Microbial strain of lachnospiraceae, drug for preventing or treating tumors and use

Abstract

Provided are a microbial strain of Lachnospiraceae, a drug for preventing and / or treating at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases and inflammatory bowel diseases, and the use. The microbial strain of Lachnospiraceae is a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae, the nucleotide sequence of 16S rDNA is as shown in SEQ ID No. 1, and the microbial strain of the Lachnospiraceae can inhibit the growth rate of tumors and can be used for the prevention and / or treatment of tumors. The tumors include at least one of the following: liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer and fibrosarcoma.

Description

Lachnospiraceae microbial strains, drugs for preventing or treating tumors, and applications thereof Technical Field

[0001] The present invention relates to the technical field of microorganisms and their applications, in particular to Lachnospiraceae microbial strains, drugs for preventing or treating tumors and their applications. Background Art

[0002] There are a large number of symbiotic microorganisms in the human intestine, and the total amount of genetic information they carry is 50 to 100 times that of the human genome. This is the "gut microbiome", also known as the "second genome" of humans. The intestinal microbiome is the largest and most direct external environment of the human body and plays an indispensable role in maintaining human health. At present, there have been research reports on the role of the intestinal microbiome in nutritional disorders, metabolic abnormalities and complex diseases (such as obesity, diabetes, inflammatory bowel disease and tumors).

[0003] In recent years, with the rapid development of molecular biology, genomics, bioinformatics analysis technology, high-throughput sequencing technology, and microbial culture technology, the impact and role of the intestinal flora on intestinal and extraintestinal diseases has become increasingly clear. The intestinal flora is more like an organ with metabolic, immune, and endocrine functions, which can affect the human digestive system, circulatory system, and nervous system, and is closely related to the occurrence of various chronic diseases (diabetes, hypertension, cardiovascular disease, brain disease, etc.) and tumors in humans. Studying the relationship between the intestinal flora and human health and disease is not only an important scientific research, but also has important significance and value in clinical diagnosis, treatment, and even translation.

[0004] Gut microbial imbalance increases the incidence of colorectal cancer, while some metabolites of intestinal microorganisms can directly slow carcinogenesis or inhibit tumor development. In addition to intestinal-related colon and rectal cancers, intestinal microbiota can also affect breast cancer, liver cancer, and other diseases. There is growing evidence that intestinal microbiota can influence tumor formation, tumor development, and tumor treatment. How to regulate the immune and inflammatory responses caused by intestinal microbiota so that it does not induce tumors while enhancing the efficacy of anti-tumor drugs is still under discussion. If these problems can be solved, it will have great significance for future innovation and transformation in the field of tumor treatment.

[0005] Michael Scharl et al. reported in Cell Host Microbe that Clostridium is associated with low tumor burden, demonstrating that a mixture of commensal Clostridium strains can be expressed through CD8 + T cells produce a strong anti-tumor effect, and by using a variety of solid tumor models, the feasibility of using specific intestinal bacteria as a single agent in cancer treatment was demonstrated. Clostridium mixture CC4 can activate CD8 +T cells simultaneously downregulate immunosuppressive factors to exert anti-cancer effects. Furthermore, CC4's ability to increase CD8+ T cell infiltration places the tumor in a highly immunogenic state, potentially improving the response rate of CRC patients to aPD-1 therapy. These findings open a new chapter in the development of gut microbiota supplementation as an independent therapeutic approach.

[0006] The short-chain fatty acids (SCFAs) valerate and butyrate can enhance the antitumor activity of cytotoxic T lymphocytes (CTLs) and chimeric antigen receptor (CAR) T cells through metabolic and epigenetic reprogramming. This SCFA is a rare bacterial metabolite produced by low-abundance commensals such as Megasphaera massiliensis, classifying it as a valerate-producing bacterial species. Interestingly, dominant commensal bacteria are unable to produce valerate. Studies have shown that in vitro treatment of CTLs and CAR T cells with valerate and butyrate increases the function of mTOR, a central cellular metabolic sensor, and inhibits class I histone deacetylase activity. This reprogramming leads to increased production of effector molecules such as CD25, IFN-γ, and TNF-α and significantly enhances the antitumor activity of antigen-specific CTLs and ROR1-targeting CAR T cells in syngeneic mouse models of melanoma and pancreatic cancer. These experimental data reveal microbial molecules that can be used to enhance cellular antitumor immunity and support the identification of valerate and butyrate as two SCFAs with therapeutic utility in cellular cancer immunotherapy.

[0007] Therefore, further discovery, exploration and research of microbial strains with the potential for tumor prevention or treatment have important application value and market prospects.

[0008] In view of this, the present invention is proposed.

[0009] Summary of the Invention

[0010] The present invention aims to provide a Lachnospiraceae microbial strain, a drug for preventing or treating tumors, and its application. The present invention has discovered that a new genus-level intestinal anaerobic microbial strain, Lachnospiraceae sp., can inhibit tumor growth and can be used for the prevention and / or treatment of tumors.

[0011] The technical solutions provided by the present invention are as follows:

[0012] In one aspect, the present invention provides the use of a Lachnospiraceae microbial strain in the preparation of a drug for preventing and / or treating tumors, wherein the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family, and the nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown in SEQ ID No. 1; the tumor includes at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer and fibrosarcoma.

[0013] In one embodiment, the use of a Lachnospiraceae microbial strain in the preparation of a drug for preventing and / or treating tumors includes Lachnospiraceae microbial strain MNH 46686, which is deposited in the Guangdong Provincial Microbial Culture Collection Center with the deposited name Lachnospiraceae sp. MNH 46686 and the strain deposit number GDMCC No: 62002. The present invention covers the use of bacterial cells of Lachnospiraceae microbial strains, extracts thereof, and metabolites (e.g., fermentation broth) in the preparation of drugs for preventing and / or treating tumors, as well as their use in the preparation of drugs for inhibiting tumor growth rate (inhibiting tumor size and dimension).

[0014] In one embodiment, the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family, and the nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown as SEQ ID No. 1.

[0015] The strain of the present invention has anti-tumor activity and can significantly inhibit the growth rate of tumors.

[0016] The Lachnospiraceae microbial strain of the present invention has the following biological characteristics:

[0017] (1) Bacterial characteristics: The strain can produce spores, has no flagella, is non-motile, and is rod-shaped, approximately 2-5 μm × 20-50 μm in length; the strain is Gram-negative;

[0018] (2) Colony characteristics: After inoculation on anaerobic blood plate culture medium and anaerobic culture at 37°C for 48 h, visible colonies are formed on the anaerobic blood plate culture medium; the colonies are round, with regular and smooth edges, about 0.5 mm in diameter, light yellow, opaque, and no secretions are formed around the colonies;

[0019] (3) Growth characteristics and physiological and biochemical characteristics: The growth temperature range is 30-42°C, with the optimum growth at 37°C; it can grow in the pH range of 7.0-9.0, with the optimum growth pH being 7.0; it will not grow on media with a NaCl content exceeding 2%; strain MNH 46686 can survive and grow in a bile salt concentration range of 0%-0.3%, but will not grow when the bile salt concentration is greater than or equal to 0.4%;

[0020] (4) Other characteristics: It is resistant to erythromycin, ciprofloxacin, co-trimoxazole, ampicillin, ceftriaxone and lincomycin; it is sensitive to antibiotics such as gentamicin, chloramphenicol, tetracycline and penicillin; it can synthesize a large amount of short-chain fatty acids during growth: acetic acid, isobutyric acid, butyric acid and isovaleric acid.

[0021] In another aspect, the present invention provides a microbial agent comprising the aforementioned Lachnospiraceae microbial strain of the present invention or a metabolite of the Lachnospiraceae microbial strain.

[0022] In another aspect, the present invention provides a drug for preventing and / or treating tumors, comprising the aforementioned Lachnospiraceae microbial strain or a metabolite of the Lachnospiraceae microbial strain.

[0023] In one embodiment, the drug further comprises a pharmaceutically acceptable carrier; preferably, the carrier is selected from one or more of a diluent, a dispersant, an excipient, a stabilizer, a lubricant, and a disintegrant; the excipients include mannitol, lactose, starch, microcrystalline cellulose, and the like. The disintegrants include polyvinyl pyrrolidone, carboxymethyl cellulose, sodium carboxymethyl cellulose, and the like. The lubricants include talc, magnesium stearate, and the like.

[0024] Preferably, the drug is in the form of a liquid preparation, a solid preparation, a capsule preparation, a sustained-release preparation, and a nanoformulation. For example, the drug is in the form of an injection, a tablet, a granule, a pill, a capsule, and the like.

