Application of Muriaculum gardroncarteri in preparation of medicine for preventing and / or treating colorectal cancer and application of Muriaculum gardroncarteri in preparation of medicine for preventing and / or treating colorectal cancer

Abstract

The invention discloses an application of Muriaculum gardroncarteri in preparation of a medicine for preventing and / or treating colorectal cancer, and belongs to the field of prevention and treatment of the colorectal cancer. The application comprises the following steps: (1) application of the Muriaculum gardroncarteri in preparation of the medicine, wherein the medicine is used for preventing and / or treating the colorectal cancer, and (2) application of the Muriaculum gardroncarteri in preparation of the medicine; (2) an application of the Muriaculum cordoncarter detection reagent in the preparation of a kit for diagnosing the colorectal cancer or evaluating the prognosis of the colorectal cancer, and an application of the Muriaculum cordoncarter detection reagent in the preparation of a kit for diagnosing the colorectal cancer; and (3) application of the Muriaculum gardroncarter detection reagent in preparation of a kit for predicting the curative effect of the anti-PD-1 (programmed death-1), and application of the Muriaculum gardroncarter detection reagent in preparation of the kit. And (4) an application of the Muriaculum gardroncarter i in the preparation of an auxiliary reagent for improving the curative effect of the anti-PD-1 on the colorectal cancer.

Description

Technical Field

[0001] The present invention relates to the field of prevention and treatment of colorectal cancer, and specifically relates to the application of Muribaculum gordoncarteri in the preparation of drugs for preventing and / or treating colorectal cancer. Background Art

[0002] According to the national cancer data statistical analysis of the National Cancer Center in 2024, among all cancer types, the incidence and mortality of colorectal cancer are both among the top. Due to the lack of obvious early symptoms, about 20% of colorectal cancer patients are diagnosed at the middle and advanced stages, and the 5-year survival rate after diagnosis is less than 20%. Colorectal cancer can be treated by surgery and non-surgical methods. Common non-surgical therapies mainly include chemotherapy, targeted therapy, and immunotherapy, etc. Developing new treatment methods can provide new effective options for the treatment of colorectal cancer. Summary of the Invention

[0003] In view of the deficiencies of the prior art, the present invention proposes the application of Muribaculum gordoncarteri in the preparation of drugs for preventing and / or treating colorectal cancer.

[0004] The object of the present invention can be achieved by the following technical solutions:

[0005] The first aspect of the present invention relates to an application, including any one of the following applications:

[0006] (1) The application of Muribaculum gordoncarteri in the preparation of a drug for preventing and / or treating colorectal cancer;

[0007] (2) The application of a Muribaculum gordoncarteri detection reagent in the preparation of a kit for diagnosing colorectal cancer or evaluating the prognosis of colorectal cancer;

[0008] (3) The application of a Muribaculum gordoncarteri detection reagent in the preparation of a kit for predicting the efficacy of anti-PD-1;

[0009] (4) The application of Muribaculum gordoncarteri in the preparation of an adjuvant reagent for enhancing the efficacy of anti-PD-1 against colorectal cancer.

[0010] According to an embodiment of the present invention, the colorectal cancer includes colon cancer and rectal cancer.

[0011] According to an embodiment of the present invention, the drug is used to inhibit the increase in the volume of colorectal cancer tumors or the proliferation of tumor cells.

[0012] According to an embodiment of the present invention, the drug is used to inhibit the increased infiltration of myeloid-derived suppressor cells (MDSC) in tumors caused by colorectal cancer.

[0013] According to an embodiment of the present invention, the drug is used to inhibit the reduction of CD8 + T cell infiltration in tumors caused by colorectal cancer.

[0014] According to an embodiment of the present invention, the drug is used to inhibit the reduction of activated CD8 + T cell infiltration in tumors caused by colorectal cancer.

[0015] According to an embodiment of the present invention, the drug is used to increase the content of the anti-tumor substance urocanic acid in the feces and / or serum of colorectal cancer mice and / or patients.

