Application of ginsenoside Rc in prevention and treatment of colorectal tumor

Abstract

The invention discloses an application of ginsenoside Rc or a salt thereof in preparation of a medicine for preventing and / or treating colorectal tumor and / or combining other tumors. The ginsenoside Rc disclosed by the invention shows a relatively good colorectal cancer treatment effect and good safety in an MC38 tumor-bearing mouse model, and no adverse reaction is found.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the use of ginsenoside Rc in preventing and treating colorectal tumors. Background Art

[0002] Colorectal cancer (CRC) is the second leading cause of cancer-related death and the third most common malignancy worldwide. It develops in the last two parts of the large intestine (i.e., the colon and rectum), accounting for approximately 10% of all cancer cases, with the majority of cases affecting individuals aged 50 and older. Previous studies have projected a 71.5% increase in CRC cases in men and a 60.0% increase in CRC cases in women by 2035. CRC has become a major public health issue due to the significant economic burden it imposes on countries worldwide.

[0003] In terms of treatment, surgical resection is considered the mainstay of treatment for resectable CRC, while standard therapies, including chemotherapy, radiotherapy, and immunotherapy, are used for unresectable cases. However, these therapies are not without drawbacks. Some of these drawbacks include nonspecificity and cytotoxicity to normal cells, leading to secondary complications. Currently, chemotherapy regimens for CRC primarily consist of a fluoropyrimidine (5-fluorouracil or capecitabine) combined with oxaliplatin and / or irinotecan and capecitabine. Adding leucovorin to this combination is also recommended, depending on the patient's condition and the chosen treatment regimen. The drug regimen used depends on multiple factors, such as disease stage (adjuvant chemotherapy is not recommended for patients with stage 0 and stage 1 disease), drug availability, and the patient's tolerance to chemotherapy. Bevacizumab, a monoclonal antibody (mAb), is currently used as first-line treatment alongside chemotherapy. Guidelines from the American Society of Clinical Oncology recommend against the routine use of adjuvant therapy for patients with stage II colon cancer who have a low risk of recurrence. However, adjuvant therapy should be offered to patients with stage IIB and stage IIC colon cancer. Adjuvant therapy may also be used for patients with stage IIA colon cancer who have high-risk features. The role of adjuvant therapy in the treatment of stage II colon cancer remains controversial, but it is still being considered for patients with high-risk disease. Indeed, 5-FU is an effective first-line chemotherapy for metastatic colon cancer. Other drugs, such as cetuximab and leucovorin, are also well-known for their efficacy in CRC. Newer regimens include combinations of several drugs to achieve better outcomes, such as leucovorin with 5-FU and irinotecan, fluorouracil and leucovorin (FOLFIRI), or oxaliplatin (FOLFOX). The combination of 5-FU and leucovorin improves progression-free survival by 20% and late-stage survival by 6%. This combination is now used as a first- and second-line treatment option for patients with stage II and III CRC. 5-FU's mechanism of action is primarily through inhibition of thymidylate synthase, interfering with cellular deoxyribonucleic acid (DNA) synthesis, thereby leading to tumor cell death. Specifically, 5-FU is converted into 5-fluorouracil deoxynucleotide within the cell, which inhibits deoxythymidylate synthase, preventing the conversion of deoxyuridine methyl groups to deoxythymidylate, thereby interfering with DNA synthesis. In addition, 5-FU can also be converted into metabolites, inhibiting the activity of purine synthase, reducing the ability of cells to synthesize purine, and causing DNA damage, thereby affecting the normal function and proliferation of cells.

