A parp1 selective protein degrader and its application in anti-tumor

Abstract

The application discloses a PARP1 selective protein degradation agent and application thereof in anti-tumor. The degradation agent comprises a structural formula as follows. Wherein 3-ethyl-7-((4-(pyridine-3-yl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one is a PARP1 target protein ligand, L is a connecting unit, and B is an E3 ubiquitin ligase ligand. The PARP1 selective protein degradation agent prepared by the application can effectively degrade PARP1 protein, inhibit cell proliferation, and induce tumor cell apoptosis. Meanwhile, when the degradation agent is combined with a chemotherapy drug, the degradation agent has the effect of enhancing the efficacy of the chemotherapy drug, and almost has no physiological toxicity. The compound is expected to provide an ideal way for improving the treatment of various diseases caused by over-activation of PARP1.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a PARP1 selective protein degradation agent and application thereof in anti-tumor. BACKGROUND

[0002] In the process of life activities, the genome of organisms is susceptible to exogenous or endogenous factors, leading to DNA damage, wherein the endogenous damage includes DNA replication errors, hydrolysis DNA damage, cell biochemical processes and excessive reactive oxygen species (RSO) production, etc., and the 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 mutation, DNA crosslinking, DNA single-strand break (SSB), DNA double-strand break (DSB) and base mismatch.

[0003] During long-term evolution, living organisms have formed 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 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 break repair (SSBR) is a mechanism for coping with single-strand breaks in the double helix chain of DNA. SSBR mainly includes base excision repair (BER), mismatch repair (MMR), and nucleotide excision repair (NER). In the process of studying this repair mechanism, it is found that single-strand broken DNA usually leads to fatal DNA double-strand breaks if it cannot be repaired in time. DNA double-strand break repair (DSBR) mechanism mainly includes homologous recombination (HR) and non-homologous end joining (NHEJ). These two types of DNA damage repair mechanisms are essential for maintaining normal cell survival. Therefore, researchers have developed a series of methods to regulate these mechanisms to treat diseases such as tumors, for example, blocking tumor cell DNA repair pathways, interfering with proteins involved in DNA repair, etc.

[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 characteristic member of the poly ADP ribose polymerase (PARP) family. As a DNA-dependent nuclease, PARP1 plays a key role in the signaling and repair of DNA damage. After DNA damage caused by cell metabolic activity, chemical poisons or ionizing radiation, PARP1 is rapidly activated and then recognizes and binds to DNA single-strand breaks (SSBs). Among them, it catalyzes the transfer of nicotinamide adenine dinucleotide (NAD +) and then transfer ADP ribose to synthesize poly-ADP ribose chains on itself or several acceptor proteins. These polymeric chains with more negative charges and larger 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 part, which is a key process of 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 an adjunct to 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 known as "synthetic lethality". So far, six PARP inhibitors, including olaparib, rucaparib, niraparib, and talazoparib, have been approved for marketing. Olaparib, as the first oral potent PARP1 / 2 inhibitor, has been approved for clinical treatment of BRCA mutant-related tumor diseases.

[0005] However, there are still multiple challenges in the use of PARPi that limit its therapeutic efficacy, including long-term drug safety and the acquisition of drug resistance. In terms of drug safety, in addition to inhibiting PARP enzyme activity, PARPi can also cause PARP trapping (PARP-Trapping) during DNA damage repair. The trapping mechanism of PARP is that when PARP protein binds to DNA, PARPi prevents PARP protein from dissociating from DNA breaks to form a stable PARP1-PARPi complex. This complex can block and damage DNA replication, so its toxicity to normal cells is greater than that of unrepaired SSBs. Researchers cannot assess the specific effects of these two pathways (PARP catalytic inhibition and trapping) on PARP inhibitor-mediated cytotoxicity. Meanwhile, PARPi inhibition of PARP2 can trigger blood system-related adverse reactions. In addition to drug safety, the drug resistance of tumors caused by long-term use of PARPi cannot be ignored. The development of PARP inhibitor resistance is a complex process, one theory being that secondary mutations in the BRCA1 / 2 gene in cells restore the function of the protein and thus develop resistance to PARP inhibitors. After the development of drug resistance, the efficacy of PARPi is significantly reduced. In order to achieve the goal of selectively blocking the function of PARP and reducing the occurrence of PARP trapping, while improving the drug 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 drug research and development field, and the proteolysis targeting chimera (PROTAC) technology is a representative technology for tumor targeted therapy. The PROTAC molecule is a bifunctional small molecule containing two ligands, and the two ligands are connected together by a linker 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, leading to the polyubiquitination of the target protein, and then the degradation is mediated by the proteasome. The PROTAC strategy has made great progress in inducing the degradation of target proteins in vitro and in vivo, such as BET, AR, ER and CDK9, which is attributed to the discovery of several high-efficiency small-molecule ligands for E3 ubiquitin ligases, such as thalidomide, pomalidomide, etc. Compared with traditional small-molecule inhibitors, PROTAC compounds have their unique advantages. Using the PROTAC technology, researchers have developed many PROTAC compounds that can efficiently degrade pathogenic proteins in vitro and in vivo, and currently, several anti-tumor drugs based on this technology have entered clinical research. The present application uses the PROTAC technology to design a PARP1 degrader, in order to overcome the defects of small-molecule PARP1 inhibitors and provide a new method for tumor targeted therapy. SUMMARY

[0007] The purpose of the embodiments of the present application is to overcome 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 overcome 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 object, the technical scheme adopted by the present application to solve its technical problems is:

[0010] A compound, the structural formula of the compound is:

[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 connecting unit, comprising a covalently coupled bond or a chemical linking moiety in the molecule of the compound, including at least one or a combination of a straight-chain diamine connecting segment, a straight-chain amino acid connecting segment, or a cyclic structure connecting segment; or a compound formed by a straight-chain amino acid of a certain length, a straight-chain diamine, piperazine, piperidine, pyrrolidine, and azetidine;

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

[0015] The structural general 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 2-10; m, s, t are each independently selected from 0, 1, 2, 3; X1, X2 are each independently selected from CH or N.

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

[0026]

[0027]

[0028] Further, the straight-chain diamine connecting segment is a connecting segment from a fatty chain diamine or a polyethylene glycol diamine.

[0029] Further, the linear diamine connecting segment is at least one from the group consisting of butanediamine, pentanediamine, hexanediamine, heptanediamine, octanediamine, nonanediamine, decanediamine, undecanediamine, dodecanediamine, and diamine containing oxygen atom.

[0030] Further, X, Y is one of H atom or fluorine atom.

[0031] Further, the connecting unit L is composed of linear amino acid connecting segment; the E3 ubiquitin ligase ligand is CRBN ligand lenalidomide; the structural formula of the compound is:

[0032]

[0033] Further, the linear amino acid connecting segment is at least one from the group consisting of glycine, beta-alanine, aminobutyric acid, aminopentanoic acid, aminohexanoic acid, aminooctanoic acid, aminononanoic acid, aminodecanoic acid, aminoundecanoic acid, aminododecanoic acid, and polyethylene glycol-containing amino acid.

[0034] Further, the connecting unit L is composed of cyclic structure connecting segment or combination of cyclic structure connecting segment and aliphatic chain connecting segment; the E3 ubiquitin ligase ligand is CRBN ligand thalidomide and its derivatives; the structural formula of the compound is:

[0035]

[0036] Further, the cyclic structure is aliphatic cyclic structure.

[0037] Further, the aliphatic cyclic structure is at least one from the group consisting of piperazine, piperidine, pyrrolidine, azetidine.

