, 90% yield, white solid). HRMS (ESI): m / z calcd for C 22 H 26 N5O3[M+H] +:408.1962,found:408.1964.

[0074] Then, compound 11 (4.1 g, 10 mmol) and LiOH (1.0 g, 40 mmol) were added to a 100 mL round-bottom flask, dissolved with 10 mL MeOH and 10 mL H2O, stirred and reacted overnight, and after the reaction was completed, the pH was adjusted to 5-6, and a large amount of solid was observed to precipitate in the reaction solution. The reaction solution was filtered, the filter cake was washed with water, and the filter cake was retained and dried to obtain product 12 (3.3 g, 85% yield, white solid). 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.38(d,J=2.0Hz,1H),7.87(d,J=8.8Hz,1H),7.69(d,J=8.0Hz,1H),7.34(dd,J=8.8,2.0Hz,1H), 7.28(s,1H),7.24(d,J=8.0Hz,1H),3.62(s,2H),3.40(m,4H),2.80(q,J=7.2Hz,2H),2.55(s,4H),1.22(t,J=7.2Hz,3H); HRMS(ESI):m / z calcd for C 21 H 23 N5O3[M+H] + :394.1801,found:394.1803.

[0075] Example 2

[0076] Preparation of compound P1

[0077] Its synthetic route is:

[0078]

[0079] General procedure A: B1 (276 mg, 1.0 mmol) and L1 (202 mg, 1.0 mmol) were added to a 25 mL reaction bottle, 10 mL of NMP was added to dissolve, and Et3N (150 mg, 1.5 mmol) was added dropwise. The reaction was carried out at 90°C for 2-4 h. TLC monitored the complete reaction of the raw materials. 10 mL of water was added to the reaction bottle, and then extracted with ethyl acetate (3×20 mL). The organic phases were combined and the solvent was evaporated under reduced pressure. Purification was performed by silica gel column chromatography (PE:EA=1:1). 30% TFA / DCM 10 mL was added to the intermediate and the reaction was carried out for 2 h. The solvent was evaporated under reduced pressure to obtain a crude intermediate Z1 (286 mg, 80% yield, yellow solid). Compound 12 (393 mg, 1.0 mmol) was added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF and placed in an ice-water bath, DIPEA (260 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added in batches under stirring, the reaction was placed in an ice-water bath for 1-2 h, and then intermediate Z1 (358 mg, 1.0 mmol) was added, and the reaction was transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of half-saturated saline was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL), the organic layers were combined and washed once with saturated sodium chloride aqueous solution, dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product. Finally, column chromatography was performed using DCM: MeOH = 10:1~5:1 system to obtain product P1 (219 mg, 30% yield, yellow solid). 1 H NMR (400MHz, CDCl3) δ11.23(s,1H),8.94(s,1H),8.08(s,1H),7.98–7.95(m,1H),7.81(t,J=6.0Hz,1H),7.71(d,J=4. 0Hz,1H),7.38(t,J=7.6Hz,1H),7.25(t,J=7.2Hz,2H),7.14(d,J=9.2Hz,1H),7.00(d,J=7.2Hz,1H),6.78(d,J=8.4Hz ,1H),6.17(s,1H),4.88(d,J=6.4Hz,1H),3.64(s,2H),3.40(d,J=3.6Hz,2H),3.31(s,4H),2.98–2.83(m,4H),2.69(d ,J=9.2Hz,2H),1.62(d,J=6.8Hz,4H),1.45(d,J=6.4Hz,2H),1.31–1.25(m,6H),0.80(d,J=6.4Hz,3H).HRMS(ESI):m / z calcd for C 39 H 43 N9O6[M+H]+ :734.3409,found:734.3410.

[0080] The L structures involved in the following embodiments are shown in the following table:

[0081]

[0082]

[0083] The structure of B is shown in the following table:

[0084]

[0085] Example 3

[0086] Preparation of compound P2

[0087] The synthesis of P2 refers to general step A, and B1, L2 and 12 are used as reaction raw materials to obtain product P2 as a yellow solid with a yield of 31%. 1 H NMR (500 MHz, DMSO-d6) δ11.03(s,1H),8.68(dd,J=4.5,1.0 Hz,1H),8.46(dd,J=8.5,1.5 Hz,1H),8.36–8.30(m,1H),8.20(d,J=2.5 Hz,1H),7.76(d,J=9.0 Hz,1H),7.63(d,J=8.0 Hz,1H),7.53–7.48(m,1H),7.44(dd,J=8.5,4.5 Hz,1H),7.19(d,J=8.5Hz,1H),7.01(d,J=8.5 Hz,1H),6.94(d,J=7.0 Hz,1H),6.45(t,J=6.0Hz,1H),4.98(dd,J=12.5,5.5 Hz,1H),3.60(s,2H),3.19(dd,J=15.5,7.0 Hz,6H),2.78–2.71(m,2H),2.53(s,4H),1.95(dd,J=12.0,6.5 Hz,2H),1.47(dd,J=15.0,12.0 Hz,6H),1.26–1.19(m,10H),0.78(t,J=7.0 Hz,3H).HRMS(ESI):m / z calcd for C 42 H 49 N9O6[M+H]+:776.3879, found:776.3877. Its chemical structure is:

[0088]

[0089] Example 4

[0090] Preparation of compound P3

[0091] The synthesis of P3 refers to general step A, and B1, L3 and 12 are used as reaction raw materials to obtain product P3 as a yellow solid with a yield of 33%. 1 H NMR (500MHz, DMSO-d6) δ12.28(s,1H),11.11(s,1H),8.34(t,J=6.0Hz,1H),8.27(s,1H),7.83(d,J=9.0Hz,1H) ,7.69(d,J=8.0Hz,1H),7.57(t,J=8.0Hz,1H),7.39(d,J=6.5Hz,1H),7.28–7.23(m,2H),7.12(d,J=8.5Hz,1H) ,7.03(d,J=7.0Hz,1H),6.60(s,1H),5.06(dd,J=12.5,5.5Hz,1H),3.61(d,J=9.5Hz,4H),3.52(t,J=10.0Hz,1 0H),3.45–3.38(m,6H),2.82–2.78(m,2H),2.58(d,J=14.0Hz,6H),1.22(t,J=7.5Hz,3H).HRMS(ESI):m / zcalcd for C 40 H 45 N9O8[M+H] + :780.3464, found:780.3465. Its chemical structure is:

[0092]