[0016] According to an embodiment of the present invention, the drug is used to increase the content of the anti-tumor substance urocanic acid in the tumors of colorectal cancer mice and / or patients.

[0017] According to an embodiment of the present invention, the drug is used to increase the anti-tumor efficacy of anti-PD-1 in colorectal cancer mice and / or patients.

[0018] According to an embodiment of the present invention, the drug is administered orally or by gavage.

[0019] According to an embodiment of the present invention, the dosage of the drug is 0.5 - 1.5×10 9 CFU Muribaculum gordoncarteri per day.

[0020] The second aspect of the present invention relates to a drug for treating colorectal cancer, comprising Muribaculum gordoncarteri and anti-PD-1.

[0021] The third aspect of the present invention relates to a dietary therapy product, comprising Muribaculum gordoncarteri.

[0022] Advantages of the present invention:

[0023] By administering Muribaculum gordoncarteri to colorectal cancer mice, the present invention can effectively relieve the symptoms of colorectal cancer. Thus, it is revealed that Muribaculum gordoncarteri has the efficacy of preventing and / or treating colorectal cancer, is safe to use, has good application prospects, and provides new ideas for the prevention and treatment of colorectal cancer clinically. Description of the Drawings

[0024] The present invention will be further described below in conjunction with the accompanying drawings.

[0025] Figure 1 It shows the experimental flow chart of subcutaneous transplanted tumor of mouse MC38 in Example 1 and the graph of tumor volume change;

[0026] Figure 2 It shows the graph of the infiltration change of MDSC in the colorectal tumor of the mouse in Example 1;

[0027] Figure 3 It shows the infiltration change graph of CD8 + T in the colorectal tumor of the mouse in Example 1;

[0028] Figure 4 It shows the infiltration change graph of activated CD8 + T in the colorectal tumor of the mouse in Example 1;

[0029] Figure 5 It shows the graph of the change in the content of urocanic acid in the feces and serum of the mouse in Example 1;

[0030] Figure 6 It shows the graph of the change in the content of urocanic acid in the colorectal tumor of the mouse in Example 1;

[0031] Figure 7 It shows the graph of the change in the abundance of Muribaculum gordoncarteri in patients with response (R) and non-response (NR) to anti-PD-1 antibody therapy in Example 2;

[0032] Figure 8 It shows the graph of the relationship between the abundance of Muribaculum gordoncarteri and the overall survival period in colorectal cancer patients in Example 2;

[0033] Figure 9 It shows the graph of the change in the tumor volume of subcutaneous transplanted tumor of mouse MC38 in Example 3;

[0034] Figure 10 It shows the procurement record of the strain Muribaculum gordoncarteri DSM108194 used in the examples of the present application. Detailed implementation manners

[0035] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts shall fall within the protection scope of the present invention.

[0036] In the embodiments of the present application, the bacterium used is Muribaculum gordoncarteri DSM 108194, which was purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ). For the specific procurement source, procurement time and procurement records, please refer to Figure 10 , it can be understood that in order to protect the privacy information of citizens, information such as citizen names, contact information, addresses, email addresses, bank accounts, etc. has been covered, while necessary information such as the procurement source and procurement time of this bacterial strain has been retained.

[0037] Example 1

[0038] Female C57BL / 6 mice aged 6 - 8 weeks and weighing 18 - 20 g were used. 18 mice were randomly divided into 3 groups: a model group (CM group), an Escherichia coli negative control group (E.coli group), and a Muribaculum gordoncarteri intervention group (M.gordoncarteri group).