[0038] Further, the aliphatic chain is linear amino acid compound.

[0039] Further, the linear amino acid is one from the group consisting of aminooctanoic acid and aminodecanoic acid.

[0040] Further, X, Y is one of H atom or 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 medicine for inhibiting tumor cell proliferation, comprising the compound and pharmaceutically acceptable salts or co-crystals, deuterium derivatives, solvates, enantiomers thereof.

[0047] A PARP1 protein degradation agent, comprising the compound and pharmaceutically acceptable salts or co-crystals, deuterium derivatives, solvates, enantiomers thereof.

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

[0049] An anti-tumor medicine, comprising the compound and pharmaceutically acceptable salts or co-crystals, deuterium derivatives, solvates, enantiomers thereof, and pharmaceutically acceptable auxiliary ingredients thereof.

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

[0051] A combined anti-tumor medicine, comprising the compound and a chemotherapeutic drug used in combination.

[0052] Further, the chemotherapeutic drug is cisplatin.

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

[0054] The PARP1 selective protein degradation agent prepared by the present application can effectively degrade PARP1 protein, inhibit cell proliferation, and induce tumor cell apoptosis. At the same time, when used in combination with a chemotherapeutic drug, it has the effect of enhancing the efficacy of the chemotherapeutic drug, and almost no physiological toxicity. The compound is expected to provide an ideal way to improve the treatment of various diseases caused by overactivation of PARP1.

[0055] It should be understood that the above general description and the following detailed description are only exemplary and explanatory, and cannot limit the present application. BRIEF DESCRIPTION OF DRAWINGS

[0056] Figure 1 The degradation effect of compounds P3 and P4 on intracellular PARP1 / 2 protein was detected;

[0057] Figure 2 The degradation effect of compounds P3 and P4 on intracellular PARP1 / 2 protein was detected; 50 The detection results;

[0058] Figure 3 The half-inhibition rate IC of compounds P3 and P4 on tumor cell line MDA-MB-436 cells was detected; 50 The detection results;

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

[0060] The exemplary embodiments will be described in detail herein with reference to the attached drawings. In the following description, like reference numerals refer to like elements, unless the context clearly dictates otherwise. The following exemplary embodiments are described in the context of the implementations described herein, but the implementations are not intended to represent all implementations consistent with the present disclosure.

[0061] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and / or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0062] It will be understood that, although the terms first, second, third, etc. can be used herein to describe various information, the information should not be limited by these terms. These terms are used only to distinguish one piece of information from another. For example, a first information could be termed a second information, and, similarly, a second information could be termed a first information without departing from the scope of the present disclosure. As used herein, the term "if' can be construed to mean "when" or "in response to determining" depending on the context.

[0063] Example 1 Preparation of PARP1 small molecule ligand

[0064] The synthetic route thereof is as follows:

[0065]

[0066]

[0067] In a 500 mL round bottom flask, 1 (19.9 g, 100 mmol), 2 (15.3 g, 110 mmol) were dissolved in 200 mL THF, and the reaction solution was cooled to 0 °C. After adding NaHCO3(9.24 g, 110 mmol), the reaction was carried out at room temperature for 12 h. TLC monitoring showed that the raw material was completely reacted. 300 mL water was added to the reaction solution. Then the reaction solution was 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 x 20 mL). The combined organic phase was evaporated under reduced pressure to obtain the intermediate crude product 3 (26.8 g, 92% yield, yellow solid). 1H NMR (400 MHz, CDC13) δ 8.90 (d, J = 1.6 Hz, 1H), 8.68 (d, J = 6.8 Hz, 1H), 8.05 (dd, J = 9.2, 1.2 Hz, 1H), 6.75 (d, J = 9.2 Hz, 1H), 4.28 (dd, J = 13.2, 6.4 Hz, 1H), 3.90 (s, 3H), 3.80 (s, 3H), 2.03 (ddd, J = 21.6, 14.4, 7.2 Hz, 2H), 1.06 (t, J = 7.6 Hz, 3H); HRMS (ESI): m / z calcd for C 13 H 16 N2O6[M+H] + :297.1008, found:297.1011.

[0068] In a 1000 mL three-necked flask, 3 (29.6 g, 100 mmol), Fe (44.6 g, 800 mmol), NH4CI (42.8 g, 800 mmol) were dissolved in 500 mL MeOH and the reaction was placed under mechanical stirring. The reaction was heated to 85 °C for 12 h. TLC monitoring showed that the starting material was completely consumed. The methanol was evaporated under reduced pressure and the residue was dissolved in 200 mL DMF and heated to 90 °C. The reaction was filtered hot and the filter cake was washed with a small amount of hot DMF. The filtrate was poured into a large amount of ice water and the filter cake was retained and dried to give the crude intermediate 4 (16.4 g, 70% yield, white solid). 1 H NMR (400 MHz, DMSO-d6) δ 7.82 - 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 for C 12 H 14 N2O3[M+H] + :235.1077, found:235.1078.

[0069] In a 250 mL round-bottom flask, 4 (23.4 g, 100 mmol) was dissolved in 100 mL 1,4-dioxane and the reaction was cooled to 10 °C. DDQ (24.9 g, 110 mmol) was added slowly in portions. After the addition was complete, the reaction was allowed to warm to room temperature and stirred for 0.5 h. TLC monitoring showed that the starting material was completely consumed. Saturated aqueous NaHC03was added to the reaction and the pH was adjusted to 7-8. The reaction was filtered and the filter cake was washed with a small amount of water. The filter cake was retained and dried to give the crude intermediate 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 in 150 mL THF, and cool the reaction solution to 0 °C. Slowly add LiAlH4 (6.0 g, 150 mmol) in portions. After completion, transfer to room temperature and react for 0.5 h. Monitor the reaction of the starting material by TLC until complete. Add a small amount of aqueous solution to the reaction solution to quench the 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 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] Add 6 (2.0 g, 10 mmol) to a 100 mL round-bottom flask, dissolve it in 20 mL of HBr / AcOH, and heat the reaction solution to 80 °C for 2 h. After completion, add ice water, and a large amount of solid precipitates in the reaction solution. Filter the reaction solution, wash the filter cake with water, retain the filter cake, and dry it to obtain intermediate crude product 7 (2.1 g, 80% yield, white solid). 1H NMR (400 MHz, DMSO-d6) δ 8.34 (t, J = 6.4 Hz, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.69 (t, J = 9.6 Hz, 1H), 7.35 (dt, J = 13.6, 6.8 Hz, 1H), 3.63 (s, 2H), 2.81 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H); HRMS (ESI): m / z calcd for C 11 H 11 BrN2O[M+H] + :267.0128, found:267.0126.

[0072] In a 500 mL round bottom flask, 8 (15.5 g, 100 mmol), 9 (20.4 g, 110 mmol) were dissolved in 100 mL THF, then the reaction was heated to 60 °C, after the addition of K2CO3 (15.2 g, 110 mmol), the reaction was carried out at room temperature for 12 h. TLC monitoring of the complete reaction of raw materials, 100 mL water was added to the reaction solution. Then the reaction was extracted with ethyl acetate (3 x 100 mL), the organic phase was combined and dried, purified by silica gel column (PE:EA = 2:1), then 30% TFA / DCM was added, the reaction was carried out for 0.5 h, the solvent was evaporated under reduced pressure, and the product 10 (18.1 g, 82% yield, white solid) was obtained. 1 H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 7.60 (m, 1H), 7.32 (m, 1H), 7.06 (t, J = 7.2 Hz, 1H), 2.07 (s, 3H), 1.40 (m, 8H); HRMS (ESI): m / z calcd for C 11 H 15 N3O2[M+H] + :222.1164, found:222.1164.

