Epigenetic drugs for enhancing the anticancer effect of Parib antitumor agents and their applications

Abstract

The present invention discloses an epigenetic drug that enhances the anticancer effects of palibronil antitumor agents and its applications. The present invention first synthesizes a compound represented by formula (I), which can inhibit the PSF-lncRNA interaction, increase the expression of p27 and p53, inhibit the activation of the HR pathway, and arrest cancer cells in the G2 / M phase, thereby exerting an anticancer effect. The compound represented by formula (I) can significantly increase the sensitivity of palibronil antitumor agents to tumors, enhancing the therapeutic effect of palibronil antitumor agents in tumors resistant to palibronil antitumor agents. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an epigenetic drug for enhancing the anti-cancer effect of a Parib anti-tumor agent and its application. Background Art

[0002] Epithelial ovarian cancer (EOC) is the leading cause of gynecologic cancer death and the fifth most common cause of death among female malignancies. EOC is divided into two subtypes: (a) Type I, which includes endometrioid carcinoma, clear cell carcinoma, mucinous carcinoma, and low-grade serous carcinoma; and (b) Type II, which includes high-grade serous ovarian cancer (HGSOC). HGSOC is the most common subtype, accounting for 70%-80% of ovarian cancer deaths. Although neoadjuvant therapy has increased in recent years, patients with ovarian cancer typically undergo debulking surgery combined with platinum-based chemotherapy and paclitaxel. Approximately 80% of patients relapse after chemotherapy, resulting in a poor clinical prognosis. Drug resistance is a significant challenge in the treatment of ovarian cancer and a major contributor to poor prognosis.

[0003] Parib anti-tumor agents are inhibitors of the poly (ADP-ribose) polymerase (PARP) enzymes PARP-1 and PARP-2. First described in 1963, PARP is a ubiquitous protein translation-modifying enzyme present in eukaryotic cells, involved in over 90% of DNA repair processes in cells. PARP1 is the most abundant of these enzymes. When DNA is damaged, PARP binds to the damaged DNA via its N-terminal zinc finger structure, causing a change in its HD conformation and activation of its C-terminal catalytic domain. Activated PARP1 binds to and catalyzes the degradation of its substrate nicotinamide (NAD+) into ADP and nicotinic acid. Using ADP-ribose as a substrate, PARP1 undergoes poly (ADP-ribose) sylation, recruiting various repair proteins to repair the damaged DNA. The PARP1-ADP-ribose polymer, dissociated from DNA, is cleaved by poly (ADP-ribose) hydrolase and reactivated to bind to DNA, repeating the DNA damage repair cycle. When a single strand of the DNA double helix breaks, a shear repair process involving PARP1 is required. Inhibition of PARP1 can lead to defects in single-strand DNA repair. At this time, normal cells will attempt to replicate DNA and repair double-stranded DNA through homologous recombination (HR). If DNA damage is not repaired in time, cell senescence or apoptosis signals are activated, and the abnormal activation of DNA damage repair (DDR) maintains the vitality of cancer cells, significantly inducing resistance to chemotherapy drugs and PARPis, affecting the treatment effect. PARP1 / 2 small molecule inhibitors (PARPi) have been approved for the treatment of breast, ovarian, prostate and pancreatic tumors that lack homologous recombination (HR). Due to intrinsic or acquired resistance, many patients do not respond to PARPi treatment.