[0039] The feeding method for the mice in the CM group was as follows: free diet, drinking antibiotic water (1 mg / mL ampicillin, 1 mg / mL colistin, 5 mg / mL streptomycin, the purpose was to remove most of the microorganisms in the intestine), and intragastrically administrating 0.2 mL of blank medium two to three times a week;

[0040] The feeding method for the mice in the E.coli group was as follows: free diet, drinking antibiotic water (1 mg / mL ampicillin, 1 mg / mL colistin, 5 mg / mL streptomycin, the purpose was to remove most of the microorganisms in the intestine), and intragastrically administrating 0.2 mL of E.coli (5×10 8 CFU) two to three times a week;

[0041] The feeding method for the mice in the M.gordoncarteri group was as follows: free diet, drinking antibiotic water (1 mg / mL ampicillin, 1 mg / mL colistin, 5 mg / mL streptomycin, the purpose was to remove most of the microorganisms in the intestine), and intragastrically administrating 0.2 mL of Muribaculum gordoncarteri (5×10 8 CFU) two to three times a week.

[0042] The behavior and health status of the mice were observed daily, and the changes in the body weight and tumor volume of the mice were recorded. The results are as Figure 1; After the experiment, the mice were sacrificed, and the tumors were collected. The tumor tissues were digested into single-cell suspensions and subjected to flow cytometry staining. The infiltration ratio of MDSC in the tumors was analyzed by flow cytometry. The results are shown in Figure 2 ; The infiltration ratio of CD8 + T cells in the tumors was analyzed by flow cytometry. The results are shown in Figure 3 ; The infiltration ratio of activated CD8 + T cells in the tumors was analyzed by flow cytometry. The results are shown in Figure 4 ; After the experiment, the mice were sacrificed by blood collection, and the mouse serum, intestinal contents, and tumors were collected. The content of urocanic acid in the mouse serum and intestinal contents was analyzed by LC-MS / MS. The results are shown in Figure 5 ; The content of urocanic acid in the mouse tumors was analyzed by LC-MS / MS. The results are shown in Figure 6 .

[0043] Figure 1 The results showed that the tumor growth rates in the CM group and the E. coli group were relatively fast, and the growth of mouse tumors could be significantly inhibited after intervention with Muribaculum gordoncarteri (P < 0.05).

[0044] Figure 2 The results showed that the infiltration ratio of MDSC in the tumors in the CM group and the E. coli group was relatively high, reaching 30%, and the infiltration ratio of MDSC in the mouse tumors could be significantly reduced after intervention with Muribaculum gordoncarteri, which could be reduced to 10% (P < 0.05).

[0045] Figure 3 The results showed that the infiltration ratio of CD8 + T cells in the tumors in the CM group and the E. coli group was relatively low, and the infiltration ratio of CD8 + T cells in the mouse tumors could be significantly increased after intervention with Muribaculum gordoncarteri (P < 0.05).

[0046] Figure 4 The results showed that the infiltration ratio of activated CD8 + T cells (IFN-γ + , GZMB + , Perforin + ) in the tumors in the CM group and the E. coli group was relatively low, and the infiltration ratio of activated CD8 + T cells (IFN-γ + , GZMB + , Perforin + ) in the mouse tumors could be significantly increased after intervention with Muribaculum gordoncarteri (P < 0.05).

[0047] Therefore Figure 3 、 Figure 4 The results of + suggest that Muribaculum gordoncarteri contributes to the activation of intratumoral CD8 + T cells, thereby enhancing the anti-tumor effect and the survival rate of colorectal cancer patients.

[0048] Figure 5 The results showed that the content of urocanic acid in the feces and serum of the CM group and the E. coli group was lower, and after intervention with Muribaculum gordoncarteri, the content of urocanic acid in the feces and serum of mice increased significantly (P<0.05).

[0049] Figure 6 The results showed that the content of urocanic acid in the tumors of mice in the CM group and the E. coli group was lower, and after intervention with Muribaculum gordoncarteri, the content of urocanic acid in the tumors of mice increased significantly (P<0.05).