[0004] New advances have been made in the field of CRC, mainly targeted therapies and immunotherapies, aiming to overcome drug resistance, improve efficacy and reduce side effects. One of the major advances in targeted therapy is targeting RAS-mutant CRC by: (1) using direct RAS inhibitors; (2) targeting downstream pathways such as the MAPK pathway; (3) using immunotherapy combinations; and (4) targeting RAS through metabolic pathways. Another approach is by targeting BRAF-mutant CRC by: (1) using a monotherapy strategy; (2) blocking BRAF and EGFR; and (3) blocking BRAF and MEK. Regarding immunotherapy, the most promising approaches under development are therapeutic cancer vaccines, novel ICIs and their combinations, bispecific T cell engagers and ACT. Overcoming resistance to targeted therapy and immunotherapy

[0005] However, both targeted therapy and immunotherapy face some unique challenges in cancer treatment, and drug resistance remains a major challenge in enhancing their therapeutic efficacy. Resistance to targeted therapy can be acquired through several mechanisms: (1) alterations in driver oncogenes; (2) activation of parallel or downstream key signaling pathways; and (3) driving pro-survival signaling through different pathways. For example, resistance to anti-EGFR therapy in CRC may be caused by mutations in genes downstream of the EGFR signaling pathway (i.e., RAS / RAF / MEK and PI3K / AKT / mTOR) or activation of EGFR compensatory pathways (i.e., ERBB2 and MET). The development of novel mAbs targeting different EGFR epitopes is a promising process for overcoming anti-EGFR resistance. As for resistance to anti-VEGF, activation of alternative signaling pathways and production of angiogenesis-related proteins may lead to resistance to anti-angiogenic therapy, and targeting CD5L is a target to overcome this type of resistance in CRC.

[0006] The main reasons for resistance to immunotherapy in CRC are: (1) low tumor mutation burden in pMMR / MSS tumors; (2) immunosuppressive shifts caused by the TME; (3) downregulation of MHC molecules on tumor cells; and (4) low TIL density in tumors due to vascular abnormalities. To this end, chemoimmunotherapy, fecal microbiota transplantation, immune adjuvants, oncolytic viruses, and dendritic cell proliferation trials are being tested to overcome CRC resistance to immunotherapy.

[0007] Therefore, drug resistance to chemotherapy, targeted therapy or immunotherapy is one of the key factors leading to low patient survival rate, and there is an urgent need to develop new drugs to improve the survival rate of colorectal cancer patients. At the same time, there are no relevant reports in the prior art on the use of ginsenoside Rc claimed in the present invention for preventing and treating colorectal tumors and / or combined with other tumors. Summary of the Invention

[0008] Based on this, the present invention provides a use of ginsenoside Rc or a salt thereof in the preparation of a medicament for preventing and / or treating colorectal tumors and / or combined with other tumors.

[0009] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising ginsenoside Rc or a salt thereof in the preparation of a medicament for preventing and / or treating colorectal tumors and / or combined with other tumors.

[0010] Furthermore, the ginsenoside Rc or its salt is used to prevent and / or treat colorectal tumors through one or both of the following: inhibiting the growth of colorectal tumor volume and reducing the mass of colorectal tumors.

[0011] Furthermore, the colorectal tumor is a benign colorectal tumor or colorectal cancer.

[0012] Furthermore, the colorectal cancer is MC38 colorectal cancer.

[0013] Further, the other tumor is not hepatocellular carcinoma and / or melanoma.

[0014] According to another aspect of the present invention, there is provided a use of ginsenoside Rc or a salt thereof in preparing a medicament for preventing and / or treating tumors, including colorectal cancer, but excluding hepatocellular carcinoma and / or melanoma.

[0015] Furthermore, the pharmaceutical composition further comprises one or more other drugs for preventing and / or treating colorectal tumors.

[0016] Furthermore, the other drugs include chemotherapy drugs and targeted drugs.

[0017] Furthermore, the chemotherapy drugs include one or more of the following: 5-fluorouracil, oxaliplatin and irinotecan.

[0018] Furthermore, the targeted drug includes one or more of the following: cetuximab, bevacizumab, regorafenib and fruquintinib.