[0093] Example 5

[0094] Preparation of compound P4

[0095] The synthesis of P4 refers to general step A, and B1, L4 and 12 are used as reaction raw materials to obtain product P4 as a yellow solid with a yield of 15%. 1H NMR (500MHz, DMSO-d6) δ12.28(s,1H),11.11(s,1H),8.34(t,J=5.5Hz,1H),8.27(s,1H),7.83(d,J=9.0Hz, 1H),7.69(d,J=8.0Hz,1H),7.57(t,J=8.0Hz,1H),7.39(d,J=6.5Hz,1H),7.28–7.23(m,2H),7.12(d,J=8.5 Hz,1H),7.03(d,J=7.0Hz,1H),6.60(s,1H),5.06(dd,J=13.0,5.5Hz,1H),3.61(d,J=9.5Hz,4H),3.52(m,1 0H),3.45–3.38(m,10H),2.82–2.78(m,2H),2.58(d,J=12.0Hz,6H),1.22(t,J=7.5Hz,3H).HRMS(ESI):m / z calcdfor C 44 H 53 N9O6[M+H] + :804.4192, found:804.4188. Its chemical structure is:

[0096]

[0097] Example 6: Preparation of Compound P5

[0098] Its synthetic route is:

[0099]

[0100] General procedure B: B2 (259 mg, 1.0 mmol) and L5 (254 mg, 1.1 mmol) were added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF and placed in an ice-water bath, DIPEA (258 mg, 2.0 mmol) was added, and HATU (570 mg, 1.5 mmol) was added in batches with stirring. The reaction was placed in an ice-water bath for 1-2 h. After the reaction was completed by TLC monitoring, 20 mL of half-saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL). The combined organic layers were washed once with saturated sodium chloride aqueous solution, added with anhydrous Na2SO4 for drying, filtered and evaporated under reduced pressure to obtain an oily crude product, and finally separated by column chromatography using DCM:MeOH=10:1~5:1 system to obtain a white solid intermediate. The obtained intermediate was added with 10 mL of 30% TFA / DCM for 2 h, and the solvent was evaporated under reduced pressure to obtain a crude intermediate Z5 (231 mg, 60% yield, white solid). Compound 12 (393 mg, 1.0 mmol) was added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF and placed in an ice-water bath, DIPEA (258 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added in batches under stirring, the reaction was placed in an ice-water bath for 1-2 h, and then intermediate Z5 (386 mg, 1.0 mmol) was added, and the reaction was transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL), the organic layers were combined and washed once with saturated sodium chloride aqueous solution, dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product, and finally separated by column chromatography using DCM: MeOH = 10:1~5:1 system to obtain product P5 (219 mg, 25% yield, white solid). 1H NMR (400MHz, DMSO-d6) δ12.30(s,1H),11.05(s,1H),9.77(s,1H),8.40(s,1H),8.27(s,1H),7.83(t,J= 6.4Hz,2H),7.70(d,J=8.0Hz,1H),7.50(t,J=7.6Hz,2H),7.39(d,J=7.2Hz,1H),7.31–7.22(m,2H),5.16 (dd,J=13.2,4.8Hz,1H),4.37(q,J=17.6Hz,2H),3.64(s,2H),3.25(d,J=6.0Hz,2H),3.07–2.70(m,6H) ,2.72–2.53(m,6H),2.43–2.22(m,4H),2.03(d,J=6.8Hz,4H),1.60(s,2H),1.49(s,3H).HRMS(ESI):m / z calcd for C 40 H 45 N9O6[M+H] + :748.3566,found:748.3565.

[0101] Example 7

[0102] Preparation of compound P6

[0103] The synthesis of P6 refers to general procedure B, and B2, L6 and 12 are used as reaction raw materials to obtain product P6 as a white solid with a yield of 34%. 1 H NMR (500MHz, CDCl3) δ11.04(s,1H),9.78(s,1H),8.46(t,J=6.0Hz,1H),8.33(s,1H),7.91–7.76(m,3H),7.54–7. 43(m,3H),7.40(s,2H),5.15(dd,J=13.5,5.0Hz,1H),4.36(q,J=17.5Hz,2H),4.04(d,J=7.0Hz,2H),3.47(m,10H ),3.23(dd,J=13.0,6.0Hz,6H),2.97–2.87(m,2H),2.84(q,J=7.5Hz,2H),2.71(d,J=11.5Hz,1H),2.61(d,J=14. 0Hz,1H),2.37–2.33(m,2H),2.09–1.99(m,2H),1.60(s,2H),1.51(d,J=6.5Hz,2H),1.39(s,3H).HRMS(ESI):m / z calcd for C 43 H 51N9O6[M+H] + :790.4035, found:790.4033. Its chemical structure is:

[0104]

[0105] Example 8

[0106] Preparation of compound P7

[0107] The synthesis of P7 refers to general procedure B, and B2, L7 and 12 are used as reaction raw materials to obtain product P7, with a white solid yield of 26%. 1 H NMR (400MHz, DMSO-d6) δ12.30(s,1H),11.05(s,1H),9.77(s,1H),8.40(s,1H),8.27(s,1H),7.83(t,J=6.4Hz,2H ),7.70(d,J=8.0Hz,1H),7.50(t,J=7.6Hz,2H),7.39(d,J=7.2Hz,1H),7.31–7.22(m,2H),5.16(dd,J=13.2,4.8Hz ,1H),4.37(q,J=17.6Hz,2H),3.64(s,2H),3.25(d,J=6.4Hz,4H),3.03–2.86(m,2H),2.82–2.76(m,4H),2.72–2.5 3(m,6H),2.43–2.22(m,2H),2.03(d,J=6.8Hz,4H),1.60(s,2H),1.49(s,3H),1.25–1.20(m,10H).HRMS(ESI):m / z calcd for C 45 H 55 N9O6[M+H] + :804.4192, found:804.4195. Its chemical structure is:

[0108]

[0109] Example 9

[0110] Preparation of compound P8

[0111] The synthesis of P8 refers to general procedure B, and B2, L8 and 12 are used as reaction raw materials to obtain product P8 as a white solid with a yield of 43%. 1H NMR(500MHz,DMSO-d6)δ12.58(s,1H),11.03(s,1H),10.42(s,1H),9.77(s,1H),8.45(s,1H) ,8.33(s,1H),7.91–7.79(m,3H),7.46(dd,J=19.0,12.0Hz,3H),5.15(dd,J=13.0,4.5Hz,1H) ,4.44(d,J=18.0Hz,2H),4.36(q,J=17.5Hz,2H),4.07(s,2H),3.30–3.08(m,8H),2.96–2.78 (m,4H),2.34(d,J=6.5Hz,2H),1.60(s,2H),1.49(s,3H),1.29–1.20(m,16H).HRMS(ESI):m / z calcd for C 46 H 57 N9O6[M+H] + :818.4384, found:818.4388. Its chemical structure is:

[0112]

