Application of ginsenoside Ra1 in preventing and treating digestive tract tumors

Abstract

The present invention discloses a use of ginsenoside Ra1 or its salt in preparing a drug for preventing and / or treating digestive tract tumors. The ginsenoside Ra1 or its salt of the present invention shows better therapeutic effect than 5-FU and Rg3 in a tumor-bearing mouse model, and no adverse reactions are found. Ginsenoside Ra1 or its salt inhibits the growth and metastasis of colorectal cancer by relying on immunomodulatory pathways.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an application of ginsenoside Ra1 in preventing and treating digestive tract tumors. Background Art

[0002] Studies have shown that the multidisciplinary team (MDT) model can improve the diagnosis and treatment of colorectal cancer. Chemotherapy based on 5-fluorouracil and oxaliplatin is a common first-line therapy for patients with colon cancer, and is also effective for patients with advanced colorectal cancer after surgery. 5-Fluorouracil (5-FU) is a DNA / RNA synthesis inhibitor that interferes with nucleotide synthesis in colorectal cancer cells by inhibiting thymine nucleotide synthetase, thereby inhibiting cancer cell proliferation. Although chemotherapy, radiotherapy, and adjuvant chemotherapy can significantly prolong the survival of patients, colorectal cancer cells often destroy the cell cycle mechanism through mutations, leading to genomic instability and chemotherapy resistance. Therefore, chemotherapy resistance is one of the key factors leading to low patient survival, and there is an urgent need to develop new drugs to improve the survival rate of patients with colorectal cancer.

[0003] According to previous survey data, about 83% of colorectal cancer patients are already in the middle and late stages when diagnosed. For middle and late stage colorectal cancer, drug treatment is critical. In recent years, immune checkpoint inhibitors have become an important treatment option for colorectal cancer. Studies have confirmed that drugs such as nivolumab ± ipilimumab, Keytruda, and Jemperli (dostarlimab-gxly) have shown outstanding effects in advanced colorectal cancer, and their targets include CTLA-4 and PD-L1 / PD-1. Professor James Allison and Professor Honjo Tasuku won the 2018 Nobel Prize in Medicine for their pioneering research on tumor immune checkpoint blockade therapy (ICI). The U.S. Food and Drug Administration and the European Medicines Agency have also approved the use of immunotherapy for melanoma, lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, and other tumors. Although immune checkpoint inhibitors are effective in treating MSI or MMR tumors, the immune response is still limited in microsatellite stable (MSS) tumors. About 95% of colorectal cancer patients are in MSS status, indicating that these patients will not benefit from immunotherapy for the time being. Some scholars believe that insufficient infiltration of immune cells in the tumor immune microenvironment is the main reason for the low immune response rate. Screening for adjuvant drugs that can enhance immunotherapy has become a current research hotspot. Therefore, although immune checkpoint inhibitor therapy is not effective for MSS colorectal cancer patients, immunotherapy is still one of the most promising therapies currently. Summary of the invention

[0004] Based on this, the present invention provides a use of ginsenoside Ra1 or a salt thereof in the preparation of a medicament for preventing and / or treating digestive tract tumors.

[0005] Furthermore, the method for preparing the solution of ginsenoside Ra1 or its salt comprises: weighing an appropriate amount of ginsenoside Ra1 or its salt, adding a reagent to dissolve and dilute it, and obtaining the solution of ginsenoside Ra1.

[0006] Furthermore, the reagent is 1% dimethyl sulfoxide and 99% physiological saline.

[0007] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising ginsenoside Ra1 or a salt thereof in the preparation of a medicament for preventing and / or treating digestive tract tumors.

[0008] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating digestive tract tumors.

[0009] Furthermore, the drug further includes one or more of the following: DNA / RNA synthesis inhibitors, platinum anticancer drugs, PD-1 / PD-L1 inhibitors, and EGFR inhibitors.

[0010] Furthermore, the DNA / RNA synthesis inhibitor is 5-fluorouracil.

[0011] Furthermore, the platinum anticancer drug is oxaliplatin.

[0012] Furthermore, the PD-1 / PD-L inhibitor includes one or more of the following: palivizumab, nivolumab, ipilimumab, and dotalimumab.

[0013] Furthermore, the EGFR inhibitor is canertinib and / or cetuximab.

[0014] According to another aspect of the present invention, there is provided a use of a pharmaceutical preparation comprising ginsenoside Ra1 or a salt thereof and a pharmaceutically acceptable excipient in the preparation of a medicament for preventing and / or treating digestive tract tumors.

[0015] 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, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0016] Furthermore, the pharmaceutical preparation is in the form of powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0017] Furthermore, the digestive tract tumor is a benign digestive tract tumor and / or a malignant digestive tract tumor.

[0018] Further, the digestive tract is selected from one or more of the following: esophagus, stomach, intestine, gallbladder, pancreas and liver.

[0019] Furthermore, the digestive tract tumor includes metastatic liver tumor.

[0020] Furthermore, the digestive tract tumor does not include an in situ liver tumor, such as in situ liver cancer.

[0021] Furthermore, the intestine is the large intestine and / or the small intestine.

[0022] Further, the large intestine is selected from one or more of the following: cecum, appendix, colon, rectum and anal canal.

[0023] Further, the small intestine is selected from one or more of the following: duodenum, jejunum and ileum.

[0024] Furthermore, ginsenoside Ra1 inhibits the growth and metastasis of digestive tract tumors through immune regulation pathways.

[0025] Furthermore, the pharmaceutical concentration of ginsenoside Ra1 is 0.1-10 mg / ml, for example, about 1 mg / ml.

[0026] Beneficial effects of the present invention:

[0027] The ginsenoside Ra1 or its salt of the present invention shows better therapeutic effect than 5-FU and Rg3 in tumor-bearing mouse model, and no adverse reaction is found. Ginsenoside Ra1 or its salt inhibits the growth and metastasis of colorectal cancer by relying on immunomodulatory pathway. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. 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.