[0025] In one embodiment, the composition further comprises a combination drug; the combination drug comprises at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0026] In one embodiment, the chemotherapy drug includes one or more of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, or epirubicin;

[0027] The photosensitizer includes one or more of boron dipyrrole, dihydrochlorin or red bengal;

[0028] The photothermal agent is one or more of indocyanine green, new indocyanine green or gold nanoparticle rods.

[0029] In one embodiment, the drug of the present invention can be used in combination with other anti-tumor drugs, such as but not limited to oxaliplatin, mitoxantrone, doxorubicin, epirubicin, etoposide, vinorelbine or methotrexate.

[0030] In one embodiment, the drug of the present invention can be used together with an immune adjuvant, such as CpG, IL-2, aluminum hydroxide, and the like.

[0031] In one embodiment, the immunotherapy drug includes PD-1 antibody, CTLA-4 antibody, PD-L1 antibody, PD-L1 inhibitor;

[0032] Preferably, the PD-L1 inhibitor is selected from durvalumab, atezolizumab or avelumab; the PD-1 antibody or PD-L1 antibody is selected from pembrolizumab or nivolumab; and the CTLA-4 antibody is selected from ipilimumab.

[0033] In addition, the present invention provides a drug for preventing or treating tumors, comprising a Lachnospiraceae microorganism and a pharmaceutically acceptable excipient or adjuvant or liquid preparation; the 16s rRNA sequence of the Lachnospiraceae microorganism is shown in SEQ ID NO: 1. The applicant has demonstrated through experiments that the drug has preventive and therapeutic effects on lung tumors, liver tumors, pancreatic cancer (cell lines: KPC and Panc02), glioma (cell line: GL261), and fibrosarcoma (cell line: WEHI164).

[0034] Specifically, after a pancreatic cancer cell line, PanO2, was inoculated into the subcutaneous tissues of mice and a tumor was grown, intervention with Lachnospiraceae microorganisms significantly inhibited the growth of pancreatic tumors.

[0035] After the GL261 glioma cell line was inoculated into the subsplenic area of ​​mice and a tumor was grown, intervention with Lachnospiraceae microorganisms had a significant inhibitory effect on glioma growth.

[0036] After the fibrosarcoma cell line WEHI164 was inoculated into the subcutaneous tissue of mice to grow tumors, intervention with microbial bacteria of the Lachnospiraceae family had a significant effect in inhibiting the growth of fibrosarcoma.

[0037] At the same time, it should be noted that the drug for preventing or treating tumors can also be a composition, which includes a microorganism of the Lachnospiraceae family and a combination drug; or includes a microorganism of the Lachnospiraceae family and an immunosuppressant; the composition obtained by combining the above-mentioned microorganism of the Lachnospiraceae family with a combination drug / immunosuppressant also has preventive and therapeutic effects on lung tumors, liver tumors, pancreatic cancer tumors (cell line is PanO2), glioma (cell line is GL261), and fibrosarcoma (cell line is WEHI164).

[0038] The combination drug includes at least one of a radiotherapy drug, a targeted drug, a chemotherapy drug, a photosensitizer, a photothermal agent, an immunotherapy drug, and an enhanced cell therapy drug, wherein:

[0039] The chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin or epirubicin; the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin or red bengal; the photothermal agent includes at least one of indocyanine green, neoindocyanine green or gold nanoparticle rods;

[0040] The immunosuppressant includes at least one of a PD-1 antibody, a CTLA-4 antibody, a PD-L1 antibody or a PD-L1 inhibitor; the PD-L1 inhibitor is selected from durvalumab, atezolizumab or avelumab; the PD-1 antibody or PD-L1 antibody is selected from pembrolizumab or nivolumab; and the CTLA-4 antibody is selected from ipilimumab.

[0041] Specifically, the combination of MNH46686 and PD-1 antibodies significantly reduced tumor volume and weight, and significantly improved the response rate of tumor treatment in mice subcutaneously inoculated with PanO2 and KPC pancreatic cancer cell lines to form an ectopic syngeneic tumor model;

[0042] The combination of MNH46686 and PD-1 antibodies significantly reduced tumor volume and weight and significantly improved the response rate of tumor treatment in mice subcutaneously inoculated with the GL261 glioma cell line to form an ectopic syngeneic tumor model;

[0043] The combination of MNH46686 and PD-1 antibody significantly reduced tumor volume and weight and significantly improved the response rate of tumor treatment in mice subcutaneously inoculated with WEHI164 fibrosarcoma cell line to form an ectopic homologous tumor model.

[0044] The combination of MNH46686 and PD-1 antibodies significantly reduced tumor volume and weight and significantly improved the response rate of tumor treatment in mice subcutaneously inoculated with liver cancer cell lines (H22, Hepal-1) to form an ectopic syngeneic tumor model;

[0045] The combination of MNH46686 and PD-1 antibodies significantly reduced tumor volume and weight and significantly improved the response rate of tumor treatment in mice subcutaneously inoculated with a lung cancer cell line (LLC1) to form an ectopic syngeneic tumor model;

[0046] Flow cytometric analysis of the combination with PD-1 showed that the combination of the strain of the present invention and the PD-1 antibody significantly improved the response rate of tumor treatment. The combination of the strain of the present invention and PD-1 can activate the immune system of mice; upregulate immune T cells such as CD45+CD3+, CD45+CD4+, CD45+CD8+, and upregulate the proportion of peripheral blood NK cells.

[0047] As used herein, "enhancing" the efficacy of a cell therapy (e.g., CAR-T) refers to the effect of a composition of the present invention on the cell therapy (e.g., in the case of CAR-T, a T cell-mediated immune response, particularly against a tumor antigen) as a result of its administration, when compared to no administration. For example, a subject treated with a composition of the present invention and a cell therapy may exhibit a greater such therapeutic effect from the cell therapy than a control subject treated with the cell therapy instead of a composition of the present invention.

[0048] The microbial strains, microbial agents, drugs, and compositions provided by the present invention, including microbial fungi of the family Lachnospiraceae and / or metabolites of microbial fungi of the family Lachnospiraceae and / or supernatants of microbial fungi of the family Lachnospiraceae, can inhibit tumor growth. Preferably, inhibiting tumor growth includes inhibiting tumor volume growth, inhibiting tumor weight increase, increasing the NK ratio in the peripheral blood of mice to activate the systemic immunity of mice, enhancing the efficacy of immunotherapeutics and CAR-T therapeutics, and improving the tumor response rate. At least one of the effects.

[0049] The content of the microbial strain in the unit dose of the oral composition includes 2×10 7 ~9×10 10 CFU / mL or 1×10 6 ~2×10 10 CFU / mg of Lachnospiraceae microbial strains; the unit dose is preferably 4×10 7 CFU / mL, 5×10 7 CFU / mL, 6×10 7 CFU / mL, 7×10 7 CFU / mL, 8×10 7 CFU / mL, 1×10 8 CFU / mL, 2×10 8 CFU / mL, 3×10 8 CFU / mL, 4×10 8 CFU / mL, 5×10 8 CFU / mL, 6×10 8 CFU / mL, 7×10 8 CFU / mL, 8×10 8 CFU / mL, 9×10 8CFU / mL, 1×10 9 CFU / mL, 2×10 9 CFU / mL, 3×10 9 CFU / mL, 4×10 9 CFU / mL, 5×10 9 CFU / mL, 6×10 9 CFU / mL, 7×10 9 CFU / mL, 8×10 9 CFU / mL, 9×10 9 CFU / mL, 2×10 10 or 1.2×10 6 CFU / mg, 4×10 6 CFU / mg, 5×10 7 CFU / mg, 6×10 7 CFU / mg, 7×10 7 CFU / mg, 8×10 7 CFU / mg, 9×10 7 CFU / mg, 1×10 8 CFU / mg, 2×10 8 CFU / mg, 3×10 8 CFU / mg, 4×10 8 CFU / mg, 5×10 8 CFU / mg, 6×10 8 CFU / mg, 7×10 8 CFU / mg, 8×10 8 CFU / mg, 9×10 8 CFU / mg.

