Application of pulsatilla saponin D in preparation of medicine for preventing or treating human prostatic hyperplasia
By using Pulsatilla saponin D to inhibit prostate cell proliferation, the problem of side effects of existing drugs has been solved, and effective treatment of benign prostatic hyperplasia has been achieved.
Patent Information
- Application Number
- CN202511381132.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-25
- Publication Date
- 2025-12-16
AI Technical Summary
Existing medications for treating benign prostatic hyperplasia (BPH) have side effects, such as orthostatic hypotension and sexual dysfunction, necessitating the development of highly effective and low-toxicity treatment options.
Using Pulsatilla saponin D as the active ingredient, a pharmaceutical composition was prepared to prevent or treat benign prostatic hyperplasia by inhibiting the proliferation of human prostate stromal cells, fibroblasts and hyperplastic epithelial cells.
Pulsatilla saponin D has shown an inhibitory effect on human prostate cells, inducing apoptosis and effectively preventing or treating benign prostatic hyperplasia without significant side effects.
Smart Images
Figure BDA0005614143230000011 
Figure BDA0005614143230000041 
Figure HDA0005614143250000011
Abstract
Description
Technical Field
[0001] This invention belongs to the field of medicine, specifically relating to new uses of Pulsatilla saponin D, particularly its use in the preparation of drugs for the prevention or treatment of benign prostatic hyperplasia in humans. Background Technology
[0002] Benign prostatic hyperplasia (BPH) is one of the most common chronic diseases of the male genitourinary system, characterized primarily by the proliferation of epithelial and stromal cells in the prostate. Currently, most BPH patients still rely on medication for treatment. However, current medications generally have side effects such as orthostatic hypotension and sexual dysfunction. Therefore, there is an urgent need to develop highly effective and low-toxicity drugs for the treatment of BPH. Summary of the Invention
[0003] This invention provides a new use for Pulsatilla saponin D (PSD) in the pharmaceutical field.
[0004] The structural formula of Pulsatilla saponin D is as follows:
[0005]
[0006] According to a first aspect of the invention, the use of Pulsatilla saponin D (PSD) or a pharmaceutical composition containing Pulsatilla saponin D is provided in the preparation of a medicament for the prevention or treatment of benign prostatic hyperplasia in humans.
[0007] In some embodiments, Pulsatilla saponin D prevents or treats benign prostatic hyperplasia (BPH) by inhibiting the proliferation of human prostate stromal cells, human prostate fibroblasts, and / or human prostatic hyperplasia epithelial cells.
[0008] In some embodiments, in particular, the human prostate stromal cells are WPMY-1, the human prostate fibroblasts are HPRF, and the human prostate hyperplasia epithelial cells are BPH-1.
[0009] According to a second aspect of the invention, the use of pulsatilla saponin D or a pharmaceutical composition containing pulsatilla saponin D in the preparation of a medicament for inhibiting the proliferation of human prostate stromal cells, human prostate fibroblasts and / or human prostate hyperplastic epithelial cells is provided.
[0010] In some embodiments, in particular, the human prostate stromal cells are WPMY-1, the human prostate fibroblasts are HPRF, and the human prostate hyperplasia epithelial cells are BPH-1.
[0011] In the first and second aspects of the invention, the pharmaceutical composition comprising Pulsatilla saponin D may include optional medically or pharmaceutically acceptable excipients.
[0012] In some embodiments, the pharmaceutical composition may be any pharmacologically acceptable dosage form, such as any one selected from tablets, powders, granules, capsules, oral solutions, injections, sustained-release formulations, and controlled-release formulations.
[0013] According to a third aspect of the invention, a pharmaceutical composition for the prevention or treatment of benign prostatic hyperplasia in humans is provided, comprising, or consisting of, pulsatilla saponin D and optional medically or pharmaceutically acceptable excipients.
[0014] In some embodiments, the pharmaceutical composition may be any pharmacologically acceptable dosage form, such as any of the following selected: tablets, powders, granules, capsules, oral solutions, injections, sustained-release formulations, and controlled-release formulations. The dosage of Pulsatilla saponin D may be a pharmacologically acceptable dosage.
[0015] According to a fourth aspect of the invention, a method for preventing or treating benign prostatic hyperplasia in humans is provided, comprising administering a therapeutically effective amount of Pulsatilla saponin D to a subject in need.
