Application of cimicifugin as targeted HTR1E agonist and in preparation of ovarian cancer drugs

Abstract

The invention discloses application of cimicifugin serving as a targeted HTR1E agonist in preparation of ovarian cancer drugs. The invention discloses application of cimicifugin in preparation of a medicine for treating ovarian cancer. The cimicifugin is used as a targeted HTR1E agonist. The cimicifugin inhibits an SRC downstream signal channel by activating HTR1E. The application comprises a combination of cimicifugin and any one or more of the following (1)-(4): (1) chemotherapeutic drugs; (2) a targeted drug; (3) an anti-angiogenesis drug; and (4) hormone drugs.

Description

Technical Field

[0001] The present invention relates to the pharmaceutical use of a compound, in particular to its application in the preparation of drugs for ovarian cancer. Background Art

[0002] Ovarian cancer is one of the female reproductive system tumors with the highest fatality rate. Since 2005, ovarian cancer has become the second leading cause of death among female reproductive system tumors in China after cervical cancer. The destruction of the functions of important organs caused by tumor metastasis is often the primary cause of death of patients. Due to the insidious onset, about 2 / 3 of ovarian cancer patients have peritoneal metastasis at the time of diagnosis, resulting in a very low 5-year survival rate. The 5-year survival rates of patients in stage III and stage IV are only 27% and 13% respectively. The combination of platinum drugs and paclitaxel is still the first choice for first-line intervention in ovarian cancer. However, this treatment plan still has traditional problems such as low tumor specificity, easy generation of drug resistance, and large side effects. Research shows that during the entire treatment process of ovarian cancer, the incidence of depression and anxiety in patients is significantly higher than that in healthy women. The incidence of ovarian cancer patients suffering from depression or anxiety at the same time is as high as 90%, far exceeding the incidence of these two mental stress diseases in non-diseased women. Moreover, the disease stage of ovarian cancer patients in a depressive state is often worse than that of ovarian cancer patients in a non-depressive state. Women suffering from depression have a 1.3-fold higher risk of developing ovarian cancer than non-depressed women. These all suggest a close connection between mental stress and the progression of ovarian cancer. There is an urgent need to break through the research on the molecular mechanism of peritoneal metastasis of ovarian cancer and the targeted intervention means developed based on this.

[0003] As a direct effector of GPCR, SRC protein participates in the signal pathways related to cell survival, proliferation, and migration mediated by the Gα subunit in membrane receptors. It can phosphorylate focal adhesion kinase (FAK) in epithelial cells under the induction of calpain, causing it to lose its docking function and thus inhibiting the downstream signal pathway; at the same time, it can also phosphorylate E-cadherin and β-catenin in epithelial cells, resulting in the weakening of cell-cell connections, and thus making cell migration and cell invasion more likely to occur. SRC protein can also promote cell proliferation by phosphorylating the Y925 site of FAK or binding to IL-3R to activate the Ras-MAPK signal pathway. Therefore, inhibiting the signal pathway mediated by SRC is an important strategy for tumor treatment.

[0004] The HTR1E belongs to the G protein-coupled receptor family and is the fourth member of the 5-HT receptor 1 subfamily (HT1R) discovered in 1989. Research has shown that after 5-HT binds to HTR1E, it inhibits the phosphorylation of SRC protein, thereby inactivating the FAK, PI3K-AKT, and MEK-ERK signaling pathways downstream of SRC, and further inhibiting the proliferation, migration, and EMT process of ovarian cancer cells; moreover, it has also been proven in a mouse orthotopic transplantation tumor model of ovarian cancer that the activation of HTR1E inhibits the formation of orthotopic tumors, intestinal metastases, and bloody ascites in mouse ovarian cancer. Through experimental verification, if the HTR1E / 1F agonist BRL54443 or the SRC inhibitor Dasatinib is given, the inhibitory effect on its downstream pro-cancer pathways can be partially restored. However, due to the complex functions of the 5-HT receptor family, the 5-HT-mediated signaling pathways can promote or inhibit the progression of various tumors. Therefore, developing small molecule drugs specifically targeting HTR1E is an important strategy and direction for ovarian cancer treatment. However, there is currently no report on small molecule drugs specifically targeting HTR1E for tumor treatment.

