Application of total glucosides of paeony in preparation of medicine for treating triple negative breast cancer
By using total paeoniflorin (TGP) to inhibit the proliferation, migration, and invasion of triple-negative breast cancer cells and induce apoptosis, the lack of effectiveness of existing therapeutic drugs is addressed, providing an effective treatment option for triple-negative breast cancer.
Patent Information
- Application Number
- CN202511529993.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-24
- Publication Date
- 2025-12-26
AI Technical Summary
Current drugs for treating triple-negative breast cancer lack effectiveness, especially endocrine and targeted therapies, which are limited, and chemotherapy has significant side effects. Immunotherapy and ADC drugs have limited efficacy and significant individual differences.
Using total paeoniflorin (TGP) as the active ingredient, in vitro and in vivo experiments have verified that it can inhibit the proliferation, migration and invasion of triple-negative breast cancer cells, induce apoptosis, and exert anti-tumor effects through mitochondrial damage and inhibition of oxidative phosphorylation mechanisms. It is prepared into oral or injectable dosage forms.
Paeonia lactiflora total glucosides significantly inhibited the proliferation, migration, and invasion of triple-negative breast cancer cells, induced apoptosis, and significantly inhibited tumor growth in in vivo experiments, providing a new and effective drug option for the treatment of triple-negative breast cancer.
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Figure CN121197261A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of medicine, specifically relating to the application of total glutasides of Paeony (TGP) in the preparation of drugs for the treatment of triple-negative breast cancer (TNBC). Background Technology
[0002] Triple-negative breast cancer (TNBC) is a subtype of breast cancer, accounting for approximately 15%-20% of all breast cancers. It is characterized by negative estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) cells, exhibiting high heterogeneity, poor differentiation, and strong invasiveness. TNBC cells are highly invasive and proliferative, easily breaching the basement membrane into the bloodstream, leading to distant metastasis. Common sites of metastasis include the lungs, liver, and brain. Statistics show that over one-third of TNBC patients experience recurrence or distant metastasis. Once metastasis occurs, the median survival is only 18 months, and the 5-year survival rate is less than 15%, significantly lower than the overall survival rate for breast cancer patients (31%). Due to its pathological characteristics, there is a lack of effective endocrine and targeted therapies, primarily relying on chemotherapy and radiotherapy, which have significant side effects. Although immunotherapy and adjuvant therapy (ADC) have been explored, their efficacy is limited and exhibits significant individual variability, restricting their clinical application.
[0003] White peony root (Paeonia lactiflora) is a plant of the Ranunculaceae family. It possesses the functions of nourishing blood and softening the liver, relieving pain, astringing yin and stopping sweating, and calming liver yang. It is commonly used to treat conditions such as blood deficiency and chlorosis, irregular menstruation, spontaneous sweating, and night sweats. Total glucosides of paeony (TGP) are a class of glycosides extracted from white peony root. The main components include paeoniflorin, paeoniflorin lactone, benzoylpaeoniflorin, and oxypaeoniflorin, which have good anti-inflammatory, antioxidant, and anti-fibrotic pharmacological effects. They also have protective effects on the liver and cardiovascular system and show significant efficacy in the clinical application of autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, psoriasis, and eczema. Studies have shown that total glucosides of paeony have broad-spectrum and highly effective antitumor activity, exerting antitumor activity against cancers such as liver cancer, sarcoma, and lung cancer. White peony root is often used as an ingredient in antitumor decoctions. Currently, the effects of total glucosides of paeony on triple-negative breast cancer are unclear. Summary of the Invention
[0004] The purpose of this invention is to provide a novel use for total paeoniflorin, namely, its application in the preparation of drugs for treating triple-negative breast cancer. This invention demonstrates through in vitro and in vivo experiments that total paeoniflorin can effectively inhibit the proliferation, migration, and invasion of triple-negative breast cancer cells, induce apoptosis, and exert anti-tumor effects through mechanisms such as mitochondrial damage and inhibition of oxidative phosphorylation.