[0019] Furthermore, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0020] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0021] Furthermore, the dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0022] Beneficial effects of the present invention:

[0023] The ginsenoside Rc of the present invention showed good colon cancer therapeutic effect and good safety in the MC38 tumor-bearing mouse model, and no adverse reactions were found. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0025] Figure 1 Schematic diagram of the effect of ginsenoside Rc on the growth of MC38 colorectal cancer subcutaneous transplanted tumor model in mice. (A) A total of 2×10 4 MC38 cells were injected subcutaneously on the right side of the back of C57 mice. The mice were randomly divided into a control group and several Chinese medicine small molecule treatment groups (5 mice in each group). The changes in MC38 tumor growth volume were measured; (B) The weight change rate of mice in each treatment group was measured and statistically analyzed; (C) The tumor inhibition rate of tumor-bearing mice was statistically analyzed; (D) The MC38 tumor weight of each treatment group was measured and statistically analyzed; (E) The tumor inhibition rate of MC38 tumor in each treatment group was statistically analyzed; Data are presented as mean ± SEM, NS represents P>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0026] Figure 2 Schematic diagram of the effect of 10 mg / kg dose of ginsenoside Rc on the growth of MC38 colorectal cancer subcutaneous transplanted tumors in mice. (A) A total of 2×10 4 MC38 colon cancer cells were injected subcutaneously into the right side of the back of C57BL / 6J mice. The mice were randomly divided into a control group and a 10 mg / kg Rc treatment group (n=5 per group). Changes in MC38 tumor volume were measured. (B) Tumor inhibition rates of tumor-bearing mice were calculated. (C) Body weight changes of mice in each treatment group were measured and analyzed. (D) Images of MC38 tumor size in mice in each treatment group were presented. (E) MC38 tumor weights were measured and analyzed. Data are expressed as mean ± SEM. NS indicates P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0027] Figure 3 Schematic diagram of the effect of 10 mg / kg dose of ginsenoside Rc on the growth of Heap1-6 liver cancer subcutaneous tumors. (A) A total of 1×10 6Heap1-6 hepatocellular carcinoma cells were injected subcutaneously into the right side of the back of C57BL / 6J mice. The mice were randomly divided into a control group, a sorafenib-treated group, and a Rc 10 mg / kg-treated group (n=3 per group). Heap1-6 hepatocellular carcinoma tumor growth was measured. (B) Tumor inhibition rates of tumor-bearing mice were calculated. (C) Body weight changes of mice in each dose-treated group were measured and analyzed. (D) Images show the size of Heap1-6 hepatocellular carcinoma tumors in each treatment group. (E) Heap1-6 hepatocellular carcinoma tumor weights were measured and analyzed. Data are expressed as mean ± SEM. NS indicates P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0028] Figure 4 Schematic diagram of the effect of 10 mg / kg dose of ginsenoside Rc on the subcutaneous tumor growth of B16F10 melanoma cells. (A) A total of 2×10 4 B16F10 melanoma cells were injected subcutaneously into the right side of the back of C57BL / 6J mice. The mice were randomly divided into a control group and a Rc 10 mg / kg treatment group (n=3 per group). B16F10 melanoma tumor growth was measured. (B) Tumor inhibition rate of tumor-bearing mice was calculated. (C) Body weight changes of mice in each treatment group were measured and analyzed. (D) Images of B16F10 melanoma tumor sizes in mice in each treatment group were presented. (E) B16F10 melanoma tumor weights were measured and analyzed in each treatment group. Data are expressed as mean ± SEM. NS indicates P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. DETAILED DESCRIPTION

[0029] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

[0030] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those skilled in the art in the field of the present invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar or equivalent to those disclosed herein can be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0031] The present invention is intended to encompass all alternatives, modifications, and equivalents that may come within the scope of the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0032] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0033] In the present invention, the term "comprising" is synonymous with "including." As used herein, the terms "comprises," "including," "having," "containing," or any other variations thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0034] As described in the background section, drug resistance to chemotherapy, targeted therapy, or immunotherapy is one of the key factors leading to low patient survival rates, and there is an urgent need to develop new drugs to improve the survival rate of colorectal cancer patients. Furthermore, there are no prior reports on the use of ginsenoside Rc, as claimed in the present invention, for preventing and treating colorectal tumors and / or other tumors. To address the above issues, the present invention provides a use of ginsenoside Rc or a salt thereof in the preparation of a medicament for preventing and / or treating colorectal tumors and / or other tumors.