[0005] Prim-O-glucosylcimifugin (POG, CAS: 80681-45-4) is a chromone extracted from the traditional Chinese medicine Radix Saposhnikovia, with the molecular formula C 22 H 28 O 11 . It has been used for a long time in traditional Chinese medicine treatment for fevers, rheumatism, etc., and previous reports have mostly focused on its anti-inflammatory effects. It also has potential value in cancer treatment. POG can reverse cisplatin resistance in lung cancer cells by downregulating the expression of the p65 subunit of NF-κB. POG can inhibit the proliferation, metabolism, and immunosuppression of polymorphonuclear myeloid-derived suppressor cells, thereby increasing the infiltration of CD8 + T cells in tumors. POG can also enhance the anti-tumor effect of PD-1 inhibitors. POG can also induce the degradation of β-tubulin and downregulate the level of phosphorylated CDK1, resulting in apoptosis of acute lymphoblastic leukemia cells and cell cycle arrest in the G2 / M phase. However, the target protein directly acted on by POG has not been identified yet, and there is no research report on the treatment of ovarian cancer with POG. The exploration of POG in ovarian cancer treatment in this invention may become another innovative discovery in the treasure trove of traditional Chinese medicine. Based on this, finding a drug compound with a targeting HTR1E agonist and then providing its application in the preparation of ovarian cancer drugs has become an urgent problem to be solved in the existing technology. Summary of the Invention

[0006] To solve the above problems existing in the prior art, we discovered the effect of Prim-O-glucosylcimifugin as a targeting HTR1E agonist in our research. Based on this,

[0007] The technical solution provided by this invention is as follows:

[0008] Application of cimifugin in the preparation of a drug for treating ovarian cancer.

[0009] The application of cimifugin in the preparation of a drug for treating ovarian cancer. Further, the cimifugin is used as a targeting HTR1E agonist.

[0010] The application of cimifugin in the preparation of a drug for treating ovarian cancer. Further, the application includes the combination of cimifugin and any one or more of the following drugs (1) to (4):

[0011] (1) Chemotherapy drugs

[0012] (2) Targeted drugs

[0013] (3) Anti-angiogenic drugs

[0014] (4) Hormone drugs.

[0015] The application of cimifugin in the preparation of a drug for treating ovarian cancer. Further, the chemotherapy drugs are selected from taxanes and platinum-based drugs; the targeted drugs are selected from PARP inhibitors and anti-angiogenic drugs;

[0016] The application of cimifugin in the preparation of a drug for treating ovarian cancer. Further, the platinum-based chemotherapy drugs are selected from carboplatin, cisplatin, and oxaliplatin; the taxane chemotherapy drugs are selected from paclitaxel, docetaxel, and cabazitaxel; the PARP inhibitors are selected from olaparib and niraparib, the anti-angiogenic drugs are selected from bevacizumab; the hormone drugs are selected from tamoxifen and letrozole.

[0017] The present invention also provides a pharmaceutical composition, which is composed of an active ingredient and at least one pharmaceutically acceptable excipient, and the active ingredient is composed of cimifugin and any one or more of the following drugs (1) to (4):

[0018] 1) Chemotherapy drugs

[0019] (2) Targeted drugs

[0020] (3) Anti-angiogenic drugs

[0021] (4) Hormone drugs.

[0022] For the pharmaceutical composition, further, the chemotherapy drugs are selected from taxanes and platinum-based drugs; the targeted drugs are selected from PARP inhibitors and anti-angiogenic drugs;

[0023] For the described drug combination, further, the platinum-based chemotherapeutic drug is selected from carboplatin, cisplatin, and oxaliplatin; the taxane-based chemotherapeutic drug is selected from paclitaxel, docetaxel, and cabazitaxel; the PARP inhibitor is selected from olaparib and niraparib, the anti-angiogenic drug is selected from bevacizumab; the hormonal drug is selected from tamoxifen and letrozole.

