Use of steviol glycoside derivatives in the preparation of a medicament for increasing the sensitivity of prostate cancer to chemotherapy

Abstract

The application belongs to the technical field of biological medicine, and relates to application of a stevioside derivative in preparation of a medicine for improving chemotherapeutic sensitivity of prostate cancer. A chemical structural formula of the stevioside derivative is: the application firstly finds that the derivative SN can not only act on lysosomes to exert a pharmacological effect of reversing docetaxel resistance, but also can significantly improve selectivity and reduce toxicity by taking the lysosome as a target.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to the application of stevioside derivatives in the preparation of drugs that improve the chemosensitivity of prostate cancer. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Prostate cancer (PCa) is a malignant tumor that seriously threatens the lives and health of middle-aged and elderly men. As a hormone-dependent tumor, endocrine therapy is the first-line treatment for prostate cancer; however, almost all patients who are sensitive to hormone therapy will develop hormone resistance after 1-2 years, progressing to castration-resistant prostate cancer (CRPC), accompanied by bone or soft tissue metastases, resulting in a poor prognosis and high mortality rate. Docetaxel (Doc), as a first-line chemotherapy drug for CRPC, is currently the only drug proven to significantly prolong patient survival. However, about 50% of CRPC patients are congenitally insensitive to docetaxel chemotherapy, while sensitive patients often develop tolerance after about two years of chemotherapy, accompanied by multidrug resistance. To date, there is no effective treatment strategy for multidrug-resistant CRPC. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the purpose of this invention is to provide the application of stevioside derivatives in the preparation of drugs that improve the chemosensitivity of prostate cancer.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows:

[0006] On the one hand, there is the application of a stevioside derivative in the preparation of drugs that inhibit prostate cancer cells;

[0007] The chemical structural formula of the steviol glycoside derivative is as follows:

[0008]

[0009] On the other hand, the application of a steviol glycoside derivative in the preparation of drugs that improve the chemosensitivity of prostate cancer; the chemical structural formula of the steviol glycoside derivative is as follows:

[0010]

[0011] Thirdly, a chemotherapy drug for prostate cancer includes docetaxel and a resistance reversal agent, wherein the chemical structural formula of the resistance reversal agent is as follows:

[0012]

[0013] Docetaxel, as a first-line chemotherapy drug for castration-resistant prostate cancer (CRPC), is currently the only drug proven to significantly prolong patient survival. However, drug resistance and toxic side effects remain major challenges in treatment. Overcoming bottlenecks in drug target selection and drug development has become a core issue in the field of CRPC drug resistance. Lysosomes, as key participants in cell metabolism, apoptosis, and autophagy, play multiple roles in tumor cell survival and drug resistance. Intervening in lysosomal function has unique advantages in reversing drug resistance. Therefore, by systematically elucidating the characteristics of SN-targeted lysosome-mediated reversal of drug resistance and the chemotherapy strategy of combining docetaxel with CRPC, this study not only provides theoretical support for the clinical application of SN as a lysosomal-targeted drug but also expands existing strategies for reversing drug resistance.

[0014] The beneficial effects of this invention are as follows:

[0015] This invention is the first to discover that the derivative SN not only acts on lysosomes to reverse docetaxel resistance, but also that targeting lysosomes significantly improves selectivity and reduces toxicity. It systematically elucidates the chemotherapy strategy of SN as a lysosomal targeted drug to kill drug-resistant cells and synergistically intervene in CRPC with docetaxel, providing a basis for its application in the field of CRPC resistance and the development of novel chemotherapy regimens.

[0016] This invention reveals for the first time the regulatory mechanism by which SN intervenes in lysosomal function to reverse drug resistance. Based on the unique biological characteristics of lysosomes in drug-resistant cells, this invention elucidates the role of SN in reversing drug resistance by elucidating how SN acts on lysosomes, inducing lysosomal membrane leakage, causing cathepsin leakage, and subsequently leading to apoptosis.

[0017] This invention elucidates for the first time the clinical application value of SN as a lysosomal targeting agent: by using homologous mouse CRPC cells to inoculate mice to construct a drug-resistant animal model, the pharmacodynamic activity of SN in killing drug-resistant cells and the therapeutic effect of SN in combination with docetaxel are elucidated at the animal level, providing theoretical support for the clinical application of SN. Attached Figure Description

[0018] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0019] Figure 1The above diagram shows the inhibitory effect of stevioside derivative SN on prostate cancer cells in the embodiments of the present invention. A is the in vitro activity curve of SN against prostate cancer cells, B is the in vitro activity bar chart of SN against drug-resistant cells, and C is the cell apoptosis detection result of Annexin V-FITC / PI double staining method.