[0073] In a 100 mL round bottom flask, 7 (2.66 g, 10 mmol), 10 (2.21 g, 10 mmol), Et3N (1.5 g, 11 mmol) were dissolved in 20 mL CH3CN, then the reaction was heated to 60 °C for 2 h, TLC detection of complete reaction, after cooling, a large amount of solid was observed in the reaction solution. The reaction was filtered, the filter cake was washed with water, 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), LiOH (1.0 g, 40 mmol) were added in a 100 mL round bottom flask, dissolved in 10 mL MeOH and 10 mL H2O, stirred overnight, after the reaction was completed, the pH was adjusted to 5-6, and a large amount of solid was observed in the reaction solution. The reaction solution was filtered, the filter cake was washed with water, the filter cake was retained, and the product 12 (3.3 g, 85% yield, white solid) was obtained by drying. 1 H NMR (400 MHz, DMSO-d6) δ 12.26 (s, 1H), 8.38 (d, J = 2.0 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.34 (dd, J = 8.8, 2.0 Hz, 1H), 7.28 (s, 1H), 7.24 (d, J = 8.0 Hz, 1H), 3.62 (s, 2H), 3.40 (m, 4H), 2.80 (q, J = 7.2 Hz, 2H), 2.55 (s, 4H), 1.22 (t, J = 7.2 Hz, 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] The synthetic route thereof is:

[0078]

[0079] General procedure A: In a 25 mL reaction flask, B1 (276 mg, 1.0 mmol), L1 (202 mg, 1.0 mmol) were added, after dissolved in 10 mL NMP, dropwise added Et3N (150 mg, 1.5 mmol), 90 °C for 2-4 h, TLC monitoring of raw materials until the reaction was complete, 10 mL water was added to the reaction flask, then extracted with ethyl acetate (3 x 20 mL), the combined organic phase was evaporated under reduced pressure, purified by silica gel column chromatography (PE:EA = 1:1), the obtained intermediate was added to 30% TFA / DCM 10 mL for 2 h, evaporated under reduced pressure to obtain the crude intermediate Z1 (286 mg, 80% yield, yellow solid). In a 25 mL reaction flask, compound 12 (393 mg, 1.0 mmol) was dissolved in 10 mL 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 portionwise with stirring, the reaction was placed in an ice water bath for 1-2 h, 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 half-saturated brine was added to dilute the reaction solution, then extracted with ethyl acetate (3 x 20 mL), the combined organic layer was washed once with saturated aqueous sodium chloride solution, dried over 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 separate the product P1 (219 mg, 30% yield, yellow solid). 1 H NMR (400 MHz, CDCl3) δ 11.23 (s, 1H), 8.94 (s, 1H), 8.08 (s, 1H), 7.98-7.95 (m, 1H), 7.81 (t, J = 6.0 Hz, 1H), 7.71 (d, J = 4.0 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 7.25 (t, J = 7.2 Hz, 2H), 7.14 (d, J = 9.2 Hz, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.78 (d, J = 8.4 Hz, 1H), 6.17 (s, 1H), 4.88 (d, J = 6.4 Hz, 1H), 3.64 (s, 2H), 3.40 (d, J = 3.6 Hz, 2H), 3.31 (s, 4H), 2.98-2.83 (m, 4H), 2.69 (d, J = 9.2 Hz, 2H), 1.62 (d, J = 6.8 Hz, 4H), 1.45 (d, J = 6.4 Hz, 2H), 1.31-1.25 (m, 6H), 0.80 (d, J = 6.4 Hz, 3H). HRMS (ESI): m / z calcd for C 39 H 43 N9O6[M+H]+ :734.3409, found:734.3410.

[0080] The structures of L involved in the following examples are listed in the table below, respectively:

[0081]

[0082]

[0083] The structure of B is listed in the table below:

[0084]

[0085] Example 3

[0086] Preparation of compound P2

[0087] P2 was synthesized according to general procedure A with B1, L2 and 12 as the reactants. The product P2 was obtained as a yellow solid in 31% yield. 1 H NMR (500 MHz, DMSO-d6) δ 11.03 (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 = 8.5 Hz, 1H), 7.01 (d, J = 8.5 Hz, 1H), 6.94 (d, J = 7.0 Hz, 1H), 6.45 (t, J = 6.0 Hz, 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] P3 was synthesized by the general procedure A with B1, L3 and 12 as the reactants to give product P3 as a yellow solid in 33% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.28 (s, 1H), 11.11 (s, 1H), 8.34 (t, J = 6.0 Hz, 1H), 8.27 (s, 1H), 7.83 (d, J = 9.0 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.57 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 6.5 Hz, 1H), 7.28 - 7.23 (m, 2H), 7.12 (d, J = 8.5 Hz, 1H), 7.03 (d, J = 7.0 Hz, 1H), 6.60 (s, 1H), 5.06 (dd, J = 12.5, 5.5 Hz, 1H), 3.61 (d, J = 9.5 Hz, 4H), 3.52 (t, J = 10.0 Hz, 10H), 3.45 - 3.38 (m, 6H), 2.82 - 2.78 (m, 2H), 2.58 (d, J = 14.0 Hz, 6H), 1.22 (t, J = 7.5 Hz, 3H). HRMS (ESI): m / z calcd 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] P4 was synthesized by the general procedure A with B1, L4 and 12 as the reactants to give product P4 as a yellow solid in 15% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.28 (s, 1H), 11.11 (s, 1H), 8.34 (t, J = 5.5 Hz, 1H), 8.27 (s, 1H), 7.83 (d, J = 9.0 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.57 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 6.5 Hz, 1H), 7.28 - 7.23 (m, 2H), 7.12 (d, J = 8.5 Hz, 1H), 7.03 (d, J = 7.0 Hz, 1H), 6.60 (s, 1H), 5.06 (dd, J = 13.0, 5.5 Hz, 1H), 3.61 (d, J = 9.5 Hz, 4H), 3.52 (m, 10H), 3.45 - 3.38 (m, 10H), 2.82 - 2.78 (m, 2H), 2.58 (d, J = 12.0 Hz, 6H), 1.22 (t, J = 7.5 Hz, 3H). HRMS (ESI): m / z calcd for C 44 H 53 N9O6[M+H] + : 804.4192, found: 804.4188. Its chemical structural formula is:

[0096]

[0097] Example 6: Preparation of compound P5

[0098] The synthetic route is:

[0099]