[0004] HR deficiency is a feature of many HGSOC cases (approximately 50%) and is considered a predictive biomarker for sensitivity to platinum drugs and PARPis. Restoration of HR pathway activity leads to acquired resistance to platinum drugs and PARPis in HR-deficient ovarian cancer patients. More than a decade ago, 53bp1 was thought to be required for BRCA1 deficiency to induce senescence and apoptosis. Soon after, researchers determined that 53bp1 inhibits HR by blocking end resection of DNA breaks, thereby promoting hypersensitivity to PARPi in BRCA1 mutant cells. The tumor suppressor gene p53 is the most commonly mutated gene in human tumors and controls cell cycle checkpoints and apoptosis by transactivating the transcription of a group of genes. These activities prevent the proliferation of cells carrying alterations in their genetic material. Studies have shown that p53 can also be directly involved in maintaining genomic stability by inhibiting homologous recombination (HR).Studies have shown that HR restoration is considered to be the main driver of PARPi resistance (see: Edwards SL. et al. Resistance to therapy caused by intragenic deletion in BRCA2. Nature 2008; 451: 1111-1115; Sakai W. et al. Secondary mutations as a mechanism of cisplatin resistance in BRCA2-mutated cancers. Nature 2008; 451: 1116-1120; Sakai W. et al. Functional restoration of BRCA2 protein by secondary BRCA2 mutations in BRCA2-mutated ovarian carcinoma. Cancer Res 2009; 69: 6381-6386; Cao L. et al. A selective requirement for 53BP1 in the biological response to genomic instabilityinduced by Brca1 deficiency. Mol. Cell. 2009; 35: 534-541; Bunting SF.et al.53BP1inhibits homologous recombination in brca1-deficient cells by blockingresection of DNA breaks.Cell2010;141:243-254;Bouwman P.et al.53BP1 lossrescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers.Nat Struct Mol Biol 2010;17:688-695).

[0005] Because the restoration of HR pathway activity leads to acquired resistance to PARPis in HR-deficient ovarian cancer patients, there is an urgent need to find a way to inhibit the activation of the HR pathway and enhance the sensitivity of ovarian cancer patients to PARPis. Summary of the Invention

[0006] To address the deficiencies of the above-mentioned prior art, the present invention provides an epigenetic drug and its application for enhancing the anticancer effects of palibrolamine antitumor agents. Compared with the prior art, the present invention first synthesizes a compound represented by formula (I), which can inhibit the PSF-lncRNA interaction, increase the expression of p27 and p53, inhibit the activation of the HR pathway, and arrest cancer cells in the G2 / M phase, thereby exerting an anticancer effect. When the compound represented by formula (I) is used in combination with palibrolamine antitumor agents, it can enhance the anticancer effects of palibrolamine antitumor agents, becoming a new anticancer sensitization strategy with great value for promotion and application.

[0007] The first object of the present invention is to provide an epigenetic drug, characterized in that it is a compound as shown in formula (I) or a pharmaceutically acceptable salt thereof; wherein the compound shown in formula (I) is named: 4-(2,4,5-trifluorobenzyl)-7,8-dihydroxycoumarin.

[0008]

[0009] The applicant has discovered that the epigenetic drug can inhibit the PSF-lncRNA interaction and inhibit the growth of cancer cells. Therefore, it can be used to prepare a drug for treating cancer. The cancer is at least one of breast cancer, prostate cancer, pancreatic cancer, ovarian cancer, progressive ovarian cancer, high-grade serous ovarian cancer (including fallopian tube cancer or primary peritoneal cancer) and metastatic cancer spread from primary ovarian cancer, small cell / non-small cell lung cancer, peritoneal cancer, prostate cancer, soft tissue sarcoma, endometrial cancer, and gastric cancer.

[0010] Preferably, the epigenetic drug is used to increase the expression of p27 and p53 proteins to change the cell cycle, arrest cancer cells in the G2 / M phase, and affect the synthesis of RNA and protein.

[0011] A second objective of the present invention is to provide the aforementioned epigenetic drug for enhancing the sensitivity of pali-type anti-tumor agents to tumors, thereby enhancing the anti-cancer effect of pali-type anti-tumor agents, and for use in the preparation of pali-type anti-tumor agent sensitizers. The pali-type anti-tumor agent is at least one of olaparib, rucaparib, niraparib, talazoparib, and niraparib tosylate.

[0012] The third object of the present invention is to provide an anticancer sensitization composition comprising a compound represented by formula (I) and a Parib antitumor agent.

[0013] Preferably, the molar ratio of the Parib antitumor agent to the compound of formula (I) is 1:1-1:4.