[0035] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising ginsenoside Rc or a salt thereof in the preparation of a medicament for preventing and / or treating colorectal tumors and / or combined with other tumors.

[0036] In a preferred embodiment, the ginsenoside Rc or its salt is used to prevent and / or treat colorectal tumors through one or both of the following: inhibiting the growth of colorectal tumor volume and reducing colorectal tumor mass.

[0037] In a preferred embodiment, the colorectal tumor is a benign colorectal tumor or colorectal cancer.

[0038] In a preferred embodiment, the colorectal cancer is MC38 colorectal cancer.

[0039] In a preferred embodiment, the other tumor is not hepatocellular carcinoma and / or melanoma.

[0040] According to another aspect of the present invention, there is provided a use of ginsenoside Rc or a salt thereof in preparing a medicament for preventing and / or treating tumors, including colorectal cancer, but excluding hepatocellular carcinoma and / or melanoma.

[0041] In a preferred embodiment, the pharmaceutical composition further comprises one or more other drugs for preventing and / or treating colorectal tumors.

[0042] In a preferred embodiment, the other drugs include chemotherapy drugs and targeted drugs.

[0043] In a preferred embodiment, the chemotherapy drug comprises one or more of the following: 5-fluorouracil, oxaliplatin and irinotecan.

[0044] In a preferred embodiment, the targeted drug includes one or more of the following: cetuximab, bevacizumab, regorafenib and fruquintinib.

[0045] In a preferred embodiment, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0046] In a preferred embodiment, the excipient is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, glidants, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

[0047] In a preferred embodiment, the pharmaceutical composition is in the form of powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0048] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or conditions recommended by the manufacturer.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0050] The above-mentioned features of the present invention or the features described in the embodiments may be combined in any combination. All features disclosed in this patent specification may be used in any combination, and each feature disclosed in the specification may be replaced by any alternative feature that can provide the same, equal, or similar purpose. Therefore, unless otherwise specified, the features disclosed are only general examples of equal or similar features.

[0051] The present invention is divided into two parts: a colon cancer-related pharmacodynamics experiment of ginsenoside Rc and a liver cancer / melanoma pharmacodynamics experiment.

[0052] Objective: To explore the preventive and therapeutic effects of ginsenoside Rc on colon cancer based on animal models.

[0053] Methods: (1) Ginsenoside Rc and other saponins were screened for their efficacy in inhibiting tumor growth in MC38 tumor-bearing mouse models, and statistical results showed that ginsenoside Rc monomer had a better inhibitory effect on tumor growth in tumor-bearing mice than other saponins; (2) Ginsenoside Rc monomer was used to treat C57BL / 6J mouse allograft orthotopic colon cancer model, and the tumor growth volume curve of mice was monitored; (3) A Heap1-6-luc subcutaneous tumor model of C57BL / 6j mice was established, and ginsenoside Rc was used for treatment, and the tumor growth size of mice was monitored; (4) A B16F10 subcutaneous melanoma model of C57BL / 6j mice was established, and ginsenoside Rc was used for treatment and tumor growth was monitored.

[0054] Results: (1) Compared with other saponin drugs, ginsenoside Rc can effectively inhibit the growth of tumors in MC38 tumor-bearing mice, and ginsenoside Rc has the strongest inhibitory effect on the growth of tumors in tumor-bearing mice; (2) Ginsenoside Rc has a significant inhibitory effect on the growth of MC38 colon cancer, without organ toxicity or adverse reactions; (3) By establishing a subcutaneous Heap1-6-luc liver cancer model in C57BL / 6j mice, it was found that ginsenoside Rc has no therapeutic effect on hepatocellular carcinoma; (4) By establishing a subcutaneous B16F10 melanoma model in C57BL / 6j mice, it was found that ginsenoside Rc has no effect on the growth of melanoma.