[0050] In the above detection results, urocanic acid is used to reflect the efficacy of PD-1 antibody (for example, patent document CN116068160A, a biomarker related to the efficacy of PD-1 antibody and its application). As Figure 5 with Figure 6 shown, the increase in urocanic acid in the feces, serum and tumors of Muribaculum gordoncarteri, Example 2

[0051] Collect the feces of colorectal cancer patients receiving anti-PD-1 treatment, divide them into a response group (R) and a non-response (NR) group according to whether the colorectal cancer patients respond to anti-PD-1, and use real-time fluorescence quantitative PCR technology to detect the abundance of Muribaculum gordoncarteri in the feces of patients. The results are as Figure 7 ; according to the high and low abundance of Muribaculum gordoncarteri in the feces of colorectal cancer patients, the patients are divided into a Muribaculum gordoncarteri low-abundance group (Low) and a Muribaculum gordoncarteri high-abundance group (High). The difference in the overall survival rate of colorectal cancer patients between the two groups is as Figure 8 .

[0052] Figure 7 The results showed that the feces of anti-PD-1 therapy-responsive patients had a higher abundance of Muribaculum gordoncarteri (P<0.05).

[0053] Figure 8The results showed that patients with a higher abundance of Muribaculum gordoncarteri in their feces had a longer survival period (P<0.05). The results of this trial implied that the intake of Muribaculum gordoncarteri could improve the survival rate and prognosis of colorectal cancer patients. On the other hand, it also showed that by detecting the content of Muribaculum gordoncarteri in the feces or intestines of colorectal cancer patients, the prognosis of colorectal cancer patients could be predicted or evaluated. Therefore, the experimental conclusion of this example revealed that detection products related to Muribaculum gordoncarteri could be used as reagents for prognostic assessment of colorectal cancer patients. On the other hand, although this application did not provide a toxicity test, the improvement in the survival rate of mice in this example also implied to a certain extent that the health status of mice was not negatively affected after ingesting Muribaculum

[0054] gordoncarteri, indicating that Muribaculum

[0055] gordoncarteri had potential medicinal value.

[0056] Example 3

[0057] Female C57BL / 6 mice aged 6 - 8 weeks with a body weight of 18 - 20 g were used. Twenty mice were randomly divided into 4 groups: a solvent control group (Control group), an anti-PD-1 single administration group (anti-PD-1 group), a Muribaculum gordoncarteri single administration group (M. gordoncarteri group), and an anti-PD-1 and Muribaculum gordoncarteri combination administration group (anti-PD-1 + M. gordoncarteri group).

[0058] The feeding method for mice in the Control group was: free diet and water, and gavage with 0.2 mL of phosphate buffer solution (PBS) two to three times a week;

[0059] The feeding method for mice in the anti-PD-1 group was: free diet and water, and intraperitoneal injection of 200 μg anti-PD-1 antibody per mouse twice a week;

[0060] The feeding method for mice in the M. gordoncarteri group was: free diet and water, and gavage with 0.2 mL of Muribaculum gordoncarteri (5×10 8 CFU) two to three times a week;

[0061] The feeding method for the mice in the anti-PD-1 + M. gordoncarteri group was as follows: they had free access to food and water, were intraperitoneally injected with 200 μg of anti-PD-1 antibody per mouse twice a week, and were gavaged with 0.2 mL of Muribaculum gordoncarteri (5×10 8 CFU) two to three times a week.

[0062] The behavior and health status of the mice were observed daily, and the changes in the body weight and tumor volume of the mice were recorded. The results of the tumor volume were as Figure 9 .

[0063] Figure 9 The results showed that Muribaculum gordoncarteri could increase the anti-colorectal cancer efficacy of anti-PD-1 (P < 0.05).

[0064] Combined with Figure 7 and Figure 9 the results, it can be seen that detecting the content of Muribaculum gordoncarteri in the intestine or feces can be used to evaluate the efficacy of anti-PD-1 or the response rate / sensitivity of patients to anti-PD-1 therapy, and thus can predict in advance or assist in decision-making whether anti-PD-1 therapy should be implemented for patients.

[0065] Of course, those skilled in the art should be aware that in the actual process of implementing the above prediction or auxiliary decision-making, it is not limited to using the Muribaculum gordoncarteri level as the only prediction / detection index. In order to improve the accuracy and specificity of prediction or evaluation, other known markers for predicting the efficacy of anti-PD-1 can also be combined.