[0005] Specifically, the present invention is verified through the following experiments: Inhibition of cell proliferation: CCK8 assay and colony formation assay showed that TGP dose- and time-dependently reduced the viability and colony formation ability of MDA-MB-231 and MDA-MB-468 cells. Figure 1-2 ).
[0006] Inhibition of migration and invasion: Scratch and Transwell assays showed that TGP dose-dependently inhibited cell migration and invasion. Figure 3-4 ).
[0007] Inhibition of epithelial-mesenchymal transition (EMT): Western blotting showed that TGP upregulated the expression of epithelioid proteins (E-cadherin, Occludin) and downregulated the expression of mesenchymal proteins (Vimentin). Figure 5 ).
[0008] Induction of apoptosis: TUNEL / DAPI staining, flow cytometry, and Western blotting confirmed that TGP dose-dependently induced apoptosis and regulated apoptosis-related proteins (Bcl-2, Bcl-xl, BAX, cleaved-caspase3, cleaved-caspase9, cleaved-PARP). Figure 6-8 ).
[0009] Induced mitochondrial damage: Mitro-Track staining and JC-1 staining showed that TGP caused mitochondrial structural damage and decreased membrane potential. Figure 9-10 ).
[0010] Inhibition of oxidative phosphorylation: RT-qPCR and ELISA showed that TGP inhibited the expression of mitochondrial respiratory chain complex enzymes (such as NDUFA9, SDHB, ATP5A1, etc.) and NAD+ / NADH levels. Figure 11-12 ).
[0011] In vivo antitumor effects: In the MDA-MB-231 xenograft nude mouse model, TGP (500 mg / kg) significantly inhibited tumor growth. Figure 13 ).
[0012] Based on the above findings, this invention proposes that total paeoniflorin can be used to prepare a drug for treating triple-negative breast cancer. The drug comprises a therapeutically effective amount of total paeoniflorin and a pharmaceutically acceptable carrier, and the dosage form may be an oral preparation (such as capsules, tablets) or an injection. Attached Figure Description
[0013] Figure 1 This is a graph showing the effect of TGP on the viability of MDA-MB-231 and MDA-MB-468 cells in Example 1; Figure 2 This is an image showing the effect of TGP on the cloning ability of MDA-MB-231 and MDA-MB-468 cells in Example 1; Figure 3 This is an image of the effect of TGP on the migration ability of MDA-MB-231 and MDA-MB-468 cells in Example 2; Figure 4 This is an image showing the effect of TGP on the invasive ability of MDA-MB-231 and MDA-MB-468 cells in Example 2; Figure 5 This is a graph showing the effect of TGP on the levels of EMT-related proteins in MDA-MB-231 and MDA-MB-468 cells in Example 3; Figure 6 This is an image showing the effect of TGP on apoptosis in MDA-MB-231 and MDA-MB-468 cells in Example 4; Figure 7 This is a graph showing the effect of TGP on different apoptosis rates in MDA-MB-231 and MDA-MB-468 cells in Example 4; Figure 8 This is a graph showing the effect of TGP on the levels of apoptosis-related proteins in MDA-MB-231 and MDA-MB-468 cells in Example 4; Figure 9 This is an image of the effect of TGP on mitochondrial damage in MDA-MB-231 and MDA-MB-468 cells in Example 5; Figure 10 This is a graph showing the effect of TGP on mitochondrial membrane potential in MDA-MB-231 and MDA-MB-468 cells in Example 5. Figure 11 This is a graph showing the effect of TGP on the expression of mitochondrial respiratory chain complex enzymes in MDA-MB-231 and MDA-MB-468 cells in Example 6; Figure 12 This is a graph showing the effect of TGP on NAD+ / NADH levels in MDA-MB-231 and MDA-MB-468 cells in Example 6; Figure 13This is a graph showing the experimental data on the effect of TGP on the growth of TNBC tumors in nude mice, as described in Example 7. Detailed Implementation
[0014] The present invention will be further described below with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and should not be used to limit the scope of protection of the present invention.
[0015] Example 1
[0016] Total glucosides of paeony inhibit the proliferation of triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. The effect of TGP on the viability of triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) was detected by CCK8 assay.