[0100] General Procedure B: In a 25 mL reaction vial was placed B2 (259 mg, 1.0 mmol), L5 (254 mg, 1.1 mmol), dissolved in 10 mL dry DMF and placed in an ice water bath, DIPEA (258 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added portion wise with stirring, the reaction was placed in an ice water bath for 1-2 h, TLC monitoring of the reaction was used to determine completion of the reaction, the reaction was diluted with 50 mL saturated brine, extracted with ethyl acetate (3 x 20 mL), the organic layers were combined and washed once with saturated aqueous sodium chloride, dried over anhydrous Na2SO4, filtered and the solvent was removed under reduced pressure to give the crude product as an oil, the crude product was purified by column chromatography using DCM:MeOH = 10:1 to 5:1 to give the intermediate as a white solid, the intermediate was dissolved in 10 mL of 30% TFA / DCM and stirred for 2 h, the solvent was removed under reduced pressure to give the crude intermediate Z5 (231 mg, 60% yield, white solid). In a 25 mL reaction vial was placed compound 12 (393 mg, 1.0 mmol), dissolved in 10 mL dry DMF and placed in an ice water bath, DIPEA (258 mg, 2.0 mmol) was added, HATU (570 mg, 1.5 mmol) was added portion wise with stirring, the reaction was placed in an ice water bath for 1-2 h, the intermediate Z5 (386 mg, 1.0 mmol) was added, the reaction was transferred to room temperature and stirred for 3 h. TLC monitoring of the reaction was used to determine completion of the reaction, the reaction was diluted with 50 mL saturated brine, extracted with ethyl acetate (3 x 20 mL), the organic layers were combined and washed once with saturated aqueous sodium chloride, dried over anhydrous Na2SO4, filtered and the solvent was removed under reduced pressure to give the crude product as an oil, the crude product was purified by column chromatography using DCM:MeOH = 10:1 to 5:1 to give the product P5 (219 mg, 25% yield, white solid). 1H NMR (400 MHz, DMSO-d6) δ 12.30 (s, 1H), 11.05 (s, 1H), 9.77 (s, 1H), 8.40 (s, 1H), 8.27 (s, 1H), 7.83 (t, J = 6.4 Hz, 2H), 7.70 (d, J = 8.0 Hz, 1H), 7.50 (t, J = 7.6 Hz, 2H), 7.39 (d, J = 7.2 Hz, 1H), 7.31 - 7.22 (m, 2H), 5.16 (dd, J = 13.2, 4.8 Hz, 1H), 4.37 (q, J = 17.6 Hz, 2H), 3.64 (s, 2H), 3.25 (d, J = 6.0 Hz, 2H), 3.07 - 2.70 (m, 6H), 2.72 - 2.53 (m, 6H), 2.43 - 2.22 (m, 4H), 2.03 (d, J = 6.8 Hz, 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 was performed according to the general procedure B using B2, L6 and 12 as starting materials to give the product P6 as a white solid in 34% yield. 1 H NMR (400 MHz, DMSO-d6) δ 12.30 (s, 1H), 11.05 (s, 1H), 9.77 (s, 1H), 8.40 (s, 1H), 8.27 (s, 1H), 7.83 (t, J = 6.4 Hz, 2H), 7.70 (d, J = 8.0 Hz, 1H), 7.50 (t, J = 7.6 Hz, 2H), 7.39 (d, J = 7.2 Hz, 1H), 7.31 - 7.22 (m, 2H), 5.16 (dd, J = 13.2, 4.8 Hz, 1H), 4.37 (q, J = 17.6 Hz, 2H), 3.64 (s, 2H), 3.25 (d, J = 6.0 Hz, 2H), 3.07 - 2.70 (m, 6H), 2.72 - 2.53 (m, 6H), 2.43 - 2.22 (m, 4H), 2.03 (d, J = 6.8 Hz, 4H), 1.60 (s, 2H), 1.49 (s, 3H). HRMS (ESI): m / z calcd for C 43 H 51N9O6[M+H] + : 790.4035, found: 790.4033. Its chemical structural formula is:

[0104]

[0105] Example 8

[0106] Preparation of compound P7

[0107] The synthesis of P7 was performed according to the general procedure B by using B2, L7 and 12 as raw materials to give product P7 as a white solid in 26% yield. 1 H NMR (400 MHz, DMSO-d6) δ 12.30 (s, 1H), 11.05 (s, 1H), 9.77 (s, 1H), 8.40 (s, 1H), 8.27 (s, 1H), 7.83 (t, J = 6.4 Hz, 2H), 7.70 (d, J = 8.0 Hz, 1H), 7.50 (t, J = 7.6 Hz, 2H), 7.39 (d, J = 7.2 Hz, 1H), 7.31 - 7.22 (m, 2H), 5.16 (dd, J = 13.2, 4.8 Hz, 1H), 4.37 (q, J = 17.6 Hz, 2H), 3.64 (s, 2H), 3.25 (d, J = 6.4 Hz, 4H), 3.03 - 2.86 (m, 2H), 2.82 - 2.76 (m, 4H), 2.72 - 2.53 (m, 6H), 2.43 - 2.22 (m, 2H), 2.03 (d, J = 6.8 Hz, 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 structural formula is:

[0108]

[0109] Example 9

[0110] Preparation of compound P8

[0111] The synthesis of P8 was performed according to the general procedure B by using B2, L8 and 12 as raw materials to give product P8 as a white solid in 43% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.58 (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, 12.0 Hz, 3H), 5.15 (dd, J = 13.0, 4.5 Hz, 1H), 4.44 (d, J = 18.0 Hz, 2H), 4.36 (q, J = 17.5 Hz, 2H), 4.07 (s, 2H), 3.30 - 3.08 (m, 8H), 2.96 - 2.78 (m, 4H), 2.34 (d, J = 6.5 Hz, 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 structural formula is:

[0112]

[0113] Example 10

[0114] Preparation of compound P9

[0115] The synthesis of P9 was performed according to the general procedure B, using B2, L9 and 12 as starting materials to give the product P9 as a white solid in 35% yield. 1H NMR (500 MHz, CDC13) δ 8.33 (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, 4.0 Hz, 1H), 4.32 (d, J = 8.0 Hz, 2H), 3.70 (ddd, J = 12.5, 8.5, 3.5 Hz, 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, 9.0 Hz, 3H). HRMS (ESI): m / z calcd for C 43 H 51 N9O9[M+H] +:838.3883, found: 838.3885. Its chemical structural formula is:

[0116]

[0117] Example 11

[0118] Preparation of compound P10

[0119] The synthesis of P10 was performed according to the general procedure B with B2, L10 and 12 as the reactants to give the product P10 as a white solid in 27% yield. 1 H NMR (500 MHz, CDC13) δ 11.04 (s, 1H), 9.86 (s, 1H), 8.35 (t, J = 5.5 Hz, 1H), 8.28 (d, J = 2.5 Hz, 1H), 7.83 (t, J = 7.0 Hz, 2H), 7.69 (d, J = 8.0 Hz, 1H), 7.53 - 7.48 (m, 2H), 7.40 (dd, J = 9.0, 2.5 Hz, 1H), 7.31 - 7.17 (m, 2H), 5.15 (dd, J = 13.5, 5.0 Hz, 1H), 4.35 (q, J = 17.5 Hz, 2H), 3.70 (t, J = 6.5 Hz, 2H), 3.64 (s, 2H), 3.49 (dd, J = 8.5, 5.5 Hz, 12H), 3.42 (dd, J = 12.0, 6.0 Hz, 4H), 3.39 (s, 2H), 3.35 (s, 6H), 2.80 (q, J = 7.5 Hz, 2H), 2.59 (dd, J = 15.5, 9.0 Hz, 6H), 1.09 (t, J = 7.0 Hz, 3H). HRMS (ESI): m / z calcd for C 45 H 55 N9O 10 [M+H] + :882.4145, found: 882.4144. Its chemical structural formula is:

[0120]

[0121] Example 12: Preparation of compound P11, whose synthetic route is as follows:

[0122]