[0050] The microbial strains, microbial agents, drugs, and compositions provided by the present invention, and / or the metabolites of the microbial bacteria of the Lachnospiraceae family and / or the supernatants of the microbial bacteria of the Lachnospiraceae family and / or the supernatants of the microbial bacteria of the Lachnospiraceae family can enhance the anti-tumor activity of cytotoxic T cells, enhance the immune response of the subject, inhibit the growth of tumor cells, inhibit the spread of tumor cells, inhibit the immune escape of tumors, regulate the levels of one or more symbiotic microorganisms in the intestine of the subject, upregulate proinflammatory cytokines, chemokines, reduce intestinal barrier permeability and / or upregulate protective immunogenic cytokines, regulate chemokines, and increase at least one of the infiltration effects of T cells in tumors.

[0051] The present invention provides an application in the preparation of a drug for treating and / or preventing a disease mediated by HDAC activity, characterized in that a Lachnospiraceae microorganism and / or a metabolite of a Lachnospiraceae microorganism and / or a supernatant of a Lachnospiraceae microorganism inhibits HDAC activity to achieve treatment or prevention of a disease mediated by HDAC activity; the disease mediated by HDAC activity includes at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel disease;

[0052] Preferably, the tumor comprises at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer and fibrosarcoma;

[0053] Preferably, HDAC activity is inhibited by regulating short-chain fatty acids / short-chain fatty acid salts; the short-chain fatty acids include one or more of acetate, butyrate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the short-chain fatty acid salts include one or more of acetate, propionate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the histone acetylase is class I, class II, class III and / or class IV histone acetylase.

[0054] The microbial strain or metabolite of the microbial strain or supernatant of the microbial strain provided by the present invention prevents and / or treats at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel disease by inhibiting the activity of histone acetylase (HDAC); preferably, HDAC activity is inhibited by regulating short-chain fatty acids / short-chain fatty acid salts; the short-chain fatty acids include one or more of acetic acid, butyric acid, and valeric acid; more preferably, the short-chain fatty acid is butyric acid; preferably, the short-chain fatty acid salt includes one or more of acetate, propionate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the histone acetylase is class I, class II, class III, and / or class IV histone acetylase.

[0055] In one aspect, the present invention also provides a method for preventing and / or treating tumors, comprising treating a patient in need thereof with an effective amount of the Lachnospiraceae microbial strain or metabolite of the present invention; or treating a patient in need thereof with a bacterial agent or drug containing the strain or metabolite of the present invention as an active ingredient. Preferably, the strain is Lachnospiraceae microbial strain MNH 46686, deposited in the Guangdong Provincial Microbial Culture Collection Center under the deposited name Lachnospiraceae sp. MNH 46686 and the strain deposit number GDMCC No: 62002.

[0056] Embodiments of the present invention include the following:

[0057] Embodiment 1. Use of a Lachnospiraceae microbial strain, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation broth or supernatant of the fermentation broth, in the preparation of a drug for preventing and / or treating tumors, characterized in that the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family, and the Lachnospiraceae microbial strain has a 16S rDNA having a nucleotide sequence as shown in SEQ ID No. 1 or having at least 90% identity thereto, for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.65%, 98.7%, or 99% identity thereto, for example, 97%, 98.7%, or 99% identity;

[0058] The tumor includes a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by viral infection or a tumor caused by bacterial infection, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Epstein-Barr virus infection, and Helicobacter pylori (HP) infection and Fusobacterium nucleatum (F. nucleatum) infection.

[0059] Embodiment 2: The use according to embodiment 1, characterized in that the Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in Guangdong Microbial Culture Collection Center with the deposit name of Lachnospiraceae sp. MNH 46686 and the deposit number of the strain is GDMCC No: 62002;

[0060] Alternatively, the Lachnospira microbial strain has an average nucleotide identity ANI value of at least 95% with the Lachnospira microbial strain MNH 46686, preferably, the average nucleotide identity ANI value is 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

[0061] Embodiment 3. A composition comprising a Lachnospiraceae microbial strain in an enteric coating and a pharmaceutically acceptable carrier, wherein the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family, and the Lachnospiraceae microbial strain has a 16S rDNA of the nucleotide sequence shown in SEQ ID No. 1 or has at least 90% identity thereto, for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.65%, 98.7%, 99% identity, for example, 97% or 98.7% or 99% identity; preferably, the composition is an oral composition; preferably, the composition further comprises a pharmaceutically acceptable carrier; preferably, the Lachnospiraceae microbial strain and the pharmaceutically acceptable carrier are present in the enteric coating.

[0062] Embodiment 4: The composition according to embodiment 3, characterized in that the Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in Guangdong Provincial Microbiological Culture Collection Center with the deposit name of Lachnospiraceae sp. MNH46686 and the strain deposit number of GDMCC No: 62002;

[0063] Alternatively, the Lachnospira microbial strain has an average nucleotide identity ANI value of at least 95% with the Lachnospira microbial strain MNH 46686, preferably, the average nucleotide identity ANI value is 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

[0064] Embodiment 5. The composition according to any one of Embodiments 3-4 is characterized in that it comprises living cells of a Lachnospiraceae microbial strain or living bacteria or attenuated bacteria or irradiated bacteria or inactivated bacteria or a culture of the Lachnospiraceae microbial strain or a metabolite of the Lachnospiraceae microbial strain or a fermentation broth of a Lachnospiraceae microbial strain or a supernatant of a fermentation broth.

[0065] Embodiment 6: The composition according to embodiment 5, characterized in that the composition further comprises a combined drug; the combined drug is selected from at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, an immunotherapy drug, and an enhanced cell therapy drug;

[0066] Preferably, the chemotherapy drugs include one or more of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin or epirubicin;

[0067] Preferably, the photosensitizer comprises one or more of boron dipyrrole, chlorin or red bengal;

[0068] Preferably, the photothermal agent is one or more of indocyanine green, new indocyanine green or gold nanoparticle rods;

[0069] Preferably, the immunotherapy drug comprises a PD-1 antibody, a CTLA-4 antibody, a PD-L1 antibody, and a PD-L1 inhibitor; more preferably, the PD-L1 inhibitor is selected from durvalumab, atezolizumab, or avelumab; the PD-1 antibody or PD-L1 antibody is selected from pembrolizumab or nivolumab; the CTLA-4 antibody is selected from ipilimumab; more preferably, the effective dose of the PD-1 inhibitor is 1 to 11 mg / kg; preferably, the effective dose of PD-1 includes: 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg;

[0070] Preferably, the enhanced cell therapy comprises CAR-T;

[0071] Preferably, the content of microbial strains in the composition per unit dose includes 2×10 7 ~9×10 10 CFU / mL or 1×10 6 ~2×10 10 CFU / mg of Lachnospiraceae microbial strains; the unit dose is preferably 4×10 7 CFU / mL, 5×10 7 CFU / mL, 6×10 7 CFU / mL, 7×10 7 CFU / mL, 8×10 7 CFU / mL, 1×10 8 CFU / mL, 2×10 8 CFU / mL, 3×10 8 CFU / mL, 4×10 8 CFU / mL, 5×10 8 CFU / mL, 6×10 8 CFU / mL, 7×10 8 CFU / mL, 8×10 8 CFU / mL, 9×10 8 CFU / mL, 1×10 9 CFU / mL, 2×10 9 CFU / mL, 3×10 9 CFU / mL, 4×10 9 CFU / mL, 5×109 CFU / mL, 6×10 9 CFU / mL, 7×10 9 CFU / mL, 8×10 9 CFU / mL, 9×10 9 CFU / mL, 2×10 10 or 1.2×10 6 CFU / mg, 4×10 6 CFU / mg, 5×10 7 CFU / mg, 6×10 7 CFU / mg, 7×10 7 CFU / mg, 8×10 7 CFU / mg, 9×10 7 CFU / mg, 1×10 8 CFU / mg, 2×10 8 CFU / mg, 3×10 8 CFU / mg, 4×10 8 CFU / mg, 5×10 8 CFU / mg, 6×10 8 CFU / mg, 7×10 8 CFU / mg, 8×10 8 CFU / mg, 9×10 8 CFU / mg.

[0072] Embodiment 7. The composition according to embodiment 5, characterized in that the composition further comprises a pharmaceutically acceptable carrier; preferably, the carrier is selected from one or more of a diluent, a dispersant, an excipient, a stabilizer, a lubricant, and a disintegrant;

[0073] Preferably, the drug is in the form of a liquid preparation, a solid preparation, a capsule preparation, a sustained-release preparation, and a nanoformulation.

[0074] Embodiment 8: The composition according to embodiment 5, wherein at least 50% or more of the bacteria are live bacteria, for example, at least 90% or more are live bacteria.