[0024] The present invention also provides the use of prim-O-glucosylcimifugin as a targeting HTR1E agonist, which inhibits the SRC downstream signaling pathway by activating HTR1E.

[0025] The beneficial effects of the present invention are as follows: The technical solution of this patent designs and screens a specific targeting agonist for the novel ovarian cancer drug target HTR1E, and it is first discovered that prim-O-glucosylcimifugin can specifically activate HTR1E and inhibit the pro-cancer signaling pathway mediated by the GPCR direct effector protein SRC. It overcomes the defect that the current HTR1E agonists 5-HT and BRL54443 lack 5-HT receptor specificity and may cause unpredictable side effects when directly applied to ovarian cancer. Based on this, the application of prim-O-glucosylcimifugin in the preparation of ovarian cancer drugs and the combined application of prim-O-glucosylcimifugin and other ovarian cancer treatment drugs are provided, and its exploration in ovarian cancer treatment may become another innovative discovery in the treasure house of traditional Chinese medicine. Description of the Drawings

[0026] Figure 1 It is the flow chart of the expression and purification of HTR1E;

[0027] In the figure, (A) The styrene-maleic anhydride polymer is hydrolyzed to styrene maleic polymer by NaOH; (B) The Western Blot experiment shows that the HeLa cell line stably overexpressing HTR1E-His is successfully established; (C) The purification flow chart of HTR1E; (D) The electron microscopy results show that the prepared Nanodisc has a diameter of about 10 nm; (E) The Western Blot experiment shows the purified HTR1E.

[0028] Figure 2 It is the schematic diagram of the binding activity of the purified HTR1E; In the figure, (A) The HPLC results show that compared with the control group, the detected 5-HT in the experimental group binding to HTR1E is reduced by half; (B) The comparison of the peak areas of the HPLC results in Figure A.

[0029] Figure 3 It is the schematic diagram of the primary screening process of the drug targeting HTR1E.

[0030] Figure 4 It is the electrophoresis and column gray scale analysis comparison diagram of the secondary screening results of the drug targeting HTR1E; In the figure, ns: p>0.05, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0031] Figure 5 Figure for comparing the effects of POG on the proliferation of ovarian cancer cells; in the figure, (A) the efficiency of stably knocking down HTR1E (shHTR1E) in SK-OV-3 cells by shRNA was detected by RT-qPCR experiment, and the shRNA targeting the bacterial lacZ gene (shlacz) was used as a control; (B) the proliferation curves of SK-OV-3 cells treated with different drugs (5 μM POG or BRL54443) or solvent control (DMSO); (C) the knockdown efficiency of HTR1E in OVCAR-8 cells was detected by RT-qPCR experiment; (D) the proliferation curves of OVCAR-8 cells treated with different drugs (5 μM POG or BRL54443) or DMSO; (E, F) representative pictures (E) and quantitative results (F) of colony formation of SK-OV-3 cells 14 days after treatment with different drugs; (G, H) representative pictures (G) and quantitative results (H) of colony formation of OVCAR-8 cells after treatment with different drugs; the statistical method for (B) and (D) was two-way ANOVA (means ± SEM, n = 3), ***p < 0.001, *p < 0.05, ns: p > 0.05; the statistical method for (F) and (H) was Student's t-test (means ± SEM, n = 3), ****p < 0.0001, **p < 0.01, ns: p > 0.05.

[0032] Figure 6 Figure for comparing the effects of POG on the migration of ovarian cancer cells; in the figure, (A) representative pictures of Transwell of SK-OV-3 cells treated with DMSO, 5 μM BRL54443, and 5 μM POG for 48 h, and the scale bar was 50 μm; (B) the quantitative results of Transwell (means ± SEM, n = 3), and the statistical method was Student's t-test, ***p < 0.001, **p < 0.01.

