Use of traditional Chinese medicine monomer juglone in preparation of medicine for treating female cancer

Abstract

The application discloses application of a traditional Chinese medicine monomer, carya cathayensis, in preparation of a medicine for treating female cancers, relates to the technical field of biological medicine, and the female cancers include cervical cancer, endometrial cancer and ovarian cancer. The application provides an effective alternative for treatment of female cancers. The carya cathayensis has obvious inhibiting effects on the proliferation ability and migration ability of cervical cancer cells, endometrial cancer cells and ovarian cancer cells from different sources, and in addition, the carya cathayensis can promote cell cycle arrest. The carya cathayensis can inhibit the malignant biological behaviors of the above female cancer cells by down-regulating the expression level of an apoptosis-related protein Bcl-2, up-regulating Bax, Cleaved caspase 9 and Cleaved caspase 3.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of the traditional Chinese medicine monomer kaempferol in the preparation of drugs for treating female cancers. Background Technology

[0002] Among female cancers, cervical cancer, endometrial cancer, and ovarian cancer are three common malignant tumors of the reproductive system, each with different pathogenesis, clinical characteristics, and prevention and treatment strategies. Cervical cancer is closely related to persistent infection with high-risk human papillomavirus (HPV) and is currently the only gynecological cancer with a clearly defined cause that is preventable and controllable. Effective prevention can be achieved through regular cervical liquid-based cytology (TCT) testing, HPV screening, and HPV vaccination. Early detection leads to a high cure rate after surgery or radiotherapy / chemotherapy. Endometrial cancer is often associated with long-term abnormally high estrogen levels (such as anovulation, obesity, diabetes, hypertension, or long-term use of estrogen alone). Typical symptoms include irregular vaginal bleeding or discharge around menopause. This noticeable sign aids in early detection. Diagnosis can be made by ultrasound examination of endometrial thickness combined with hysteroscopic biopsy. Since most cases are in the early stages, the overall prognosis is good. Ovarian cancer, due to the deep location of both ovaries in the pelvic cavity and their relatively small size, often lacks specific symptoms in its early stages. It frequently presents with easily overlooked atypical symptoms such as abdominal distension, abdominal discomfort, and decreased appetite. Furthermore, the lack of efficient and convenient early screening methods means that approximately 70% of patients are diagnosed at an advanced stage. Therefore, developing highly effective and low-toxicity drugs that can simultaneously target multiple types of female cancers is of significant clinical importance.

[0003] In recent years, innovations in separation, purification, and structural analysis techniques have propelled the development of natural medicines. Plant-derived active ingredients, due to their unique chemical diversity and multi-pathway regulatory capabilities, have become key target resources for anti-tumor drug screening. Currently, there are no reports on the use of pecan extract in the treatment of female cancers (including cervical cancer, endometrial cancer, and ovarian cancer). Summary of the Invention

[0004] The purpose of this invention is to provide the application of the traditional Chinese medicine monomer caryatin in the preparation of medicaments for treating female cancers, including cervical cancer, endometrial cancer, and ovarian cancer.

[0005] The molecular structural formula of pecan extract described in this invention is as follows:

[0006]

[0007] In a first aspect, the invention provides the use of pecanin in the preparation of a medicament for treating female cancers, preferably cervical cancer, endometrial cancer, or ovarian cancer.

[0008] Furthermore, in the aforementioned applications, pecanin inhibits the proliferation, migration, and cell cycle arrest of cervical cancer cells, endometrial cancer cells, or ovarian cancer cells.

[0009] Furthermore, in the aforementioned applications, pecan extract downregulates the expression levels of apoptosis-related proteins Bcl-2 and upregulates the expression levels of Bax, Cleaved caspase 9, and Cleaved caspase 3. These effects are applicable to cervical cancer, endometrial cancer, and ovarian cancer cells.

[0010] This invention also provides the application of pecanin in the preparation of combination drugs, wherein the combination drug is pecanin combined with at least one marketed female cancer treatment drug, and the combination preparation is used to prepare drugs for the prevention and / or treatment of female cancers; wherein the female cancers include at least one of breast cancer, ovarian cancer, cervical cancer, endometrial cancer, fallopian tube cancer, and vaginal cancer.