[0029] Figure 1The schematic diagram shows the cytotoxicity results of ginsenoside Ra1 and 5-FU on mouse colorectal cancer cell lines CT26, MC38 and human colorectal cancer cell lines HCT116, Lovo, and HT29. Among them, (A) the cell viability of CT26 mouse colorectal cancer cell line after being treated with ginsenoside Ra1 at a concentration gradient of 5000μM to 9.76μM for 72 hours; (B) the cell viability of MC38 mouse colorectal cancer cell line after being treated with ginsenoside Ra1 at a concentration gradient of 5000μM to 9.76μM for 72 hours; (C) the cell viability of HCT16 mouse colorectal cancer cell line after being treated with ginsenoside Ra1 at a concentration gradient of 5000μM to 9.76μM for 72 hours; (D) the cell viability of Lovo mouse colorectal cancer cell line after being treated with ginsenoside Ra1 at a concentration gradient of 5000μM to 9.76μM for 72 hours. (E) Cell viability of HT29 mouse colorectal cancer cell line treated with ginsenoside Ra1 at concentrations ranging from 5000 μM to 9.76 μM for 72 h. Data are expressed as mean ± SEM, **P < 0.01, ****P < 0.0001.

[0030] Figure 2 Schematic diagram of the effect of ginsenoside Ra1 on the growth of MC38 colorectal cancer subcutaneous transplanted tumor model in mice. 4 MC38 cells were injected subcutaneously into the right side of the back of C57 mice. The mice were randomly divided into a control group and a Ra1 10 mg / kg treatment group (3 mice in each group), and the MC38 tumor growth was measured; (B) The tumor growth inhibition rate of the drug on tumor-bearing mice was statistically analyzed; (C) The MC38 tumor weight of each treatment group was measured and statistically analyzed; (D) The weight change rate of mice in each treatment group was measured and statistically analyzed; (E) The picture of the MC38 tumor size of mice in each treatment group was displayed. The data are expressed as mean ± SEM, NS is P>0.05, *P<0.05,

[0031] **P<0.01, ***P<0.001, ****P<0.0001.

[0032] Figure 3 Schematic diagram of the effect of 10 mg / kg dose of ginsenoside Ra1 on the growth of MC38 colorectal cancer subcutaneous transplanted tumors in mice. 4MC38 cells were injected subcutaneously into the right side of the back of C57 mice. The mice were randomly divided into a control group and a Ra1 10 mg / kg treatment group (9 mice in each group), and the MC38 tumor growth was measured; (B) The MC38 tumor weight of the treatment group was measured and counted; (C) The weight change rate of mice in each different dose treatment group was measured and counted; (D) The tumor growth inhibition rate of the drug on tumor-bearing mice was counted; (E) The MC38 tumor size of mice in each treatment group was displayed in pictures. Data are expressed as mean ± SEM, NS is P>0.05,

[0033] *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0034] Figure 4 Schematic diagram of the effect of the same dose of ginsenoside Ra1 and Rg3 on the growth of MC38 colorectal cancer subcutaneous transplanted tumors in mice. 4 MC38 cells were injected subcutaneously into the right side of the back of C57 mice. The mice were randomly divided into a control group and a Ra1 10mg / kg treatment group (5 mice in each group), and the MC38 tumor growth was measured; (B) The MC38 tumor weight of the treatment group was measured and counted; (C) The tumor inhibition rate of the drug on tumor growth in tumor-bearing mice was counted; (D) The weight change rate of mice in each different dose treatment group was measured and counted; (E) The picture of the MC38 tumor size of mice in each treatment group was displayed. The data are expressed as mean ± SEM, NS is P>0.05,

[0035] *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0036] Figure 5 Schematic diagram of the effect of ginsenoside Ra1 on the growth of HT29 colorectal cancer subcutaneous transplanted tumors in Babl / c nude mice. 6 HT29 cells were injected subcutaneously into the right side of the back of Babl / c nude mice. The mice were randomly divided into a control group and a Ra1 10 mg / kg administration group (5 mice in each group), and the MC38 tumor growth was measured; (B) The weight change rate of mice in each different dose administration group was measured and counted; (C) The weight of MC38 tumors in the administration group was measured and counted; (D) The tumor inhibition rate of the drug on tumor growth in tumor-bearing mice was counted; (E) The pictures of the MC38 tumor size of mice in each administration group were displayed. Data are expressed as mean ± SEM, NS is P>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0037] Figure 6Schematic diagram of the effect of ginsenoside Ra1 on the growth of homologous MC38-luc colorectal cancer orthotopic transplanted tumors in C57 mice. 6 MC38-luc cells were injected subcutaneously into the right side of the back of C57BL / 6J mice. 3 The tumor mass was orthotopically transplanted into the branch of the colonic mesenteric vessels at the lower end of the cecum of C57BL / 6J mice to establish an orthotopic colon transplant tumor model. The mice were randomly divided into a control group and a Ra1 10 mg / kg treatment group (6 mice in each group), and the growth of MC38-luc orthotopic tumors was monitored by in vivo fluorescence imaging; (B) The changes in the fluorescence size of the orthotopic tumors in the mice in the Ra1 treatment group were measured and statistically analyzed; (C) The image of the orthotopic tumor size of the MC38-luc in mice was displayed; (D) The weight of the orthotopic tumors in the treatment group and the solvent group was measured and statistically analyzed; (E) The fluorescence value statistics showed the effect of drugs on the tumor growth of orthotopic tumor-bearing mice at different time points. The data are expressed as mean ± SEM, NS is P>0.05, *P<0.05, **P<0.01, ***P<0.001,

[0038] ****P<0.0001.