[0033] Figure 7Figure for comparing the effects of POG on inhibiting the in-situ growth and peritoneal dissemination of ovarian cancer in mice; in the figure, (A) Schematic diagram of the drug administration cycle for the mouse ovarian cancer model. Ten days after injecting SK-OV-3 cells into the ovarian bursa of mice, 50 mg / kg and 100 mg / kg of POG and normal saline in the control group were intraperitoneally administered respectively, and the mice were collected 46 days later; (B) Pictures of in-situ tumors of three groups of mice, and the black circles represent the mice that died during the experiment; (C) Statistical analysis of the weights of in-situ tumors of three groups of mice (means±SEM, n = 4-6), and the statistical method was Student’s t-test, **p<0.01, *p<0.05; (D) H&E staining of tumors, and the scale bar was 20 μm; (E, F) Pictures of ascites formation in the abdomen of mice (E) and statistical analysis of the ascites volume (F, means±SEM, n = 4-6), and the statistical method was Student’s t-test, ***p<0.001, *p<0.05; (G, H) Display (G) and quantity statistics (H, means±SEM, n = 4-6) of the metastatic foci of mice (marked by yellow arrows), and the statistical method was Student’s t-test, ***p<0.001, **p<0.01; (I) H&E staining of metastatic foci, and the scale bars were 20 μm and 50 μm.

[0034] Figure 8 Figure for comparing the effects of POG on not inhibiting the in-situ growth and peritoneal dissemination of mice in the shHTR1E group; in the figure, (A / B) Ten days after injecting SK-OV-3 shHTR1E cells into the ovarian bursa of mice, 100 mg / kg of POG and normal saline were intraperitoneally administered respectively, and the mice were collected 46 days later. The in-situ tumors were displayed and weighed, and the statistical method was Student’s t-test (means±SEM, n = 5), ns: p>0.05; (C) H&E staining of in-situ tumors, and the scale bar was 20 μm; (D / E) Display and volume statistics of ascites in mice, and the statistical method was Student’s t-test (means±SEM, n = 5), ns: p>0.05; F / G. Display and quantity statistics of metastatic foci of mice, and the statistical method was Student’s t-test (means±SEM, n = 5), ns: p>0.05.

[0035] Figure 9Comparison diagrams of the effects of POG on inhibiting SRC phosphorylation and its downstream signaling pathways; in the figure, (A) Western Blot results of proteins collected from SK-OV-3 cells in the shlacz group and shHTR1E group after treatment with 5 μM BRL54443, 5 μM POG, and control DMSO; (B) Verification of the inhibition of the SRC downstream signaling pathway by collecting proteins from SK-OV-3 cells in the shlacz group and shHTR1E group after treatment with 5 μM BRL54443, 5 μM POG, and control DMSO; (C) Immunofluorescence staining of in-situ tumor sections of mice in the normal saline group, 50 mg / kg POG group, and 100 mg / kg POG group; (D) Statistical results of Figure C (means ± SEM, n = 6), the statistical method is Student's t-test, ****p < 0.0001, *p < 0.05.

[0036] Figure 10 Comparison diagrams of the effects of POG on inhibiting cancer cell proliferation and the EMT process in a mouse ovarian cancer model; in the figure, (A) Immunohistochemical staining of in-situ tumor sections of mice in the normal saline group, 50 mg / kg POG group, and 100 mg / kg POG group, the scale bar is 50 μm; (B) Statistical results of Figure A (means ± SEM, n = 6), the statistical method is Student's t-test, ****p < 0.0001, **p < 0.01. Detailed implementation manners

[0037] The present invention will be further described below in conjunction with embodiments, but the present invention is not limited to the following embodiments.

[0038] Example 1 In vitro purification of active HTR1E

[0039] The specific steps / results are as follows: First, a stable cell line stably overexpressing HTR1E with His6-tag was constructed in HeLa cells, and the successful construction of the cell line was verified by Western Blot experiment ( Figure 1 B); then, HTR1E with a discoidal phospholipid membrane structure was prepared by the aforementioned method, and Ni-NTA was used to enrich Nanodisc-HTR1E with His6-tag ( Figure 1 C). The electron microscopy imaging results of Nanodisc are as shown in Figure 1 D, and its particle size is about 10 nm, which is consistent with the expectation. Finally, we collected the protein samples during the purification process and detected them by Western Blot experiment; as shown in Figure 1 E, it was verified that the cell membrane protein HTR1E was successfully obtained.

