Application of ezetimibe derivative in preparation of anti-prostatic cancer drugs

Abstract

The invention relates to an application of an ezetimibe derivative in preparation of an anti-prostatic cancer drug, and belongs to the technical field of medicinal chemistry. The general formula structure of the ezetimibe derivative L14 is # imgabs0. The ezetimibe derivative L14 can be used for preparing new drugs for treating CRPC and NEPC without depending on an androgen receptor pathway, the clinical curative effect is improved, and the clinical drug selection is increased. The micromolecule L14-8 (R1 = R2 =-C (O) CH = CH2) can specifically inhibit proliferation of prostate cancer cells in vitro and can also inhibit growth of prostate tumors in vivo; in the aspect of drug combination, the AR antagonist and the low-dose small molecule L14-8 are combined for use, so that the sensitivity of CRPC and NEPC to the AR antagonist can be remarkably improved, the drug resistance induced by the AR antagonist can be possibly inhibited, and the clinical benefits of the AR antagonist can be improved.

Description

Technical Field

[0001] The invention relates to application of ezetimibe derivatives in preparing anti-prostate cancer drugs, and belongs to the technical field of pharmaceutical chemistry. Background Art

[0002] Prostate cancer (PCa) is one of the most common malignant tumors in the male urinary system, posing a great threat to the health of male residents. Currently, the main methods for clinical treatment of prostate cancer include surgery or androgen deprivation therapy (ADT). However, prostate cancer will develop into castration-resistant prostate cancer (CRPC) or even progress to neuroendocrine prostate cancer (NEPC), leading to the death of most prostate cancer patients.

[0003] As the most effective drug for the treatment of CRPC, androgen receptor signaling inhibitors (ARSIs) can effectively inhibit the increase of extratesticular androgens, intratumoral androgen receptor (AR) amplification and intratumoral AR overexpression after ADT treatment. However, based on mechanisms such as AR transcriptional reprogramming, AR mutation, AR alternative splicing and AR-independent lineage plasticity, patients will develop resistance to ARSIs.

[0004] The incidence of NEPC has increased in the era of new ARSIs, and NEPC is an even more lethal stage of prostate cancer. In this stage, AR loss and activation of neuronal features often occur simultaneously, leading to treatment-induced neuroendocrine prostate cancer (t-NEPC). Because the AR signaling pathway is blocked, patients are unable to effectively respond to ARSIs, and the development of NEPC increases mortality.

[0005] Due to the limitations of clinical drug ARSIs, it is necessary to develop a new therapeutic drug to treat CRPC and NEPC, but its effect is not limited to prostate cancer. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides the use of ezetimibe derivative L14 in the preparation of anti-prostate cancer drugs. The technical problem to be solved by the present invention is to provide a new drug for treating CRPC and NEPC, and to find a suitable small molecule L14 among many small molecules, which has a clear target for inhibiting prostate tumors.

[0007] The present invention provides the use of an ezetimibe derivative L14 in the preparation of an anti-prostate cancer drug, wherein the general structural formula of the ezetimibe derivative L14 is as follows:

[0008] Where: R 1 ,R 2 Each independently selected from H, n=0,1,2,3,4,5,6; R is selected from C1-C8 alkyl, C2-C8 olefin, halogenated thiophene, etc.

[0009] Furthermore, in the above technical solution, the R 1 and R 2 Selected from the following specific structures:

[0010]

[0011] Furthermore, in the above technical solution, the ezetimibe derivative L14 is L14-8, and the structural formula is as follows:

[0012]

[0013] Furthermore, in the above technical solution, the prostate cancer is selected from castration-resistant prostate cancer CRPC and neuroendocrine prostate cancer NEPC. In particular, it has an effect on inhibiting the proliferation of hormone-independent prostate cancer cells and has low toxicity.

[0014] Furthermore, in the above technical solution, the prostate cancer is selected from prostate cancer PC3, 22RV2 or C4-2B cells.