[0017] 1) Experimental protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 96-well plates (0.3 × 10⁻⁶). 4 Cells were placed in wells (number per well), with PBS solution added around the outer edge. The cells were then placed in a cell culture incubator for 24 hours to allow adhesion. Different concentrations of TGP (0.1, 0.2, 0.4, 0.8, and 1.6 mg / ml) were added, and cell morphology was observed after 24 and 48 hours of incubation. The original culture medium was discarded, and serum-free culture medium and 10 μL CCK8 solution were added in the dark. After incubation for 2 hours, absorbance was measured at 450 nm using a microplate reader. Cell viability was calculated using the following formula. Results are shown below. Figure 1 .
[0018] Cell viability = [OD value of experimental group - OD value of blank group] / [OD value of control group - OD value of blank group] × 100%.
[0019] 2) Experimental results: such as Figure 1 As shown, the viability of MDA-MB-231 and MDA-MB-468 cells decreased with increasing TGP concentration and duration, indicating that TGP can inhibit the viability of MDA-MB-231 and MDA-MB-468 cells.
[0020] 2. Clonogenesis assay to detect the effect of TGP on the proliferation of triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1) Experimental protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates (500 cells / well) and cultured in a cell culture incubator for 48 hours. The culture medium was changed and different concentrations of TGP (0, 0.2, 0.4, and 0.8 mg / ml) were added. The culture was continued, and the medium was changed every three days for 2 weeks. After fixation with 4% paraformaldehyde, the cells were stained with crystal violet and photographed. The results are shown in the figure. Figure 2 .
[0021] 2) Experimental results: such as Figure 2 As shown, TGP can reduce the clonogenic ability of MDA-MB-231 and MDA-MB-468 cells in a dose-dependent manner.
[0022] Example 2
[0023] Total glucosides of paeony inhibit the migration and invasion of triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. Scratch assay to detect the effect of TGP on the migration of MDA-MB-231 and MDA-MB-468 cells. 1) Experimental Protocol: Using a marker pen, draw three evenly spaced horizontal lines on the back of a 12-well plate. Evenly seed logarithmically growing triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) into the 12-well plate and incubate for 24 hours to allow adhesion. Using a sterile 200μL pipette tip, make vertical scratches perpendicular to the black horizontal lines in the wells. Wash the cells in the wells three times with PBS solution, then replace with serum-free 1640 medium, adding TGP to final concentrations of 0, 0.2, 0.4, and 0.8 mg / ml. Observe and photograph the scratch width at 0 h under a microscope. After culturing for 48 h, observe and photograph the scratch width again. See the results below. Figure 3 .
[0024] 2) Experimental results: such as Figure 3 As shown, in MDA-MB-231 and MDA-MB-468 cells, the control group migrated to the scratch edge relatively quickly, while TGP treatment inhibited cell migration.
[0025] 2. Transwell assay to detect the effect of TGP on the invasive ability of MDA-MB-231 and MDA-MB-468 cells. 1) Experimental protocol: A layer of matrix gel was placed in a Transwell chamber. Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were taken and resuspended in serum-free 1640 blank medium, adjusting the cell concentration to 5 × 10⁻⁶ cells / mL. 4Cells / mL. Add 600 μL of 1640 complete culture medium to a 24-well plate, placing the chamber at the top. Add 200 μL of cell suspension to each chamber of the control and experimental groups. Add TGP to the drug treatment groups at final concentrations of 0, 0.2, 0.4, and 0.8 mg / mL. Incubate for 48 hours, then remove the 24-well plate. Carefully aspirate the culture medium from the middle and lower layers of the chambers. Wash the chambers with PBS and fix the cells with 4% paraformaldehyde for 15 min. Add crystal violet and stain for 30 min. Wash away excess staining solution. Gently wipe away the cells from the inner layer of the chambers with a cotton swab. Place the cells back into a clean 24-well plate and observe and photograph under a microscope. Results are shown below. Figure 4 .