[0039] Figure 7 Schematic diagram of the effect of ginsenoside Ra1 on the growth of homologous CT26-luc colorectal cancer orthotopic transplanted tumors in Babl / c mice. 6 CT26-luc cells were injected into the right side of the back of Babl / c mice. 3 The tumor mass was orthotopically transplanted into the confluence of the colonic mesenteric vessels at the lower end of the cecum of Babl / c mice to establish a colon orthotopic liver metastasis transplant tumor model. The mice were randomly divided into a control group and a Ra1 10 mg / kg administration group (5 mice in each group), and the growth of CT26-luc orthotopic tumors was monitored by in vivo fluorescence imaging; (B) The changes in the fluorescence size of the orthotopic tumors in the mice in the Ra1 administration group were measured and statistically analyzed; (C) The image of the orthotopic tumor size of CT26-luc in mice was displayed; (D) The survival time of Babl / c mice in the CT26 homologous transplant tumor model; (E) The weight of the orthotopic tumors of CT26-luc in the administration group and the solvent group was measured and statistically analyzed; (F) Tumor metastasis in the liver of Babl / c mice in the CT26 homologous transplant tumor model. The mouse liver was sampled after dissection and fixed in 4% formaldehyde solution; (G) The mouse liver was sampled after dissection, fixed with formaldehyde, embedded in paraffin, and then sectioned and stained with H&E. The data are expressed as mean ± SEM, NS is P>0.05, *P<0.05,

[0040] **P<0.01, ***P<0.001, ****P<0.0001.

[0041] Figure 8 Schematic diagram of the effect of ginsenoside Ra1 on the growth of homologous melanoma in C57 mice. 5 B16F10 cells were injected subcutaneously into the right side of the back of C57 mice. The mice were randomly divided into a control group and a Ra15mg / kg administration group (3 mice in each group), and the MC38 tumor growth was measured; (B) The tumor weight of the administration group was measured and counted; (C) The weight change rate of mice in each different dose administration group was measured and counted; (D) The tumor growth inhibition rate of the drug on tumor-bearing mice was counted; (E) The picture of the B16F10 tumor size of mice in each administration group was displayed. Data are expressed as mean ± SEM, NS is P>0.05, *P<0.05, **P<0.01,

[0042] ***P<0.001, ****P<0.0001.

[0043] Fig. 9 Schematic diagram of the effect of ginsenoside Ra1 on the growth of homologous Heap1-6-luc orthotopic liver cancer in C57 mice. 5 Heap1-6-luc cells were injected into the liver of C57BL / 6J mice to establish an orthotopic liver cancer growth model. On the third day, mice were randomly divided into a control group and a Ra1 5mg / kg administration group (3 mice in each group) according to the fluorescence value, and the growth of Heap1-6-luc orthotopic tumors was monitored by in vivo fluorescence imaging; (B) Pictures of the orthotopic Heap1-6-luc tumor size of mice were displayed; (C) The changes in the fluorescence size of the orthotopic tumors of mice in the Ra1 administration group were counted; (D) The ratio of Heap1-6-luc liver weight to mouse body weight in the administration group and the solvent group was measured and counted; (E) Data are expressed as mean ± SEM, NS is P>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. DETAILED DESCRIPTION

[0044] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0045] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0046] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in 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 can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0047] 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.

[0048] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed 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.

[0049] As described in the background technology section, current existing colorectal cancer treatment drugs have problems such as easy to produce chemotherapy resistance and poor efficacy. In order to solve the above problems, the present invention provides a use of ginsenoside Ra1 or its salt in the preparation of a drug for preventing and / or treating digestive tract tumors.

[0050] In the present invention, the term "treatment" also includes "prevention", unless there is a specific description to the contrary. The terms "treatment" and "therapeutically" should be understood accordingly.

[0051] In the present invention, the term "treatment" includes alleviating, inhibiting or improving the symptoms or conditions of a disease; inhibiting the occurrence of complications; improving or preventing potential metabolic syndrome; inhibiting the occurrence of a disease or symptom, such as controlling the development of a disease or condition; alleviating a disease or symptom; reducing a disease or symptom; alleviating complications caused by a disease or symptom, or preventing or treating signs caused by a disease or symptom. As used herein, a pharmaceutical composition or a pharmaceutical preparation, after administration, can improve a disease, symptom or condition, especially improve its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Whether fixed or temporary administration, continuous or intermittent administration, can be attributed to or related to the administration.

[0052] In the present invention, the term "salt", "acid addition salt" or "pharmaceutically acceptable salt" includes all acceptable salts. Examples of acid salts are hydrochloride, hydrobromide, hydrofluoride, perchlorate, nitrate, sulfate, phosphate, formates, acetate, trifluoroacetate, propionate, malate, succinate, malonate, methanesulfonate, etc. The salt form of triazatriazine (naphthyridine) is characterized by being protonated by one or more nitrogen atoms. Complete salts consist of anions linked to a monoprotonated or polyprotonated macrocycle.

[0053] In a preferred embodiment, the method for preparing the solution of ginsenoside Ra1 or its salt comprises: weighing an appropriate amount of ginsenoside Ra1 or its salt, adding a reagent to dissolve and dilute it, and obtaining the solution of ginsenoside Ra1.

[0054] In a preferred embodiment, the reagent is 1% dimethyl sulfoxide and 99% physiological saline.

[0055] According to another aspect of the present invention, there is provided a use of a pharmaceutical composition comprising ginsenoside Ra1 or a salt thereof in the preparation of a medicament for preventing and / or treating digestive tract tumors.

[0056] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating digestive tract tumors.

[0057] In a preferred embodiment, the drug further includes one or more of the following: DNA / RNA synthesis inhibitors, platinum anticancer drugs, PD-1 / PD-L1 inhibitors, EGFR inhibitors.

[0058] In a preferred embodiment, the DNA / RNA synthesis inhibitor is 5-fluorouracil.

[0059] In a preferred embodiment, the platinum anticancer drug is oxaliplatin.

[0060] In a preferred embodiment, the PD-1 / PD-L inhibitor includes one or more of the following: palivizumab, nivolumab, ipilimumab, and dotalizumab.

[0061] In a preferred embodiment, the EGFR inhibitor is canertinib and / or cetuximab.