[0015] The present invention also provides a pharmaceutical composition containing the above-mentioned ezetimibe derivative L14, wherein the active ingredients thereof include the above-mentioned ezetimibe derivative L14 and a pharmaceutically acceptable salt thereof.

[0016] Furthermore, in the above technical solution, the pharmaceutically acceptable salt includes a salt formed by the ezetimibe derivative L14 and an organic acid or an inorganic acid. The organic acid is selected from one or more of malic acid, lactic acid, camphorsulfonic acid, citric acid, fumaric acid or oxalic acid, and the inorganic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

[0017] The invention also provides the use of AR antagonists and ezetimibe derivative L14 in the preparation of anti-prostate cancer drugs.

[0018] Furthermore, in the above technical solution, the AR antagonist is selected from Darolutamide or Enzalutamide.

[0019] Furthermore, in the above technical solution, the ezetimibe derivative is L14-8.

[0020] Beneficial effects of the present invention

[0021] 1. The present invention provides a new use of the small molecule L14, which is an ezetimibe derivative, and can be used to prepare new drugs for the treatment of CRPC and NEPC that are independent of the androgen receptor pathway, thereby improving clinical efficacy and increasing clinical drug selection. The small molecule L14-8 can specifically inhibit the proliferation of prostate cancer cells in vitro and can also inhibit the growth of prostate tumors in vivo.

[0022] 2. Combination therapy: The combined use of AR antagonists and low-dose small molecule L14-8 can significantly increase the sensitivity of CRPC and NEPC to AR antagonists, thereby possibly inhibiting AR antagonist-induced drug resistance and improving its clinical benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The classification of the small molecule library, the chemical formula structure of Ezetimibe and L14 small molecules, and the CCK-8 results of Ezetimibe and L14 small molecules in treating normal prostate cells and prostate cancer cells;

[0024] Figure 2 The optimized chemical structure of L14 small molecule and the results of treating prostate cancer cells with CCK-8, the results of treating normal prostate cells and prostate cancer cells with CCK-8 by Ezeti mibe and L14-8, the results of treating prostate cancer cells with CCK-8 by AR inhibitor Daro lutamide, AR inhibitor Enzalutamide, L14-8, and L14-8 combined with two AR inhibitors;

[0025] Figure 3 Comparison of tumor size differences among mice in each group after 3 weeks of drug administration and Ki67 staining results of mouse tumor tissues;

[0026] Figure 4 Comparison of HE pathological morphology of mouse organ tissues and blood biochemical indexes of mice in each group after 3 weeks of drug administration;

[0027] Figure 5Transcriptome analysis, KEGG enrichment analysis, differential gene enrichment analysis (D GEA), gene set enrichment analysis (GSEA), C11 BODIPY (581 / 591) detection of prostate cancer cells flow cytometry and confocal microscopy observation results were performed after L14-8 treatment;

[0028] Figure 6 To predict the target PLK1 view, molecular docking simulation view of L14-8 and PLK1, after treating prostate cancer cells with L14-8, the protein was treated with step temperature and immunoblotting experiment results were performed (different gradient temperature views), immunoblotting experiment results after treating prostate cancer cells with different concentrations of L14-8, DAPI and TP53 probe observation results under confocal microscope after treating prostate cancer cells with different concentrations of L14-8, and CCK-8 results after knocking out TP53 in prostate cancer cells and treating them with different concentrations of L14-8. DETAILED DESCRIPTION

[0029] In order to enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods.

[0030] 1. Screening of small molecule L14

[0031] First, a small molecule library containing 461 compounds was constructed. The library includes seven categories with different core structures: tosylhydrazone, alkenyl, pyrazoles, isoxazoles, cyclopropanes, ethers / thioethers, aryl compounds and amines. Small molecules in the library were obtained by using commercial reagents without purification, and the reactions were carried out under argon atmosphere (except for hydrazone synthesis), and water was excluded using standard techniques for handling air-sensitive compounds. Unless otherwise stated, all reactions were carried out in an inert atmosphere of dry argon in oven-dried glassware with magnetic stirring. Small molecules with various core structures were rapidly constructed, and many drug analogs were successfully obtained through late-stage modifications, including those derived from ketoprofen (anti-inflammatory drug), sertraline (antidepressant), fenofibrate (lipid-lowering drug), lurasidone (antipsychotic drug), fendiline (anti-anginal drug), and bifonazole (antifungal drug).