[0016] The experiments of this invention show that Pulsatilla saponin D has an inhibitory effect on human prostate cell lines WPMY-1, HPRF and BPH-1, and can induce apoptosis to a certain extent, thus showing the efficacy of preventing or treating benign prostatic hyperplasia. Attached Figure Description
[0017] Figure 1 The effect of different concentrations of Pulsatilla saponin D on the viability of human prostate stromal cells WPMY-1 is shown in the figure. Compared with the solvent control group, *P<0.05, ***P<0.001.
[0018] Figure 2 The effect of different concentrations of Pulsatilla saponin D on the activity of human prostatic fibroblasts (HPRF) is shown in the figure. Compared with the solvent control group, *P<0.05, **P<0.01, ***P<0.001.
[0019] Figure 3 The effect of different concentrations of Pulsatilla saponin D on the viability of human prostatic hyperplasia epithelial cells BPH-1 is shown in the figure. Compared with the solvent control group, ***P<0.001.
[0020] Figure 4 The diagram shows the inhibitory effect of Pulsatilla saponin D on human prostate stromal cells WPMY-1, human prostate fibroblasts HPRF, and human prostate hyperplasia epithelial cells BPH-1 in the examples.
[0021] Figure 5The results of the apoptosis detection experiment induced by Pulsatilla saponin D in WPMY-1 cells are shown (left) and the bar chart of statistical analysis based on the apoptosis detection experiment results (right). Compared with the solvent control group, *P<0.05, ***P<0.001.
[0022] Figure 6 The results of the apoptosis detection experiment induced by Pulsatilla saponin D in HPRF cells are shown (left) and the bar chart of statistical analysis based on the apoptosis detection experiment results (right). Compared with the solvent control group, *P<0.05, ***P<0.001.
[0023] Figure 7 The results of the apoptosis detection experiment induced by Pulsatilla saponin D in BPH-1 cells are shown (left) and the bar chart of statistical analysis based on the apoptosis detection experiment results (right). Compared with the solvent control group, *P<0.05, **P<0.01. Detailed Implementation
[0024] To enable those skilled in the art to better understand the present invention, the present invention will be further described below in conjunction with embodiments and accompanying drawings.
[0025] As those skilled in the art know, WPMY-1 is an immortalized myofibroblast cell line that provides a novel and useful model for studying the role of paracrine matrix epithelial interactions in benign prostatic hyperplasia (BPH) as well as carcinogenesis and tumor progression; HPRF is one of the important primary cell models for studying the prostate stromal microenvironment and its role in BPH and carcinogenesis; BPH-1 is a prostatic hyperplasia cell line that is commonly used in BPH studies to investigate the promoting or inhibiting effects of various compounds on prostatic hyperplasia.
[0026] The following examples use WPMY-1, HPRF, and BPH-1 cells as cell models to demonstrate the efficacy of Pulsatilla saponin D on benign prostatic hyperplasia.
[0027] Unless otherwise specified, the raw materials, reagents, methods, and equipment used in this application are conventional or commercially available in the field.
[0028] Pulsatilla saponin D (PSD): Purchased from MedChemExpress, purity determined by high performance liquid chromatography (HPLC) was 98.60%. A 10 mM stock solution was prepared using dimethyl sulfoxide (DMSO) and stored at -80°C. It was diluted to the working concentration with culture medium before each use. The positive control drug, doxazosin (Doxa), was prepared as a 100 mM stock solution using DMSO and stored at -20°C. It was diluted to the working concentration with culture medium before each use.
[0029] Cell lines: The WPMY-1 cell line used in the examples was purchased from Shanghai Zhongqiao Xinzhou, HPRF was purchased from Sciencell, and BPH-1 cell line was purchased from Shanghai Yansheng.
[0030] Example 1: Pulsatilla saponin D inhibits the growth of prostate cells.
[0031] Prostate cells in logarithmic growth phase, WPMY-1, HPRF, and BPH-1, were seeded in 96-well plates at seeding numbers of WPMY-1, HPRF, and BPH-1 (48h: 4000 cells / well; 72h: 3000 cells / well). After incubation at 37°C for 24 hours, the cells were treated with 500 nM-4 μM PSD, respectively. Doxazosin (40 μM) was used as a positive control. After incubation for 48 or 72 hours, 10 μL of CCK-8 reagent (Shanghai Beyotime Biotechnology Co., Ltd.) was added to each well. After incubation in the dark for 4 hours, the absorbance (OD450) at 450 nm was measured using a microplate reader, and cell viability was calculated. The results are shown in Figures 1-3. Figure 1-4 And in Table 1.