[0026] 2) Experimental results: such as Figure 4 As shown, in MDA-MB-231 and MDA-MB-468 cells, compared with the control group, the number of cells crossing the chamber membrane decreased after TGP treatment, indicating that TGP inhibited cell invasion.
[0027] Example 3
[0028] Total glucosides of paeony inhibit epithelial-mesenchymal transition (EMT) in triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. Western blotting assay to detect the effect of TGP on epithelial-mesenchymal transition-related proteins in MDA-MB-231 and MDA-MB-468 cells. 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. The cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. The culture medium was discarded, and the cells were slowly washed once with PBS. 100 μL of Western blotting and IP cell lysis buffer was added to each well of the 6-well plate, and the cells were lysed on ice for 10 min. Cells were scraped off with a cell scraper and collected in 1.5 ml EP tubes. The tubes were centrifuged at 15000 rpm for 10 min in a pre-chilled centrifuge at 4°C. The supernatant was carefully aspirated and transferred to EP tubes. Protein concentration was quantified according to the instructions of the BCA kit. The total protein concentration was set at 30 μg. The corresponding volume of 5× buffer was added to prepare the protein sample, which was then denatured by boiling in a metal bath at 100°C for 10 min. The samples were then aliquoted and stored at -80°C. Protein samples were separated by SDS-PAGE electrophoresis, transferred to PVDF membranes, and blocked on a shaker. The membranes were incubated overnight at 4°C with protein-specific primary antibodies, washed, and then incubated with anti-rabbit or anti-mouse secondary antibodies at room temperature for 1 hour. After washing, the membranes were developed with ECL working solution and photographed. GAPDH was used as an internal control. Results are shown in the figure below. Figure 5 .
[0029] 2) Experimental results: such as Figure 5 As shown, in MDA-MB-231 and MDA-MB-468 cells, compared with the control group, TGP treatment downregulated the expression level of the mesenchymal-like protein Vimentin and upregulated the expression of the epithelial-like proteins E-cadherin and Occludin. The results suggest that TGP can inhibit epithelial-mesenchymal transition.
[0030] Example 4
[0031] Paeonia lactiflora total glucosides induced apoptosis in triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. The TUNEL / DAPI apoptosis detection kit was used to detect TGP-induced apoptosis in MDA-MB-231 and MDA-MB-468 cells.
[0032] 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. The cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. After a single wash with PBS, the cells were fixed with 4% paraformaldehyde for 30 minutes, washed once with PBS, permeabilized with 0.3% Triton X-100 PBS, incubated at room temperature for 5 minutes, washed twice with PBS, and covered with TUNEL assay solution (TdT enzyme: fluorescent labeling solution = 1:50). The cells were incubated at 37°C in the dark for 1 hour, followed by three washes with PBS for 5 minutes each. The cells were counterstained with DAPI solution and incubated at room temperature in the dark for 5 minutes. Cell fluorescence was photographed under a fluorescence microscope using red and blue light, and the images were collected. The results are shown in the figure below. Figure 6 .
[0033] 2) Experimental results: such as Figure 6 As shown, the nuclei of viable cells stained blue by DAPI, while the nuclei of TUNEL-positive cells are stained green. The number of TUNEL-positive cells can reflect the degree of apoptosis. With increasing TGP concentration, the number of TUNEL-positive cells increases, and TGP can induce apoptosis in MDA-MB-231 and MDA-MB-468 cells in a dose-dependent manner.
[0034] 2. Flow cytometry analysis of TGP-induced apoptosis in MDA-MB-231 and MDA-MB-468 cells. 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. Cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. Cells were digested with EDTA-free trypsin, centrifuged, resuspended in 1× Binding Buffer, stained with Annexin V-FITC and PI, gently vortexed, and incubated at room temperature in the dark for 5 minutes. The number of apoptotic cells was statistically analyzed using flow cytometry. Results are shown below. Figure 7 .
[0035] 2) Experimental results: such as Figure 7 As shown, TGP can induce apoptosis in MDA-MB-231 and MDA-MB-468 cells, and the number of apoptotic cells increases with increasing TGP concentration. In MDA-MB-231 cells, the number of apoptotic cells changed from 20.82% to 40.74%, and in MDA-MB-468 cells, the number of apoptotic cells changed from 4.28% to 37.15%.