[0032] A high-throughput screening of the antitumor activity of the library small molecules was performed in AR-positive C4-2B and AR-negative PC3 prostate cancer cells to evaluate the antitumor potential of these compounds.

[0033] The specific operation is as follows: prostate cancer cells PC3 and C4-2B are cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin in a 37°C, 5% CO2 cell culture incubator. Cells in the logarithmic growth phase are digested and counted and plated on 96-well plates. The treatment conditions are 461 small molecules in the control group and the drug group (drug concentration is 10μM), with 5 replicates in each group; drug administration is performed 24h after plating, and cell counting reagent (CCK-8) is used to detect cell viability 72h after cell administration.

[0034] The L14 small molecule was screened out as an analogue derived from Ezetimibe. The inhibitory effects of the L14 small molecule and the original drug Ezetimibe on prostate cancer cells were compared.

[0035] The specific operation is as follows: prostate cancer cells PC3 and C4-2B are cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C and 5% CO2 in a cell culture incubator. Cells in the logarithmic growth phase are digested and counted and plated on 96-well plates. The treatment conditions are control group, drug-treated group L14 and drug-treated group Ezetimibe (drug concentration is 10 μM), with 5 replicates per group; drug administration is performed 24 hours after plating, and cell viability is detected using a cell counting reagent (CCK-8) 72 hours after cell administration.

[0036] 2. The structure of the above L14 small molecule was optimized to improve the efficacy

[0037] First, it was confirmed that L14 showed better antitumor activity compared with the original drug and showed the importance of modifying the hydroxyl group of ezetimibe. Therefore, seven monosubstituted L14-1 to L14-7 were synthesized, including different α,β-unsaturated esters and saturated esters to determine the necessity of additional groups. However, a single ester group resulted in lower activity than L14. Subsequently, disubstituted ezetimibe derivatives L14-7 to L14-12 incorporating various Michael acceptors were synthesized by esterification of aliphatic hydroxyl groups.

[0038] Furthermore, the above 12 optimized small molecules and Ezetimibe were validated in two prostate cancer cell lines, C4-2B and PC3, respectively.

[0039] The specific operation is as follows: prostate cancer cells PC3 and C4-2B are cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin in a 37°C, 5% CO2 cell culture incubator. Cells in the logarithmic growth phase are digested and counted and plated on 96-well plates. The treatment conditions are control group, Ezetimibe, L14-1 to L14-12 drug administration group (drug concentration is 10 μM), 5 replicates per group; drug administration is performed 24 hours after plating, and cell viability is detected using a cell counting reagent (CCK-8) 72 hours after cell administration.

[0040] Furthermore, the most potent small molecule L14-8 for inhibiting prostate cancer cells was screened out above, and L14-8 and Ezetimibe were respectively applied to two prostate cancer cell lines, C4-2B and PC3, for comparison.

[0041] The specific operation is as follows: prostate cancer cells PC3 and C4-2B are cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin in a 37°C, 5% CO2 cell culture incubator. Cells in the logarithmic growth phase are digested and counted and plated on 96-well plates. The treatment conditions are control group, Ezetimibe, and L14-8 administration group (drug concentration is 10 μM), with 5 replicates in each group; 24 hours after plating, the drug is administered, and 72 hours after the cell administration, the cell counting reagent (CCK-8) is used to detect cell viability.

[0042] Furthermore, the toxicity of the L14-8 small molecule in normal prostate cells was verified.