[0075] Embodiment 9. The composition of embodiment 5, wherein the composition is in the form of a powder, microencapsulated powder, capsule, tablet, lozenge, granule, emulsion, suspension, suppository, beverage, food, medicine or nutritional product, food additive, dietary supplement or dairy product.

[0076] Embodiment 10. Use of the composition described in any one of Embodiments 3-9 in the preparation of a medicament for preventing and / or treating at least one of metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel diseases.

[0077] Embodiment 11, use of the composition according to any one of Embodiments 3-9 in the preparation of a drug for anti-tumor or tumor growth inhibition,

[0078] Preferably, inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid, propionic acid, butyric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one of acetic acid, propionic acid, butyric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect;

[0079] The tumor includes a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by a virus or a tumor caused by bacteria, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Helicobacter pylori infection (HP), Epstein-Barr virus infection, and tumors caused by Helicobacter pylori (HP) infection and Fusobacterium nucleatum (F.nucleatum) infection.

[0080] Embodiment 12, use of the composition of any one of Embodiments 3-9 in the preparation of a medicament for treating and / or preventing a disease mediated by HDAC activity, preferably, the disease mediated by HDAC activity includes at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel disease; preferably, the tumor includes a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor; preferably, the tumor is selected from liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, or ovarian cancer. At least one of ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by viral infection or a tumor caused by bacterial infection, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Epstein-Barr virus infection, and Helicobacter pylori (HP) infection, Fusobacterium nucleatum (F.nucleatum) infection.

[0081] Embodiment 13, a Lachnospiraceae microbial strain or a culture of the Lachnospiraceae microbial strain or a metabolite of the Lachnospiraceae microbial strain or a fermentation broth of the Lachnospiraceae microbial strain or a supernatant of the fermentation broth, characterized in that the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family (Lachnospiraceae), and the nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown in SEQ ID No. 1.

[0082] Embodiment 14. The Lachnospiraceae microbial strain or the culture of the Lachnospiraceae microbial strain or the metabolite of the Lachnospiraceae microbial strain or the fermentation broth of the Lachnospiraceae microbial strain or the supernatant of the fermentation broth according to Embodiment 13, characterized in that the Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in the Guangdong Provincial Microbial Culture Collection Center with the preservation name of Lachnospiraceae sp. MNH 46686 and the strain preservation number of GDMCC No: 62002.

[0083] Embodiment 15. A method for treating or preventing tumors or diabetes, comprising the step of administering to a subject in need thereof an effective amount of the Lachnospiraceae microbial strain described in Embodiment 13 or 14, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation broth of the Lachnospiraceae microbial strain, or a supernatant of the fermentation broth, or the composition described in any one of Embodiments 3-9;

[0084] Preferably, the tumor comprises a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from one or more of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a viral tumor or a bacterial tumor, such as a tumor caused by human papillomavirus infection, hepatitis B infection or hepatitis C infection, human immunodeficiency virus infection, Helicobacter pylori infection, Epstein-Barr virus infection or Fusobacterium nucleatum infection;

[0085] Preferably, the strain prevents and / or treats tumors by inhibiting tumor growth, and inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one short-chain fatty acid or short-chain fatty acid salt of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect.

[0086] The Lachnospiraceae microbial strain or the culture of the Lachnospiraceae microbial strain or the metabolite of the Lachnospiraceae microbial strain or the fermentation broth or the supernatant of the fermentation broth of the Lachnospiraceae microbial strain as described in embodiment 16, embodiment 13 or 14, or the composition as described in any one of embodiments 3-9, for preventing and / or treating tumors or diabetes;

[0087] Preferably, the tumor comprises a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from one or more of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a viral tumor or a bacterial tumor, such as a tumor caused by human papillomavirus infection, hepatitis B infection or hepatitis C infection, human immunodeficiency virus infection, Helicobacter pylori infection, Epstein-Barr virus infection or Fusobacterium nucleatum infection;

[0088] Preferably, the strain prevents and / or treats tumors by inhibiting tumor growth, and inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one short-chain fatty acid or short-chain fatty acid salt of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect.

[0089] Embodiments of the present invention also include the following:

[0090] Embodiment 1: Use of a Lachnospiraceae microbial strain in the preparation of a drug for preventing and / or treating tumors, characterized in that the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family, and the nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown in SEQ ID No. 1; the tumor includes at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer and fibrosarcoma.

[0091] Embodiment 2: The use according to embodiment 1 is characterized in that the Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in Guangdong Provincial Microbiological Culture Collection Center with the deposited name Lachnospiraceae sp. MNH 46686 and the strain deposit number GDMCC No: 62002.

[0092] Embodiment 3, the Lachnospiraceae microbial strain is characterized in that the Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family (Lachnospiraceae), and the nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown in SEQ ID No. 1.

[0093] Embodiment 4: The Lachnospiraceae microbial strain according to Embodiment 3 is characterized in that the Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in the Guangdong Provincial Microbial Culture Collection Center with the deposited name Lachnospiraceae sp. MNH 46686 and the strain deposit number GDMCC No: 62002.

[0094] Embodiment 5: A microbial agent, characterized in that the microbial agent comprises the Lachnospiraceae microbial strain described in Embodiment 3 or Embodiment 4, or a metabolite of the Lachnospiraceae microbial strain, or a supernatant of the Lachnospiraceae microbial strain.

[0095] Embodiment 6: A drug for preventing and / or treating tumors, characterized in that it comprises the Lachnospiraceae microbial strain according to embodiment 3 or embodiment 4, or a metabolite of the Lachnospiraceae microbial strain, or a supernatant of the Lachnospiraceae microbial strain;

[0096] Preferably, the unit dose of the drug comprises 9×10 8 ~2×10 9 CFU / mL or 1×10 6 ~2×10 9 CFU / mg of Lachnospira microbial strain; the unit dose is preferably 9×10 8 CFU / mL or 1×10 9 CFU / mg, 9.2×10 9 CFU / mL or 1.2×10 9 CFU / mg, 9.3×10 9 CFU / mL or 1.4×10 9 CFU / mg, 9.4×10 9 CFU / mL or 1.1×10 9 CFU / mg, 1.2×10 9 CFU / mL or 1.8×10 9 CFU / mg and 1.4×10 9 CFU / mL or 1.6×10 9 CFU / mg, 1.7×10 9 CFU / mL or 1.2×10 10 CFU / mg, 1.8×10 9 CFU / mL or 1.4×10 9 CFU / mg, 1.6×10 9 CFU / mL or 1.6×10 10 CFU / mg, 1.8×10 10 CFU / mL or 1.8×1010 CFU / mg and 2×10 10 CFU / mL or 2×10 10 CFU / mg.

[0097] Embodiment 7. The drug according to embodiment 6, characterized in that the drug further comprises a pharmaceutically acceptable carrier; preferably, the carrier is selected from one or more of a diluent, a dispersant, an excipient, a stabilizer, a lubricant, and a disintegrant;

[0098] Preferably, the drug is in the form of a liquid preparation, a solid preparation, a capsule preparation, a sustained-release preparation, and a nanoformulation.

[0099] Embodiment 8. A pharmaceutical composition, characterized in that the pharmaceutical composition comprises the microbial strain of the Lachnospiraceae family according to any one of Embodiments 3-4 and / or the microbial agent according to Embodiment 5 and / or the drug according to any one of Embodiments 6-7; the pharmaceutical composition further comprises a combined drug; the combined drug comprises at least one of a chemotherapeutic drug, a photosensitizer, a photothermal agent, an immunotherapy drug, and an enhanced cell therapy drug;

[0100] Preferably, the chemotherapy drugs include one or more of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin or epirubicin;

[0101] Preferably, the photosensitizer comprises one or more of boron dipyrrole, chlorin or red bengal;

[0102] Preferably, the photothermal agent is one or more of indocyanine green, new indocyanine green or gold nanoparticle rods;

[0103] Preferably, the immunotherapy drug comprises a PD-1 antibody, a CTLA-4 antibody, a PD-L1 antibody, or a PD-L1 inhibitor; more preferably, the PD-L1 inhibitor is selected from durvalumab, atezolizumab, or avelumab; the PD-1 antibody or PD-L1 antibody is selected from pembrolizumab or nivolumab; and the CTLA-4 antibody is selected from ipilimumab.