[0113] Example 10

[0114] Preparation of compound P9

[0115] The synthesis of P9 refers to general procedure B. The product P9 was obtained by using B2, L9 and 12 as the reaction raw materials. The product was a white solid with a yield of 35%. 1H NMR (500MHz, CDCl3) δ8.33 (s, 1H), 8.19–8.06 (m, 2H), 7.99 (d, J = 9.0 Hz, 1H), 7.71 (d, J = 8.0 Hz, 1H), 7.45–7.41 (m, 1H), 7.25–7.11 (m, 3H), 7.06 (d, J = 7.0 Hz, 1H), 6.84 (d, J = 8.5 Hz, 1H), 6.46 (t, J = 5.5 Hz, 1H), 5. 18(dd,J=12.0,4.0Hz,1H),4.32(d,J=8.0Hz,2H),3.70(ddd,J=12.5,8.5,3.5Hz,14H),3.42–3.33(m,4H ),2.93–2.68(m,8H),2.16–1.99(m,2H),1.35–1.20(m,4H),0.87(dd,J=16.0,9.0Hz,3H).HRMS(ESI):m / z calcdfor C 43 H 51 N9O9[M+H] +:838.3883, found:838.3885. Its chemical structure is:

[0116]

[0117] Embodiment 11

[0118] Preparation of compound P10

[0119] The synthesis of P10 refers to general procedure B, and B2, L10 and 12 are used as reaction raw materials to obtain product P10 as a white solid with a yield of 27%. 1 H NMR (500MHz, CDCl3) δ11.04(s,1H),9.86(s,1H),8.35(t,J=5.5Hz,1H),8.28(d,J=2.5Hz,1H),7.83(t,J=7.0Hz,2H),7 .69(d,J=8.0Hz,1H),7.53–7.48(m,2H),7.40(dd,J=9.0,2.5Hz,1H),7.31–7.17(m,2H),5.15(dd,J=13.5,5.0Hz,1H), 4.35(q,J=17.5Hz,2H),3.70(t,J=6.5Hz,2H),3.64(s,2H),3.49(dd,J=8.5,5.5Hz,12H),3.42(dd,J=12.0,6.0Hz,4H) ,3.39(s,2H),3.35(s,6H),2.80(q,J=7.5Hz,2H),2.59(dd,J=15.5,9.0Hz,6H),1.09(t,J=7.0Hz,3H).HRMS(ESI):m / z calcd for C 45 H 55 N9O 10 [M+H] + :882.4145, found:882.4144. Its chemical structure is:

[0120]

[0121] Example 12: Preparation of compound P11, the synthetic route is:

[0122]

[0123] General procedure C: B3 (276 mg, 1.0 mmol) and L2 (202 mg, 1.0 mmol) were added to a 25 mL reaction bottle, and 10 mL of NMP was added to dissolve the mixture. Et3N (150 mg, 1.5 mmol) was then added dropwise. The mixture was reacted at 90 °C for 2 h. TLC monitored the complete reaction of the raw materials. 10 mL of water was added to the reaction bottle, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined and the solvent was evaporated under reduced pressure. The mixture was purified by silica gel column chromatography (PE: EA = 1: 1). 10 mL of 30% TFA / DCM was added to the intermediate and the reaction was continued for 2 h. The solvent was evaporated under reduced pressure to obtain a crude intermediate Z11 (286 mg, 80% yield, yellow solid). Compound 12 (393 mg, 1.0 mmol) was added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF and placed in an ice-water bath, DIPEA (260 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added in batches under stirring, the reaction was placed in an ice-water bath for 1-2 h, and then intermediate Z11 (358 mg, 1.0 mmol) was added, and the reaction was transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of half-saturated saline was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL), the organic layers were combined and washed once with saturated sodium chloride aqueous solution, dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product. Finally, the product P11 (204 mg, 28% yield, yellow-green solid) was separated by column chromatography using DCM: MeOH = 10:1~5:1 system. 1 H NMR (400MHz, DMSO) δ12.27(s,1H),11.06(s,1H),8.43–8.33(m,2H),8.27(s,1H),7.83(d,J=7.2Hz ,1H),7.69(d,J=7.2Hz,1H),7.56(d,J=8.4Hz,1H),7.38(t,J=10.3Hz,2H),7.25(d,J=8.4Hz,2H), 7.10(s,1H),5.03(dd,J=12.8,5.2Hz,1H),3.62(s,6H),3.26(s,6H),3.13(d,J=15.2Hz,2H),2.80 (dd,J=14.4,7.2Hz,4H),2.54(d,J=10.0Hz,10H),1.60–1.47(m,4H),1.22(s,3H).HRMS(ESI):m / z calcd for C 42 H 49 N9O6[M+H] + :776.3879,found:776.3877.

[0124] Example 13: Preparation of Compound P12

[0125] The synthesis of P12 refers to general step C, and B3, L1 and 12 are used as reaction raw materials to obtain product P12 as a yellow solid with a yield of 26%. 1 H NMR(400MHz,DMSO-d6)δ12.58(s,1H),11.03(s,1H),8.39(s,1H),8.02(s,1H),7 .88(d,J=8.8Hz,2H),7.81–7.63(m,2H),7.31(dd,J=28.0,9.0Hz,3H),5.18(d,J =8.2Hz,1H),3.82(s,4H),3.63(s,2H),3.41(m,4H),3.05–2.77(d,J=6.4Hz,4H) ,2.56(m,6H),1.23(dd,J=8.4,4.4Hz,6H),1.21(t,J=4.4Hz,3H).HRMS(ESI):m / z calcd forC 42 H 49 N9O6[M+H] + :734.3409, found:734.3408. Its chemical structure is:

[0126]

[0127] Embodiment 14

[0128] Preparation of compound P13

[0129] The synthesis of P13 refers to general step C, and B3, L4 and 12 are used as reaction raw materials to obtain product P13 as a yellow solid with a yield of 22%. 1H NMR (400MHz, DMSO-d6) δ12.57(s,1H),11.06(s,1H),8.44(d,J=5.8Hz,2H),8.34(s,1H),7.90(d,J= 8.8Hz,2H),7.84(d,J=8.4Hz,2H),7.56(t,J=8.0Hz,2H),6.95(s,1H),6.85(d,J=8.2Hz,1H),5.03(d t,J=22.0,11.0Hz,1H),3.27(dd,J=11.2,6.4Hz,4H),3.16(m,6H),2.91(s,2H),2.86–2.81(m,4H),2 .75(d,J=8.4Hz,2H),1.57(d,J=4.0Hz,2H),1.51(s,4H),1.25(m,12H),1.23(s,3H).HRMS(ESI):m / z calcd for C 44 H 53 N9O6[M+H] + :804.4192, found:804.4189. Its chemical structure is:

[0130]

[0131] Embodiment 15

[0132] Preparation of compound P14

[0133] The synthesis of P14 refers to general step A, and B1, L3 and 12 are used as reaction raw materials to obtain product P14 as a yellow solid with a yield of 23%. 1H NMR (500MHz, DMSO-d6) δ12.28(s,1H),11.11(s,1H),8.34(t,J=6.0Hz,1H),8.27(s,1H),7.83(d,J=9.0Hz,1H ),7.69(d,J=8.0Hz,1H),7.57(t,J=8.0Hz,1H),7.39(d,J=6.5Hz,1H),7.28–7.23(m,2H),7.12(d,J=8.5Hz,1 H),7.03(d,J=7.0Hz,1H),6.60(s,1H),5.06(dd,J=12.5,5.5Hz,1H),3.61(d,J=9.5Hz,4H),3.52(t,J=10.0H z,6H),3.45–3.38(m,10H),2.82–2.78(m,2H),2.58(d,J=14.0Hz,6H),1.22(t,J=7.5Hz,3H).HRMS(ESI):m / z calcd for C 40 H 45 N9O8[M+H] + :780.3464, found:780.3465. Its chemical structure is:

[0134]

[0135] Example 16: Preparation of compound P15, the synthetic route is:

[0136]

[0137] General procedure D: Add B2 (259 mg, 1.0 mmol) and L11 (192 mg, 1.1 mmol) to a 25 mL reaction bottle, dissolve with 10 mL of anhydrous DMF and place in an ice-water bath, add DIPEA (258 mg, 2.0 mmol), add HATU (570 mg, 1.5 mmol) in batches with stirring, place the reaction in an ice-water bath for 1-2 h, and after the reaction is completed by TLC monitoring, add 20 mL of half-saturated brine to dilute the reaction solution, and then extract with ethyl acetate (3×20 mL), combine the organic layers, wash once with saturated sodium chloride aqueous solution, add anhydrous Na2SO4 to dry, filter and evaporate the solvent under reduced pressure to obtain an oily crude product, and finally use DCM:MeOH=10:1~5:1 system for column chromatography to obtain a white solid intermediate, and the obtained intermediate is then added with 30% TFA / DCM 10mL was reacted for 2h, and the solvent was evaporated under reduced pressure to obtain the crude intermediate Z15 (198mg, 60% yield, white solid). Subsequently, Z15 (330mg, 1.0mmol) and L5 (254mg, 1.1mmol) were added to a 25mL reaction bottle, dissolved in 10mL of anhydrous DMF and placed in an ice-water bath, DIPEA (258mg, 2.0mmol) was added, and HATU (570mg, 1.5mmol) was added in batches under stirring, and the reaction was placed in an ice-water bath for 1-2h. After the reaction was completed by TLC monitoring, 20mL of half-saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20mL), the organic layers were combined and washed once with saturated sodium chloride aqueous solution, dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product, and finally separated by column chromatography using DCM:MeOH=10:1~5:1 system to obtain a white solid intermediate, and the obtained intermediate was then added with 30% TFA / DCM 10mL was reacted for 2h, and the solvent was evaporated under reduced pressure to obtain a crude intermediate (222mg, 50% yield, white solid). The intermediate was added to a 25mL reaction bottle and compound 12 (393mg, 1.0mmol) was added, dissolved with 10mL anhydrous DMF and placed in an ice-water bath, DIPEA (258mg, 2.0mmol) was added, and HATU (570mg, 1.5mmol) was added in batches under stirring. The reaction was placed in an ice-water bath for 1-2h, and then transferred to room temperature for 3h. After the reaction was completed by TLC monitoring, 50mL of half-saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20mL), the organic layers were combined and washed once with saturated sodium chloride aqueous solution, dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product, and finally separated by column chromatography using DCM:MeOH=10:1~5:1 system to obtain product P15 (241mg, 30% yield, white solid). 1H NMR (400MHz, DMSO-d6) δ11.95(s,1H),10.96(d,J=16.4Hz,1H),8.39(t,J=7.2Hz,1H),7.89(t,J=8.0Hz,1H),7.73–7.63(m,1H),7.36(dd,J= 8.8,2.8Hz,1H),7.27(d,J=9.6Hz,1H),7.22(t,J=7.6Hz,1H),6.94(d,J=7.2Hz,1H),6.83(d,J=7.6Hz,1H),6.74(d,J=2.0Hz,1H),5.12(dt, J=16.4,8.0Hz,1H),4.19(dd,J=9.2,6.8Hz,2H),3.83(s,2H),3.61(d,J=4.2Hz,2H),3.45–3.38(m,2H),2.94(ddd,J=23.2,13.2,6.0Hz,2H) ,2.85–2.77(m,2H),2.69–2.54(m,2H),2.21(t,J=7.2Hz,2H),1.54–1.43(m,2H),1.38(d,J=11.6Hz,10H),1.32–1.16(m,3H).HRMS(ESI):m / z calcd for C 42 H 48 N 10 O7[M+H] + :805.3780,found:805.3781.

[0138] Example 17: Preparation of Compound P16

[0139] The synthesis of P16 refers to general step D, and B2, L5, L12 and 12 are used as reaction raw materials to obtain product P16 as a white solid with a yield of 14%. 1H NMR (400MHz, DMSO-d6) δ12.17(s,1H),11.54(s,1H),8.39(d,J=14.9Hz,1H),7.92(d,J=8.8Hz,1H),7.74(d,J=8.0Hz,1H),7.39(d,J=7 .6Hz,1H),7.35–7.28(m,2H),7.24(t,J=7.6Hz,2H),6.97(d,J=7.2Hz,2H),6.86(d,J=7.6Hz,2H),5.18–5.13(m,1H),4.22(d,J=12.0H z,2H),3.86(s,4H),3.67(s,2H),3.05(dd,J=17.2,10.4Hz,2H),2.93(dd,J=18.0,12.4Hz,4H),2.85(d,J=7.2Hz,2H),2.37(m,2H),2. 22(d,J=6.8Hz,2H),2.06(d,J=6.0Hz,2H),1.52(dd,J=18.8,11.6Hz,4H),1.27(t,J=6.8Hz,6H),1.02(t,J=6.0Hz,3H).HRMS(ESI):m / z calcd for C 43 H 50 N 10 O7[M+H] + :819.3937, found:819.3938. Its chemical structure is:

[0140]

[0141] Example 18: Preparation of Compound P17

[0142] The synthesis of P17 refers to general step D, and B2, L6, L11 and 12 are used as reaction raw materials to obtain product P17 as a white solid with a yield of 16%. 1H NMR (400MHz, DMSO-d6) δ12.26(s,1H),11.96(s,1H),10.97(d,J=6.4Hz,1H),8.39(s,1H),7.89(d,J=8.4Hz,1H),7.70(d,J=7.6Hz ,1H),7.35(d,J=8.0Hz,1H),7.31–7.23(m,2H),7.20(d,J=6.8Hz,1H),6.93(d,J=6.8Hz,1H),6.82(d,J=7.2Hz,1H),6.71(d,J=6. 8Hz,1H),5.37(d,J=8.0Hz,2H),5.09(t,J=23.2Hz,1H),4.18(dd,J=19.2,6.8Hz,2H),3.82(m,2H),3.64(m,2H),3.42(m,4H),2.8 6(dd,J=14.8,5.6Hz,6H),2.57(s,4H),2.33(d,J=12.8Hz,2H),2.20(m,2H),1.25(m,10H),1.05(t,J=4.0Hz,3H).HRMS(ESI):m / z calcd for C 45 H 54 N 10 O7[M+H] + :847.4205, found:847.4206. Its chemical structure is:

[0143]

[0144] Example 19: Preparation of Compound P18

[0145] The synthesis of P18 refers to general step D, and B2, L6, L12 and 12 are used as reaction raw materials to obtain product P18 as a white solid with a yield of 13%. 1H NMR (400MHz, DMSO-d6) δ12.16(s,1H),11.82(s,1H),8.36(d,J=8.4Hz,1H),7.89(d,J=8.4Hz,1H),7.70(d,J=8.0Hz,1H),7.35(d,J=7.6Hz,1H),7 .30–7.23(m,2H),7.21(t,J=7.2Hz,1H),6.93(d,J=7.2Hz,1H),6.88(s,1 H),6.82(d,J=7.8Hz,1H),6.74(s,1H),5.42(s,2H),5.12(d,J=8.8Hz,1H ),4.27–4.12(m,2H),3.82(m,3H),3.63(m,2H),3.01(d,J=7.2Hz,2H),2. 94(dd,J=11.2,5.2Hz,4H),2.82(d,J=7.2Hz,2H),2.67(d,J=5.2Hz,2H), 2.56(m,2H),2.29(t,J=7.6Hz,2H),2.17(t,J=6.8Hz,2H),2.07(s,1H),1.46(d,J=4.4Hz,4H),1.25(m,8H),0.97(t,J=6.0Hz,3H).HRMS(ESI):m / z calcd forC 46 H 56 N 10 O7[M+H] + :861.4406, found:861.4408. Its chemical structure is:

[0146]

[0147] Example 20: Preparation of Compound P19

[0148] The synthesis of P19 was carried out according to the general procedure A. B4, L2 and 12 were used as the raw materials to obtain the product P19 as a white solid with a yield of 15%. HRMS (ESI): m / z calculated for C 41 H 51 N9O5[M+H] + :750.4086, found:750.4087. Its chemical structure is:

[0149]

[0150] Example 21: Preparation of Compound P20

[0151] The synthesis of P20 was carried out according to the general procedure A. B5, L2 and 12 were used as the raw materials to obtain the product P20 as a white solid with a yield of 21%. HRMS (ESI): m / z calculated for C 41 H 50 FN9O5[M+H] + :768.3992, found:768.3991. Its chemical structure is:

[0152]

[0153] Example 22: Preparation of Compound P21

[0154] The synthesis of P21 was carried out according to the general procedure A. B6, L2 and 12 were used as the reaction materials to obtain the product P21, a white solid, with a yield of 17%. HRMS (ESI): m / z calculated for C 41 H 50 FN9O5[M+H] + :768.3992, found:768.3988. Its chemical structure is:

[0155]

[0156] Example 23: Preparation of Compound P22

[0157] The synthesis of P22 was carried out according to the general procedure A. B4, L3 and 12 were used as the raw materials to obtain the product P22 as a white solid with a yield of 15%. HRMS (ESI): m / z calculated for C 39 H 47 N9O7[M+H] + :754.3671, found:754.3673. Its chemical structure is:

[0158]

[0159] Embodiment 24:

[0160] Preparation of compound P23

[0161] The synthesis of P23 was carried out according to the general procedure A. B5, L3 and 12 were used as the raw materials to obtain the product P23 as a white solid with a yield of 13%. HRMS (ESI): m / z calculated for C 39 H 46 FN9O7[M+H] + :772.3577, found:772.3578. Its chemical structure is:

[0162]

[0163] Example 25: Preparation of Compound P24

[0164] The synthesis of P24 was carried out according to the general procedure A. B6, L3 and 12 were used as the raw materials to obtain the product P24 as a white solid with a yield of 11%. HRMS (ESI): m / z calculated for C 39 H 46 FN9O7[M+H] + :772.3577, found:772.3577. Its chemical structure is:

[0165]

[0166] Example 26: Preparation of Compound P25

[0167] The synthesis of P25 was carried out according to the general procedure A. B4, L4 and 12 were used as the raw materials to obtain the product P25 as a white solid with a yield of 22%. HRMS (ESI): m / z calculated for C 43 H 55 N9O5[M+H] + :778.4399, found:778.4400. Its chemical structure is:

[0168]

[0169] Example 27: Preparation of Compound P26

[0170] The synthesis of P26 was carried out according to the general procedure A. B5, L4 and 12 were used as the raw materials to obtain the product P26 as a white solid with a yield of 23%. HRMS (ESI): m / z calculated for C 43 H 54 FN9O5[M+H] + :796.4305, found:796.4306. Its chemical structure is:

[0171]

[0172] Example 28: Preparation of Compound P27

[0173] B4 (250 mg, 1.0 mmol) and N-Boc-piperazine (185 mg, 1.0 mmol) were added to a 25 mL reaction bottle. After dissolving in 10 mL NMP, Et3N (150 mg, 1.5 mmol) was added dropwise. The reaction was carried out at 90 °C for 2-4 h. TLC monitored the complete reaction of the raw materials. 10 mL of water was added to the reaction bottle, and then extracted with ethyl acetate (3×20 mL). The organic phases were combined and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography (PE:EA=1:1). 10 mL of 30% TFA / DCM was added to the intermediate and the reaction was carried out for 2 h. The solvent was evaporated under reduced pressure to obtain a crude intermediate Z27 (252 mg, 80% yield, white solid). Compound L13 (159 mg, 1.0 mmol) was added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF and placed in an ice-water bath, DIPEA (260 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added in batches under stirring, the reaction was placed in an ice-water bath for 1-2 h, and then intermediate Z27 (252 mg, 0.8 mmol) was added, and the reaction was transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of half-saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL), the organic layers were combined, washed once with saturated brine, dried over anhydrous Na2SO4, filtered, and the solvent was evaporated under reduced pressure to obtain an oily crude product. Finally, the intermediate was separated by column chromatography using a DCM: MeOH = 10:1 to 5:1 system, and then 10 mL of 30% TFA / DCM was added to react for 2 h. The solvent was evaporated under reduced pressure to obtain a crude intermediate Z27' (137 mg, 30% yield, white solid). Compound 12 (393 mg, 1.0 mmol) was added to a 25 mL reaction bottle, dissolved in 10 mL of anhydrous DMF, and placed in an ice-water bath. DIPEA (260 mg, 2.0 mmol) was added, and HATU (570 mg, 1.5 mmol) was added in batches under stirring. The reaction was placed in an ice-water bath for 1-2 h, and then the intermediate Z27 (358 mg, 1.0 mmol) was added, and the reaction was transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of half-saturated brine was added to the reaction solution for dilution, and then extracted with ethyl acetate (3×20 mL). The organic layers were combined and washed once with a saturated sodium chloride aqueous solution, dried by adding anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain an oily crude product. Finally, column chromatography was performed using a DCM:MeOH=10:1~5:1 system to obtain intermediate Z27' (83 mg, 11% yield, white solid). HRMS (ESI): m / z calcd for C 45 H 56 N 10 O6[M+H] + :833.4457,found:833.4455.