[0011] The marketed female cancer treatment drugs are selected from one or more of the following clinically approved chemotherapy drugs, targeted drugs, endocrine therapy drugs, and immunotherapy drugs; among them, chemotherapy drugs include marketed chemotherapy preparations such as paclitaxel, platinum, anthracyclines, and vincristine alkaloids; endocrine drugs include marketed preparations such as aromatase inhibitors, selective estrogen receptor modulators, and gonadotropin-releasing hormone agonists; targeted drugs include approved targeted preparations such as HER2 inhibitors, PARP inhibitors, and CDK4 / 6 inhibitors; and immunotherapy drugs are marketed anti-tumor drugs of the PD-1 / PD-L1 inhibitor class.

[0012] The combined use method is as follows: pecan extract and marketed drugs can be prepared into formulations independently and administered simultaneously or sequentially; the two can be packaged separately into independent drug formulations to form a compound drug box, or they can be combined to prepare a single compound formulation.

[0013] The beneficial effects of this invention are:

[0014] 1. This invention provides an effective treatment option for female cancers (especially cervical cancer, endometrial cancer, and ovarian cancer). Hickory extract significantly inhibits the proliferation and migration of cervical cancer cells, endometrial cancer cells, and ovarian cancer cells from different sources. Furthermore, hoecrin can promote cell cycle arrest. Hickory extract may inhibit the malignant biological behavior of these female cancer cells by downregulating the expression levels of apoptosis-related proteins Bcl-2 and upregulating the expression levels of Bax, Cleaved caspase 9, and Cleaved caspase 3.

[0015] 2. This invention uses natural products, making it safe to use. Attached Figure Description

[0016] Figure 1 The effect of pecan extract on the proliferative capacity of cervical cancer, endometrial cancer, and ovarian cancer cells;

[0017] Figure 2 The effect of pecan extract on the migration ability of cervical cancer cells (scratch assay).

[0018] Figure 3 The effect of pecan extract on the migration ability of endometrial cancer cells (scratch assay).

[0019] Figure 4 The effect of pecan extract on the migration ability of ovarian cancer cells (scratch assay).

[0020] Figure 5 The effect of pecan extract on the migration ability of cervical cancer cells (Transwell migration assay).

[0021] Figure 6 The effect of pecan extract on the migration ability of endometrial cancer cells (Transwell migration assay).

[0022] Figure 7 The effect of pecan extract on the migration ability of ovarian cancer cells (Transwell migration assay).

[0023] Figure 8 The effect of pecan extract on cell cycle arrest in cervical cancer cells;

[0024] Figure 9 The effect of pecan extract on cycle arrest in endometrial cancer cells;

[0025] Figure 10 The effect of pecan extract on cycle arrest in ovarian cancer cells;

[0026] Figure 11 The effect of pecan extract on the expression of Bax and Bcl-2 proteins in cervical cancer cells;

[0027] Figure 12 Effects of pecan extract on the expression of Bax and Bcl-2 proteins in endometrial cancer cells;

[0028] Figure 13 Effects of pecan extract on the expression of Bax and Bcl-2 proteins in ovarian cancer cells;

[0029] Figure 14 Effects of pecan extract on the expression of Caspase 9 and Caspase 3 proteins in cervical cancer cells;

[0030] Figure 15 Effects of pecan extract on the expression of Caspase 9 and Caspase 3 proteins in endometrial cancer cells;

[0031] Figure 16 Effects of pecan extract on the expression of Caspase 9 and Caspase 3 proteins in ovarian cancer cells; Detailed Implementation

[0032] The pecanin described in this invention is a known compound, and its structure and acquisition method have been reported in the literature (Fel B, Baudouin A, Fache F, et al. Caryatin and 3'-O-methylcaryatin contents inedible yams (Dioscorea spp.)[J]. Journal of Food Composition and Analysis, 2021, 102: 104010.). Those skilled in the art can obtain the pecanin used in this invention based on the literature or conventional methods in the field.