[0123] General procedure C: In a 25 mL reaction flask, B3 (276 mg, 1.0 mmol), L2 (202 mg, 1.0 mmol) were added, after dissolved in 10 mL NMP, Et3N (150 mg, 1.5 mmol) was added dropwise, 90 °C for 2 h, TLC monitoring of the complete reaction of raw materials, 10 mL water was added to the reaction flask, then extracted with ethyl acetate (3 x 20 mL), the combined organic phase was evaporated under reduced pressure, purified by silica gel column chromatography (PE:EA = 1:1), the obtained intermediate was added to 30% TFA / DCM 10 mL for 2 h, evaporated under reduced pressure to obtain the crude intermediate Z11 (286 mg, 80% yield, yellow solid). In a 25 mL reaction flask, compound 12 (393 mg, 1.0 mmol) was 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 portionwise with stirring, the reaction was placed in an ice water bath for 1-2 h, then intermediate Z11 (358 mg, 1.0 mmol) was added, and the reaction was transferred to room temperature for 3 h. After TLC monitoring of the completion of the reaction, 50 mL of half-saturated brine was added to dilute the reaction solution, then extracted with ethyl acetate (3 x 20 mL), the combined organic layer was washed once with saturated aqueous sodium chloride solution, dried over 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 separate the product P11 (204 mg, 28% yield, yellow-green solid). 1 H NMR (400 MHz, DMSO) δ 12.27 (s, 1H), 11.06 (s, 1H), 8.43 - 8.33 (m, 2H), 8.27 (s, 1H), 7.83 (d, J = 7.2 Hz, 1H), 7.69 (d, J = 7.2 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.38 (t, J = 10.3 Hz, 2H), 7.25 (d, J = 8.4 Hz, 2H), 7.10 (s, 1H), 5.03 (dd, J = 12.8, 5.2 Hz, 1H), 3.62 (s, 6H), 3.26 (s, 6H), 3.13 (d, J = 15.2 Hz, 2H), 2.80 (dd, J = 14.4, 7.2 Hz, 4H), 2.54 (d, J = 10.0 Hz, 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 follows general procedure C, using B3, L1, and 12 as reactants to obtain product P12, 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 structural formula is:

[0126]

[0127] Example 14

[0128] Preparation of compound P13

[0129] The synthesis of P13 follows general step C, using B3, L4, and I2 as reactants to obtain product P13, a yellow solid, in 22% yield. 1H NMR (400 MHz, DMSO-d6) δ 12.57 (s, 1H), 11.06 (s, 1H), 8.44 (d, J = 5.8 Hz, 2H), 8.34 (s, 1H), 7.90 (d, J = 8.8 Hz, 2H), 7.84 (d, J = 8.4 Hz, 2H), 7.56 (t, J = 8.0 Hz, 2H), 6.95 (s, 1H), 6.85 (d, J = 8.2 Hz, 1H), 5.03 (dt, J = 22.0, 11.0 Hz, 1H), 3.27 (dd, J = 11.2, 6.4 Hz, 4H), 3.16 (m, 6H), 2.91 (s, 2H), 2.86 - 2.81 (m, 4H), 2.75 (d, J = 8.4 Hz, 2H), 1.57 (d, J = 4.0 Hz, 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] Example 15

[0132] Preparation of compound P14

[0133] The synthesis of P14 was performed according to the general procedure A using B1, L3 and 12 as the starting materials to give the product P14 as a yellow solid in 23% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.28 (s, 1H), 11.11 (s, 1H), 8.34 (t, J = 6.0 Hz, 1H), 8.27 (s, 1H), 7.83 (d, J = 9.0 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.57 (t, J = 8.0 Hz, 1H), 7.39 (d, J = 6.5 Hz, 1H), 7.28 - 7.23 (m, 2H), 7.12 (d, J = 8.5 Hz, 1H), 7.03 (d, J = 7.0 Hz, 1H), 6.60 (s, 1H), 5.06 (dd, J = 12.5, 5.5 Hz, 1H), 3.61 (d, J = 9.5 Hz, 4H), 3.52 (t, J = 10.0 Hz, 6H), 3.45 - 3.38 (m, 10H), 2.82 - 2.78 (m, 2H), 2.58 (d, J = 14.0 Hz, 6H), 1.22 (t, J = 7.5 Hz, 3H). HRMS (ESI): m / z calcd for C 40 H 45 N9O8[M+H] + : 780.3464, found: 780.3465. Its chemical structural formula is:

[0134]

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

[0136]

[0137] General Step D: Add B2 (259 mg, 1.0 mmol) and L11 (192 mg, 1.1 mmol) to a 25 mL reaction flask, dissolve in 10 mL of anhydrous DMF, and place in an ice-water bath. Add DIPEA (258 mg, 2.0 mmol), and add HATU (570 mg, 1.5 mmol) in portions while stirring. Incubate the reaction in an ice-water bath for 1-2 h. After the reaction is complete as monitored by TLC, dilute the reaction solution with 20 mL of semi-saturated saline solution, and extract with ethyl acetate (3 × 20 mL). Combine the organic layers, wash once with saturated sodium chloride aqueous solution, dry with anhydrous Na2SO4, filter, and evaporate the solvent under reduced pressure to obtain an oily crude product. Finally, perform column chromatography using a DCM:MeOH system of 10:1 to 5:1 to obtain a white solid intermediate. Add 30% TFA / DCM to the obtained intermediate. The reaction was carried out in 10 mL for 2 h. The solvent was removed by vacuum distillation to obtain crude intermediate Z15 (198 mg, 60% yield, white solid). Then, Z15 (330 mg, 1.0 mmol) and L5 (254 mg, 1.1 mmol) were added to a 25 mL reaction flask, 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 portions with stirring. The reaction was carried out in an ice-water bath for 1-2 h. After the reaction was completed by TLC monitoring, 20 mL of semi-saturated saline solution was added to the reaction solution for dilution, followed by extraction with ethyl acetate (3 × 20 mL). The combined organic layers were washed once with saturated sodium chloride aqueous solution, dried over anhydrous Na2SO4, filtered, and the solvent was removed by vacuum distillation to obtain an oily crude product. Finally, column chromatography was performed using a DCM:MeOH system of 10:1–5:1 to obtain a white solid intermediate. The obtained intermediate was then added to 30% TFA / DCM. The reaction mixture was reacted in 10 mL for 2 h. The solvent was removed by vacuum distillation to obtain a crude intermediate (222 mg, 50% yield, white solid). This intermediate was added to a 25 mL reaction flask, followed by compound 12 (393 mg, 1.0 mmol). The mixture was 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 portions with stirring. The reaction was carried out in an ice-water bath for 1-2 h, and then transferred to room temperature for 3 h. After the reaction was completed by TLC monitoring, 50 mL of semi-saturated saline solution was added to the reaction solution for dilution, followed by extraction with ethyl acetate (3 × 20 mL). The combined organic layers were washed once with saturated sodium chloride aqueous solution, dried over anhydrous Na2SO4, filtered, and the solvent was removed by vacuum distillation to obtain an oily crude product. Finally, the product P15 (241 mg, 30% yield, white solid) was obtained by column chromatography using a DCM:MeOH system of 10:1 to 5:1. 1H NMR (400 MHz, DMSO-d6) δ 11.95 (s, 1H), 10.96 (d, J = 16.4 Hz, 1H), 8.39 (t, J = 7.2 Hz, 1H), 7.89 (t, J = 8.0 Hz, 1H), 7.73 - 7.63 (m, 1H), 7.36 (dd, J = 8.8, 2.8 Hz, 1H), 7.27 (d, J = 9.6 Hz, 1H), 7.22 (t, J = 7.6 Hz, 1H), 6.94 (d, J = 7.2 Hz, 1H), 6.83 (d, J = 7.6 Hz, 1H), 6.74 (d, J = 2.0 Hz, 1H), 5.12 (dt, J = 16.4, 8.0 Hz, 1H), 4.19 (dd, J = 9.2, 6.8 Hz, 2H), 3.83 (s, 2H), 3.61 (d, J = 4.2 Hz, 2H), 3.45 - 3.38 (m, 2H), 2.94 (ddd, J = 23.2, 13.2, 6.0 Hz, 2H), 2.85 - 2.77 (m, 2H), 2.69 - 2.54 (m, 2H), 2.21 (t, J = 7.2 Hz, 2H), 1.54 - 1.43 (m, 2H), 1.38 (d, J = 11.6 Hz, 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 was performed according to the general procedure D using B2, L5, L12 and 12 as starting materials to give the product P16 as a white solid in 14% yield. 1H NMR (400 MHz, DMSO-d6) δ 12.17 (s, 1H), 11.54 (s, 1H), 8.39 (d, J = 14.9 Hz, 1H), 7.92 (d, J = 8.8 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H), 7.35 - 7.28 (m, 2H), 7.24 (t, J = 7.6 Hz, 2H), 6.97 (d, J = 7.2 Hz, 2H), 6.86 (d, J = 7.6 Hz, 2H), 5.18 - 5.13 (m, 1H), 4.22 (d, J = 12.0 Hz, 2H), 3.86 (s, 4H), 3.67 (s, 2H), 3.05 (dd, J = 17.2, 10.4 Hz, 2H), 2.93 (dd, J = 18.0, 12.4 Hz, 4H), 2.85 (d, J = 7.2 Hz, 2H), 2.37 (m, 2H), 2.22 (d, J = 6.8 Hz, 2H), 2.06 (d, J = 6.0 Hz, 2H), 1.52 (dd, J = 18.8, 11.6 Hz, 4H), 1.27 (t, J = 6.8 Hz, 6H), 1.02 (t, J = 6.0 Hz, 3H). HRMS (ESI): m / z calcd for C 43 H 50 N 10 O7[M+H] + : 819.3937, found: 819.3938. Its chemical structural formula is:

[0140]

[0141] Example 18: Preparation of compound P17

[0142] The synthesis of P17 was refered to the general procedure D, using B2, L6, L11 and 12 as raw materials, the product P17 was obtained, white solid, yield 16%. 1H NMR (400 MHz, DMSO-d6) δ 12.26 (s, 1H), 11.96 (s, 1H), 10.97 (d, J = 6.4 Hz, 1H), 8.39 (s, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.35 (d, J = 8.0 Hz, 1H), 7.31 - 7.23 (m, 2H), 7.20 (d, J = 6.8 Hz, 1H), 6.93 (d, J = 6.8 Hz, 1H), 6.82 (d, J = 7.2 Hz, 1H), 6.71 (d, J = 6.8 Hz, 1H), 5.37 (d, J = 8.0 Hz, 2H), 5.09 (t, J = 23.2 Hz, 1H), 4.18 (dd, J = 19.2, 6.8 Hz, 2H), 3.82 (m, 2H), 3.64 (m, 2H), 3.42 (m, 4H), 2.86 (dd, J = 14.8, 5.6 Hz, 6H), 2.57 (s, 4H), 2.33 (d, J = 12.8 Hz, 2H), 2.20 (m, 2H), 1.25 (m, 10H), 1.05 (t, J = 4.0 Hz, 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 was refered to the general procedure D, using B2, L6, L12 and 12 as raw materials. The product P18 was obtained as white solid in 13% yield. 1H NMR (400 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.82 (s, 1H), 8.36 (d, J = 8.4 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 7.6 Hz, 1H), 7.30 - 7.23 (m, 2H), 7.21 (t, J = 7.2 Hz, 1H), 6.93 (d, J = 7.2 Hz, 1H), 6.88 (s, 1H), 6.82 (d, J = 7.8 Hz, 1H), 6.74 (s, 1H), 5.42 (s, 2H), 5.12 (d, J = 8.8 Hz, 1H), 4.27 - 4.12 (m, 2H), 3.82 (m, 3H), 3.63 (m, 2H), 3.01 (d, J = 7.2 Hz, 2H), 2.94 (dd, J = 11.2, 5.2 Hz, 4H), 2.82 (d, J = 7.2 Hz, 2H), 2.67 (d, J = 5.2 Hz, 2H), 2.56 (m, 2H), 2.29 (t, J = 7.6 Hz, 2H), 2.17 (t, J = 6.8 Hz, 2H), 2.07 (s, 1H), 1.46 (d, J = 4.4 Hz, 4H), 1.25 (m, 8H), 0.97 (t, J = 6.0 Hz, 3H). HRMS (ESI): m / z calcd for C 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 refered to the general procedure A, using B4, L2 and 12 as the raw materials, the product P19 was obtained as a white solid in 15% yield. HRMS (ESI): m / z calcd 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 product P20 was obtained as white solid in 21% yield by the general procedure A with B5, L2 and 12 as the starting materials. HRMS (ESI): m / z calcd 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 product P21 was obtained as white solid in 17% yield by the general procedure A with B6, L2 and 12 as the starting materials. HRMS (ESI): m / z calcd 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 product P22 was obtained as white solid in 15% yield by the general procedure A with B4, L3 and 12 as the starting materials. HRMS (ESI): m / z calcd for C 39 H 47 N9O7[M+H] + :754.3671, found: 754.3673. Its chemical structure is:

[0158]

[0159] Example 24:

[0160] Preparation of compound P23

[0161] The product P23 was obtained as white solid in 13% yield by the general procedure A with B5, L3 and 12 as the starting materials. HRMS (ESI): m / z calcd 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 performed according to the general procedure A using B6, L3 and 12 as the reactants to give the product P24 as a white solid in 11% yield. HRMS (ESI): m / z calcd 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 performed according to the general procedure A using B4, L4 and 12 as the reactants to give the product P25 as a white solid in 22% yield. HRMS (ESI): m / z calcd 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 performed according to the general procedure A using B5, L4 and 12 as the reactants to give the product P26 as a white solid in 23% yield. HRMS (ESI): m / z calcd 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] Add B4 (250 mg, 1.0 mmol) and N-Boc-piperazine (185 mg, 1.0 mmol) to a 25 mL reaction flask, dissolve in 10 mL of NMP, then add Et3N (150 mg, 1.5 mmol) dropwise. React at 90 °C for 2–4 h. Monitor the reaction of the starting material by TLC until complete. Add 10 mL of water to the reaction flask and extract with ethyl acetate (3 × 20 mL). Combine the organic phases and evaporate the solvent under reduced pressure. Purify by silica gel column chromatography (PE:EA = 1:1). Add 10 mL of 30% TFA / DCM to the obtained intermediate and react for 2 h. Evaporate the solvent under reduced pressure to obtain crude intermediate Z27 (252 mg, 80% yield, white solid). Compound L13 (159 mg, 1.0 mmol) was added to a 25 mL reaction flask, 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 portions with stirring. The reaction was carried out in an ice-water bath for 1-2 h. Then, intermediate Z27 (252 mg, 0.8 mmol) was added, and the mixture was transferred to room temperature and reacted for 3 h. After the reaction was completed by TLC monitoring, 50 mL of semi-saturated saline solution was added to the reaction solution for dilution, and the mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed once with saturated saline solution, dried over anhydrous Na2SO4, filtered, and the solvent was removed by vacuum evaporation to obtain an oily crude product. Finally, column chromatography was performed using a DCM:MeOH system of 10:1 to 5:1 to separate the intermediate. 10 mL of 30% TFA / DCM was added, and the reaction proceeded for 2 h. The solvent was removed under reduced pressure to obtain crude intermediate Z27' (137 mg, 30% yield, white solid). Compound 12 (393 mg, 1.0 mmol) was added to a 25 mL reaction flask, 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 portions with stirring. The reaction was carried out in an ice-water bath for 1-2 h, followed by the addition of intermediate Z27 (358 mg, 1.0 mmol), and the reaction was carried out at room temperature for 3 h. After the reaction was completed as monitored by TLC, 50 mL of semi-saturated saline solution was added to the reaction solution for dilution, followed by extraction with ethyl acetate (3 × 20 mL). The combined organic layers were washed once with saturated sodium chloride aqueous solution, dried over anhydrous Na₂SO₄, filtered, and the solvent was removed under reduced pressure to obtain an oily crude product. Finally, column chromatography was performed using a DCM:MeOH system of 10:1 to 5:1 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] The synthesis of P28 was carried out according to reference example 28, using B5, L13, N-Boc-piperazine and 12 as the starting materials to give the product P28 as a white solid in 25% yield. 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] The synthesis of P29 was carried out according to reference example 28, using B4, L7, N-Boc-piperazine and 12 as the starting materials to give the product P29 as a white solid in 11% yield. HRMS (ESI): m / z calcd 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] The synthesis of P30 was carried out according to reference example 28, using B5, L7, N-Boc-piperazine and 12 as the starting materials to give the product P30 as a white solid in 16% yield. HRMS (ESI): m / z calcd 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] P31 was synthesized according to Reference Example 28 using B4, L14 and 12 as the starting materials. The product P31 was obtained as a white solid in 33% yield. HRMS (ESI): m / z calcd 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] P32 was synthesized according to Reference General Example 28 using B5, L14 and 12 as the starting materials. The product P32 was obtained as a white solid in 27% yield. HRMS (ESI): m / z calcd 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] P33 was synthesized according to Reference General Example 28 using B4, L15 and 12 as the starting materials. The product P33 was obtained as a white solid in 23% yield. HRMS (ESI): m / z calcd 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] P34 was synthesized according to Reference Example 28 using B5, L15 and 12 as the starting materials. The product P34 was obtained as a white solid in 18% yield. HRMS (ESI): m / z calcd 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 refered to general example 28, using B5, L16 and 12 as the reactants to give the product P35 as a white solid in 16% yield. 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] The synthesis of P36 was refered to example 28, using B6, L16 and 12 as the reactants to give the product P36 as a white solid in 14% yield. HRMS (ESI): m / z calcd 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] The synthesis of P37 was refered to example 28, using B4, L17 and 12 as the reactants to give the product P37 as a white solid in 25% yield. HRMS (ESI): m / z calcd 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] The synthesis of P38 was refered to example 28, using B5, L17 and 12 as the reactants to give the product P38 as a white solid in 16% yield. HRMS (ESI): m / z calcd 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] The synthesis of P39 was carried out according to reference example 28, using B4, L18 and 12 as the starting materials to give the product P39 as a white solid in 16% yield. HRMS (ESI): m / z calcd 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] The synthesis of P40 was carried out according to reference example 28, using B5, L18 and 12 as the starting materials to give the product P40 as a white solid in 19% yield. HRMS (ESI): m / z calcd 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] The synthesis of P41 was carried out according to reference example 28, using B4, L19 and 12 as the starting materials to give the product P41 as a white solid in 12% yield. HRMS (ESI): m / z calcd 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] The product P42 was obtained as white solid in 15% yield by the reaction of B5, L19 and 12 in Reference Example 28. HRMS (ESI): m / z calcd for C 41 H 47 N 10 O5[M+H] + :779.3788, found: 779.3789. Its chemical structure was:

[0219]

[0220] Example 44: Preparation of compound P43

[0221] The product P43 was obtained as white solid in 22% yield by the reaction of B4, L20 and 12 in Reference Example 28. HRMS (ESI): m / z calcd for C 42 H 50 N 10 O5[M+H] + :775.4038, found: 775.4039. Its chemical structure was:

[0222]

[0223] Example 45: Preparation of compound P44

[0224] The product P44 was obtained as white solid in 11% yield by the reaction of B5, L20 and 12 in Reference Example 28. HRMS (ESI): m / z calcd for C 42 H 49 FN 10 O5[M+H] + :793.3944, found: 793.3940. Its chemical structure was:

[0225]

[0226] Example 46: Preparation of compound P45

[0227] The product P45 was obtained as white solid in 18% yield by the reaction of B4, L21 and 12 in Reference Example 27. HRMS (ESI): m / z calcd for C 47 H 59 N 11 O5[M+H] + :858.4773, found: 858.4772. Its chemical structure was:

[0228]

[0229] Example 47:

[0230] P46 was synthesized according to Reference Example 28, using B4, L22 and 12 as the starting materials. The product P46 was obtained as a white solid in 12% yield. HRMS (ESI): m / z calcd for C 48 H 61 N 11 O5[M+H] + : 872.4930, found: 872.4927. Its chemical structure is:

[0231]

[0232] Example 48:

[0233] P47 was synthesized according to Reference Example 28, using B4, L23 and 12 as the starting materials. The product P47 was obtained as a white solid in 19% yield. HRMS (ESI): m / z calcd for C 49 H 63 N 11 O5[M+H] + : 886.5086, found: 886.5082. Its chemical structure is:

[0234]

[0235] Example 49: PARP1 / 2 protein degradation experiment of compounds P1-P18

[0236] Preparation of main solutions: Preparation of test compounds: weigh a certain amount of test compound powder, dissolve in DMSO to make the final concentration 10 mmol / L, 10 μL / tube, -20 ℃ light-protected storage, thaw before use, 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), and the cells were cultured in a 37 ℃ constant temperature incubator containing 100% air. The cells were observed regularly for passage to carry out subsequent experiments. The cells in the logarithmic growth phase were used for experiments, and 2*10^5 cells were inoculated in each well of a 12-well plate.

[0238] Dosing: After the inoculated cells adhered, the medium was replaced with serum-free medium, and the cells were treated with compounds P1 to P18 at 10 μM for 24 hours. The medium was then removed, the protein was extracted, and Western blot analysis was performed. Semi-quantitative analysis was performed using Image J.

[0239] Detect the protein expression of cells by immunoblotting analysis:

[0240] 1) Protein extraction: Prepare cell lysate (add 1% protease and 1% protein phosphatase inhibitor to RIPA lysate, purchased from Selleck, and prepare the lysate immediately before use). After the end of drug treatment, remove the cell culture solution in the culture plate, wash twice with PBS, and remove the residual PBS as much as possible. Add an appropriate amount of lysate and place it on ice for 30 minutes. During this period, use a cell scraper to scrape the cells and collect the lysate into a 1.5 mL EP tube. Place it on ice and vortex every 5 minutes for a total of three to five times. Continue to lyse on ice. Place the EP tube in a pre-cooled 4°C centrifuge at 13500 rpm for 15 minutes. Transfer the supernatant to a new 1.5 mL EP tube and determine the protein concentration by the BCA method. According to the measured protein concentration, equalize the volume of the required protein, and supplement the volume of the smaller sample with RIPA lysate to ensure equal volume and quality of the same batch of protein samples. Add 5x loading buffer to the protein sample to make its working concentration 1x. After vortexing, place it in a boiling water bath for 5-7 minutes to denature the protein.

[0241] 2) Electrophoresis: Take an equal amount of denatured protein sample and load it into 5% concentrated gel and 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, and TEMED). The amount of protein loaded per well is about 40μg. According to the different colors of the positive and negative electrodes (black and red), place the electrodes correctly and cover the electrophoresis tank cover. Turn on the power of the electrophoresis instrument and adjust the voltage to 80V to start electrophoresis. When the protein enters the separation gel, the protein marker will be layered. Adjust the voltage to 120V to continue electrophoresis. When the bromophenol blue runs out of the gel plate, turn off the electrophoresis instrument and end the electrophoresis.