[0036] 3. Western blotting assay to detect the effect of TGP on apoptosis-related proteins in MDA-MB-231 and MDA-MB-468 cells. 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. The cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. The culture medium was discarded, and the cells were slowly washed once with PBS. 100 μL of Western blotting and IP cell lysis buffer was added to each well of the 6-well plate, and the cells were lysed on ice for 10 min. Cells were scraped off with a cell scraper and collected in 1.5 ml EP tubes. The tubes were centrifuged at 15000 rpm for 10 min in a pre-chilled centrifuge at 4°C. The supernatant was carefully aspirated and transferred to EP tubes. Protein concentration was quantified according to the instructions of the BCA kit. The total protein concentration was set at 30 μg. The corresponding volume of 5× buffer was added to prepare the protein sample, which was then denatured by boiling in a metal bath at 100°C for 10 min. The samples were then aliquoted and stored at -80°C. Protein samples were separated by SDS-PAGE electrophoresis, transferred to PVDF membranes, and blocked on a shaker. The membranes were incubated overnight at 4°C with protein-specific primary antibodies, washed, and then incubated with anti-rabbit or anti-mouse secondary antibodies at room temperature for 1 hour. After washing, the membranes were developed with ECL working solution and photographed. GAPDH was used as an internal control. Results are shown in the figure below. Figure 8 .
[0037] 2) Experimental results: such as Figure 8 As shown, in MDA-MB-231 and MDA-MB-468 cells, compared with the control group, TGP treatment downregulated the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-xl, upregulated the expression level of the pro-apoptotic protein BAX, and gradually upregulated the expression levels of apoptosis-related proteins cleaved-caspase3, cleaved-caspase9, and cleaved-PARP. These results suggest that TGP can induce apoptosis in cells.
[0038] Example 5
[0039] Paeonia lactiflora total glucosides induce mitochondrial damage in triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. Mitro-Track staining to detect TGP-induced mitochondrial damage in MDA-MB-231 and MDA-MB-468 cells.
[0040] 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. Cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. Mitro-Track detection solution was added to cover the cells, and the plates were incubated at 37°C in the dark for 30 minutes. Cells were washed three times with PBS solution for 5 minutes each time. Cell fluorescence was observed under a fluorescence microscope, and images were collected. Results are shown in the figure below. Figure 9 .
[0041] 2) Experimental results: such as Figure 9 As shown, Mitro-Track stains normal mitochondria red. With increasing TGP concentration, the mitochondria change from a normal filamentous structure to a sheet-like structure, and the fluorescence weakens. The results suggest that TGP can induce mitochondrial damage in MDA-MB-231 and MDA-MB-468 cells in a dose-dependent manner.
[0042] 2. Effect of JC-1 staining on the effect of TGP on mitochondrial membrane potential changes in MDA-MB-231 and MDA-MB-468 cells. 1) Experimental Protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. The cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. The culture medium was discarded, and the cells were rinsed once with PBS. 1 mL of culture medium was added, followed by 1 mL of JC-1 working solution. The mixture was gently agitated and incubated at 37°C for 20 minutes. After incubation, the cells were washed twice with 1×JC-1 staining buffer. 1 mL of culture medium was added to each well, and the cells were collected. The fluorescence intensity of the cells was detected using flow cytometry. The results are shown in the figure below. Figure 10 .
[0043] 2) Experimental results: such as Figure 10 As shown, in MDA-MB-231 and MDA-MB-468 cells, with increasing TGP concentration, JC-1 monomers (green fluorescence) increased, multimers (red fluorescence) decreased, and mitochondrial membrane potential decreased, indicating damage. The results suggest that TGP can cause mitochondrial damage in MDA-MB-231 and MDA-MB-468 cells.
[0044] Example 6
[0045] Paeonia lactiflora total glucosides inhibit oxidative phosphorylation in triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468). 1. RT-qPCR detection of TGP inhibiting the expression of mitochondrial respiratory chain enzymes in MDA-MB-231 and MDA-MB-468 cells.