[0062] According to another aspect of the present invention, there is provided a use of a pharmaceutical preparation comprising ginsenoside Ra1 or a salt thereof and a pharmaceutically acceptable excipient in the preparation of a medicament for preventing and / or treating digestive tract tumors.

[0063] In the present invention, the term "pharmaceutically acceptable" refers to a substance, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound and is relatively non-toxic, i.e., a substance that does not cause undesirable biological effects or interact in a deleterious manner with any of its components when administered to an individual.

[0064] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with subjects and active ingredients (i.e., capable of inducing the desired therapeutic effect without causing any undesirable local or systemic effects), which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995).

[0065] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0066] Those skilled in the art will know how to select specific chemical substances within the scope of the above-mentioned auxiliary material category.For example, the dispersant can be selected from one or more of the following: cross-linked sodium carboxymethyl cellulose, sodium starch glycolate and pre-gelatinized corn starch.The wetting agent can be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, cetyl alcohol, glycerol fatty acid ester (such as triacetin, glyceryl monostearate and the like), polyoxyethylene fatty acid ester, polyethylene glycol fatty acid ester, sodium lauryl sulfate, sorbitol fatty acid ester, sucrose fatty acid ester, polyoxyethylene ether, benzalkonium chloride, polyoxyethylene castor oil and docusate sodium.The adhesive can be selected from one or more of the following: polyvinyl pyrrolidone, hydroxypropyl cellulose, polyethylene glycol and methylcellulose. The diluent may be selected from one or more of the following: powdered sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate and calcium carbonate. The glidant is micropowder silica gel. The lubricant may be selected from one or more of the following: calcium stearate, talc, magnesium stearate, stearic acid and colloidal silicon dioxide. The sustained-release agent may be selected from one or more of the following: sodium carboxymethylcellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, hydroxyethyl cellulose, gum arabic, gelatin and shellac. The plasticizer may be dibutyl sebacate and / or various citrates. The disintegrant may be selected from one or more of the following: carboxymethyl cellulose, carboxymethyl cellulose calcium salt and sodium carboxymethyl cellulose. The inclusion agent is hydroxypropyl beta-cyclodextrin. The flavoring agent may be selected from one or more of the following: sorbitol, glucose, mannose, sucrose and lactose. The opacifier is titanium dioxide. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium metabisulfite, sodium sulfite and sodium thiosulfite.

[0067] These excipients are preferably pharmaceutically inert, or may have a synergistic or additive effect to enhance the therapeutic activity of the pharmaceutical composition, and the above-mentioned excipients are only listed. The excipients actually used in the present invention are not limited to the above-mentioned excipients and can be adjusted according to actual conditions to achieve the effects of the present invention.

[0068] In a preferred embodiment, the pharmaceutical preparation is a powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0069] In a preferred embodiment, the digestive tract tumor is a benign digestive tract tumor and / or a malignant digestive tract tumor.

[0070] In a preferred embodiment, the digestive tract is selected from one or more of the following: esophagus, stomach, intestine, gallbladder, pancreas and liver.

[0071] In a preferred embodiment, the digestive tract tumor includes a metastatic liver tumor.

[0072] In a preferred embodiment, the digestive tract tumor does not include an in situ liver tumor, such as in situ liver cancer.

[0073] In a preferred embodiment, the intestine is the large intestine and / or the small intestine.

[0074] In a preferred embodiment, the large intestine is selected from one or more of the following: cecum, appendix, colon, rectum and anal canal.

[0075] In a preferred embodiment, the small intestine is selected from one or more of the following: duodenum, jejunum and ileum.

[0076] In a preferred embodiment, the ginsenoside Ra1 inhibits the growth and metastasis of digestive tract tumors through an immunomodulatory pathway.

[0077] In a preferred embodiment, the pharmaceutical concentration of ginsenoside Ra1 is 0.1-10 mg / ml, such as about 1 mg / ml.

[0078] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0079] The present invention further provides a method for preventing and / or treating a digestive tract tumor in a subject, comprising administering an effective amount of the above composition or the above pharmaceutical preparation to the subject.

[0080] The present invention further provides the above composition or the above pharmaceutical preparation for preventing and / or treating digestive tract tumors in a subject.

[0081] In the present invention, the term "subject" is a mammal. The mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse or a cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects representing digestive tract tumor models. Preferably, the subject is a human.

[0082] The "effective amount" of the pharmaceutical composition or preparation used in the present invention can obtain the desired treatment and / or prevention effect. The amount effective for this purpose will depend on, for example, the pharmaceutical composition, the mode of administration, the stage and severity of the disease to be treated, the individual's body weight and overall health, and the judgment of the prescribing physician. The dosage can be given once a week, or once every two days or every day, or even several times a day. The dosage unit can be given in a short period of time (e.g., weeks to months) or longer time periods (months to years).

[0083] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0084] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0085] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0086] The present invention is divided into two parts: in vitro pharmacodynamics experiment and in vivo pharmacodynamics experiment. Mouse and human colorectal cancer cell lines were used to explore the cytotoxicity of ginsenoside Ra1 in vitro. In vivo experiments used colorectal cancer cell lines for subcutaneous injection to form tumors, homologous mouse colon cancer in situ model and immunodeficient mouse subcutaneous tumor model, and used ginsenoside Ra1 for treatment to observe tumor growth and metastasis.

[0087] Example In vitro toxicity study of ginsenoside Ra1 on colorectal cancer cells

[0088] 1.1 Experimental Materials

[0089] Mouse colorectal cancer cell lines MC38 and CT26, and human colorectal cancer cell lines HT29, lovo, and HCT116 (maintained in our laboratory).