[0014] As a preference, when the epigenetic drug is used in combination with a Pareto anti-tumor agent, the IC of the Pareto anti-tumor agent can be significantly reduced in Caov-3 ovarian cancer cells. 50, and obtain better anti-tumor effects.

[0015] The anticancer sensitization composition further comprises one or more pharmaceutically acceptable carriers, including conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, and the like in the pharmaceutical field.

[0016] The anti-cancer sensitization composition can be used orally or parenterally, for example, by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation, such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue; or can be mixed or encapsulated with other substances and then introduced into the body.

[0017] For oral administration, it can be prepared into conventional solid preparations such as tablets, powders, granules, capsules, ointments, creams, etc.; it can be prepared into liquid preparations such as aqueous or oily suspensions or other liquid preparations such as oral liquids, etc. For parenteral administration, it can be prepared into solutions for injection, aqueous or oily suspensions, etc.

[0018] The beneficial effects of the present invention are:

[0019] 1. The present invention first synthesized a compound as shown in formula (I), which can inhibit the PSF-lncRNA interaction, increase the expression of p27 and p53, inhibit the activation of the HR pathway, and block cancer cells in the G2 / M phase, thereby exerting an anti-cancer effect.

[0020] 2. The compound represented by formula (I) can significantly increase the sensitivity of Parib anti-tumor agents to tumors and enhance the therapeutic effect of Parib anti-tumor agents on tumors resistant to Parib anti-tumor agents. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The compound represented by formula (I) 1 H NMR spectrum;

[0022] Figure 2 The compound represented by formula (I) 13 C NMR spectrum;

[0023] Figure 3 is the HR-MS spectrum of the compound represented by formula (I);

[0024] Figure 4 This is the Western blot protein band diagram of the RNA-pulldown experimental results;

[0025] Figure 5The effect of the compound represented by formula (I), the lead compound 10-3, and compound A on the Caov-3 cell cycle; wherein, blue: G0 / G1 phase, green: S phase, and red: G2 / M phase;

[0026] Figure 6 The effects of the compound represented by formula (I), the lead compound 10-3, and compound A on the expression of p27 and p53 proteins in Caov-3 cells;

[0027] Figure 7 Graph showing the anti-tumor effects of the compound represented by formula (I), the lead compound 10-3, compound A, the paly anti-tumor agent niraparib tosylate, and their combination therapy. DETAILED DESCRIPTION

[0028] The present invention will be further described below with reference to specific examples, but this does not limit the present invention.

[0029] Unless otherwise specified, the contents of the following components are expressed in weight percentages; the experimental methods used are conventional methods; and the reagents and biological materials used are all commercially available.

[0030] The structural formula of the lead compound 10-3 (4-(4'-methoxyphenyl)-7,8-dihydroxycoumarin) used in the present invention is as follows:

[0031]

[0032] The structural formula of the lead compound A (4-(3'-trifluoromethylbenzyl)-7,8-dihydroxycoumarin, see CN2024106623279) used in the present invention is as follows:

[0033]

[0034] The pali antitumor agent used in the present invention is niraparib tosylate, and its structural formula is shown below:

[0035]

[0036] Example 1: Preparation of epigenetic drugs represented by formula (I)

[0037] (I) The specific preparation process of compound I (4-(2,4,5-trifluorobenzyl)-7,8-dihydroxycoumarin) represented by formula (I) is as follows:

[0038] The synthetic route is:

[0039]

[0040] Among them, a: oxalyl chloride, DMF, DCM, 37-40°C; b: Michaelis' acid (cycloisopropyl malonate), triethylamine, DCM, 0°C-rt; c: ethanol, 70-100°C; d: trifluoroacetic acid, sulfuric acid, DCM, 0°C-rt.

[0041] (1) In a 100 mL single-necked reaction flask, add 2,4,5-trifluorophenylacetic acid (5 mmol, 1 eq), oxalyl chloride (30 mmol, 6 eq), 5-6 drops of DMF and 50 mL of anhydrous DCM, place in a heated magnetic stirrer, and reflux and stir at 37-40°C for 1-3 h. After the reaction is completed, spin dry to obtain the crude 2,4,5-trifluorophenylacetyl chloride, which is directly carried out to the next step.