[0174]

[0175] Example 29: Preparation of Compound P28

[0176] Synthesis of P28 Refer to Example 28, using B5, L13, N-Boc-piperazine and 12 as reaction raw materials to obtain product P28, a white solid, with a yield of 25%. HRMS (ESI): m / z calcd for C 45 H 55 FN 10 O6[M+H] + :851.4363, found:851.4364. Its chemical structure is:

[0177]

[0178] Example 30: Preparation of Compound P29

[0179] Synthesis of P29 Refer to Example 28, using B4, L7, N-Boc-piperazine and 12 as reaction raw materials to obtain product P29, a white solid, with a yield of 11%. HRMS (ESI): m / z calculated for C 47 H 60 N 10 O6[M+H] + :861.4770, found:861.4771. Its chemical structure is:

[0180]

[0181] Example 31: Preparation of Compound P30

[0182] Synthesis of P30 Referring to Example 28, B5, L7, N-Boc-piperazine and 12 were used as reaction raw materials to obtain product P30 as a white solid with a yield of 16%. HRMS (ESI): m / z calculated for C 47 H 59 FN 10 O6[M+H] + :879.4676, found:879.4677. Its chemical structure is:

[0183]

[0184] Example 32: Preparation of Compound P31

[0185] Synthesis of P31 Refer to Example 28, using B4, L14 and 12 as reaction raw materials to obtain product P31, a white solid, with a yield of 33%. HRMS (ESI): m / z calculated for C 37 H 41 N9O5[M+H] + :692.3303, found:692.3305. Its chemical structure is:

[0186]

[0187] Example 33: Preparation of Compound P32

[0188] The synthesis of P32 was carried out in accordance with General Example 28. B5, L14 and 12 were used as the reaction raw materials to obtain the product P32 as a white solid with a yield of 27%. HRMS (ESI): m / z calculated for C 37 H 41 FN9O5[M+H] + :710.3209, found:710.3210. Its chemical structure is:

[0189]

[0190] Example 34: Preparation of Compound P33

[0191] The synthesis of P33 was carried out by referring to General Example 28, and the product P33 was obtained by using B4, L15 and 12 as reaction materials, as a white solid, with a yield of 23%. HRMS (ESI): m / z calculated for C 38 H 43 N9O5[M+H] + :706.3460, found:706.3461. Its chemical structure is:

[0192]

[0193] Example 35: Preparation of Compound P34

[0194] Synthesis of P34 Refer to Example 28, using B5, L15 and 12 as reaction raw materials to obtain product P34, a white solid, with a yield of 18%. HRMS (ESI): m / z calculated for C 38 H 43 FN9O5[M+H] + :724.3366, found:724.3367. Its chemical structure is:

[0195]

[0196] Example 36: Preparation of Compound P35

[0197] The synthesis of P35 was carried out by referring to General Example 28, and the product P35 was obtained by using B5, L16 and 12 as reaction materials, as a white solid, with a yield of 16%. HRMS (ESI): m / z calcd for C 39 H 45 N9O5[M+H] + :720.3616, found:720.3617. Its chemical structure is:

[0198]

[0199] Example 37: Preparation of Compound P36

[0200] Synthesis of P36 Refer to Example 28, using B6, L16 and 12 as reaction raw materials to obtain product P36, a white solid, with a yield of 14%. HRMS (ESI): m / z calculated for C 39 H 44 FN9O5[M+H] + :738.3522, found:738.3521. Its chemical structure is:

[0201]

[0202] Example 38: Preparation of Compound P37

[0203] Synthesis of P37 Refer to Example 28, using B4, L17 and 12 as reaction materials to obtain product P37, a white solid, with a yield of 25%. HRMS (ESI): m / z calculated for C 43 H 52 N 10 O5[M+H] + :789.4195, found:789.4194. Its chemical structure is:

[0204]

[0205] Example 39: Preparation of Compound P38

[0206] Synthesis of P38 Refer to Example 28, using B5, L17 and 12 as reaction materials to obtain product P38, a white solid, with a yield of 16%. HRMS (ESI): m / z calculated for C 43 H 51 FN 10 O5[M+H]+ :807.4101, found:807.4103. Its chemical structure is:

[0207]

[0208] Example 40: Preparation of Compound P39

[0209] Synthesis of P39 Refer to Example 28, using B4, L18 and 12 as reaction raw materials to obtain product P39, a white solid, with a yield of 16%. HRMS (ESI): m / z calculated for C 43 H 52 N 10 O5[M+H] + :789.4195, found:789.4166. Its chemical structure is:

[0210]

[0211] Example 41: Preparation of Compound P40

[0212] Synthesis of P40 Refer to Example 28, using B5, L18 and 12 as reaction raw materials to obtain product P40, a white solid, with a yield of 19%. HRMS (ESI): m / z calculated for C 43 H 51 FN 10 O5[M+H] + :807.4101, found:807.4137. Its chemical structure is:

[0213]

[0214] Example 42: Preparation of Compound P41

[0215] Synthesis of P41 Refer to Example 28, using B4, L19 and 12 as reaction raw materials to obtain product P41, a white solid, with a yield of 12%. HRMS (ESI): m / z calculated for C 41 H 48 N 10 O7[M+H] + :761.3882, found:761.3881. Its chemical structure is:

[0216]

[0217] Example 43: Preparation of Compound P42

[0218] Synthesis of P42 Refer to Example 28, using B5, L19 and 12 as reaction materials to obtain product P42, a white solid, with a yield of 15%. HRMS (ESI): m / z calculated for C 41 H 47 N 10 O5[M+H] + :779.3788, found:779.3789. Its chemical structure is:

[0219]

[0220] Example 44: Preparation of Compound P43

[0221] Synthesis of P43 Refer to Example 28, using B4, L20 and 12 as reaction raw materials to obtain product P43, a white solid, with a yield of 22%. HRMS (ESI): m / z calculated for C 42 H 50 N 10 O5[M+H] + :775.4038, found:775.4039. Its chemical structure is:

[0222]

[0223] Example 45: Preparation of Compound P44

[0224] Synthesis of P44 Refer to Example 28, using B5, L20 and 12 as reaction materials to obtain product P44, a white solid, with a yield of 11%. HRMS (ESI): m / z calculated for C 42 H 49 FN 10 O5[M+H] + :793.3944, found:793.3940. Its chemical structure is:

[0225]

[0226] Example 46: Preparation of Compound P45

[0227] Synthesis of P45 Refer to Example 27, using B4, L21 and 12 as reaction raw materials to obtain product P45, a white solid, with a yield of 18%. HRMS (ESI): m / z calculated for C 47 H 59 N 11 O5[M+H] + :858.4773, found:858.4772. Its chemical structure is:

[0228]

[0229] Embodiment 47:

[0230] Synthesis of P46 Refer to Example 28, using B4, L22 and 12 as reaction materials to obtain product P46, a white solid, with a yield of 12%. HRMS (ESI): m / z calculated for C 48 H 61 N 11 O5[M+H] + :872.4930, found:872.4927. Its chemical structure is:

[0231]

[0232] Embodiment 48:

[0233] Synthesis of P47 Refer to Example 28, using B4, L23 and 12 as reaction materials to obtain product P47, a white solid, with a yield of 19%. HRMS (ESI): m / z calculated for C 49 H 63 N 11 O5[M+H] + :886.5086, found:886.5082. Its chemical structure is:

[0234]

[0235] Example 49: Experiment on the degradation of PARP1 / 2 protein by compounds P1-P18

[0236] Preparation of main solutions: Preparation of test compounds: Weigh an appropriate amount of test compound powder and dissolve it in DMSO to make the final concentration of 10 mmol / L, 10 μL / tube, store at -20°C away from light, thaw before use, and dilute to the required concentration.

[0237] Cell culture and plating: MDA-MB-436 cells were cultured in L15 medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody). The cells were cultured in a 37°C constant temperature incubator containing 100% air. The cells were regularly observed and subcultured for subsequent experiments. Cells in the logarithmic growth phase were used for the experiment, and 2*10^5 cells / well were inoculated in a 12-well plate.

[0238] Administration: After the seeded cells adhered to the wall, the medium was replaced with serum-free medium. After treatment with compounds P1 to P18 10 μM for 24 hours, the medium was removed, the protein was extracted, and immunoblot analysis was performed, and semi-quantitative analysis was performed using Image J.

[0239] Immunoblotting was used to detect the protein expression of cells:

[0240] 1) Protein extraction: Prepare cell lysis buffer (1% protease and 1% protein phosphatase inhibitor are added to RIPA lysis buffer, purchased from Selleck, and the lysis buffer must be prepared before use). After the drug treatment is completed, the cell culture medium in the culture plate is aspirated, washed twice with PBS, and the residual PBS is aspirated as much as possible. After adding an appropriate amount of lysis buffer, it is placed on ice for full lysis for 30 minutes. During this period, cells can be scraped off with a cell scraper, and the lysis buffer is collected into a 1.5mL EP tube, placed on ice, and vortexed once every 5 minutes for a total of three to five times, and lysis is continued on ice; the EP tube is placed in a pre-cooled 4°C centrifuge and centrifuged at 13500rpm for 15 minutes; the supernatant is transferred to a new 1.5mL EP tube, and the protein concentration is determined by the BCA method; the required protein volume is normalized according to the measured protein concentration, and the small volume sample is supplemented with RIPA lysis buffer to ensure that the same batch of protein samples has the same volume and quality. Add 5× loading buffer to the protein sample to make the working concentration 1×, vortex to mix, and place in a boiling water bath for 5-7 minutes to denature the protein.

[0241] 2) Electrophoresis: Take an equal amount of the denatured protein sample and load it on a 5% stacking gel and a 10% separation gel (mainly

[0242] The ingredients are ddH2O, 30% polyacrylamide, 1.5M Tris-HCl (pH=8.8) or 1.0M Tris-HCl (pH=6.8), 10% SDS, 10% ammonium persulfate, TEMED), and the amount of protein loaded in each well is about 40μg. According to the different colors of the positive and negative electrodes (black and red), the electrodes are matched and the electrophoresis tank cover is covered. After turning on the power of the electrophoresis instrument, adjust the voltage to 80V to start electrophoresis. After the protein enters the separation gel and the protein marker is visible in the layer, adjust the voltage to 120V and continue electrophoresis until bromophenol blue runs out of the gel plate. At this time, turn off the electrophoresis instrument and end the electrophoresis.

[0243] 3) Transfer: Use wet transfer, cut the NC membrane to the appropriate size, pour the pre-cooled transfer buffer into a specific tray, and open the transfer clamp; gently pry the gel glass plate along one side, carefully remove the thin glass plate to expose the gel, and cut off the unnecessary part; place the sponge pad, 3 layers of filter paper, gel, NC membrane, 3 layers of filter paper, and finally the sponge pad on the transfer clamp from cathode to anode, and clamp it after all bubbles are removed; put the clamp into the transfer tank in the correct direction, add 1L transfer buffer, and then put in an ice bag to cool it down, cover the lid, and place it in an ice bath; 250mA constant current transfer, transfer time 2 hours.

[0244] 4) Blocking: After transfer, remove the NC membrane and block it with 5% BSA or skim milk at room temperature for 1 hour.

[0245] 5) Antibody incubation: Dilute the antibody with a specific primary antibody diluent according to the antibody instructions. After the NC membrane is cut into a suitable size according to the protein molecular weight, place it in the corresponding primary antibody diluent and incubate it on a shaker at 4°C overnight; take it out the next day, recover the antibody, add an appropriate amount of 1×TBST to the box where the NC membrane is located, and wash it on a shaker at room temperature for 3 times, 5 minutes each time; dilute the fluorescent secondary antibody with PBS at a ratio of 1:10000, add an appropriate amount to the washed NC membrane, and incubate it on a shaker at room temperature in the dark for 1 hour. After the secondary antibody incubation is completed, remove the secondary antibody incubation solution, add an appropriate amount of 1×TBST to the container where the NC membrane is located, and wash it on a shaker at room temperature in the dark for 3 times, 5 minutes each time.

[0246] 6) Scanning of experimental results: Using the Odyssey CLx infrared dual-color image analysis system of the Zhejiang University City College technology platform, select the appropriate scanning channel, set the optimal scanning intensity to identify and scan the fluorescent signal, and save and analyze the scanning results.

[0247] Results: After the MDA-MB-436 cells were treated with the obtained compounds P1 to P18 for 24 hours, the protein immunoblotting results showed that the degradation effect of compounds P1-P18 on PARP1 protein at a concentration of 10 μM was observed. The results showed that the 18 compounds P1-P18 in the present invention all had a certain degradation effect on PARP1, but had no degradation effect on PARP2. Among them, the degradation effect of compounds P3 to P4 on PARP1 protein in MDA-MB-436 cell line at 10 μM was as follows: Figure 1 shown.

[0248] Example 50: Half-maximal degradation concentration DC of compound P1-P18 for PARP1 degradation activity 50 Determination

[0249] In the MDA-MB-436 cell line, compounds P1 to P18 of the present invention were treated with the method of Example 49 for 72 hours, and then PARP1 and PARP2 of the cells were detected by immunoblotting. Semi-quantitative analysis was performed using Image J, and DC was calculated using GraphPad Prism. 50 After 72 hours of treatment with compounds P1-P18 obtained in Examples 2-19, the results of quantitative analysis of protein immunoblotting showed that most compounds had excellent PARP1 degradation ability. The DC obtained by semi-quantitative analysis of the degradation ability of the obtained compounds 50The data are divided into four gradients, as shown in Table 1. The degradation effects of compounds P3 and P4 on PARP1 protein in MDA-MB-436 cell line at 0.05 nM, 0.1 nM, 0.5 nM, 1 nM, and 5 nM, as well as their half-maximal degradation concentration DC 50 like Figure 2 shown.