[0040] To verify that the purified HTR1E protein has binding activity, an experiment was conducted using the only natural ligand 5-HT in vivo. 1 μM of 5-HT was incubated with NandiscHTR1E immobilized on a nickel column and a blank nickel column for 25 min respectively. Then the supernatant containing 5-HT that did not bind to HTR1E or the empty nickel column was extracted, and the content of 5-HT in the supernatant was measured using a high-performance liquid chromatograph. The detected peak area results showed that compared with the control group (blank nickel column group), the 5-HT in the supernatant of the NanodiscHTR1E group decreased by approximately half ( Figure 2 A, B). It was inferred therefrom that the reduced part of 5-HT was bound to HTR1E. This result proved that the purified HTR1E protein has binding activity and can interact with its natural ligand 5-HT, enabling subsequent small molecule drug screening.

[0041] Example 2 Drug Screening Identified Prim-O-glucosylcimifugin as a Potential HTR1E Agonist

[0042] The specific steps / results are as follows: We grouped 160 1 μM small molecule compounds and incubated them with HTR1E immobilized on a nickel column, and added 10 μM of 5-HT to each group for competitive binding. The operation process was as Figure 3 . After removing the supernatant, that is, the drugs that did not bind to Nanodisc on the nickel column HTR1E , then protein denaturation was used to release the bound part of the small molecules for liquid chromatography-tandem mass spectrometry detection, and 16 small molecule compounds including Prim-O-glucosylcimifugin were identified as candidate drugs.

[0043] Screening based on affinity cannot determine whether a drug exerts an agonistic or inhibitory effect after binding to HTR1E. Therefore, the established HTR1E activation mechanism, that is, its activation can inhibit the phosphorylation of the downstream direct effector protein SRC, was further used for drug function screening. First, the drug molecules obtained previously by affinity screening were added to the SK-OV-3 cell line expressing HTR1E, and the concentration gradients were set as 1 μM, 3 μM, and 5 μM. At the same time, the HTR1E natural ligand 5-HT and the existing 5-HT receptor agonist BRL54443 were used as positive controls. Western Blot experiments were performed, and gray scale analysis was carried out on the exposure results. The electrophoresis and gray scale analysis were compared as Figure 4 . The statistical method was Student’s t-test, and the software for making column charts was graphpad.

[0044] Western Blot experiments showed that after adding drugs at different concentrations, the phosphorylation level of SRC in some drug-treated groups of cells changed significantly. Among them, the four small molecules with drug numbers 4A, 6D, 7F (i.e., POG), and 7H could significantly inhibit the phosphorylation of SRC in SK-OV-3 cells at multiple different concentrations (). This indicates that these small drug molecules may have the effect of activating HTR1E and then inhibiting the phosphorylation of its downstream molecule SRC.

[0045] To further analyze the binding ability of the candidate small molecules to HTR1E, we used Discovery Studio software to perform a simulated docking of the crystal structure (PDB: 3E33) of the above-screened drug molecules and HTR1E to evaluate the interaction of these drug molecules with HTR1E.

[0046] Considering that BRL54443 is an agonist of HTR1E / 1F, and its affinity for HTR1E (pKi = 8.2) is slightly stronger than that of the natural ligand 5-HT (pKi = 8.7) ( https: / / pdsp.unc.edu / pdspweb / ). Therefore, we used the docking score of the docking result of BRL54443 and HTR1E as a benchmark. The docking scores showed that the binding abilities of the two small drug molecules numbered 6D (score 54.1478) and 7F (score 61.5644) to HTR1E were stronger than that of BRL54443 (score 50.0832), which suggested that they might have stronger potential to activate HTR1E. Combining with the Western Blot results in the previous round of functional screening, 7F, that is, POG, was finally selected as the object for subsequent research.