[0090] Table 1 Experimental instruments

[0091]

[0092]

[0093] Table 2 Experimental reagents

[0094] Experimental reagents Part Number factory Origin RPMI 1640 medium MA0559 Meilun Dalian, China CCK-8 Kit BS350B White Shark Wuhan, China 1× PBS buffer BL302A White Shark Wuhan, China Dimethyl sulfoxide (Shanghai trial brand) 30072418 Sinopharm Chemical Reagent Co., Ltd. Shanghai, China Anhydrous methanol (Shanghai test brand) 10014108 Sinopharm Chemical Reagent Co., Ltd. Shanghai, China Trypan blue ST798 Blue Sky Shanghai, China Ginsenoside Ra1 BP1846 Chengdu Aifa Biotechnology Co., Ltd. Sichuan, China 5-Fluorouracil B25419 Yuanye Biotechnology Co., Ltd. Germany Pancreatin 25200056 Gibco USA Fetal bovine serum 04-001-1ACS BI USA

[0095] Drug preparation: According to the needs of the cell experiment, 11.2 mg of ginsenoside Ra1 was weighed on an electronic balance and evenly dissolved in 100 μl of 1% dimethyl sulfoxide and 99% saline solution to prepare a 100 mM ginsenoside Ra1 drug stock solution for gradient dilution in the experiment. The positive control group was 5-FU, and a 100 μM 5-FU drug stock solution was prepared for gradient dilution in the experiment. The negative control group was added with 1% dimethyl sulfoxide and 99% saline solvent at the same concentration as the dilution gradient of the drug administration group to eliminate the interference of the solvent on the cell experiment.

[0096] 1.2 Experimental Methods

[0097] Cytotoxicity assay (IC 50 )

[0098] Take the colorectal cancer cell line in the logarithmic growth phase, use trypsin to digest the cells, centrifuge at 1000RPM for 3 minutes, resuspend the cells in RPMI1640 medium, and count them using a cell counting plate after appropriate dilution. Add 100μl of deionized water or PBS to the outer circle of wells in the 96-well plate to reduce the evaporation of the culture medium in the inner circle and affect the drug concentration. Inoculate the cell suspension in a 96-well plate, with a volume of approximately 100μl / 2000 cells per well, and 3 replicate wells. The drug concentration gradient is set to a 1:1 dilution ratio, and 5-Fu is diluted in a 4-fold gradient, such as: 2.5 / 5 / 10 / 20 / 40 / 80 / 160μM. At least 8 effective concentration gradients need to be set to reasonably fit the IC 50 Curve. Place the 96-well plate in a CO2 constant temperature incubator and incubate overnight or for 2-6 hours (37°C, 5% CO2). When the cells are in good condition and growing rapidly, the time required for attachment is shorter. Avoid direct contact between the gun tip and the bottom of the well throughout the experiment to avoid cell loss caused by scratches, which will affect the accuracy of the experiment. After incubation overnight, aspirate the culture medium. Add 100 μl of culture medium containing different concentration gradient drugs to each well of the 96-well plate. Place the 96-well plate in an incubator and incubate for 72 hours. Aspirate the culture medium and add culture medium containing 10% CCK-8. Avoid bubbles during the addition process to avoid affecting the absorbance reading. Place the culture plate in an incubator and incubate for 2 hours. Measure the absorbance at 450nm with an enzyme marker. Data processing: Cell viability (%) = [A (drug added) - A (blank)] / [A (0 drug added) - A (blank)] × 100%. Use Graphpad to input the activity value experimental data, use XY table format for 3 replicate wells, the X axis is the drug concentration, and the Y axis is the name of the drug group. For example: Ra1, 5-FU. Select the fitting method of the four-parameter Loginhibitor in the toolbar Analyze-Transform for the experimental data to obtain the fitting curve and obtain the logarithm for IC 50 Value calculation.

[0099] 1.3 Experimental Results

[0100] The results are as follows Figure 1 The cytotoxicity of ginsenoside Ra1 was evaluated on mouse colorectal cancer cell lines CT26 and MC38 and human colorectal cancer cell lines HCT116, Lovo, and HT29. IC 50 The values ​​were 0.8501μM, 1.484μM, 4.718μM, 3.370μM, and 4.613μM; the IC50 of Ra1 for CT26 cell line was 1961μM; the IC 50 The IC value of Ra1 for human HCT116 cell line is 1828 μM. 50 The IC value of Lovo cell line is 1471μM. 50 The IC value of HT29 cell line is 438.1μM. 50 The concentration of ginsenoside Ra1 was 1827 μM. Ginsenoside Ra1 had no cytotoxicity to various colorectal cancer cell lines. Compared with the positive control drug 5-FU, ginsenoside Ra1 had no direct killing effect on colorectal cancer cells.

[0101] Example 2 Effect of ginsenoside Ra1 on the growth of colorectal tumors in mice

[0102] 2.1 Experimental Materials

[0103] Mouse colorectal cancer cell line MC38 (maintained in our laboratory). 24 C57BL / 6J mice, male, SPF grade, 5 weeks old, weighing 20±2g. Purchased from Spaif (certificate number SCXK(Beijing)2019-0010). All mice were given free access to food. All animal experiments were carried out in strict accordance with the relevant regulations on animal use of 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).

[0104] Table 3 Experimental instruments

[0105]

[0106] Table 4 Experimental reagents

[0107]

[0108]

[0109] 2.2 Experimental Methods

[0110] 2.2.1 Preparation of test drugs

[0111] The required amount of test sample was calculated based on the most recent animal body weight before administration. Before administration, an appropriate amount of test sample was weighed, diluted with solvent and fixed to the required volume, and a solution of the required concentration was prepared (established configuration scheme: mother solution 400 mg ginsenoside Ra1 powder + 980 μl DMSO = 400 mg / ml, working solution: 3366 μl NS + 34 μl mother solution = 4 mg / ml, 1000 μl NS + 1000 μl 4 mg / ml colorless transparent solution Ra1 = 2 mg / ml, 1500 uL NS + 500 ul 4 mg / ml colorless transparent solution Ra1 = 1 mg / ml, 5-FU 3.6 mg powder + 1764 μl NS = 2 mg / ml.

[0112] The intraperitoneal administration was carried out at 10 μl / g, and the dosage was adjusted later according to the experimental changes).