[0104] Preferably, the enhanced cell therapy comprises CAR-T;

[0105] Preferably, the unit dose of the drug comprises 9×10 8 ~2×10 9 CFU / mL or 1×10 6 ~2×10 9 CFU / mg of Lachnospiraceae microbial strains; the unit dose is preferably 9×10 8 CFU / mL or 1×109 CFU / mg, 9.2×10 9 CFU / mL or 1.2×10 9 CFU / mg, 9.3×10 9 CFU / mL or 1.4×10 9 CFU / mg, 9.4×10 9 CFU / mL or 1.1×10 9 CFU / mg, 1.2×10 9 CFU / mL or 1.8×10 9 CFU / mg and 1.4×10 9 CFU / mL or 1.6×10 9 CFU / mg, 1.7×10 9 CFU / mL or 1.2×10 10 CFU / mg, 1.8×10 9 CFU / mL or 1.4×10 9 CFU / mg, 1.6×10 9 CFU / mL or 1.6×10 10 CFU / mg, 1.8×10 10 CFU / mL or 1.8×10 10 CFU / mg and 2×10 10 CFU / mL or 2×10 10 CFU / mg.

[0106] Embodiment 9, the microbial strain according to any one of embodiments 3-4, the microbial agent according to embodiment 5, the drug according to any one of embodiments 6-7, and the pharmaceutical composition according to embodiment 8, characterized in that the integrity of the butyrate production pathway of the Lachnospiraceae microbial strain is 100% and / or the strain is a high-producing butyrate and / or acetate strain; preferably, the microbial strain or the metabolite of the microbial strain or the supernatant of the microbial strain prevents and / or treats at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel diseases by inhibiting HDAC activity; preferably, HDAC activity is inhibited by regulating short-chain fatty acids / short-chain fatty acid salts; the short-chain fatty acids include one or more of acetic acid, butyric acid, and valeric acid; more preferably, the short-chain fatty acid is butyric acid; preferably, the short-chain fatty acid salts include one or more of acetate, propionate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the histone acetylase is a class I, class II, class III, and / or class IV histone acetylase.

[0107] Embodiment 10, the microbial strain as described in any one of embodiments 3-4 and 9, the microbial agent as described in any one of embodiments 5 to 9, the drug as described in any one of embodiments 6-7 and 9, and the pharmaceutical composition as described in any one of embodiments 8 to 9, characterized in that the microbial bacteria of the Lachnospiraceae family and / or the metabolites of the microbial bacteria of the Lachnospiraceae family and / or the supernatant of the microbial bacteria of the Lachnospiraceae family can inhibit tumor growth; preferably, inhibiting tumor growth includes inhibiting tumor volume growth, inhibiting tumor weight increase, increasing the NK ratio in peripheral blood to activate mouse systemic immunity, enhancing the efficacy of immunotherapeutics, CAR-T therapeutics, and improving tumor response rate. At least one of the effects.

[0108] Embodiment 11, the microbial strain according to any one of embodiments 3-4, 9 to 10, the microbial agent according to any one of embodiments 5 to 10, the drug according to any one of embodiments 6-7, 9 to 10, and the pharmaceutical composition according to any one of embodiments 8 to 10, characterized in that the microbial bacteria of the Lachnospiraceae family and / or the metabolites of the microbial bacteria of the Lachnospiraceae family and / or the supernatant of the microbial bacteria of the Lachnospiraceae family can enhance the anti-tumor activity of toxic T cells, enhance the immune response of the subject, inhibit the growth of tumor cells, inhibit the spread of tumor cells, inhibit the immune escape of tumors, regulate the level of one or more commensal microorganisms in the intestine of the subject, upregulate proinflammatory cytokines, chemokines, reduce intestinal barrier permeability and / or upregulate protective immunogenic cytokines, regulate chemokines, and increase at least one of the infiltration effects of T cells in tumors.

[0109] Embodiment 12: Use of the microbial strain according to any one of Embodiments 3-4 and 9 to 11, the microbial agent according to any one of Embodiments 5 to 11, the drug according to any one of Embodiments 6-7 and 9 to 11, or the pharmaceutical composition according to any one of Embodiments 8 to 11 in the preparation of a drug for treating and / or preventing a disease mediated by HDAC activity, characterized in that the Lachnospiraceae microorganism and / or its metabolites and / or the supernatant of Lachnospiraceae microorganism inhibit HDAC activity to achieve the treatment or prevention of the disease mediated by HDAC activity; the disease mediated by HDAC activity comprises at least one of tumors, metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel diseases; preferably, the tumor comprises at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, and fibrosarcoma;

[0110] Preferably, HDAC activity is inhibited by regulating short-chain fatty acids / short-chain fatty acid salts; the short-chain fatty acids include one or more of acetate, butyrate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the short-chain fatty acid salts include one or more of acetate, propionate, and valerate; more preferably, the short-chain fatty acid is butyrate; preferably, the histone acetylase is class I, class II, class III and / or class IV histone acetylase.

[0111] In this article, strain MNH 46686, and MNC-686 appearing hereinafter are equivalent to MNH 46686;

[0112] Related content on bacterial taxonomy

[0113] Application of 16s rRNA in taxonomy:

[0114] Judgment criteria: When the similarity between the 16S rRNA gene sequences of two strains is less than 97% to 98.65%, they can be judged to belong to different species.

[0115] How "Consistency" or "Consistency" is calculated:

[0116] "Identity" between the nucleic acid sequences of two nucleic acid molecules can be determined as a percentage of identity using known computer algorithms, such as the "FASTA" program, using default parameters such as those described in Pearson et al. (Other programs include the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387 (1984)), BLASTP, BLASTN, FASTA). For example, the BLAST function of the NCBI database, other commercially or publicly available programs include the DNAStar "MegAlign" program.

[0117] In some embodiments, the method further comprises administering a prebiotic to the subject. In some embodiments, the prebiotic is fructooligosaccharide, galacto-oligosaccharide, trans-galacto-oligosaccharide, xylo-oligosaccharide, chito-oligosaccharide, soy oligosaccharide, gentio-oligosaccharide, isomaltooligosaccharide, mannose oligosaccharide, malto-oligosaccharide, mannose oligosaccharide, lactulose, lactose sucrose, palatinose, glycosyl sucrose, guar gum, gum arabic, tagatose, amylose, amylopectin, pectin, xylan or cyclodextrin.

[0118] "Administration" or "administering" generally refers to the route by which a composition (e.g., composition) is administered to a subject. Examples of routes of administration include oral administration, rectal administration, topical administration, inhalation (nasal), or injection. Injection administration includes intravenous (IV), intramuscular (IM), intratumoral (IT), and subcutaneous (SC) administration. The compositions or pharmaceutical compositions described herein can be administered in any form by any effective route, including but not limited to intratumoral, oral, parenteral, enteral, intravenous, intraperitoneal, topical, transdermal (e.g., using any standard patch), intradermal, intraocular, (intranasal, topical, non-oral, such as aerosol, inhalation, subcutaneous, intramuscular, buccal, sublingual, rectal, vaginal, intraarterial, and intrathecal. - The compositions described herein are administered orally, rectally, intratumorally, topically, intravesically, by injection into or near a draining lymph node, intravenously, by inhalation or aerosol, or subcutaneously. In another preferred embodiment, the compositions described herein are administered orally, intratumorally, or intravenously. - The compositions described herein are administered orally, rectally, intratumorally, topically, intravesically, by injection into or near a draining lymph node, intravenously, by inhalation or aerosol, or subcutaneously. In another preferred embodiment, the compositions described herein are administered orally, intratumorally, or intravenously.

[0119] "Identity" between the nucleic acid sequences of two nucleic acid molecules can be determined as a percentage of identity using a known computer algorithm, such as the "FASTA" program, using default parameters such as those described in Pearson et al. (Other programs include the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387 (1984)), BLASTP, BLASTN, FASTA). For example, the BLAST function of the NCBI database, other commercially or publicly available programs include the DNAStar "MegAlign" program.

[0120] The term Average Nucleotide Identity (ANI) in the present invention is an indicator for comparing the genetic relationship between two genomes at the nucleotide level. ANI is defined as the average base similarity between homologous fragments of two microbial genomes.

[0121] The term "increase" or "elevate" or "enhance" refers to a change such that the difference is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 2-fold, 4-fold, 10-fold, 100-fold, 10^3-fold, 10^4-fold, 10^5-fold, 10^6-fold and / or 10^7-fold after treatment compared to the pre-treatment state, as appropriate. Possible increased properties include the number of immune cells, bacterial cells, stromal cells, myeloid-derived suppressor cells, fibroblasts, metabolites; cytokine levels; or other physical parameters (e.g., tumor size).