[0047] Example 3: Cimifugin targets and activates HTR1E to inhibit the proliferation and migration of ovarian cancer cells

[0048] The specific steps / results are as follows: To study the effect of cimifugin on the malignant phenotype of ovarian cancer cells by targeting HTR1E, human ovarian cancer SK-OV-3 with knocked-down HTR1E was established ( Figure 5 A), and the cell proliferation curves of SK-OV-3 with non-knocked-down HTR1E (control group) and knocked-down HTR1E after treatment with 5 μM concentration of POG were measured respectively. The results showed that compared with BRL54443 at the same concentration, POG could more significantly inhibit cell proliferation ( Figure 5 B, shlacz group), while POG lost the effect of inhibiting proliferation on cells with knocked-down HTR1E ( Figure 5 B, shHTR1E group). This result was also replicated in OVCAR-8 cells, a type of HGSOC ovarian cancer cells ( Figure 5 C, D).

[0049] We further determined the effect of POG on the malignant proliferation ability of ovarian cancer cells through colony formation assays. Compared with BRL54443, POG exhibited stronger activity in inhibiting the colony formation of SK-OV-3 ( Figure 5 E, F), and this difference was more obvious in OVCAR-8 cells ( Figure 5 G, H). However, in ovarian cancer cells with HTR1E knockdown, POG lost its ability to inhibit colony formation. These results indicate that POG shows higher activity than BRL54443 in inhibiting cell malignant proliferation, and this activity depends on HTR1E.

[0050] To detect the effect of POG on the motility of ovarian cancer cells, we treated SK-OV-3 shlacz cells and SK-OV-3 shHTR1E cells with the drug for 48 h, and then evaluated the migration ability of these cells using the Transwell cell migration assay. The results showed that 9 h after cell perforation, compared with BRL54443, POG had a more obvious inhibitory effect on cell migration. This indicates that POG can inhibit the migration of ovarian cancer cells. ( Figure 6 )

[0051] Example 4

[0052] To verify the inhibitory effect of POG on ovarian cancer in vivo and its mechanism of action, we successfully established a NOD-SCID mouse orthotopic ovarian transplantation tumor model using SK-OV-3 shlacz cells and SK-OV-3 shHTR1E cells. Starting from the tenth day after tumor inoculation, we treated the mice with different drugs ( Figure 7 A). For the mice inoculated with the SK-OV-3 shlacz cell line, we divided them into three groups and administered normal saline, 50 mg / kg of POG, and 100 mg / kg of POG, respectively. Intraperitoneal administration was performed twice a week, and the mouse tumors were collected on the 46th day ( Figure 7 B), and the tumor weight ( Figure 7 C), ascites volume ( Figure 7 E, F), and the number of metastatic foci ( Figure 7 G, H) were counted. From the experimental results, POG had a significant inhibitory effect on the growth of orthotopic tumors and peritoneal dissemination of mouse ovarian cancer, and this inhibitory effect showed a dose-dependence. This finding is consistent with our results in vitro experiments, further demonstrating the potential application value of POG in the treatment of ovarian cancer.

[0053] Next, to verify whether the inhibitory effect of POG is achieved through HTR1E, we conducted similar experiments on mice inoculated with SK-OV-3 shHTR1E cells. These mice were divided into two groups and given normal saline and 100 mg / kg of POG, respectively. On the 46th day, although the tumors of some mice had not shown obvious peritoneal dissemination, the growth of the orthotopic tumors was significantly faster than that of the shacz group. And because the tumors were inoculated at the orthotopic ovarian site in the abdominal cavity, it was difficult to judge their size by external observation. After dissection, it was found that the size of the orthotopic tumors had approached or slightly exceeded the range required by animal ethics for experiments (explanation has been made to the ethics committee). By statistically analyzing the size of the orthotopic tumors ( Figure 8 A), the volume of ascites ( Figure 8 B), and the number of metastatic foci ( Figure 8 C), we found that there was almost no difference between the normal saline group and the POG group. This result indicates that POG cannot inhibit tumor growth when HTR1E is knocked down. This further proves that POG exerts its tumor-suppressive effect by targeting HTR1E.