[0243] 3) Membrane 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, the gel, the NC membrane, 3 layers of filter paper, and finally the sponge pad in the transfer clamp from the cathode to the anode, and clamp tightly after removing all air bubbles. Place the clamp in the correct direction in the transfer slot, add 1L transfer buffer, and place an ice bag to cool down. Cover the cover and place it in an ice bath. Transfer at 250mA constant current for 2 hours.

[0244] 4) Blocking: After the end of the transfer, the NC membrane was taken out and blocked with 5% BSA or skimmed milk at room temperature for 1 hour.

[0245] 5) Antibody incubation: According to the antibody instructions, the specific primary antibody diluent was used for antibody dilution. The NC membrane was cut to the appropriate size according to the protein molecular weight, and then placed in the corresponding primary antibody diluent, and incubated overnight at 4°C on a shaking table. The next day, the antibody was recovered, and an appropriate amount of 1x TBST was added to the NC membrane-containing box and placed on a shaking table at room temperature for 3 times, 5 minutes each time. The fluorescent secondary antibody was diluted with PBS at a ratio of 1:10,000, and an appropriate amount was added to the washed NC membrane, which was incubated at room temperature in the dark for 1 hour on a shaking table. After the completion of the secondary antibody incubation, the secondary antibody incubation solution was removed, and an appropriate amount of 1x TBST was added to the NC membrane-containing container and placed on a shaking table at room temperature for 3 times, 5 minutes each time.

[0246] 6) Experimental result scanning: The Odyssey CLx infrared dual-color image analysis system of Zhejiang City University Technology Platform was used to select the appropriate scanning channel and set the optimal scanning intensity to identify and scan the fluorescent signal, and the scanning results were saved and analyzed.

[0247] Results: After the MDA-MB-436 cells were treated with the obtained compounds P1 to P18 for 24 hours, Western blotting results showed that compounds P1-P18 had a degradation effect on PARP1 protein at a concentration of 10 μM. The results showed that the 18 compounds P1-P18 in the application 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 the MDA-MB-436 cell line at 10 μM is shown in Figure 1 .

[0248] Example 50: Half-maximal degradation concentration DC 50 of the degradation activity of compounds P1-P18 on PARP1

[0249] After the compounds P1 to P18 in the application were treated for 72 hours in the MDA-MB-436 cell line, Western blotting analysis was used to detect the PARP1 and PARP2 of the cells. Semi-quantitative analysis was performed using Image J, and the DC 50 value was calculated using GraphPad Prism. After the compounds P1-P18 obtained in Examples 2-19 were treated for 72 hours, the quantitative analysis results of the Western blotting results showed that most of the compounds had excellent PARP1 degradation ability. After semi-quantitative analysis of the degradation ability of the obtained compounds, the DC 50The data were divided into four gradients, as shown in Table 1. Among them, compounds P3 and P4 had degradation effects on PARP1 protein in MDA-MB-436 cell lines at 0.05 nM, 0.1 nM, 0.5 nM, 1 nM, and 5 nM, and their half-maximal degradation concentrations DC 50 As shown in Table 1. Figure 2

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

[0251] Group 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: Half-inhibition rate IC 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-inhibition rate activity of the compounds on human breast cancer cells. The corresponding cells were cultured in "L15 culture solution" containing 10% fetal bovine serum and 1% penicillin-streptomycin. The cells were cultured in a 37°C, 100% air-saturated humidity incubator, and cells in the logarithmic growth phase were used for the experiment. Cells in the logarithmic growth phase were inoculated in a 96-well culture plate at 4x10 4 μL per well, 180 μL per well. The experimental group was added with 20 μL of compounds prepared in Examples 2-19 at concentration gradients (100 μM, 10 μM, 1 μM, 0.1 μM, 0.01 μM), and the positive control drug Olaparib (purchased from MCE) was added. The negative control group was not added with drugs, and each group had 3 parallel holes. Four 96-well plates were repeated, and incubated at 37°C for 5 days, 7 days, 10 days, and 14 days. 10 μL of enhanced CCK-8 solution was added to each well. Incubate for 2 hours in the cell culture box. Measure the absorbance at 450 nm. Calculate the cell growth inhibition rate [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD x 100%] according to the absorbance, and calculate the IC 50 value using GraphPad Prism, and the average value was taken for 3 repeated experiments. The half-inhibition rate IC of the obtained compounds on tumor cell line MDA-MB-436 cells was analyzed. 50 , and the results are shown in Table 2. Among them, compounds P3 and P4 had IC 50 As shown in Table 1. Figure 3 From the results in the figure, the IC 50 value is similar to that of the positive control drug Olaparib.

[0254] Table 2: Cell growth inhibition rate of compounds in Examples 2-19 on MDA-MB-436 cells (7 days) ​

[0255] Group 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: The half-inhibition rate IC50 of compounds P3 and P4 combined with cisplatin on tumor cell line MDA-MB-436 cells 50 Determination

[0257] Safety concentration of cisplatin administration in MDA-MB-436 cells was selected for the experiment: In order to test the effect of the compound on the sensitivity of cisplatin administration, the safety concentration of cisplatin administration in MDA-MB-436 cells was selected. Cells in the logarithmic growth phase were inoculated in a 96-well culture plate at 4x10 4 4 / ml, 180 μL per well. The experimental group was added with 20 μL of cisplatin at a concentration gradient (100 uM, 10 uM, 1 uM, 0.1 uM, 0.01 uM), and the negative control group was not added with drugs, 3 parallel holes were set in each group, and 37°C was cultured for 7 days. 10 microliters of enhanced CCK-8 solution was added to each well. Incubate for 2 hours in a cell incubator. Measure the absorbance at 450 nm. Calculate the cell growth inhibition rate [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD x 100%] according to the absorbance, and select the safety concentration according to the results Figure 4 The experimental group was added with 20 μL of cisplatin (0.5 μM) prepared at a safety concentration combined with a concentration gradient (0.5 nM, 1 nM, 5 nM) of the 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 drugs, 3 parallel holes were set in each group, and 37°C was cultured for 7 days and 14 days. 10 microliters of enhanced CCK-8 solution was added to each well. Incubate for 2 hours in a cell incubator. Measure the absorbance at 450 nm. Calculate the cell growth inhibition rate [cell growth inhibition rate = (control group OD-experimental group OD) / control group OD x 100%] according to the absorbance, the experiment was repeated 3 times, and the average value was taken, and the results are shown in Figure 4 The results showed that the proliferation inhibition activity of compounds P3 and P4 combined with cisplatin on tumor cells was significantly better than that of the positive drug Olaparib combined with cisplatin.

Claims

1. A compound, characterized in that, The compound has one of the following structures: 。 2. The compound of claim 1, wherein The compound has one of the following structures: 。 3. A medicament for inhibiting proliferation of tumor cells, characterized by, The compound of any one of claims 1-2, and pharmaceutically acceptable salts thereof.

4. A PARP1 protein degrading agent, characterized by, The compound of any one of claims 1-2, and pharmaceutically acceptable salts thereof.

5. An antitumor agent, characterized by comprising a compound of the formula (I) or a pharmaceutically acceptable salt thereof. The compound of any one of claims 1-2, and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable auxiliary ingredients.

6. A combination antitumor drug, characterized in that, The compound of any one of claims 1-2, and a chemotherapeutic drug used in conjunction therewith.

7. The combination antitumor drug according to claim 6, wherein The chemotherapeutic drug is cisplatin.

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