[0066] On the other hand, Muribaculum gordoncarteri can be used as a sensitizer or auxiliary reagent and combined with anti-PD-1 to obtain better efficacy. Therefore, Muribaculum gordoncarteri can be used as a potential auxiliary component of PD-1 drugs and added to the anti-PD-1 drug treatment plan to form a more effective drug combination.

[0067] Combined with the above embodiments, those skilled in the art should be aware that in addition to being used as a drug component, Muribaculum gordoncarteri should also be able to be used as a probiotic by being incorporated into food therapy products,

[0068] After being ingested by the human body, it can achieve the effects of treating colorectal cancer, enhancing the efficacy of anti-PD-1, or preventing colorectal cancer. For example, Muribaculum gordoncarteri can combine with known traditional Chinese medicine or Western medicine ingredients that are helpful for anti-tumor, functional ingredients beneficial to recovery or prognosis, ingredients that can relieve the side effects of chemotherapy / radiotherapy, etc., to form a dietary therapy product. Of course, these dietary therapy products can be a solid food or powder, or a drink.

[0069] In summary, the colonization of Muribaculum gordoncarteri effectively inhibited the growth of colorectal cancer in mice, reduced the infiltration of MDSC in tumors, and increased the infiltration of CD8 + T cells and activated CD8 + T cells (IFN-γ + , GZMB + , Perforin + ). It increased the content of urocanic acid in the feces, serum, and tumors of colorectal cancer mice. Muribaculum gordoncarteri had a high abundance in the feces of anti-PD-1 therapy-responsive patients. Colorectal cancer patients with a high abundance of Muribaculum gordoncarteri in their feces had a longer overall survival. Muribaculum gordoncarteri could increase the anti-colorectal cancer efficacy of anti-PD-1. The above results provide a theoretical basis for Muribaculum gordoncarteri as a probiotic for the prevention and treatment of colorectal cancer.

[0070] In the description of this specification, the description referring to terms such as "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0071] The above shows and describes the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. What is described in the above embodiments and the specification only illustrates the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed.

Claims

1. An application, characterized in that, Comprising any one of the following applications: (1) Application of Muribaculum gordoncarteri in the preparation of a drug for preventing and / or treating colorectal cancer; (2) Application of a Muribaculum gordoncarteri detection reagent in the preparation of a kit for diagnosing colorectal cancer or evaluating the prognosis of colorectal cancer; (3) Application of a Muribaculum gordoncarteri detection reagent in the preparation of a kit for predicting the efficacy of anti-PD-1; (4) Application of Muribaculum gordoncarteri in the preparation of an auxiliary reagent for enhancing the efficacy of anti-PD-1 against colorectal cancer.

2. The application according to claim 1, wherein The colorectal cancer includes colon cancer and rectal cancer.

3. The application according to claim 1, wherein The drug is used to inhibit the increase in the volume of colorectal cancer tumors or the proliferation of tumor cells.

4. The application according to claim 1, wherein The drug is used to inhibit the increase in the infiltration of myeloid-derived suppressor cells in tumors caused by colorectal cancer.

5. The application according to claim 1, characterized in that, The drug is used to inhibit the reduction of CD8 + T cell infiltration in the tumor caused by colorectal cancer.

6. The application according to claim 1, wherein The drug is used to inhibit the reduction of infiltrating activated CD8 + T cells in tumors caused by colorectal cancer.

7. The application according to claim 1, characterized in that, The drug is used to increase the content of urocanic acid in the feces, serum, and / or tumors of mice and / or patients with colorectal cancer.

8. The application according to claim 1, characterized in that The dosage of the drug is 0.5 to 1.5×10 9 CFUMuribaculum gordoncarteri per day.

9. A drug for treating colorectal cancer, characterized in that, Comprising Muribaculum gordoncarteri and anti-PD-1.

10. A diet therapy product, characterized in that, Comprising Muribaculum gordoncarteri.

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