[0055] Conclusion: Ginsenoside Rc showed good therapeutic effect and good safety in the MC38 tumor-bearing mouse model of colorectal cancer, and no adverse reactions were found.

[0056] Example 1 Effects of different ginsenoside monomers on colorectal tumor growth in mice

[0057] 1.1 Experimental Materials

[0058] Mouse colorectal cancer cell line MC38 (maintained in our laboratory) was used. Twenty-four male, SPF-grade, 5-week-old C57BL / 6J mice weighing 20 ± 2 g were purchased from Spefoc (certificate number SCXK (Beijing) 2019-0010). All mice were provided with free access to food. All animal experiments were conducted in strict accordance with the relevant regulations on animal use at the Animal Center of Shanghai University of Traditional Chinese Medicine and approved by the Experimental Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval number SHZY-202103311).

[0059] The experimental instruments and reagents are shown in Table 1 and Table 2, respectively.

[0060] Table 1 Experimental instruments

[0061]

[0062]

[0063] Table 2 Experimental reagents

[0064]

[0065] 1.2 Experimental Methods

[0066] 1.2.1 Preparation of test drugs

[0067] Calculate the required amount of test sample based on the most recent animal body weight before administration. Weigh an appropriate amount of test sample before administration, dilute with solvent and make up to the required volume to prepare a solution of the required concentration (established configuration: mother solution 21 mg ginsenoside Rc powder + 210 μl DMSO = 100 mg / ml, working solution: 990 μl NS + 10 μl mother solution = 1 mg / ml colorless transparent solution. Intraperitoneal administration is performed at 10 μl / g, and the dosage is adjusted later according to the experimental changes).

[0068] 1.2.2 Experimental System

[0069] Animals: 56 male C57BL / 6J mice, SPF grade, 6-8 weeks old, weighing 20 ± 2 g, purchased from Zhejiang Weitonglihua Laboratory Animal Technology Co., Ltd., certificate number SCXK(Zhe)2019-0001. Cell line: Murine colon cancer cell line MC38, maintained in our laboratory, was subcultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U / L penicillin, and 100 mg / L streptomycin at 37°C in a humidified incubator with 5% CO2. Cells in the logarithmic growth phase were used for experiments.

[0070] 1.2.3 Model establishment

[0071] Construction of tumor-bearing mouse model: MC38 mouse colon cancer cells in logarithmic growth phase were collected and 10% matrigel was added to adjust the cell suspension concentration to 5×10 5 The cells were inoculated subcutaneously in the right back of nude mice under sterile conditions (0.2 ml / mouse) (1×10 cells / mouse). 4 The SPF grade mice were raised for 7-11 days and the tumor volume was 80-100 mm. 3 Tumor-bearing mice were randomly divided into groups.

[0072] 1.2.4 Experimental Grouping and Drug Administration

[0073] Grouping: Mice with qualified tumor volumes were randomly divided into 14 groups (3-5 mice per group): Group 1 was the model control group, and groups 2-14 were the drug-administered groups. The specific regimen is shown in Table 3.

[0074] Table 3 Experimental groups and drug administration methods

[0075]

[0076]

[0077] Drug administration: After grouping, drug administration began immediately. The drug was prepared with DMSO: normal saline = 1:99. For the model control group, DMSO: normal saline = 1:99. QD was once a day for 20 consecutive doses. The tumor volume of a single mouse exceeded 2000mm. 3 The mice were euthanized, and the mean tumor volume of each group of mice reached 2000 mm 3 The experiment was terminated when .