[0042] (2) At 0°C, place cycloisopropyl malonate (5mmol, 1eq), triethylamine (12.5mmol, 2.5eq) and 30mL of anhydrous DCM in a 100mL single-necked reaction flask, stir for 10-20min, and then slowly add a solution of 2,4,5-trifluorophenylacetyl chloride in anhydrous DCM. After the addition is complete, slowly raise the temperature to room temperature. Allow to react overnight, extract the triethylamine with 0.1mol / L citric acid solution, dry the organic phase, spin dry, add 60mL of ethanol, and reflux at 70-100°C for 3-4h. After the reaction is completed by TLC monitoring, spin dry to obtain the crude product of ethyl 2,4,5-trifluorophenylacetyl acetate, separate and purify it by column chromatography (filler: silica gel powder) (eluent: by volume, petroleum ether: ethyl acetate = 15:1), and then concentrate under reduced pressure to obtain an oily pure product (1.05g);

[0043] (3) At 0°C, H2SO4 (2mmol, 2eq) and CF3COOH (2mmol, 2eq) were stirred and mixed in a 100mL single-necked reaction flask. 30mL of anhydrous DCM, ethyl 2,4,5-trifluorophenylacetoacetate (1mmol, 1eq) and pyrogallol (1.2mmol, 1.2eq) were added in sequence. The temperature was slowly raised to room temperature and the reaction was continued with stirring. After 1 hour, 1-2mL of methanol was added to increase the solubility of the system. The reaction was allowed to proceed for 4-5 hours. After the reaction was completed by TLC monitoring, the reaction solution was extracted with saturated NaHCO3 solution and the organic phase was dried and spin-dried. Recrystallization was performed using ethyl acetate / petroleum ether (volume ratio 1:5). After the solid precipitate was precipitated, it was cooled and allowed to stand, filtered, and the filter cake was dried in a vacuum drying oven to obtain 4-(2,4,5-trifluorobenzyl)-7,8-dihydroxycoumarin (0.21g) with a yield of 65%.

[0044] Example 2: Structural Identification

[0045] The structure of the compound was identified using HPLC-MS and BRUKER 800M NMR. Figure 1-3The data are as follows: Spectrum and data of the compound 4-(2,4,5-trifluorobenzyl)-7,8-dihydroxycoumarin represented by formula (I):

[0046] 1 H NMR (800MHz, DMSO-d6): δ7.62-7.56(m,1H),7.52(ddd,J=11.8,9.6,7.2Hz,1H),7.16(d,J=8.9Hz,1H),6.81(d,J=8.8Hz,1H),5.77(s,1H),4.12(s,2H).

[0047] 13 C NMR(201MHz,DMSO-d6)δ160.15,156.26,156.21,155.05,155.00,153.95,149.71,149.0 9,149.02,148.96,147.86,147.79,147.72,146.78,146.76,146.70,145.57,145.55,145 .49,143.47,132.45,121.11,121.09,121.06,121.02,120.99,120.97,119.36,119.33,119.26,119.23,115.16,112.31,111.59,110.13,106.28,106.17,106.13,106.02,29.79.

[0048] MS: m / z (%) [M+H] + =323.0407.

[0049] Example 3: Sensitization activity assay

[0050] 1. Cell Culture

[0051] Caov-3 (human ovarian cancer cells) cells were selected and cultured in RPMI1640 medium containing 10% fetal bovine serum.

[0052] HCT116 (colon cancer cell) cells were selected and cultured in DMEM medium containing 10% serum.

[0053] All cells were purchased from Jinan Boshang Company.