[0033] Example 1

[0034] Phenotypic studies of caryatin in the treatment of cervical cancer, endometrial cancer, and ovarian cancer cells:

[0035] 1.1 Effects of pecan extract on cell proliferation:

[0036] To investigate the effects of kaempferol on the proliferation of cervical cancer, endometrial cancer, and ovarian cancer cells, we selected two cervical cancer cell lines (SiHa, HeLa), two endometrial cancer cell lines (HEC-1-A, Ishikawa), and two ovarian cancer cell lines (A2780, Ovcar-3) as research subjects. These cells were treated with different concentrations (25 μM, 50 μM, 75 μM, 100 μM, 150 μM, 200 μM, 400 μM) of kaempferol for 24 h, 48 h, and 72 h, with 0.1% DMSO as a control. CCK8 results showed that kaempferol significantly inhibited the proliferation of cervical cancer, endometrial cancer, and ovarian cancer cells in a concentration-dependent manner. Figure 1 ).

[0037] 1.2 Effects of pecan extract on the migration ability of cervical cancer cells:

[0038] Cervical cancer cells (SiHa, HeLa), endometrial cancer cells (HEC-1-A, Ishikawa), and ovarian cancer cells (A2780, Ovcar-3) in logarithmic growth phase were seeded into 6-well plates for scratch assays. Scratch widths were recorded at 0 h and 48 h after drug administration. Results are as follows: Figures 2-4As shown in the figure, Cary-L, Cary-M, and Cary-H represent low, medium, and high concentrations of kaempferol, respectively, and pos represents the positive control group. Kaempferol inhibits scratch healing in a concentration-dependent manner. As the concentration of the drug increases, the scratch healing rate gradually decreases, indicating that kaempferol can effectively inhibit the migration ability of the above three types of cancer cells. Specifically, compared with their respective control groups: in cervical cancer cells, kaempferol (25 µM, 50 µM, 100 µM) and the positive control cisplatin (CDDP: 6.25 µM) significantly reduced cell migration; in endometrial cancer cells, kaempferol (50 µM, 100 µM, 150 µM) and cisplatin (6.25 µM) also significantly inhibited migration; in ovarian cancer cells, kaempferol (A2780: 50 µM, 100 µM, 150 µM; Ovcar-3: 25 µM, 50 µM, 100 µM) and the positive control paclitaxel (PTX: 100 nM) also significantly reduced cell migration.

[0039] Transwell experimental results are as follows: Figures 5-7 As shown, kaempferol significantly reduced the number of cells that penetrated the cell membrane in each group in a concentration-dependent manner. Specifically, compared with their respective control groups: in cervical cancer cells (SiHa, HeLa), treatment with kaempferol (25µM, 50µM, 100µM) and the positive control cisplatin (6.25µM) significantly reduced the number of cells that penetrated the ventricular membrane; in endometrial cancer cells (HEC-1-A, Ishikawa), kaempferol (50µM, 100µM, 150µM) and cisplatin (6.25µM) also significantly reduced the number of cells that penetrated the membrane; in ovarian cancer cells (A2780, Ovcar-3), kaempferol (A2780: 50µM, 100µM, 150µM; Ovcar-3: 25µM, 50µM, 100µM) and the positive control paclitaxel (100 nM) also significantly inhibited the cell's ability to penetrate the membrane. The above results indicate that pecan extract can effectively inhibit the migration ability of cervical cancer, endometrial cancer, and ovarian cancer cells.

[0040] 1.3 Effects of pecan extract on cell cycle arrest in cervical cancer cells:

[0041] Cervical cancer cells (SiHa and HeLa), endometrial cancer cells (HEC-1-A and Ishikawa), and ovarian cancer cells (A2780 and Ovcar-3) in the synaptic growth phase were seeded in 6-well plates. After 24 h, the culture medium was replaced with the drug-treated medium. After 48 h, the cells were digested with trypsin, and pre-chilled 75% ethanol was added. The samples were fixed at -20°C for 12 h. Propidium iodide (PI) staining solution was prepared, and 0.5 mL of PI staining solution was added to each tube of cells. The cells were resuspended and incubated at room temperature in the dark for 20 min before analysis. Figures 8-10 As shown, compared with the control group, the number of cells in the G0 / G1 phase increased in the treatment group, while the proportion of cells in the S phase and G2 / M phase decreased. This observation suggests that the ability of pecan extract to inhibit the growth of cervical cancer, endometrial cancer, and ovarian cancer cells may be related to cell cycle disruption.