[0250] Table 1 Degradation ability of the compounds in Examples 2-19 on PARP1 (72 hours)

[0251] Group <![CDATA[DC 50 Range (nM)]]> Compound No. 1 <1.00 P3,P4,P9,P10,P11,P13,P16,P18 2 1.00-10.00 P2,P8,P14,P15,P17 3 10.00-20.00 P5,P7 4 20.00-200 P1,P6,P12

[0252] Example 51: IC50 inhibition rate of compounds P1-P18 on tumor cell line MDA-MB-436 cells 50 Determination

[0253] The human breast cancer cell line MDA-MB-436 was selected for the experiment to investigate the half-inhibitory activity of the compound on human breast cancer cells. The corresponding cells were cultured in "L15 culture medium" containing 10% fetal bovine serum and 1% penicillin-streptomycin. The cells were cultured in an incubator at 37°C and 100% air saturated humidity, and cells in the logarithmic growth phase were used for the experiment. Cells in the logarithmic growth phase were taken and 4×10 4 / mL were inoculated in a 96-well culture plate, 180μL per well. 20μL of the compound with a concentration gradient (100μM, 10μM, 1μM, 0.1μM, 0.01μM) prepared in Example 2-19 and the positive control drug Olaparib (purchased from MCE) were added to the experimental group, and no drug was added to the negative control group. Three parallel wells were set up in each group, and four 96-well plates were repeated and cultured at 37°C for 5 days, 7 days, 10 days and 14 days. 10μL of enhanced CCK-8 solution was added to each well. Incubation continued in the cell culture incubator for 2 hours. The absorbance was measured at 450nm. The cell growth inhibition rate was calculated according to the absorbance [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD×100%], and the IC was calculated using GraphPad Prism 50 The experiment was repeated 3 times and the average value was taken to analyze the half inhibition rate IC of the obtained compounds on the tumor cell line MDA-MB-436 cells. 50 , divided into four gradients, the results are shown in Table 2. Among them, P3 and P4 compounds have an IC 50 like Figure 3 , from the results in the figure, we can see that its IC 50 The value was similar to that of the positive control drug Olaparib.

[0254] Table 2 Inhibitory effect of the compounds in Example 2-19 on the cell growth of MDA-MB-436 cells (7 days)

[0255] Group <![CDATA[IC 50 Range (μM)]]> Compound No. 1 <10.00 P3,P4,P8,P9,P10,P11,P13,P18 2 10.00-20.00 P5,P8,P14,P15,P12,P16,P17 3 20.00-200 P1,P2,P7,P6 Olaparib 9.39 Positive Control

[0256] Example 52: Half inhibition rate IC of compound P3 and P4 combined with cisplatin on tumor cell line MDA-MB-436 cells 50 Determination

[0257] The experiment selected the safe concentration of cisplatin in MDA-MB-436 cells: In order to test the effect of the compound on the sensitivity of cisplatin administration, the safe concentration of cisplatin in MDA-MB-436 cells was selected. Cells in the logarithmic growth phase were taken and 4×10 4 Cells / ml were inoculated in a 96-well culture plate, 180 μL per well. 20 μL of cisplatin with a concentration gradient (100uM, 10uM, 1uM, 0.1uM, 0.01uM) was added to the experimental group, and no drug was added to the negative control group. Each group had 3 parallel wells and was cultured at 37°C for 7 days. 10 microliters of enhanced CCK-8 solution was added to each well. Incubate in the cell culture incubator for 2 hours. Measure the absorbance at 450nm. Calculate the cell growth inhibition rate based on the absorbance [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD×100%], and according to the results Figure 4 Select a safe concentration. The experimental groups were added with 20 μL of the prepared safe concentration of cisplatin (0.5 μM) combined with a concentration gradient (0.5 nM, 1 nM, 5 nM) compound, the cisplatin administration group (0.5 μM), the cisplatin combined with the positive control drug Olaparib group (5 nM), and the negative control group without drug addition. Each group had 3 parallel wells and cultured at 37°C for 7 and 14 days. Add 10 microliters of enhanced CCK-8 solution to each well. Continue incubating in the cell culture incubator for 2 hours. Measure the absorbance at 450 nm. Calculate the cell growth inhibition rate based on the absorbance [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD×100%]. The experiment was repeated 3 times and the average value was taken. The results are as follows Figure 4 As shown in the figure, the data show that the inhibitory activity of compounds P3 and P4 in combination with cisplatin on tumor cell proliferation is significantly better than the combined activity of positive drug Olaparib and cisplatin.

Claims

1. A compound, characterized in that The structural formula of the compound is: Wherein, L is a linker unit; B is a CRBN E3 ubiquitin ligase ligand.

2. The compound according to claim 1, characterized in that The general structural formula of the E3 ubiquitin ligase ligand is: wherein Z is one of -CH2- and -C(=O)-, and X and Y are each independently one of H or F; The connecting unit L is one of the following structures: -NHCH2(CH2)nCH2NH-;-NH(CH2 CH2O)n(CH2) m NH-; -NHCH2(CH2)n CH2C(=O)-;-NH(CH2 CH2O)n(CH2) m C(=O)-; -NH(CH2 CH2O)n(CH2) m C(=O)NH(CH2) m C(=O)-; The n is 2 to 10; m, s, and t are each independently selected from 0, 1, 2, and 3; X1 and X2 are each independently selected from CH or N.

3. The compound according to claim 1, characterized in that The E3 ubiquitin ligase ligand is selected from the following structures: The connecting unit L is selected from the following structures:

4. The compound according to claim 1, characterized in that The compound has one of the following structures: Wherein, X and Y are one of H or F; in, Selected from:

5. The compound according to claim 1, characterized in that The structural formula of the compound is:

6. A drug for inhibiting tumor cell proliferation, characterized in that: The invention comprises the compound according to any one of claims 1 to 5 and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate and enantiomer.

7. A PARP1 protein degradation agent, characterized in that: The invention comprises the compound according to any one of claims 1 to 5 and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate and enantiomer.

8. An anti-tumor drug, characterized in that: The invention comprises the compound according to any one of claims 1 to 5 and its pharmaceutically acceptable salt or cocrystal, deuterated substance, solvate, and pharmaceutically acceptable auxiliary ingredients.

9. A combined anti-tumor drug, characterized in that: The invention comprises the compound according to any one of claims 1 to 5, and a chemotherapeutic drug used in combination therewith.

10. The combined anti-tumor drug according to claim 9, characterized in that: The chemotherapy drug is cisplatin.

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