[0078] 1.2.5 Experimental evaluation indicators and testing

[0079] 1) Tumor diameter: Tumor diameter is the main detection indicator, and it is measured every 3-4 days. The relative tumor growth rate is calculated based on the measurement results to dynamically observe the anti-tumor effect of the test substance. Tumor volume (TV), TV = 1 / 2 × a × b 2 (where a and b are the longest and shortest diameters, respectively); relative tumor volume (RTV), RTV = Vt / V0 (where V0 is the tumor volume measured on d0 during group administration, and Vt is the tumor volume at each measurement). The anti-tumor effect was evaluated using the relative tumor growth index (TGI) (%), TGI (%) = (1-(TRTV / CRTV)) × 100% (where TRTV is the RTV of the treatment group and CRTV is the RTV of the model control group). P < 0.05 was considered effective after statistical analysis.

[0080] 2) Tumor Weight: After the experiment, animals were sacrificed, tumors were dissected, weighed, and photographed. If the average tumor weight in the model control group was less than 1 g, or if 20% of the tumors weighed less than 400 mg, it indicated poor tumor growth and the experiment was discarded. Tumor growth inhibition rate was calculated based on the weight results: tumor growth inhibition rate = [(average tumor weight in the model control group - average tumor weight in the treatment group) / average tumor weight in the model control group] × 100%. A P < 0.05 was considered statistically significant.

[0081] 3) Body weight: The animals were weighed every 3-4 days to observe the effects of the test drugs on their body weight.

[0082] 1.2.6. Collect samples

[0083] At the end of the experiment, tumor tissues of 3 mice were collected from each of the best treatment groups, group 1 Blank and group 1 with the best ginsenoside treatment effect. The tumors were about the size of soybeans and then made into single-cell suspensions for CYTOF experiments. In addition, tumors of the remaining 3 experimental animals in each group were collected and stored in a -80℃ refrigerator for RNA-seq experiments. Spleens of 3 mice in each group were collected and placed on ice for flow cytometry detection of mCD4+, mCD8+ and mIFN-γ experiments.

[0084] 1.2.7 Experimental data processing method

[0085] Each experimental group in the cell experiment provided at least 3 replicates, and the in vivo animal experiment data provided at least 5 independent data. Each experimental result is expressed as the mean ± SEM. Statistical analysis was performed using Graphpad Prism 9 (GraphPad, San Diego, California). To test the significance of each experimental data, an unpaired T test was used. A P value of 0.05 or above was not significant; a P value of 0.05 or lower was considered to be significantly different (tumor volume and tumor weight in the Rc group were significantly different from those in the control group). Data are expressed as mean ± SEM, NS is P>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0086] 1.3 Experimental Results

[0087] To verify the anti-colorectal cancer effects of different ginsenoside monomers, the present invention set up 7 groups of drugs to treat C57 mouse MC38 colorectal cancer model. During the 16-day treatment process, ginsenoside Rc was able to significantly inhibit the progression of colorectal cancer subcutaneous transplanted tumors. The results are as follows Figure 1 As shown. Compared with the solvent control group, the ginsenoside Rc group had the most significant therapeutic effect. After the end of treatment, the tumor tissues of the mice were dissected and weighed. It was found that the tumor weight of the mice in the ginsenoside Rc group was significantly reduced (P < 0.05, TGI% = 51.19%), while other saponins showed no relevant effect (P > 0.05). Some saponins, such as Panax notoginseng saponin Fe (TGI% = -19.49%), Vietnamese ginsenoside RB (TGI% = -2.88%), ginsenoside Rg4 (TGI% = -35.33%), ginsenoside F3 (TGI% = -39.59%), and ginsenoside Rb2 (TGI% = -19.91%), even had the side effect of promoting tumor growth. These results suggest that ginsenoside Rc has an anti-colorectal cancer effect.