[0043] The specific operation is as follows: normal prostate epithelial cells RWPE-1 were cultured in a 37°C, 5% CO2 cell culture incubator using K-SFM complete medium supplemented with 1% gentamicin. Cells in the logarithmic growth phase were digested and counted and plated on a 96-well plate. The treatment conditions were a control group and an L14-8 administration group (drug concentration 25 μM), with 5 replicates per group; the drug was administered 24 hours after plating, and cell viability was detected using a cell counting reagent (CCK-8) 72 hours after cell administration.

[0044] Furthermore, the L14-8 small molecule and AR antagonists that are currently used frequently in clinical practice (such as Enzalutamide and Darolutamide) were used to act on prostate cancer cells to conduct cell proliferation experiments.

[0045] The specific operation is as follows: prostate cancer cells PC3 and 22RV2 are cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin in a 37°C, 5% CO2 cell culture incubator. Cells in the logarithmic growth phase are digested and counted and plated on 96-well plates. The treatment conditions are control group, Enzalutamide administration group, L14-8 small molecule administration group and Enzalutamide combined with L14-8 small molecule administration group (all at a concentration of 10 μM), with 5 replicates per group; 24 hours after plating, the drug is administered, and 72 hours after the cell administration, the cell counting reagent (CCK-8) is used to detect cell viability.

[0046] 3. Administer L14-8 in animals to verify its efficacy and toxicity

[0047] The specific operation was as follows: 15 3-week-old SPF male nude mice were adaptively raised for one week and then injected subcutaneously with C4-2B cells (1×10 6 / mouse), were randomly divided into three groups, 5 mice in each group, divided into a control group (equal volume of pure water), a low-dose group (L14-8 normal dose 10 mg / kg) and a high-dose group (L14-8 normal dose 20 mg / kg twice). Drug administration began one week after modeling, and was administered by intraperitoneal injection every day. The tumor size was measured with a caliper every 2 days. After 25 days of treatment, the mice were euthanized. After the experiment, blood was collected from the apex of the nude mice, specimens were collected, and tissues were frozen to evaluate tumor quality, Ki67 staining, and potential organ toxicity. Paraffin sections of mouse tumor tissue were taken for Ki-67 immunohistochemical staining and observed under a microscope.

[0048] 4. After L14-8 was administered to animals, biochemical indicators and histopathological morphology of animal tissues and blood were analyzed.

[0049] The specific operation is: take the mouse heart, liver, spleen, lung, and kidney for HE paraffin sections and observe under a microscope. The serum samples obtained from the animal experiment were tested for normal blood routine and biochemical indexes, and the relative contents of platelets (PLT), white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and creatinine (CREA) in serum were determined.

[0050] 5. Verification of L14-8's effect on ferroptosis pathway

[0051] First, transcriptome analysis, KEGG enrichment analysis, differential gene enrichment analysis (DGEA) and gene set enrichment analysis (GSEA) were performed after L14-8 treatment. The analysis showed that the tumor growth inhibition mediated by L14-8 was related to the ferroptosis pathway. The ferroptosis activity after L14-8 treatment was further verified using C11BODIPY (581 / 591), a fluorescent probe that indicates lipid peroxidation and antioxidant efficacy of ferroptosis in living cells.

[0052] The specific operation is as follows: PC3 cells were cultured normally. 5 The cells were inoculated in 6-well plates at a density of 100 / well, and each well was repeated 3 times, incubated in 2 mL of culture medium for 24 h, L14-8 (dosage dose) was treated for 24 h, and the cells were incubated with a final concentration of 5 μM BODIPY-C11 probe at 37 ° C for 30 minutes. For flow cytometry analysis, the cells were digested with trypsin, collected, and then resuspended in 500 μL PBS, transferred to a test tube, and fluorescence was measured using a CytoFLEX-3 cell counter (Beckman Coulter), where dye oxidation moved the fluorescence emission peak from 590 nm to 510 nm. For confocal microscopy, cells were imaged at 40 times oil magnification using a Leica SP 5II confocal laser scanning microscope, and image analysis was performed using NIS Elements Viewer.