[0046] 1) Experimental protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. Cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. Cells were then collected for RT-qPCR detection. Results are shown in the figure below. Figure 11 .
[0047] 2) Experimental results: such as Figure 11 As shown, TGP inhibited the levels of mitochondrial respiratory chain complex enzymes in MDA-MB-231 and MDA-MB-468 cells, including inhibiting the expression of complex I (NDUFA9 and NDUFB8), complex II (SDHB and SDHA), complex III (ATP5A1), complex IV (COX4), and complex V (UQCRC1). These results suggest that TGP inhibits oxidative phosphorylation in MDA-MB-231 and MDA-MB-468 cells.
[0048] 2. ELISA was used to detect the NAD+ / NADH levels in TGP-inhibited MDA-MB-231 and MDA-MB-468 cells. 1) Experimental protocol: Triple-negative breast cancer cells (MDA-MB-231 and MDA-MB-468) in logarithmic growth phase were seeded into 6-well plates and cultured in a cell culture incubator for 24 hours to allow them to adhere. Cells were then treated with TGP at final concentrations (0, 0.2, 0.4, and 0.8 mg / ml) for 48 hours. Cells were then collected for ELISA detection. Results are shown in the figure below. Figure 12 .
[0049] 2) Experimental results: such as Figure 12 As shown, TGP can inhibit NAD+ / NADH in MDA-MB-231 and MDA-MB-468 cells. These results suggest that TGP inhibits oxidative phosphorylation in MDA-MB-231 and MDA-MB-468 cells.
[0050] Example 7
[0051] Effects of total glucosides of paeony on tumor cell growth in nude mice with MDA-MB-231 cell xenografts 1) Experimental protocol: MDA-MB-231 cells in logarithmic growth phase were used, and the concentration was adjusted to 2×10⁻⁶ cells / mL using serum-free medium. 7 MDA-MB-231 cells were injected subcutaneously into BALB / c nude mice to induce tumor growth (0.1 ml / mL). Once the tumor reached approximately 100 mm³, mice were randomly divided into two groups: a control group and a paeoniflorin-treated group (n=5 per group). The treatment group received TGP (500 mg / kg) via gavage twice a week, while the control group received the same volume of physiological saline. Tumor volume was measured twice weekly. After three weeks of treatment, the mice were sacrificed, the tumor nodules were dissected, and weighed. The tumor volume was calculated using the formula: V = 1 / 2 × a × b², where a and b represent length and width, respectively. See the results figure below. Figure 13 .
[0052] 2) Experimental results: such as Figure 13 As shown in Figure b: Tumor volume growth curve in nude mice. Figure c: Macroscopic view of the xenograft at the experimental endpoint. Figure d: Tumor weight at the experimental endpoint (n = 5). The results indicate that TGP administration can inhibit the growth of triple-negative breast cancer tumors in nude mice.
[0053] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. Use of total paeony glycosides in the preparation of a drug for treating triple-negative breast cancer.
2. Use according to claim 1, wherein The total paeony glycosides can inhibit the proliferation of triple-negative breast cancer cells.
3. The use according to claim 1, wherein The total paeony glycosides can inhibit the migration and invasion of triple-negative breast cancer cells.
4. The use according to claim 1, wherein The total paeony glycosides can inhibit the epithelial-mesenchymal transition of triple-negative breast cancer cells.
5. The use according to claim 1, wherein the compound is ###0002### The total paeony glycosides can induce apoptosis of triple-negative breast cancer cells.
6. The use according to claim 1, wherein The total paeony glycosides can induce mitochondrial damage of triple-negative breast cancer cells.
7. The use according to claim 1, wherein The total paeony glycosides can inhibit oxidative phosphorylation of triple-negative breast cancer cells.
8. The use according to claim 1, wherein The total paeony glycosides comprise a therapeutically effective amount of total paeony glycosides and a pharmaceutically acceptable carrier.
9. Use according to claim 8, wherein the compound is ###0002### The dosage form of the total paeony glycosides is oral preparation or injection.