[0113] 2.2.2 Experimental system

[0114] Animals: 24 C57BL / 6J mice, male, SPF grade, 6-8 weeks, weighing 20±2g, purchased from Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd., certificate number SCXK(Zhe)2019-0001. Cell line: Murine colon cancer cell MC38, preserved by our laboratory, subcultured in DMEM culture medium containing 10% fetal bovine serum, 100U / L penicillin and 100mg / L streptomycin at 37°C, 5% CO2 and saturated humidity in an incubator, and cells in the logarithmic growth phase were used for experiments.

[0115] 2.2.3 Model establishment

[0116] Construction of tumor-bearing mouse model: MC38 mouse colon cancer cells in logarithmic growth phase were collected and 10% matrix gel 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 routinely raised for 7-11 days and the tumor volume was 80-100 mm. 3 Tumor-bearing mice were randomly divided into groups.

[0117] 2.2.4 Experimental groups and drug administration

[0118] Grouping: The mice with qualified tumor volume were randomly divided into 8 groups (3 mice in each group): Group 8 was the model control group, and groups 1, 2, 3, 4, 5, 6, and 7 were the drug-administered groups. The specific scheme is shown in Table 5.

[0119] Table 5 Experimental groups and drug administration methods

[0120]

[0121] Drug administration: Drug administration began immediately after grouping. The drug was prepared with DMSO: saline = 1:99. The model control group DMSO: saline = 1:99. The drug was administered once a day for 20 consecutive times. 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 2000mm 3 The experiment was terminated.

[0122] 2.2.5 Experimental evaluation indicators and testing

[0123] 1) Tumor diameter: Tumor diameter is the main detection index, and it is measured once every 3-4 days. The relative tumor proliferation rate is calculated based on the measurement results, and the anti-tumor effect of the test substance is dynamically observed. Tumor volume (TV), TV = 1 / 2 × a × b 2 (where a and b are the longest diameter and the shortest diameter, respectively); relative tumor volume (RTV), RTV = Vt / V0 (where V0 is the tumor volume measured at d0 during group administration, and Vt is the tumor volume at each measurement). The anti-tumor effect evaluation index is the relative tumor proliferation rate 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). TGI (%) < 60% is ineffective; TGI (%) ≥ 60%, and P < 0.05 after statistical analysis is effective.

[0124] 2) Tumor weight: After the experiment, the animals were killed, the tumors were dissected and removed, the tumor weight was weighed, and then the tumors were photographed. If the average tumor weight of the model control group was less than 1g, or 20% of the tumor weight was less than 400mg, it means that the tumor growth was poor and the experiment was invalid. The tumor growth inhibition rate was calculated based on the weighing results. Tumor growth inhibition rate = [(average tumor weight of the model control group - average tumor weight of the treatment group) / average tumor weight of the model control group] × 100%. TGI (%) < 60% is invalid; TGI (%) ≥ 60%, and P < 0.05 after statistical processing is effective.

[0125] 3) Body weight: The animals were weighed every 3-4 days to observe the effect of the test drug on the animal body weight.

[0126] 2.2.6. Collect samples

[0127] At the end of the experiment, tumor tissues of 3 mice were collected from each of the best treatment groups, Group 1 Blank and Ginsenoside Ra1. The tumor size was about soybean size, and then made into single-cell suspension for CYTOF experiment. In addition, tumors of the remaining 3 experimental animals in each group were collected and stored in a -80℃ refrigerator for RNA-seq experiment. Spleens of 3 mice in each group were collected and placed on ice for flow cytometry detection of mCD4+, mCD8+ and mIFN-γ.

[0128] 2.2.7 Experimental data processing method

[0129] 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). In order to test the significance of each experimental data, an unpaired T test was used. P values ​​of 0.05 or above were not significant; P values ​​of 0.05 or less were considered to be significantly different.

[0130] 2.3 Experimental Results 1

[0131] In order 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 22-day treatment process, ginsenoside Ra1 was able to significantly inhibit the progression of subcutaneous colorectal cancer transplanted tumors. The results are as follows Figure 2 Compared with the solvent control group, the ginsenoside Ra1 group had a significant therapeutic effect. After the treatment, the tumor tissues of the mice were dissected and weighed, and it was found that the tumor weight of the mice in the high-concentration ginsenoside Ra1 group was significantly reduced. This result suggests that ginsenoside Ra1 has an anti-colorectal cancer effect.

[0132] 2.4 Experimental Results 2

[0133] In order to repeatedly verify the anti-colorectal cancer effect of ginsenoside monomer, the present invention sets Ra1 drug to treat C57 mouse MC38 colorectal cancer model, and the specific scheme is shown in Table 6. During the 22-day treatment process, ginsenoside Ra1 can significantly inhibit the progression of subcutaneous colorectal cancer transplanted tumors. The results are shown in Figure 3 Compared with the solvent control group, the ginsenoside Ra1 group had a significant therapeutic effect. After the treatment, the tumor tissues of the mice were dissected and weighed, and it was found that the tumor weight of the mice in the ginsenoside Ra1 group was significantly reduced. This result suggests that ginsenoside Ra1 has an anti-colorectal cancer effect.

[0134] Table 6 Experimental groups and drug administration methods II

[0135]

[0136] Example 3 Comparison of the efficacy of ginsenoside Ra1 and ginsenoside Rg3 in inhibiting the growth of colorectal tumors in mice

[0137] 3.1 Experimental Materials

[0138] The experimental materials are the same as those in Example 2.

[0139] 3.2 Experimental Methods

[0140] The experimental method is the same as that of Example 2.

[0141] 3.3 Experimental Results

[0142] The results are as follows Figure 4 As shown in the figure, the experiment found that the same dose of ginsenoside Rg3 had a weaker inhibitory effect on tumor growth than ginsenoside Ra1 monomer. Tumors were taken at the end of the experiment and found that the tumor weight of the ginsenoside Ra1 group was significantly different from that of the ginsenoside Rg3 group and the 5-Fu treatment group. At the same dose, ginsenoside Ra1 was more effective than ginsenoside Rg3 in inhibiting MC38 tumor growth.