[0053] 6.L14-8 binds to PLK1 and promotes its degradation, thereby enhancing TP53 transcriptional activity by increasing TP53 protein phosphorylation and expression, thereby inducing cell ferroptosis.

[0054] Furthermore, in order to find the direct binding targets of L14-8 involved in TP53-mediated ferroptosis in prostate cancer, the predicted targets of L14-8 were first retrieved from the SwissTargetPrediction portal. The overlap of the top 50 targets with the highest probability and TP53-interacting proteins was analyzed to analyze possible targets. Then, the possible target PLK1 was analyzed for poor overall survival (OS) and disease-free survival (DFS). Molecular docking was used to calculate the binding affinity between L14-8 and PLK1. Comparison of the interaction between L14-8 treatment and endogenous PLK1 demonstrated that L14-8 treatment increased the thermal stability of endogenous PLK1 in the prostate cancer cell model.

[0055] The specific operation is as follows: the cells were treated with L14-8 or DMSO for 6 hours and then exposed to 37°C, 41°C, 44°C, 47°C, 50°C, 53°C, 56°C, 59°C, 63°C and 67°C for 3 minutes each. The cells were then lysed for three cycles using liquid nitrogen and a 25°C heater. Proteins were collected by centrifugation at 12000rpm and 4°C for 15 minutes, and Western blot assays were performed to evaluate PLK1 and ACTB expression. Given that PLK 1 is known to directly bind to TP 53 and inhibit its transcriptional activity by reducing its protein stability and phosphorylation, PLK 1 and TP 53 mRNA and protein expression levels were further evaluated.

[0056] In addition, TP53 was knocked out using CRISPR-Cas 13 to observe the sensitivity of cells to L14-8.

[0057] The specific operation is as follows: Five guide RNA (gRNA) sequences targeting CDC25B were designed for CRISPR-Cas 13 and cloned into the gRNA backbone. The gRNA plasmid was transfected into the cells together with the CRISPR-Cas13 plasmid using a transfection reagent. After transfection, RNA was extracted from the cells for RT-qPCR to evaluate the efficiency of gRNA deletion, and two gRNAs were selected for further determination. Prostate cancer cells C4-2B were cultured in RPMI 1640 complete medium with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a 5% CO2 cell culture incubator. Cells in the logarithmic growth phase were digested, counted and plated on 96-well plates. The treatment conditions were as follows: gRNA plasmids (including empty gDummy, TP53-g1, TP53-g3) were transfected into cells together with CRISPR-Cas13 plasmids using a transfection reagent. After 6 hours of changing the medium and culturing for 24 hours, two concentrations of drugs (5 and 7.5 μM) were added, with 5 replicate wells in each group. Drugs were given 24 hours after plating, and cell viability was detected using a cell counting reagent (CCK-8) 72 hours after drug administration.

[0058] Experimental Conclusion

[0059] 1.L14 small molecule has a significant inhibitory effect on prostate cancer cells.

[0060] Compared with the other 460 compounds in the established small molecule library, L14 small molecule significantly inhibited the growth of two prostate cancer cell lines PC3 and C4-2B, while having the least effect on the growth of normal prostate cell line RWPE 1. Cell proliferation experiments were performed on AR-positive C4-2B and AR-negative PC 3 prostate cancer cells with Enzalutamide, an AR antagonist that is currently used frequently in clinical practice ( Figure 1Left 1 and left 2 are the chemical structures of Ezetimibe and L14 small molecules respectively). The data show that L14 small molecule has a significant cell inhibitory effect ( Figure 1 Center right 1).

[0061] 2. The L14 small molecule was optimized and the L14-8 small molecule was screened out, which has the strongest inhibitory effect on the growth of prostate cancer cells and the weakest effect on the growth of normal prostate cells.