[0088] Example 2 Pharmacodynamic Evaluation of Ginsenoside Rc on Murine MC38 Colorectal Cancer Cells in a Syngeneic Transplanted Tumor Model in C57BL / 6J Mice

[0089] To replicate the anti-colorectal cancer effects of ginsenoside monomers, we used Rc to treat a C57 mouse model of MC38 colorectal cancer. Over 22 days of treatment, ginsenoside Rc significantly inhibited the progression of subcutaneous colorectal cancer xenografts, as shown in the figure below. Compared to the solvent control group, the ginsenoside Rc group showed significantly greater efficacy. Following treatment, tumor tissue was dissected and weighed, revealing a significant reduction in tumor weight in the ginsenoside Rc-treated group. This finding suggests the anti-colorectal cancer effects of ginsenoside Rc.

[0090] 2.1 Experimental Materials

[0091] The experimental equipment was the same as in Example 1. Ten female C57BL / 6J mice, SPF grade, 5-6 weeks old, weighing 20 ± 2 g were purchased from Spafor (certificate number SCXK (Beijing) 2019-0010).

[0092] 2.2 Experimental Methods

[0093] The experimental system is the same as that of Example 1.

[0094] 2.3 Experimental evaluation indicators and testing

[0095] The experimental evaluation criteria are the same as those in Example 1.

[0096] 2.4 Experimental data processing method

[0097] The experimental data processing method is the same as that in Example 1.

[0098] 2.5 Experimental Results

[0099] The results are as follows Figure 2 As shown, 10 mg / kg dose of ginsenoside Rc significantly inhibited the growth of MC38 colorectal cancer subcutaneous transplanted tumors in mice.

[0100] Example Pharmacodynamic evaluation of ginsenoside Rc on murine Heap-6 liver cancer cells in a homologous transplanted tumor model in C57BL / 6J mice

[0101] 3.1 Experimental Materials

[0102] The experimental equipment was the same as in Example 1. The mouse hepatocellular carcinoma cell line Heap1-6 (maintained in our laboratory) was cultured. Fifteen female C57BL / 6J mice, SPF grade, 5-6 weeks old, weighing 20 ± 2 g, were purchased from Spafor (certificate number SCXK (Beijing) 2019-0010).

[0103] 3.2 Experimental Methods

[0104] The experimental system was the same as that in Example 1. 1×10 6 mouse Heap1-6 cells were subcutaneously inoculated. The sorafenib dosage was 30 mg / kg.

[0105] 3.3 Experimental evaluation indicators and testing

[0106] The experimental evaluation criteria are the same as those in Example 1.

[0107] 3.4 Experimental data processing method

[0108] The experimental data processing method is the same as that in Example 1.

[0109] 3.5 Experimental Results

[0110] The results are as follows Figure 3 As shown in the experiment, it was found that the same dose of chemotherapy drug sorafenib had a significant inhibitory effect on the growth of hepatocellular carcinoma tumors, but ginsenoside Rc monomers such as Figure 3 D, on the other hand, promoted the growth of subcutaneous hepatocellular carcinoma tumors. Tumor removal at the end of the experiment revealed significant differences in tumor weight between the ginsenoside Rc group and the vehicle group and the chemotherapy drug sorafenib group at the same dose. Ginsenoside Rc had no therapeutic effect on heap1-6 hepatocellular carcinoma.

[0111] Ginsenoside Rc at a dose of 10 mg / kg had no inhibitory effect on the growth of subcutaneous tumors of Heap1-6 liver cancer cells.

[0112] Example 4 Pharmacodynamic Evaluation of Rc Drug on Murine Melanoma Cell B16-F10 Syngeneic Transplanted Tumor Model in C57BL / 6J Mice

[0113] 4.1 Experimental Materials

[0114] The experimental equipment was the same as in Example 1. The mouse melanoma cell line B16F10 (maintained in our laboratory) was cultured. Ten female C57BL / 6J mice, SPF grade, 5-6 weeks old, weighing 20 ± 2 g, were purchased from Spafor (certificate number SCXK (Beijing) 2019-0010).