[0143] Example 4 Exploration of the efficacy of ginsenoside Ra1 in inhibiting tumor growth in the HT29 tumor immunodeficient mouse model

[0144] 4.1 Experimental Materials

[0145] The experimental equipment is the same as that in Example 2, and the human colorectal cancer cell line HT29 (preserved in this laboratory) was used. 12 Babl / c-nunu mice, female, SPF grade, 5-6 weeks old, weighing 20±2g were purchased from Sibeifu Company (certificate number SCXK (Beijing) 2019-0010).

[0146] 4.2 Experimental Methods

[0147] The experimental method was the same as in Example 2, and the number of human HT29 cells inoculated was 1×10 6 indivual.

[0148] 4.3 Experimental Results

[0149] The present invention also established a tumor-bearing model of humanized HT29 cells in immunodeficient mice. 6 The cells were inoculated into the right back of nude mice, and group dosing began when the tumor was measurable. Figure 5 As shown in the results, the experimental results showed that 10mg / kg ginsenoside Ra1 had no effect on the growth of colorectal tumors after three weeks of continuous intraperitoneal administration, suggesting that the inhibitory effect of ginsenoside Ra1 on tumor growth mainly depends on the body's immune system. Ginsenoside Ra1 had no inhibitory effect on tumor growth in the HT29 immunodeficient mouse model.

[0150] Example 5 Effect of ginsenoside Ra1 on the growth of orthotopic tumors in syngeneic transplanted tumors in C57BL / 6J mice

[0151] 5.1 Experimental Materials

[0152] The experimental equipment was the same as in Example 2, and the mouse colorectal cancer cell line MC38-luc (maintained in this laboratory) was used. 12 C57BL / 6J mice, female, SPF grade, 5-6 weeks old, weighing 20±2g. They were purchased from Spaif (certificate number SCXK (Beijing) 2019-0010).

[0153] 5.2 Experimental Methods

[0154] The experimental method was the same as in Example 2, and the number of mouse MC38 cells inoculated subcutaneously was 1×10 6 indivual.

[0155] 5.3 Experimental Results

[0156] The occurrence and development of colon cancer is affected by many factors. In order to better simulate the occurrence and development of colon cancer in clinic, the present invention established an in situ colon cancer tumor model. Figure 6 As shown, ginsenosides have a significant inhibitory effect on orthotopic tumors. The results showed that compared with the control group, the tumors in the treated group were smaller in vivo fluorescence imaging, and the tumor weights of the treated group and the control group were significantly different at the end of the experiment. Ginsenoside Ra1 significantly inhibited the growth of orthotopic tumors in MC38 homologous transplanted tumors.

[0157] Example 6 Effect of ginsenoside Ra1 on liver metastasis of orthotopic tumors transplanted in Babl / c mice

[0158] 6.1 Experimental Materials

[0159] The experimental equipment is the same as that in Example 2, and the mouse colorectal cancer cell line CT-26-luc (preserved in this laboratory) is used. 10 Babl / c mice, female, SPF grade, 5-6 weeks old, weighing 20±2g. Purchased from Sibeifu Company (certificate number SCXK (Beijing) 2019-0010).

[0160] 6.2 Experimental Methods

[0161] The experimental method was the same as in Example 2, and the number of mouse CT26 cells inoculated subcutaneously was 1×10 6 indivual.

[0162] 6.3 Experimental Results

[0163] Tumor metastasis refers to the spread of tumor cells from the original site to the distant end, and is the main cause of tumor treatment failure and patient death. The molecular mechanism of tumor metastasis is complex, involving multiple steps, multiple stages, and changes in multiple genes. The liver is the most important target organ for hematogenous metastasis of colorectal cancer. Colorectal liver metastasis is one of the key points and difficulties in the treatment of colorectal cancer. 15%-25% of colorectal cancer patients have liver metastasis at the time of diagnosis, and another 15%-25% of patients may develop liver metastasis after radical resection of the primary colorectal cancer lesion. The vast majority (80%-90%) of liver metastases cannot be radically removed initially. Liver metastasis is also the main cause of death in patients with colorectal cancer. The liver is the most common distant metastasis organ of the colorectal cancer, which is related to the unique anatomical structure of the liver.

[0164] The portal vein system of the liver collects venous blood from the abdominal organs. The blood velocity of venous blood is slow when passing through the liver. Tumor cells can easily invade blood vessels to form tumor thrombi and stay and grow. The structure of the liver provides good conditions for cancer cells to implant. The liver metastasis rate of tumors in different parts of the colorectum is different. The liver metastasis rate of transverse colon cancer and sigmoid colon cancer is higher than that of other parts. The process of liver metastasis can be divided into several consecutive steps. First, cancer cells in the primary tumor focus secrete angiogenic factors to cause vasodilation. After the local action of activated proteases, cancer cells enter thin-walled veins and lymphatic vessels and then enter the blood circulation. Most circulating cancer cells are attacked and destroyed by the immune system and blood flow shear force in this process. Surviving cancer cells or clumps form emboli to block the capillary bed of distant organs. Finally, cancer cells extravasate into the liver parenchyma and form micrometastases.

[0165] The present invention established an allogeneic colorectal tumor liver metastasis model on the BABL / c mouse background. After two weeks of continuous administration, it was found that ginsenoside Ra1 not only significantly inhibited the growth of the in situ tumor, but also inhibited the metastasis of the colorectal tumor to the liver. The mouse liver was fixed in 4% formaldehyde solution and then stained with HE. The results are as follows Figure 7 As shown. Ginsenoside Ra1 significantly inhibits liver metastasis of colorectal cancer.

[0166] Example 7 Study on the effect of ginsenoside Ra1 on melanoma tumor model

[0167] 7.1 Experimental Materials

[0168] The experimental equipment is the same as that in Example 2, and the mouse colorectal cancer cell line B16F10 (maintained in this laboratory). 10 C57BL / 6j mice, female, SPF grade, 5-6 weeks old, weighing 20±2g. Purchased from Sibeifu Company (certificate number SCXK (Beijing) 2019-0010).