[0054] 2. Cytotoxicity Test

[0055] Caov-3 cells were plated in a 96-well plate at a density of 5,000-7,000 cells / 100 μL per well and cultured overnight in a cell culture incubator. After cells adhered, 100 μL of drug-treated culture medium at varying concentrations was added and treated for a further 48 hours. Niraparib tosylate concentrations were applied in a gradient of 0.78, 1.56, 3.125, 6.25, 12.5, and 25 μM, respectively. Compound I concentrations ranged from 1 to 10 μM, while lead compound 10-3 and lead compound A concentrations were 5 μM. After 48 hours, 10 μL of MTT was added to each well and incubated at 37°C for 4 hours in the dark. The culture medium was discarded, and 150 μL of DMSO was added to each well. The cells were shaken for 10 minutes until the blue-purple crystals completely dissolved. Absorbance at 490 nm was measured using an enzyme-linked immunosorbent assay (ELISA). Cell viability (100%) = (mean OD value of the drug group / mean OD value of the normal control group) * 100%.

[0056] 3. Results Analysis

[0057] When niraparib tosylate is used alone: ​​IC 50 =1.211±0.017μM;

[0058] When compound I is used alone: ​​IC 50 =4.215±0.450μM;

[0059] When niraparib tosylate was used in combination with compound Ⅰ (concentration of 5 μM): IC 50 =0.405±0.015μM;

[0060] When niraparib tosylate was used in combination with the lead compound 10-3 (at a concentration of 5 μM): IC 50 =1.012±0.013μM;

[0061] When niraparib tosylate was used in combination with lead compound A (concentration of 5 μM): IC 50 =0.905±0.012μM.

[0062] It can be seen that compared with the single-drug application of niraparib tosylate and the combination with the lead compounds 10-3 and A, the compound I of the present invention has a better anti-cancer sensitization effect on niraparib tosylate.

[0063] Example 4: RNA-pulldown experiment

[0064] Using Pierce TMThe Magnetic RNA Protein Pull-Down Kit was used to validate the mechanism of RNA pull-down experiments. The kit provides streptavidin magnetic beads and reagents for effectively enriching RNA-binding proteins. In the RNA pull-down assay, in vitro transcribed RNA is first bound to the magnetic beads. The RNA-bead complex is then incubated with cell lysate. This allows proteins bound to the target RNA to be adsorbed to the beads. After the RNA-protein complex is eluted from the beads with an elution buffer, specific bound proteins are detected by Western blotting.

[0065] 1. Experimental Plan

[0066] Binding of magnetic beads to labeled RNA: resuspend 50 μL of magnetic beads in a sterile, enzyme-free 1.5 mL centrifuge tube, place on a magnetic separation rack, and discard the supernatant; wash the magnetic beads twice with 50 μL of 20 mM Tris; resuspend the magnetic beads in 50 μL of 1× RNA Capture buffer, then add 50-100 pmol of avidin-labeled RNA (purchased from Gene Pharma), mix gently, and incubate at room temperature with shaking for 30 min.

[0067] Binding of labeled RNA to PSF protein: Place the above centrifuge tube on a magnetic separation rack, collect the magnetic beads from the side, and discard the supernatant; wash the magnetic beads twice with 50 μL of 20 mM Tris; dilute 10× protein-RNA binding buffer to 1× with ultrapure water, and resuspend the magnetic beads in 100 μL of 1× protein-RNA binding buffer; prepare the RNA-protein binding reaction premix according to the table below (Table 1); collect the magnetic beads from the side of the magnetic separation rack, discard the supernatant, and add 100 μL of the RNA-protein binding reaction premix to the magnetic beads. Incubate at 4°C with shaking for 60 minutes. For the drug-treated group: Incubate the protein lysate with the compound (1 nM) at room temperature with shaking for 1 hour.

[0068] Table 1 Premix formula for RNA-protein binding reaction

[0069]

[0070] Elution of RNA-protein complexes: Place the centrifuge tube on a magnetic separation rack, collect beads on the side, and aspirate the supernatant for analysis. Wash the beads twice with 100 μL of 1× Wash buffer, retaining the supernatant for analysis. Resuspend the beads in 50 μL of 1× Wash buffer, gently mix, and incubate at 37°C with shaking for 30 minutes. Place the centrifuge tube on a magnetic separation rack and retain the supernatant. Add an equal volume of 2× Loading Buffer (purchased from Solebold) to the supernatant from the final Elution Buffer step, mix thoroughly, and boil in a 95°C metal bath for 10-15 minutes. Detect the pulled-down PSF protein by Western blot.