[0115] 4.2 Experimental Methods

[0116] The experimental system was the same as that in Example 1. 5×10 5 murine melanoma B16F10 cells were subcutaneously inoculated into C57BL / 6j mice, and after tumor formation, 10 mg / kg of cells were intraperitoneally injected daily.

[0117] 4.3 Experimental evaluation indicators and testing

[0118] The experimental evaluation criteria are the same as those in Example 1.

[0119] 4.4 Experimental data processing method

[0120] The experimental data processing method is the same as that in Example 1.

[0121] 4.5 Experimental Results

[0122] The results are as follows Figure 4 As shown in the experiment, it was found that the same dose of 10mg / kg ginsenoside Rc had no therapeutic effect on melanoma B16F10 subcutaneous tumors. At the end of the experiment, the tumor was weighed and the ginsenoside Rc monomer was found to be Figure 4 D. There was no difference in tumor size compared with the solvent group. The experiment showed that Rc had no therapeutic effect on B16F10 melanocytoma.

[0123] Ginsenoside Rc at a dose of 10 mg / kg had no inhibitory effect on the growth of subcutaneous tumors of B16 F10 melanoma cells.

[0124] Results: (1) Compared with many other saponin drugs, Rc can effectively inhibit the growth of tumors in MC38 tumor-bearing mouse models and has the strongest inhibitory effect on tumor growth in tumor-bearing mice; (2) Ginsenoside Rc has a significant inhibitory effect on the growth of MC38 colon cancer without organ toxicity or adverse reactions; (3) By establishing a subcutaneous Heap1-6-luc liver cancer model in C57BL / 6j mice, it was found that ginsenoside Rc has no therapeutic effect on hepatocellular carcinoma; (4) By establishing a subcutaneous B16F10 melanoma model in C57BL / 6j mice, it was found that ginsenoside Rc has no effect on the growth of melanoma.

[0125] Conclusion: Rc showed good therapeutic efficacy and safety in the tumor-bearing mouse model, and no adverse reactions were found.

[0126] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only intended to help understand the method of the present invention and its core concept. At the same time, changes or modifications made by those skilled in the art based on the concept of the present invention, the specific implementation methods of the present invention, and the scope of application are all within the scope of protection of the present invention. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. Use of ginsenoside Rc or its salt in the preparation of a medicament for preventing and / or treating colorectal tumors and / or combined with other tumors.

2. Use of a pharmaceutical composition comprising ginsenoside Rc or a salt thereof in the preparation of a medicament for preventing and / or treating colorectal tumors and / or combined with other tumors.

3. The use according to claim 1 or 2, characterized in that The ginsenoside Rc or its salt is used to prevent and / or treat colorectal tumors through one or both of the following: inhibiting the growth of colorectal tumor volume and reducing the mass of colorectal tumors.

4. The use according to any one of claims 1 to 3, characterized in that The colorectal tumor is a benign colorectal tumor or colorectal cancer; Preferably, the colorectal cancer is MC38 colorectal cancer; More preferably, the other tumor is not hepatocellular carcinoma and / or melanoma.

5. Use of ginsenoside Rc or its salt in the preparation of a medicament for preventing and / or treating tumors, characterized in that: The tumor includes colorectal cancer, and the tumor does not include hepatocellular carcinoma and / or melanoma.

6. The use according to claim 2, characterized in that The pharmaceutical composition further comprises one or more other drugs for preventing and / or treating colorectal tumors; Preferably, it is characterized in that the other drugs include chemotherapy drugs and targeted drugs.

7. The use according to claim 6, characterized in that The chemotherapy drugs include one or more of the following: 5-fluorouracil, oxaliplatin and irinotecan; Preferably, the targeted drugs include one or more of the following: cetuximab, bevacizumab, regorafenib and fruquintinib.

8. The use according to claim 2, characterized in that The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

9. The use according to claim 8, characterized in that The auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant.

10. The use according to claim 2, characterized in that The dosage form of the pharmaceutical composition is powder, tablet, dripping pill, capsule, film, lozenge, granule, injection or oral solution.

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