[0169] 7.2 Experimental Methods

[0170] The experimental method was the same as in Example 2. C57BL / 6j mice were subcutaneously inoculated with 5×10 mouse melanoma B16F10 cells. 5 After tumor formation, the drug was injected into the tail vein at a dose of 5 mg / kg per day.

[0171] 7.3 Experimental Results

[0172] In order to explore the effect of Ra1 on melanoma growth, a tumor-bearing mouse model was established, and LLC-luc mouse lung cancer cells and B16F10 mouse melanoma cells in the logarithmic growth phase were collected. The cell suspension concentration was adjusted to 5×10 6 The cells were inoculated into the right side of the back of C57BL / 6J mice by subcutaneous injection at a volume of 0.10 ml per mouse (5 × 10 cells per mouse) under sterile conditions. 5 The SPF animals were routinely raised for 5 to 10 days, and the tumor volume of the experimental animals was 50-100 mm. 3 After randomization and administration, the growth of subcutaneous tumors was monitored. Figure 8 As shown, the results showed that ginsenoside Ra1 had no significant effect on melanoma growth.

[0173] Example 8 Study on the Anti-liver Cancer Effect of Ginsenoside Ra1

[0174] 8.1 Experimental Materials

[0175] The experimental equipment was the same as in Example 2, and the mouse colorectal cancer cell line Heap1-6-luc (preserved in this laboratory) was used. 10 C57BL / 6j mice, female, SPF grade, 5-6 weeks old, weighing 20±2g. They were purchased from Spaif (certificate number SCXK (Beijing) 2019-0010).

[0176] 8.2 Experimental Methods

[0177] The experimental method was the same as in Example 2. C57BL / 6j mice were subcutaneously inoculated with 3×10 Heap1-6-luc cells of murine hepatocellular carcinoma 5 indivual.

[0178] 8.3 Experimental Results

[0179] After tumor formation, C57BL / 6J mice were subjected to fluorescence in vivo imaging. Before imaging, mice were anesthetized by intraperitoneal injection of 4.16 mg / mL of Shutai 50 and 0.416 mg / mL of xylazine hydrochloride, with a dosing volume of 10 mL / kg. After intraperitoneal injection of 150 mg / mL of fluorescein potassium salt substrate, with a dosing volume of 10 mL / kg, fluorescence in vivo imaging was performed 10 minutes after the injection of fluorescein potassium salt substrate, and the fluorescence numerical analysis at 30 seconds of imaging time was statistically analyzed. The experimental results are shown in Fig. 9 As shown, ginsenoside Ra1 had no effect on the progression of lung cancer.

[0180] Results: (1) Ra1 had no direct cytotoxic effect on human and mouse colorectal cancer cell lines in in vitro cell experiments; (2) Compared with other saponin drugs, Ra1 can effectively inhibit tumor growth in MC38 tumor-bearing mouse model, and Ra1's ability to inhibit tumor growth in tumor-bearing mice is better than other saponin drugs; (3) Compared with ginsenoside Rg3, ginsenoside Ra1 has a more significant inhibitory effect on the growth of colorectal tumors, and has no organ toxicity and no adverse reactions; (4) In the HT29 immunodeficient nude mouse tumor model, the use of Ra1 treatment found that tumor growth was not affected by the drug, suggesting that Ra1 relies on immune pathways to prevent and treat digestive diseases. (5) Ginsenoside Ra1 was found to significantly inhibit the growth of colorectal tumors by administering the drug to the orthotopic tumor inoculation model of C57BL / 6J syngeneic transplant tumors; (6) Ginsenoside Ra1 was found to significantly inhibit the growth of colorectal tumors and the occurrence of liver metastasis by administering the drug to the orthotopic tumor liver metastasis model of Babl / c mouse syngeneic transplant tumors; (7) By establishing a B16F10 subcutaneous melanoma model in C57BL / 6j mice, it was found that ginsenosides had no effect on the growth of melanoma; (8) By establishing a Heap1-6-luc liver cancer orthotopic model in C57BL / 6j mice, it was found that ginsenosides had no effect on the progression of orthotopic liver cancer.

[0181] 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 used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. Use of ginsenoside Ra1 or its salt as an active ingredient in the preparation of a medicament for preventing and / or treating colorectal cancer.

2. Use of ginsenoside Ra1 or its salt as an active ingredient in the preparation of a drug for inhibiting liver metastasis of colorectal cancer.

3. The use according to claim 1 or 2, characterized in that: The ginsenoside Ra1 or its salt is a solution of ginsenoside Ra1 or its salt. The method for preparing the solution of ginsenoside Ra1 or its salt comprises: weighing an appropriate amount of ginsenoside Ra1 or its salt, adding a reagent to dissolve and dilute it, and obtaining the solution of ginsenoside Ra1 or its salt.

4. The use according to claim 3, characterized in that The reagents are 1% dimethyl sulfoxide and 99% physiological saline.

5. Use of a pharmaceutical preparation comprising ginsenoside Ra1 or a salt thereof and a pharmaceutically acceptable excipient in the preparation of a medicament for preventing and / or treating colorectal cancer.

6. Use of a pharmaceutical preparation comprising ginsenoside Ra1 or a salt thereof and a pharmaceutically acceptable excipient in the preparation of a drug for inhibiting liver metastasis of colorectal cancer.

7. The use according to claim 5 or 6, 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.

8. The use according to claim 5 or 6, characterized in that: The pharmaceutical preparation is in the form of powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

9. The use according to claim 1, 2, 5 or 6, characterized in that The pharmaceutical concentration of the ginsenoside Ra1 is 0.1-10 mg / ml.

10. The use according to claim 9, characterized in that The pharmaceutical concentration of ginsenoside Ra1 is about 1 mg / ml.

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