[0032] Table 1. IC50 values of PSD's inhibitory effect on prostate cells.
[0033]
[0034] The results showed that different concentrations of PSD in this invention had varying degrees of inhibitory effects on human prostate cells WPMY-1, HPRF, and BPH-1. Table 1 shows that after 48 hours of PSD treatment, the half-maximal inhibitory rates (IC50) for WPMY-1, HPRF, and BPH-1 were 2.649 μM, 1.201 μM, and 4.816 μM, respectively; after 72 hours of treatment, the IC50 values for WPMY-1, HPRF, and BPH-1 were 2.511 μM, 1.192 μM, and 4.315 μM, respectively. Figure 4 The diagram shows the morphological changes of WPMY-1, HPRF and BPH-1 cells after 48 hours of PSD treatment. It can be seen that PSD at 2 μM can induce significant shrinkage and apoptosis in WPMY-1 and HPRF cells, and at 4 μM can induce significant shrinkage and apoptosis in BPH-1 cells.
[0035] Prostate cells in logarithmic growth phase (WPMY-1, HPRF, and BPH-1) were seeded in six-well plates at a density of 2.5 × 10⁵ cells / well and incubated at 37°C for 24 h. The cells were then treated with 500 nM–4 μM PSD. After 48 h of incubation, cells were collected for Annexin VFITC-PI staining (Yisheng Biotechnology (Shanghai) Co., Ltd.), and apoptosis rate was detected by flow cytometry.
[0036] Reference for apoptosis detection experimental results Figure 5 , Figure 6 and Figure 7 The results showed that, compared with the solvent control group, 500 nM-4 μM PSD led to an increase in the apoptosis rate of WPMY-1 and HPRF cells. Specifically, 1-4 μM PSD significantly induced apoptosis in both WPMY-1 and HPRF cells (*P < 0.05). 2 μM PSD significantly induced apoptosis in BPH-1 cells (**P < 0.01).
[0037] As confirmed by the above experiments, Pulsatilla saponin D has an inhibitory effect on human prostate cells WPMY-1, HPRF and BPH-1. Experiments show that different concentrations can induce significant shrinkage and apoptosis of WPMY-1, HPRF and BPH-1 cells, and within a certain concentration range, it leads to an increase in the apoptosis rate of WPMY-1, HPRF and BPH-1 cells, thereby inhibiting prostate hyperplasia.
Claims
1. The use of Pulsatilla saponin D or a pharmaceutical composition containing Pulsatilla saponin D in the preparation of a medicament for the prevention or treatment of benign prostatic hyperplasia in humans.
2. The application according to claim 1, wherein, Pulsatilla saponin D inhibits the proliferation of human prostate stromal cells, human prostate fibroblasts, and / or human prostate hyperplastic epithelial cells.
3. The application according to claim 2, wherein, Human prostate stromal cells are WPMY-1, human prostate fibroblasts are HPRF, and human prostate hyperplastic epithelial cells are BPH-1.
4. The use of Pulsatilla saponin D or a pharmaceutical composition containing Pulsatilla saponin D in the preparation of formulations that inhibit the proliferation of human prostate stromal cells, human prostate fibroblasts and / or human prostate hyperplastic epithelial cells.
5. The application according to claim 4, wherein, Human prostate stromal cells are WPMY-1, human prostate fibroblasts are HPRF, and human prostate hyperplastic epithelial cells are BPH-1.
6. The application according to any one of claims 1-5, wherein, The pharmaceutical composition containing pulsatilla saponin D contains optional pharmaceutically acceptable excipients.
7. The application according to any one of claims 1-5, wherein, The pharmaceutical composition containing Pulsatilla saponin D is in any pharmacologically acceptable dosage form, such as any one selected from tablets, powders, granules, capsules, oral liquids, injections, sustained-release formulations, and controlled-release formulations.
8. A pharmaceutical composition for treating benign prostatic hyperplasia in humans, comprising pulsatilla saponin D and optional pharmaceutically acceptable excipients.
9. The pharmaceutical composition according to claim 8, comprising Pulsatilla saponin D and optionally pharmaceutically acceptable excipients.
10. The pharmaceutical composition according to claim 8 or 9, wherein, The pharmaceutical composition is in the form of tablets, powders, granules, capsules, oral liquids, injections, sustained-release agents, or controlled-release agents.