[0122] The term "reduced" or "decreased" or "decreased" refers to a change such that the difference is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 1 / 100, 1 / 1000, 1 / 10,000, 1 / 100,000, 1 / 1,000,000, or undetectable after treatment, as compared to the pre-treatment state, as appropriate. Properties that may be reduced include the number of immune cells, bacterial cells, stromal cells, myeloid-derived suppressor cells, fibroblasts, metabolites; cytokine levels; or other physical parameters (e.g., ear thickness (e.g., in a DTH animal model) or tumor size).

[0123] As used herein, "metabolite" refers to a compound, composition, molecule, ion, cofactor, catalyst, or nutrient from any cellular or microbial metabolic reaction that is used as a substrate or as a product in any cellular or microbial metabolic reaction.

[0124] "Strain" refers to a member of a bacterial species that has genetic characteristics that allow it to be distinguished from closely related members of the same bacterial species. The genetic characteristic can be the total or partial absence of at least one gene, the total or partial absence of at least one regulatory region (e.g., promoter, terminator, riboswitch, ribosome binding site), the absence ("cure") of at least one native plasmid, the presence of at least one recombinant gene, the presence of at least one mutated gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one mutated regulatory region (e.g., promoter, terminator, riboswitch, ribosome binding site), the presence of at least one non-native plasmid, the presence of at least one antibiotic resistance cassette, or a combination thereof. Genetic characteristics between different strains can be identified by PCR amplification, optionally followed by DNA sequencing of the genomic region of interest or the entire genome. In the event that one strain (compared to another strain of the same species) gains or loses antibiotic resistance or gains or loses a biosynthetic capacity (e.g., an auxotrophic strain), the strains or nutrients / metabolites can be distinguished by selection or counterselection using antibiotics.

[0125] "Culture" within the meaning of the present invention refers to a culture obtained under anaerobic culture conditions using a liquid culture medium; preferably, the liquid culture medium is MM01 liquid culture medium; preferably, the culture temperature is 37°C, preferably, the culture time is 24-72 hours, preferably 36-60 hours, and more preferably 48 hours.

[0126] The components of MM01 liquid medium are as follows:

[0127] The liquid MM01 culture medium involved in this article has the following components: peptone 5g / L, trypticase 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K2HPO4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, hemoglobin 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1uL / L, inorganic salt solution 8ml / L, each 1L of inorganic salt solution includes (calcium chloride 0.25g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, sodium bicarbonate 10g, sodium chloride 2g).

[0128] The solid MM01 culture medium involved in this article has the following components: peptone 5g / L, trypticase 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K2HPO4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, hemoglobin 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1uL / L, inorganic salt solution 8ml / L, each 1L of inorganic salt solution includes (calcium chloride 0.25g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, sodium bicarbonate 10g, sodium chloride 2g), agar 15g / L.

[0129] "Fermentation broth" within the meaning of the present invention refers to the filtrate of the culture broth of the bacterial strain of the present invention obtained by culturing bacteria under anaerobic culture conditions using a liquid culture medium; "supernatant" or "supernatant" refers to the supernatant part of the culture broth of the bacterial strain according to the present invention, which contains compounds and / or cell debris of the strain, and / or metabolites and / or molecules secreted by the strain.

[0130] The term "subject" or "patient" refers to any mammal. A subject or patient described as "in need thereof" refers to a person in need of treatment (or prevention) of a disease. Mammals (i.e., mammals) include humans, laboratory animals (e.g., primates, rats, mice), livestock (e.g., cattle, sheep, goats, pigs), and household pets (e.g., dogs, cats, rodents). The subject can be a human. The subject can be a non-human mammal, including but not limited to dogs, cats, cattle, horses, pigs, donkeys, goats, camels, mice, rats, guinea pigs, sheep, camels, monkeys, gorillas, or chimpanzees. The subject or patient may be healthy or may suffer from a metabolic disease at any stage of development.

[0131] As used herein, the term "treating" a disease in a subject or "treating" a subject having or suspected of having a disease refers to administering a drug therapy, such as one or more pharmaceutical agents, to the subject to reduce or prevent worsening of at least one symptom of the disease. Thus, in one embodiment, "treating" refers, inter alia, to delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing the efficacy of an alternative therapy, or reducing resistance to an alternative therapy, or a combination thereof.

[0132] As used herein, the term "prevention" is art-recognized and, when used in connection with conditions such as local recurrence, is well known in the art and includes administration to reduce the occurrence of symptoms of a medical condition in a subject relative to a subject that has not received the composition or to delay the onset of such symptoms. Thus, prevention of cancer includes, for example, reducing the number of detectable tumors in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable tumors in a treated population relative to an untreated control population, e.g., by a statistically and / or clinically significant amount. The compositions of the present invention can be administered as a food, e.g., a nutritional supplement. Generally, the compositions of the present invention are used to treat humans, but they can be used to treat animals, including monogastric mammals, such as poultry, pigs, cats, dogs, horses, or rabbits. The compositions of the present invention can be used to enhance the growth and performance of animals. If administered to animals, oral gavage can be used.

[0133] The compositions or pharmaceutical compositions provided herein may include pharmaceutically acceptable excipients, diluents, or carriers. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art. Examples of suitable carriers include lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, or sorbitol. Examples of suitable diluents include ethanol, glycerol, and water. The choice of pharmaceutical carrier, excipient, or diluent can be selected based on the intended route of administration and standard pharmaceutical practice. The composition may include any suitable binder, lubricant, suspending agent, coating agent, or solubilizing agent as a carrier, excipient, or diluent, or in addition to a carrier, excipient, or diluent. Examples of suitable binders include starch, gelatin, natural sugars, and natural or synthetic gums. Natural sugars include glucose, anhydrous lactose, free-flowing lactose, β-lactose, or corn sweeteners. Natural or synthetic gums include gum arabic, gum tragacanth, sodium alginate, carboxymethyl cellulose, or polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, or sodium chloride. Preservatives, stabilizers, dyes, and even flavorings may be provided in the composition. Examples of preservatives include sodium benzoate, sorbic acid, or parabens. Antioxidants and suspending agents may also be used.

[0134] In certain embodiments, the bacterial strains described in the above-mentioned compositions provided by the present invention are freeze-dried. In certain embodiments, the bacterial strains described in the above-mentioned compositions provided by the present invention are spray-dried. In certain embodiments, the bacterial strains described in the above-mentioned compositions provided by the present invention are freeze-dried or spray-dried and wherein they are alive. In certain embodiments, the bacterial strains described in the above-mentioned compositions provided by the present invention are freeze-dried or spray-dried and wherein they are capable of partially or completely colonizing the intestine. In certain embodiments, in some cases, the freeze-dried bacterial strains are reconstituted prior to administration. In some cases, reconstitution is carried out by using a diluent as described herein.

[0135] In some embodiments, the compositions provided herein are administered orally. Oral administration may involve swallowing, whereby the compound enters the gastrointestinal tract, and / or oral, lingual, or sublingual administration, whereby the compound enters the bloodstream directly from the oral cavity. Pharmaceutical dosage forms suitable for oral administration include solid suppositories, solid microparticles, semisolids, and liquids (including multiphase or dispersed systems) such as tablets; soft or hard capsules containing multi- or nanoparticles, liquids (e.g., aqueous solutions), emulsions, or powders; lozenges (including liquid fillers); chewables; gels; rapidly dispersing dosage forms; ovules; sprays; and buccal / mucosal adhesive patches.

[0136] In some embodiments, the pharmaceutical formulation is an enteric formulation, i.e., a gastro-resistant formulation (e.g., resistant to gastric pH) suitable for delivering the composition of the invention to the intestine by oral administration. Enteric formulations may be particularly useful when the bacteria or another component of the composition is acid-sensitive, e.g., susceptible to degradation under gastric conditions.

[0137] In some embodiments, the enteric formulation comprises an enteric coating. In some embodiments, the formulation is an enteric-coated dosage form. For example, the formulation can be an enteric-coated tablet or an enteric-coated capsule, etc. The enteric coating can be a conventional enteric coating, such as a conventional coating for tablets, capsules, etc. for oral delivery. The formulation can comprise a film coating, such as a film layer of an enteric polymer, such as an acid-insoluble polymer. In some embodiments, the enteric formulation is enteric in nature, such as gastrointestinal and does not require an enteric coating. In some embodiments, the formulation is an inherently enteric-coated capsule (e.g., from Capsugel).