[0062] After optimizing the L14 small molecule, L14-1 to L14-12 ( Figure 2 The data of CCK-8 validation of these 12 small molecules and Ezetimibe in two prostate cancer cell lines, C4-2B and PC3, showed that L14-8 showed the most significant prostate cancer cell growth inhibition ( Figure 2 ), and the IC 50 of L14-8 in both prostate cancer cell lines was less than 10 μM ( Figure 2 ), however, at a high dose of 25 μM, L14-8 had no significant effect on the growth of normal prostate cells ( Figure 2 ), highlighting the specificity of the optimized small molecule in the treatment of prostate cancer.

[0063] Furthermore, the above L14-8 small molecule was used in combination with the AR antagonist Enzalutamide to detect cell resistance. The data showed that the L14-8 small molecule can significantly enhance the anti-tumor activity in the AR-positive prostate cancer cell model ( Figure 2 ). It was verified that the combination of L14-8 and low-dose AR antagonists can resolve AR antagonist-induced drug resistance.

[0064] 3. The results of L14-8 drug experiments in animals prove that the L14-8 small molecule can effectively inhibit the growth of prostate cancer.

[0065] C4-2B xenograft mice were treated with L14-8 at daily doses of 10 mg / kg and 20 mg / kg. Figure 3 As shown, the tumors in the drug-treated group were significantly smaller than those in the control group. Ki 67 staining (a marker of cell proliferation) data revealed that Ki 67 values ​​of tumors treated with L14-8 were significantly reduced, demonstrating its antiproliferative effect in vivo ( Figure 3 ).

[0066] 4. The results of L14-8 drug experiments in animals proved that the L14-8 small molecule effectively inhibited the growth of prostate cancer while having no significant toxicity to normal prostate cells.

[0067] In the above-mentioned in vivo animal experiments, compared with the control group, H&E staining of the main organs of mice treated with high-dose L14-8, such as the heart, kidney, liver, lung, and spleen, showed no significant damage changes ( Figure 4 In addition to the decreased ALP in the high-dose L14-8 group, liver and kidney function indices, including alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (CREA), were similar between L14-8-treated mice and control-treated mice ( Figure 4 ). The results showed that L14-8 could effectively inhibit prostate cancer growth in an in vivo model without significant toxicity.

[0068] 5.L14-8 acts on the ferroptosis pathway and causes cell apoptosis.

[0069] like Figure 5 As shown in the Figure 2, transcriptome analysis, KEGG enrichment analysis, differential gene enrichment analysis (DGEA), and gene set enrichment analysis (GSEA) after L14-8 treatment indicated that the tumor growth inhibition mediated by L14-8 was associated with the ferroptosis pathway. The significant activation of ferroptosis after L14-8 treatment was further verified using C11 BODIPY (581 / 591), a fluorescent probe that indicates lipid peroxidation and antioxidant efficacy of ferroptosis in living cells. Figure 5 Flow cytometry analysis revealed that significant apoptosis was observed only at higher doses of L14-8 in various prostate cancer cell models ( Figure 5 ). These results suggest that low doses of L14-8 inhibit cancer cell growth mainly through ferroptosis, whereas higher doses of L14-8 trigger both ferroptosis and programmed cell death.

[0070] 6. L14-8 binds to PLK1 and promotes its degradation, thereby enhancing TP53 transcriptional activity by increasing TP53 protein phosphorylation and expression, thereby inducing cell ferroptosis.

[0071] The predicted targets of L14-8 were retrieved from the SwissTargetPrediction portal, and the top 50 targets with the highest probability were analyzed for overlap with TP53-interacting proteins, showing that PLK1 was the only target that intersects with TP53 ( Figure 6 PLK1 is highly expressed in prostate tumors compared with adjacent tissues, and its elevated expression is associated with poor overall survival (OS) and disease-free survival (DFS) in various cohorts of prostate cancer patients ( Figure 6 Molecular docking showed that the binding affinity between L14-8 and PLK1 was -5.07 kJ / mol ( Figure 6By comparing the interaction between L14-8 treatment and endogenous PLK1, it was demonstrated that L14-8 treatment increased the thermostability of endogenous PLK1 in a prostate cancer cell model ( Figure 6 ).