[0054] Example 5 POG inhibits the SRC downstream signaling pathway by activating HTR1E

[0055] Previously, the laboratory had deeply explored the mechanism by which HTR1E inhibits the progression of ovarian cancer, that is, 5-HT / HTR1E effectively inhibits the processes of ovarian cancer cell proliferation and epithelial-mesenchymal transition (EMT) by inhibiting the activation of the SRC pro-cancer signal. In this study, we treated ovarian cancer SK-OV-3 cells with the same concentration of BRL54443 and POG for 24 hours respectively, and then collected proteins to detect the effects of the drugs on the downstream signaling pathways of ovarian cancer cells. The results of Western Blot experiments showed that POG inhibited the phosphorylation of SRC in SK-OV-3 cells, and compared with BRL54443, the inhibitory effect of POG on SRC phosphorylation was stronger, and the inhibitory effects on the pro-cancer signaling pathways such as PI3K / AKT, FAK, and EMT downstream of SRC were also more obvious ( Figure 9 A, B). To verify this result at the in vivo level, we performed immunofluorescence staining on sections of mouse ovarian cancer orthotopic tumor tissues. We observed that in mouse ovarian cancer orthotopic tumors, POG had a strong inhibitory effect on the activation of SRC and could also significantly inhibit the expression of SRC protein ( Figure 9 C, D). This result corroborates the in vitro experiments and further supports the mechanism by which POG exerts its anti-ovarian cancer effect by inhibiting the SRC signaling pathway.

[0056] At the same time, we verified by immunohistochemical staining on sections of mouse orthotopic tumors that compared with the normal saline control group, the expression of Ki67 in the POG group was down-regulated, proving that the treatment of mouse ovarian cancer in situ with POG inhibited cell proliferation. Figure 10A, B); We also verified that the expression of the mesenchymal cell marker Vimentin was downregulated and the expression of Claudin-1 was upregulated in the POG group ( Figure 10 A, B), demonstrating that POG inhibited the EMT process in murine ovarian cancer.

Claims

1. Use of cimifugin in the preparation of a drug for treating ovarian cancer.

2. The application of cimifugin in the preparation of a drug for treating ovarian cancer according to claim 1, wherein The cimifugin is used as a targeted HTR1E agonist.

3. The use of cimifugin in the preparation of a medicament for treating ovarian cancer according to claim 2, wherein, The use includes a combination of cimifugin and any one or more of the drugs in the following items (1) to (4): (1) Chemotherapeutic drugs (2) Targeted drugs (3) Anti-angiogenic drugs (4) Hormone drugs.

4. The use of cimifugin in the preparation of a medicament for treating ovarian cancer according to claim 3, wherein The chemotherapeutic drugs are selected from taxanes and platinum-based drugs; the targeted drugs are selected from PARP inhibitors.

5. The use of cimifugin in the preparation of a drug for treating ovarian cancer according to claim 4. Further, the platinum-based chemotherapeutic drugs are selected from carboplatin, cisplatin, and oxaliplatin; the taxane chemotherapeutic drugs are selected from paclitaxel, docetaxel, and cabazitaxel; the PARP inhibitors are selected from olaparib and niraparib; the anti-angiogenic drugs are selected from bevacizumab; and the hormone drugs are selected from tamoxifen and letrozole.

6. A pharmaceutical composition, which is composed of an active ingredient and at least one pharmaceutically acceptable excipient, is characterized in that The active ingredient consists of cimifugin and any one or more of the drugs in the following items (1) to (4): (1) Chemotherapeutic drugs (2) Targeted drugs (3) Anti-angiogenic drugs (4) Hormone drugs.

7. The pharmaceutical composition according to claim 6, characterized in that The chemotherapeutic drugs are selected from taxanes and platinum-based drugs; the targeted drugs are selected from PARP inhibitors.

8. The pharmaceutical composition according to claim 7, wherein The use of cimifugin in the preparation of a drug for treating ovarian cancer. The platinum-based chemotherapeutic drugs are selected from carboplatin, cisplatin, and oxaliplatin; the taxane chemotherapeutic drugs are selected from paclitaxel, docetaxel, and cabazitaxel; the PARP inhibitors are selected from olaparib and niraparib; and the anti-angiogenic drugs are selected from bevacizumab.

9. Use of cimifugin as a targeted HTR1E agonist.

10. The application according to claim 9, characterized in that Cimifugin inhibits the SRC downstream signaling pathway by activating HTR1E.

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