[0071] 2. Results Analysis

[0072] according to Figure 4 It can be seen that formula (I), 10-3 and compound A can all significantly inhibit the binding of lncRNA to PSF protein, and the inhibitory effect of compound I is significantly better than that of the lead compound 10-3 and compound A.

[0073] Example 5: Cell cycle detection

[0074] 1. Experimental Plan

[0075] Preparation of cell samples: When the growth rate was about 80%, HCT116 cells (Jinan Boshan Company) were seeded in a six-well plate. After the cells adhered to the wall, different compounds (25μM) were added to each well and cultured in a cell culture incubator for 48h. The culture medium was removed and washed with PBS. 300μL of trypsin was added to each well for about 1min. The digestion was stopped and the cells were collected. The cells were centrifuged at 1000rpm for 3-5min and the supernatant was discarded. The cells were washed twice with 2mL of pre-cooled PBS and centrifuged again to discard the supernatant. The cells were resuspended in 250μL of cold PSB, 750μL of pre-cooled anhydrous ethanol was added dropwise, mixed gently, and placed at 4℃ for fixation overnight.

[0076] Staining: Prepare the staining solution in the dark. After fixation, centrifuge the cells at 1000 rpm for 3-5 minutes, remove the fixative, rinse the cells with PBS, and centrifuge again to discard the supernatant. Add 500 μL of the prepared staining solution, mix gently, and incubate at 37°C in the dark for 30 minutes.

[0077] Flow cytometry: Red fluorescence was detected by flow cytometry at an excitation wavelength of 488 nm, and light scattering was detected simultaneously.

[0078] 3. Results Analysis

[0079] like Figure 5As shown, compared with the blank control group, Compound I, lead compound 10-3, and Compound A significantly reduced the number of HCT116 cells in the G0 / G1 phase and significantly increased the number of cells in the G2 / M phase. This means that the drug-treated groups were able to arrest cells in the G2 / M phase and affect RNA and protein synthesis.

[0080] Example 6: Western Blot

[0081] 1. Experimental Plan

[0082] Preparation of Cell Protein Samples: When Caov-3 cells reached approximately 80% growth, they were seeded in six-well plates. After cells adhered, different compounds (25 μM) were added to each well and cultured in a cell culture incubator for 48 hours. Cells were observed under a microscope, and the culture medium from wells showing minimal changes in cell status was discarded. The cells were rinsed once with 1 mL of PBS. Then, 150-250 μL of RIPA lysis buffer (PMSF was added to the lysis buffer several minutes earlier to a final concentration of 1 mM) was added to each well. Gently agitate to allow for thorough contact between the lysis buffer and cells, and lyse on ice for 5-10 minutes. The lysed cells were scraped off with a scraper, transferred to a centrifuge tube, and centrifuged at 12,000 rpm for 5-10 minutes at 4°C. The supernatant was retained. The culture medium from wells with significant cell death was retained in a centrifuge tube, centrifuged, and the supernatant discarded. RIPA lysis buffer (containing 1 mM PMSF) was added, gently mixed, and lysed on ice, followed by centrifugation. Protein concentration was determined using a BCA protein assay kit. Add an equal volume of 2× protein loading buffer, mix well, boil in a 95°C metal bath for 10-15 min, and store the sample in a -20°C refrigerator for later use.

[0083] Western blot was used to detect the expression level of the target protein: 20 μg of protein was loaded, the voltage was adjusted to 80 V, and the electrophoresis was stopped when the bromophenol blue just ran out of the bottom of the separation gel. Then the PAGE gel was placed on a transfer membrane instrument (BioRad, USA), and the membrane was transferred at a constant current of 200 mA for 1 hour, blocked with 5% milk powder for 2 hours, and washed with TBST buffer 3 times, each time for 10 minutes. The primary antibodies diluted in antibody diluent (p27 antibody, 1:1000; p53 antibody, 1:5000) were added respectively, and incubated overnight at 4°C. After washing with TBST, the secondary antibody labeled with horseradish peroxidase (HRP-IgG, 1:4000) was added, and the membrane was incubated at room temperature for 2 hours. After washing with TBST, ECL developer was added, and the membrane was exposed and developed on a chemiluminescence imaging system (Thermo Fisher) and its grayscale value was analyzed using the gel image processing system (see attached figure). Figure 6 shown).