[0072] Given that PLK1 is known to directly bind to TP 53 and inhibit its transcriptional activity by reducing its protein stability and phosphorylation, the PLK1 and TP 53 mRNA and protein expression levels were further evaluated. RT-qPCR results showed that after treatment with various doses of L14-8, PLK1 and TP 53 mRNA levels remained unchanged. At the same time, L14-8 treatment led to an increase in TP 53 and phosphorylated TP 53 (p-TP 53) protein levels, while PLK1 protein levels decreased ( Figure 6 ). In addition, knockout of TP53 using CRISPR-Cas 13 significantly desensitized cells to L14-8 ( Figure 6 These results suggest that L14-8 binds to PLK1 and promotes its degradation, thereby enhancing TP53 transcriptional activity by increasing TP53 protein phosphorylation and expression, thereby inducing ferroptosis.

[0073] L14-8 has the effect of inhibiting the proliferation, migration and colony formation of castration-resistant prostate cancer in vitro, and has the effect of inhibiting tumor growth in vivo. The combined use of AR antagonists and low-dose L14-8 can not only avoid the potential toxicity of L14-8, but also significantly improve the sensitivity of CRPC to AR antagonists and the inhibitory effect of the two drugs alone; the significant killing effect on prostate cancer cells suggests that L14-8 has the potential to be used as an anti-prostate cancer drug skeleton, but not limited to anti-prostate cancer drugs.

[0074] The above-described embodiments are merely descriptions of the implementation methods of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by those skilled in the art should be included in the protection scope determined by the claims of the present invention.

Claims

1. Use of the ezetimibe derivative L14 in the preparation of an anti-prostate cancer drug, characterized in that: The general structural formula of the ezetimibe derivative L14 is as follows: Wherein: R1, R2 are each independently selected from H, n=0,1,2,3,4,5,6; R is selected from C1-C8 alkyl, C2-C8 olefin, halogenated thiophene.

2. The use of the ezetimibe derivative L14 according to claim 1 in the preparation of an anti-prostate cancer drug, characterized in that: The R1 and R2 are selected from the following:

3. The use of the ezetimibe derivative L14 according to claim 1 in the preparation of an anti-prostate cancer drug, characterized in that: The ezetimibe derivative L14 is selected from L14-8.

4. The use of the ezetimibe derivative L14 according to claim 1 in the preparation of an anti-prostate cancer drug, characterized in that: The prostate cancer is selected from castration-resistant prostate cancer CRPC and neuroendocrine prostate cancer NEPC.

5. The use of the ezetimibe derivative L14 according to claim 4 in the preparation of an anti-prostate cancer drug, characterized in that: The prostate cancer cells are selected from prostate cancer PC3, 22RV2 or C4-2B cells.

6. A pharmaceutical composition containing the above-mentioned ezetimibe derivative L14, wherein the active ingredients thereof include the above-mentioned ezetimibe derivative L14 and a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition containing the ezetimibe derivative L14 according to claim 6, characterized in that: The pharmaceutically acceptable salt includes a salt formed by the ezetimibe derivative L14 and an organic acid or an inorganic acid. The organic acid is selected from one or more of malic acid, lactic acid, camphorsulfonic acid, citric acid, fumaric acid or oxalic acid, and the inorganic acid is selected from one or more of phosphoric acid, hydrohalic acid, sulfuric acid or nitric acid.

8. Application of AR antagonist and ezetimibe derivative L14 in the preparation of anti-prostate cancer drugs.

9. Use of the AR antagonist and the ezetimibe derivative L14 according to claim 8 in the preparation of an anti-prostate cancer drug, characterized in that: The AR antagonist is selected from Darolutamide or Enzalutamide.

10. Use of the AR antagonist and the ezetimibe derivative L14 according to claim 8 in the preparation of an anti-prostate cancer drug, characterized in that: The ezetimibe derivative is L14-8.

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