[0138] In certain embodiments, the preparation is a soft capsule. Soft capsules are capsules with certain elasticity and pliability due to the interpolation of the softening agent present in the capsule shell, for example, glycerol, sorbitol, maltitol and polyethylene glycol. Soft capsules can be produced, for example, on the basis of gelatin or starch. Soft capsules based on gelatin are commercially available from various suppliers. According to the method of administration, for example, oral or rectal administration, soft capsules can have various shapes, and they can be, for example, circular, oval, elliptical or torpedo-shaped. Soft capsules can be produced by conventional processes, for example, by Scherer process, Accogel process or droplet or blow molding process.

[0139] In certain embodiments, the compositions of the invention are administered to the gastrointestinal tract through a tube, e.g., a nasogastric tube, an orogastric tube, a gastric tube, a jejunostomy tube (J-tube), a percutaneous endoscopic gastrostomy (PEG), or a port, e.g., a chest wall port leading to the stomach, jejunum, and other suitable access ports.

[0140] In certain embodiments of the present invention, treatment according to the present invention is accompanied by an assessment of the patient's intestinal microbiota. If delivery of the strain of the present invention and / or partial or complete colonization is not achieved, and thus no efficacy is observed, the treatment can be repeated. If delivery and / or partial or complete colonization is successful and efficacy is observed, the treatment can be stopped.

[0141] Biodeposit Information:

[0142] The strain MNH 46686 of the present invention has been deposited with the Guangdong Provincial Microbial Culture Collection under the name Lachnospiraceae sp. MNH 46686 and the GDMCC No. 62002. The deposit date is November 4, 2021. The deposit address is: Institute of Microbiology, Guangdong Academy of Sciences, Building 59, 5th Floor, No. 100 Xianlie Middle Road, Guangzhou. The proposed taxonomic name is Lachnospiraceae sp. The deposited strain was identified as viable by the collection center. Beneficial effects:

[0143] The Lachnospiraceae microbial strain provided by the present invention does not exist in existing reports and belongs to a new genus and a new species;

[0144] The Lachnospiraceae microbial strain of the present invention has a complete butyrate production pathway and can synthesize a large amount of acetic acid, isobutyric acid, butyric acid and isovaleric acid during growth;

[0145] The Lachnospiraceae microbial strain of the present invention can inhibit the growth of tumors (such as liver cancer) and can significantly reduce the size and weight of tumors compared with the control group;

[0146] The strain of the present invention and its metabolites can be used to prepare a composition for alleviating, preventing or treating cancer (or tumor), and have very broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0147] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0148] FIG1 is a photograph of the colony morphology of the strain MNH 46686 of the present invention cultured on an anaerobic blood plate for 24 hours;

[0149] FIG2 is a microscopic photograph of the strain MNH 46686 of the present invention;

[0150] FIG3 is a microscopic photograph of the strain MNH 46686 of the present invention;

[0151] FIG4 is a Gram-stained microscopic photograph of the strain MNH 46686 of the present invention;

[0152] FIG5 is a Gram-stained microscopic photograph of the spores of strain MNH 46686 of the present invention;

[0153] FIG6 shows the tolerance of strain MNH 46686 of the present invention to different pH values;

[0154] FIG7 shows the tolerance of the strain MNH 46686 of the present invention to different concentrations of NaCl;

[0155] FIG8 shows the tolerance results of the strain MNH 46686 of the present invention to different bile salts;

[0156] FIG9 shows the results of biochemical identification of strain MNH 46686 of the present invention by API 20A;

[0157] FIG10 is a phylogenetic tree constructed by comparing the strains of the present invention with strains related to the Lachnospiraceae family;

[0158] Figure 11 shows the regulation of macrophage immune activity by strain MNH 46686;

[0159] Figure 12 shows the regulation of Primary PBMC immune activity by strain MNH 46686;

[0160] Figure 13A shows that the IFNβ gene reporter system constructed in this study has a good response to the positive drug MSA-2, and Figure 13B shows that MNH46686 can significantly promote IFNβ transcriptional activity;

[0161] Figure 14 is a comparison of tumor tissue sizes after autopsy in an animal experiment on liver cancer prevention;

[0162] Figures 15(A) to 15(D) show the therapeutic effect of MNH 46686 on lung cancer.

[0163] Figure 16 shows the proportion of immune cells in mouse tissues;

[0164] FIG17 is a correlation analysis between immune cells and tumor endpoint volume in mouse tissues;

[0165] FIG18 is a correlation analysis between immune cells and tumor weight in mouse tissues;

[0166] FIG19 is a graph showing mouse tumor volume in an experiment combining MNH46686 and PD-1;

[0167] FIG20 is a graph comparing the tumor volumes of mice at the end point of the MNH46686 and PD-1 combination experiment;

[0168] FIG21 is a graph comparing mouse tumor weights at the end point of the MNH46686 and PD-1 combination experiment;

[0169] FIG22 is a graph showing the mouse response rate at the experimental endpoint in the MNH46686 and PD-1 combination experiment;

[0170] FIG23 shows the inhibition of histone deacetylase (HDAC) by strain MNH46686;

[0171] Figure 24A-D shows the proportion of immune cells in mouse tissues. Data were analyzed using CyExpert and statistically processed using Graphpad Prism V9. Statistical analysis was performed using the Student's T test. *p<0.05, **p<0.01, ***p<0.001.

[0172] Figures 25A-D show the correlation analysis between immune cells and tumor weight in mouse tissues. Data were analyzed using CyExpert and statistically processed using Graphpad Prism V9. Statistical analysis was performed using Correlation Analysis. *p<0.05, **p<0.01, ***p<0.001.

[0173] FIG26 is a graph showing mouse tumor volume. Data are presented as mean ± standard deviation (mean ± SD). Statistical analysis was performed using two-way ANOVA with Dunnett's multiple comparisons. Significant differences are indicated by *, *p < 0.05.

[0174] FIG27 is a graph comparing tumor volumes in mice at the end point of the experiment. Data are presented as mean ± standard deviation (mean ± SD). Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test. Significant differences are indicated by *, *p < 0.05, **p < 0.01, and no significant differences are indicated.

[0175] FIG28 is a graph comparing mouse tumor weights at the end point of the experiment. Data are presented as mean ± standard deviation (mean ± SD). Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test. Significant differences are indicated by *, *p < 0.05, **p < 0.01, and no significant differences are indicated.

[0176] FIG29 is a graph showing the comparison of tumor growth inhibition rates in mice at the experimental endpoint. Data in the graph are presented as mean ± standard deviation (mean ± SD). Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test. Significant differences are indicated by *, *p < 0.05, **p < 0.01.

[0177] Figure 30 is a graph showing the mouse response rate at the end of the experiment, where the data is based on the tumor volume of a single mouse (mm 3 )show. DETAILED DESCRIPTION

[0178] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0179] Example 1

[0180] 1.1 Isolation of strain MNH 46686

[0181] The intestinal strain Lachnospiraceae sp. MNH 46686 screened in the present invention was isolated from a fecal sample of a healthy female volunteer in Guangzhou City, Guangdong Province.

[0182] Specifically, the strain isolation method is as follows:

[0183] The donor collects 2 to 5 grams of fresh feces, places them in a sample collection and preservation tube, shakes and homogenizes them, places the processed fecal sample in an ice box, and delivers it to the laboratory for strain isolation within 24 hours.

[0184] Physiological saline was dispensed in a biosafety cabinet, 9 mL / tube. Anaerobic blood agar plates (Huankai Microbiology Technology Co., Ltd.) for strain isolation were prepared and transferred to the anaerobic workstation 24 h in advance. Sample information, culture medium type, isolation date, etc. were marked.

[0185] Take fresh fecal samples and place them in an anaerobic operating station (Don Whitley Scientific H35) and shake them on a vortex shaker for 1 min. Pipette 1 mL of the sample into 9 mL of normal saline and mix for 10 min. -1 dilution, and then serially diluted to 10 -6 Dilution solution, set aside.

[0186] Take 10 -6Drop the diluted solution into the separation medium (anaerobic blood agar plate, Columbia blood agar plate and chocolate agar plate) at a drop volume of 100 μL / dish. Spread evenly. After the surface of the plate is dry, turn the plate upside down and culture at 37°C for 3 to 5 days.

[0187] 1.2 Purification of strains

[0188] Observe the growth of the strain in the isolation culture medium and pick a single colony with a sterilized toothpick for strain purification. The purified strain is placed at 37°C for anaerobically culture.

[0189] The pure culture strain was prepared into 20% glycerol / water-bacteria solution and stored at -86°C.