[0042] Example 2

[0043] Molecular mechanism experiments of caryatin in the treatment of cervical cancer, endometrial cancer, and ovarian cancer cells:

[0044] 2.1 Effects of pecan extract on the expression of apoptosis marker proteins Bax and Bcl-2 in cervical cancer, endometrial cancer, and ovarian cancer cells;

[0045] Bax and Bcl-2 are two key molecules with opposite functions in the Bcl-2 protein family, jointly regulating the endogenous mitochondrial apoptosis pathway in cells. Bcl-2 is the main anti-apoptotic protein, protecting cell survival by stabilizing the mitochondrial membrane and inhibiting the release of apoptotic factors such as cytochrome C; while Bax is a pro-apoptotic protein, translocating to the mitochondrial membrane and forming pores after receiving death signals, prompting the release of apoptotic factors and initiating the apoptosis cascade. Whether a cell undergoes apoptosis largely depends on the expression ratio of Bcl-2 to Bax: an increased Bcl-2 / Bax ratio favors cell survival, while a decreased ratio promotes apoptosis. In the evaluation of antitumor drugs, if a candidate drug can downregulate Bcl-2, upregulate Bax, and reduce the Bcl-2 / Bax ratio, it can serve as important molecular evidence that it induces tumor cell apoptosis through the mitochondrial pathway. Protein expression results are as follows: Figures 11-13 As shown, with the increase of pecan concentration, Bax expression was upregulated and Bcl-2 expression was downregulated, demonstrating that pecan can promote apoptosis of cervical cancer, endometrial cancer and ovarian cancer cells by acting on the Bcl-2 / Bax pathway.

[0046] 2.2 Effects of pecan extract on the expression of apoptosis marker proteins Caspase 9 and Caspase 3 in cervical cancer, endometrial cancer, and ovarian cancer cells;

[0047] Caspase 9 and Caspase 3 are two crucial executor molecules in the apoptosis pathway, belonging to the initiator caspase and effector caspase, respectively. Caspase 9 is the initiator of the endogenous mitochondrial apoptosis pathway. When mitochondria release cytochrome c, caspase 9 binds to the apoptosis complex and is cleaved and activated, forming an enzymatically active cleaved caspase 9, which then transmits death signals downstream. Activated caspase 9 subsequently cleaves and activates the effector caspase, caspase 3, converting it from its inactive prozymogen form into its activated cleaved caspase 3. Activated caspase 3, as the final executor of apoptosis, can cleave various cellular structural proteins and DNA repair enzymes, ultimately leading to cytoskeleton disintegration, DNA breakage, and apoptosis. In antitumor drug research, elevated expression levels of cleaved caspase 9 and cleaved caspase 3 are key molecular evidence demonstrating that drugs induce tumor cell death through the endogenous apoptosis pathway. Protein expression results are as follows: Figures 14-16 As shown, with increasing pecan concentration, the expression of cleaved Caspase 9 and cleaved Caspase 3 was upregulated, demonstrating that pecan can promote apoptosis in cervical cancer, endometrial cancer, and ovarian cancer cells by acting on cleaved Caspase 9 and cleaved Caspase 3.

[0048] In summary, the traditional Chinese medicine compounds selected in this invention have significant advantages in terms of efficacy, and therefore can be used to prepare drugs for the treatment of cervical cancer, endometrial cancer, and ovarian cancer.

Claims

1. The application of a traditional Chinese medicine monomer, kaempferol, in the preparation of a drug for treating female cancer.

2. The application of the traditional Chinese medicine monomer kaempferol according to claim 1 in the preparation of a drug for treating female cancer, characterized in that, The cancers mentioned are cervical cancer, endometrial cancer, or ovarian cancer.

3. The application of the traditional Chinese medicine monomer kaempferol according to claim 1 in the preparation of a drug for treating female cancer, characterized in that, Hickory extract inhibits the proliferation, migration, and cell cycle arrest of cervical cancer cells, endometrial cancer cells, or ovarian cancer cells.

4. The application of the traditional Chinese medicine monomer kaempferol according to claim 1 in the preparation of a drug for treating female cancer, characterized in that, Hickory extract downregulates the expression levels of apoptosis-related proteins Bcl-2 and upregulates the expression levels of Bax, Cleaved caspase 9, and Cleaved caspase 3.

5. The application of the traditional Chinese medicine monomer kaempferol according to claim 1 in the preparation of a drug for treating female cancer, characterized in that, The drug is either a single-agent drug or a combination drug of pecan extract; the combination drug is composed of pecan extract combined with one or more clinically marketed drugs for the treatment of female cancers.

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