[0020] Figure 2 The images shown are the results of the combined application of stevioside derivative SN and docetaxel in the embodiments of the present invention. A is the result of the MTT assay, and B is the result of the Annexin V-FITC / PI double staining assay for apoptosis detection.

[0021] Figure 3 The following are the test results of the SN intervention of lysosomal function by the stevioside derivative in the embodiments of the present invention: A is the result of fluorescence microscopy after staining with lysosomal red fluorescent probe (Lyso-tracker Red); B is the result of flow cytometry after staining with Lyso-tracker Red; C is the result of electron microscopy; and D is the result of cell apoptosis detection by Annexin V-FITC / PI double staining.

[0022] Figure 4 The following are the results of the in vivo pharmacodynamic effects of the stevioside derivative SN in the embodiments of the present invention. A is the result of SN affecting the body weight of mice in the RM-1 group, B is the result of SN affecting the body weight of mice in the RM-1 / Doc group, and C is the result of SN alone or in combination with docetaxel affecting the tumor weight of mice in the RM-1 group and the RM-1 / Doc group. Detailed Implementation

[0023] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0024] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0025] Given that there is currently no effective treatment strategy for castration-resistant prostate cancer, this invention proposes the application of stevioside derivatives in the preparation of drugs that improve the chemosensitivity of prostate cancer.

[0026] One typical embodiment of the present invention provides the application of a stevioside derivative in the preparation of a drug for inhibiting prostate cancer cells;

[0027] The chemical structural formula of the steviol glycoside derivative is as follows:

[0028]

[0029] In some embodiments, the prostate cancer cells are prostate cancer cell lines PC3 and / or prostate cancer cell line DU145.

[0030] In some embodiments, the prostate cancer cells are drug-resistant prostate cancer cells, such as nedocetaxel-resistant prostate cancer cells.

[0031] Another embodiment of the present invention provides the application of a steviol glycoside derivative in the preparation of a drug for improving the chemosensitivity of prostate cancer; the chemical structural formula of the steviol glycoside derivative is:

[0032]

[0033] In some embodiments, drugs that improve the chemosensitivity of prostate cancer are drugs that intervene in lysosomal function.

[0034] In some embodiments, the drug that improves the sensitivity of prostate cancer to chemotherapy is a drug resistance reversal agent.

[0035] A third embodiment of the present invention provides a chemotherapy drug for prostate cancer, comprising docetaxel and a drug resistance reversal agent, wherein the chemical structural formula of the drug resistance reversal agent is as follows:

[0036]

[0037] In some embodiments, pharmaceutical excipients are included.

[0038] In one or more embodiments, the pharmaceutical excipient is a pharmaceutical carrier and / or excipient. The amount of pharmaceutical excipient can be 1% to 99% of the total weight of the drug. The carrier includes, but is not limited to, glycerol, lecithin, phosphate, ethylene glycol, sodium carboxymethyl cellulose, aluminum stearate, serum albumin, etc. The excipient includes, but is not limited to, binders, fillers, thickeners, disintegrants, etc. The binder is, for example, gum arabic, astragalus gum, etc. The filler is, for example, corn starch, calcium phosphate, etc. The thickener is, for example, sodium alginate, pectin, etc. The disintegrant is, for example, potato starch, etc.

[0039] In some embodiments, the dosage form is an injection, tablet, granule, capsule, pill, etc.

[0040] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0041] The steviol glycoside derivatives used in the following examples have the following chemical structural formulas:

[0042] The value is denoted as SN and obtained according to the method described in the literature (Zhaomin Lin, Yanxia Guo, Yanhui Gao, Shuqi Wang, Xiaoning Wang, Zhiyu Xie, Huanmin Niu, Wenqiang Chang, Lei Liu, Huiqing Yuan, and Hongxiang Lou. ent-Kaurane Diterpenoids from Chinese Liverworts and Their Antitumor Activities through Michael Addition As Detected in Situ by a Fluorescence Probe. J. Med. Chem. 2015, 58, 9, 3944–3956).

[0043] 1. Discovery of highly effective and selective drug resistance reversal agents:

[0044] (1) Constructing a docetaxel-resistant cell model of prostate cancer PC3 / Doc

[0045] The drug-resistant cell line PC3 / Doc was obtained through screening by gradually increasing the concentration of docetaxel. PC3 cells were cultured in a medium with a final concentration of 5 nM docetaxel for 48 hours, then replaced with fresh medium (docetaxel-free) until the cells were in good condition. The concentration of docetaxel was then gradually increased until the cells could grow normally in the final concentration of docetaxel medium. Finally, they were cultured in a medium containing 1 nM docetaxel.