[0084] 2. Results Analysis

[0085] like Figure 6As shown in the results, compared with the blank control group, the expression levels of p27 and p53 proteins of the compound represented by formula (I) and compound A were significantly increased. The changes in the expression of these cell cycle-related proteins may be the reason for the above-mentioned cell cycle experimental results (the number of G0 / G1 phase cells was significantly reduced, and the number of G2 / M phase cells was significantly increased).

[0086] Example 7: In vivo animal experiment

[0087] 1. Experimental Methods

[0088] Epithelial ovarian cancer Caov-3 (1×10 6 The Caov-3 animal model was established by inoculating the forelimbs of 4-6-week-old male mice with 10-3 (10-3 / mouse) subcutaneously. One week later, the mice were randomly divided into six groups: a control group, a group receiving compound 1 alone, a group receiving niraparib tosylate alone, a group receiving 10-3 plus niraparib tosylate, a group receiving compound A plus niraparib tosylate, and a group receiving compound 1 plus niraparib tosylate, with five mice in each group.

[0089] The compound 1 alone group was intraperitoneally injected with compound 1 twice a week (5 mg / kg each time).

[0090] Niraparib tosylate alone group: Niraparib tosylate was intraperitoneally injected twice a week (5 mg / kg each time).

[0091] 10-3 + Niraparib Tosylate group: Compound 10-3 was intraperitoneally injected twice a week (5 mg / kg each time); Niraparib Tosylate group was intraperitoneally injected twice a week (5 mg / kg each time).

[0092] Compound A+niraparib tosylate group: Compound A was intraperitoneally injected twice a week (5 mg / kg each time); Niraparib tosylate group was intraperitoneally injected twice a week (5 mg / kg).

[0093] Compound 1 + niraparib tosylate group: Compound 1 was intraperitoneally injected twice a week (5 mg / kg each time); Niraparib tosylate group was intraperitoneally injected twice a week (5 mg / kg each time).

[0094] Control group: injected with the same dose of normal saline.

[0095] The tumor volume was measured with a vernier caliper every 3 days. After 21 days of administration, the mice were sacrificed and the tumors were removed.

[0096] 2. Results Analysis

[0097] like Figure 7As shown, after two weeks of group treatment, there was no significant difference between the niraparib tosylate group and the control group, indicating that this tumor model is insensitive to niraparib tosylate treatment. However, the combination of Compound I and niraparib tosylate can significantly inhibit tumor growth, resulting in a decrease in tumor volume and tumor weight, indicating that Compound I can enhance the killing effect of niraparib tosylate on its resistant tumor model. The tumor inhibition effect of Compound 1 + niraparib tosylate is greater than that of the 10-3 + niraparib tosylate group and the Compound A + niraparib tosylate group. Among them, the tumor inhibition rate of the combined action of Compound I and niraparib tosylate is greater than the sum of the tumor inhibition rate of Compound I and the tumor inhibition rate of niraparib tosylate, indicating that Compound I and niraparib tosylate have a synergistic effect.

Claims

1. An epigenetic drug, characterized in that it is A compound represented by formula (I) or a pharmaceutically acceptable salt thereof; （I）。 2. Use of the epigenetic drug according to claim 1 in the preparation of a niraparib tosylate sensitizer.

3. An anticancer sensitization composition, characterized in that: The invention comprises the compound represented by formula (I) according to claim 1 and niraparib tosylate.

4. The anticancer sensitization composition according to claim 3, wherein: The molar ratio of the niraparib tosylate to the compound represented by formula (I) is 1:1-1:

4.

5. Use of the anticancer sensitization composition according to claim 3 or 4 in the preparation of a drug for treating ovarian cancer.

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