[0046] (2) Cell proliferation activity was detected by the MTT assay. Cells were treated with different concentrations of SN (0.5, 1, 2, 5, 10 μM) for 48 h to evaluate the in vitro activity of SN against prostate cancer cells and drug-resistant cells. It was confirmed that SN had strong inhibitory activity against both PC3 and DU145 prostate cancer cell lines, but its toxicity to normal prostate epithelial cells RWPE1 was significantly reduced. Figure 1 As shown in Figure A, the inhibitory activity of SN on PC3 / Doc cells was significantly higher than that on PC3, such as... Figure 1 As shown in B.

[0047] Apoptosis was detected using Annexin V-FITC / PI double staining. Different concentrations of SN (0.5, 1 μM) were applied to PC3 and PC3 / Doc cells for 24 h, and the apoptosis rate was analyzed by flow cytometry. The results showed that SN had a stronger apoptosis-inducing ability on PC3 / Doc cells, such as... Figure 1 As shown in C.

[0048] (3) MTT assay showed that the combined application of SN and docetaxel to treat PC3 and PC3 / Doc cells produced a synergistic effect, with SN significantly enhancing the cell-inhibiting effect of docetaxel. Figure 2 As shown in Figure A. Annexin V-FITC / PI cell apoptosis detection: PC3 / Doc cells were treated with SN and docetaxel alone or in combination for 24 h, and the apoptosis rate was analyzed by flow cytometry. The results showed that SN significantly enhanced the killing effect of docetaxel on PC3 / Doc cells, such as... Figure 2 As shown in B.

[0049] 2. SN intervention in lysosomal function:

[0050] Solution:

[0051] (1) Analysis of the SN-targeted lysosomal effect of stevioside derivatives. Microscopic observation and flow cytometry analysis after Lyso-tracker Red staining showed that SN treatment for 2 hours induced an increase in the number, size, and aggregation of lysosomes. With prolonged exposure, the fluorescence gradually diffused and disappeared, indicating that lysosomes initially undergo compensatory formation after damage, but ultimately, due to the continued impact of the damage, the lysosomal system is completely inactivated. Figure 3 A, Figure 3 As shown in B; electron microscopy results further reveal the role of SN in lysosomes, such as Figure 3 As shown in C.

[0052] (2) Cells were pretreated with the cathepsin inhibitors E64d and CA074Me, and then SN-induced cell death was detected by Annexin V-FITC / PI. The results showed a reduction in the number of dead cells, confirming that SN caused leakage of lysosomal cathepsins, leading to cell death. Figure 3 As shown in D.

[0053] 3. Pharmacodynamic effects of SN in vivo:

[0054] RM-1 / Doc, docetaxel-resistant cells from mouse prostate cancer cells RM-1, were constructed. Both cell lines were then inoculated into C56BL / 6 mice to further establish a mouse model of prostate cancer and its drug resistance model. The RM-1 or RM-1 / Doc model groups were further randomly divided into four groups of five mice each: a solvent control group, a SN treatment group (30 mg / kg), a docetaxel treatment group (Doc, 5 mg / kg), and a combination group (SN+Doc). The drugs were administered intraperitoneally every other day for 12 consecutive days. Body weight was measured every two days. At the end of the experiment, the mice were weighed, and their tumors were removed and weighed.

[0055] Animal experiments confirmed that SN alone significantly inhibited tumor growth in the RM-1 and RM-1 / Doc groups, and that its combination with docetaxel enhanced the antitumor efficacy. Simultaneously, SN did not significantly inhibit mouse body weight, indicating that SN has low in vivo toxicity. Figure 4 As shown.

[0056] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of a stevioside derivative in the preparation of a drug to improve the chemosensitivity of docetaxel-resistant prostate cancer; the chemical structural formula of the stevioside derivative is: 。 2. The use of the stevioside derivative as described in claim 1 in the preparation of a drug for improving the chemosensitivity of docetaxel-resistant prostate cancer, characterized in that, Drugs that improve the chemosensitivity of docetaxel-resistant prostate cancer are drugs that intervene in lysosomal function.

3. The use of the stevioside derivative as described in claim 1 in the preparation of a drug to improve the chemosensitivity of docetaxel-resistant prostate cancer, characterized in that, Drugs that improve the chemosensitivity of prostate cancer patients resistant to docetaxel are resistance reversal agents.

4. A chemotherapy drug for prostate cancer, comprising docetaxel and a resistance reversal agent, characterized in that, The chemical structural formula of the drug resistance reversal agent is: 。 5. The chemotherapy drug for prostate cancer as described in claim 4, characterized in that, Including pharmaceutical excipients.

6. The chemotherapy drug for prostate cancer as described in claim 5, characterized in that, The pharmaceutical excipients are pharmaceutical carriers.

7. The chemotherapy drug for prostate cancer as described in claim 5, characterized in that, The pharmaceutical excipients are excipients.

8. The chemotherapy drug for prostate cancer as described in claim 4, characterized in that, Dosage forms include injections, tablets, granules, capsules, or pills.

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