[0190] Example 2. Morphological and physiological and biochemical characteristics of strain MNH 46686

[0191] 2.1 Morphological characteristics of strain MNH 46686

[0192] Strain MNH 46686 was inoculated onto anaerobic blood agar plates and incubated anaerobically at 37°C for 48 hours. Visible colonies formed on the agar plates. These colonies were round, with regular, smooth edges, approximately 0.5 mm in diameter, pale yellow, and opaque. No secretions were observed around the colonies. The strain was Gram-negative. Microscopic observation revealed the absence of flagella, the absence of motility, and rod-shaped colonies, approximately 2-5 μm x 20-50 μm in length. A photograph of the colony morphology of strain MNH 46686 after 48 hours of anaerobic blood agar plate incubation is shown in Figure 1. Microscopic photographs of strain MNH 46686 are shown in Figures 2 (overall) and 3 (partial).

[0193] The Gram-stained microscopic morphology of strain MNH 46686 is shown in FIG4 , and the spore-stained microscopic morphology of strain MNH 46686 is shown in FIG5 ( FIG4 and FIG5 are grayscaled; the original staining images appear red). The strain can produce spores and is Gram-negative.

[0194] 2.2 Physiological and biochemical characteristics of strain MNH 46686

[0195] Strain MNH 46686 has a growth temperature range of 30-42°C, with an optimal growth temperature of 37°C. It can grow in the pH range of 7.0 to 9.0, with an optimal pH of 7.0 (see Figure 6 for the strain's tolerance to different pH values). It will not grow on media containing more than 2% NaCl (see Figure 7 for the strain's tolerance to different NaCl concentrations). Strain MNH 46686 can survive and grow in bile salt concentrations between 0% and 0.3%, but will not grow at bile salt concentrations greater than or equal to 0.4% (see Figure 8 for the strain's tolerance to different bile salt concentrations).

[0196] 2.3 Results of biochemical identification of strain MNH 46686 using API 20A

[0197] The test was performed using API 20A reagent strips (BioMérieux) according to the manufacturer's instructions. Culture conditions were 37°C, anaerobic. The experimental results are shown in Table 1 and Figure 9.

[0198] Table 1. Test results of strain MNH 46686API 20A

[0199] 2.4 Antibiotic susceptibility testing of strain MNH 46686 was performed using the disk diffusion method.

[0200] The results of antibiotic susceptibility testing of MNH 46686 are shown in Table 2. Strain MNH 46686 was resistant to erythromycin, ciprofloxacin, co-trimoxazole, ampicillin, ceftriaxone, and lincomycin, but was sensitive to gentamicin, chloramphenicol, tetracycline, and penicillin.

[0201] Table 2. Antibiotic susceptibility test results of strain MNH 46686

[0202] Example 3. Identification of strain MNH 46686

[0203] 3.1 16S rRNA gene amplification

[0204] Fresh culture of strain MNH 46686 was used to extract genomic DNA, which was then used as a template for 16S rRNA gene amplification.

[0205] The primer pairs used in the 16S rRNA gene PCR of the present invention are:

[0206] 27F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID No. 2)

[0207] 1492R: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID No. 3).

[0208] The PCR reaction procedure is as follows:

[0209] Initial denaturation: 94°C, 4 min; denaturation: 94°C, 50 sec; annealing: 52°C, 40 sec; extension: 72°C, 70 sec; final extension: 72°C, 10 min. (Note: 36 cycles).

[0210] 3.2 16S rRNA gene sequencing

[0211] After PCR amplification, the PCR product was purified and sent to GENEWIZ for 16S rRNA gene sequencing to obtain the 16S rRNA gene sequence (1383 bp).

[0212] The 16S rRNA gene sequence of strain MNH 46686 is shown in SEQ ID No. 1:

[0213] 3.3 Strain identification results

[0214] The strain 16S rRNA gene sequence returned by sequencing was submitted to the NCBI Basic Local Alignment Search Tool for strain 16S rRNA gene analysis to confirm the strain classification information.

[0215] The measured sequences were compared with the data in GenBank using BLAST analysis. The alignment results showed that the strains with the highest similarity to MNH 46686 were Lachnoclostridium pacaense (93.77%) and Enterocloster citroniae (93.77%). The 16S rRNA gene sequence similarity between strain MNH46686 and other strains was less than 93.7%. The low (<95%) 16S rRNA gene sequence similarity indicated that strain MNH 46686 may be a new genus and species of the Lachnospiraceae family.

[0216] The 16S rRNA gene sequences of MNH 46686 were compared with those of Lachnospiraceae-related strains retrieved from databases such as GenBank to construct a phylogenetic tree.

[0217] A multiple sequence alignment was performed between MNH 46686 and the sequences of model strains with high 16S rRNA gene sequence similarity in the NCBI database. A phylogenetic tree was then constructed using the software MEGA 5 (maximum likelihood method). In the figure, only nodes with bootstrap values ​​greater than 50% are displayed. The superscript "T" indicates the model strain.

[0218] From the phylogenetic tree (Figure 10), it can be seen that strain MNH 46686 did not cluster together with strains of other genera in the Lachnospiraceae family, but formed a separate branch (Bootstrap support value was 90). Phylogenetic tree analysis supported the classification of strain MNH 46686 as a new genus and species in the Lachnospiraceae family.

[0219] Example 4. Genome analysis of strain MNH 46686

[0220] The genome of the original strain MNH46686 was fragmented using ultrasound to fragments of approximately 350 bp. An Illumina sequencing library was then constructed using a standard DNA library construction kit (NEB Ultra™). The constructed library was sequenced using a NovaSeq (Illumina) with paired-end 150-bp sequencing. The sequencing yielded 1.34 Gbp of data, of which Q20 accounted for 97.325%.

[0221] The raw sequencing data were filtered using fastp (version 0.20.0) with the filtering parameters: "--poly_g_min_len 10--poly_x_min_len 10--q 15--u 40--n 5--l 50". The filtered raw data were assembled using SPAdes (version v3.14.0) with the assembly parameters "--isolate--cov-cutoff 10". The assembled genome yielded a total gene length of 3.34 Mbp, an N50 length of 174.9 kbp, and a GC content of 51.93%.

[0222] Genomic gene prediction and analysis were performed using the prokaryotic analysis software Genome Annotation Pipeline prokka (version 1.14.5) with the parameters "--gcode 11 --evalue 1e-09". A total of 3052 CDS sequences were predicted, with an average CDS sequence length of 977 bp.

[0223] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version 4.2.2), using the CARD (version 3.0.0, https: / / card.mcmaster.ca / analyze / rgi) database. Detailed comparison information is shown in Table 3.

[0224] Table 3. List of drug resistance gene information

[0225] The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to compare against the virulence factor database (VFDB, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). Detailed comparison results are shown in Table 4.

[0226] Table 4. List of potential toxic genes of MNH-46686

[0227] AntiSMASH5 (version 5.1.1) was used to analyze the potential secondary metabolism gene clusters in the genome. Detailed alignment results are shown in Table 5.

[0228] Table 5. List of potential secondary metabolic gene clusters of MNH-46686

[0229] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version 1.0.0). Detailed alignment results are shown in Table 6.

[0230] Table 6. List of potential primary metabolic gene clusters of MNH-46686

[0231] The butyrate production capacity of this strain was evaluated. The genes related to the butyrate production pathway in the article (Vital M, Howe C, Tiedje M. Revealing the Bacterial Butyrate Synthesis Pathways by Analyzing (Meta) genomic Data [J]. Mbio, 2014, 5 (2): 1-11) were used as a reference database. The genome sequence of this strain was compared with the reference database using NCBI blastp (version: 2.7.1+). The detailed comparison results are shown in Table 7. The integrity of the butyrate production pathway was then calculated. The calculation showed that the integrity of the butyrate production pathway of this strain was 100%.

[0232] Table 7. List of potential butyrate-producing genes in MNH-46686

[0233] Example 5. Fatty acid composition analysis of strain MNH 46686

[0234] Strain MNH 46686 was inoculated on TSA plates and cultured anaerobically at 37°C for 48 h. The cells were then harvested for fatty acid extraction and methylation. The fatty acid composition of strain MNH 46686 was analyzed using the Microbial ID, Inc., Newark, Del. (Kroppenstedt, 1985; Meier et al., 1993) automated bacterial identification system.

[0235] The fatty acid composition of the experimental strain MNH 46686 is shown in the table below.

[0236] The results showed that the major fatty acids (>10%) in MNH 46686 were hexadecene saturated fatty acids (C16:0 34.61%), 18:1CIS 9FAME (18.30%), 18:1c11 / t9 / t6FAME (12.74%), and octadecene saturated fatty acids (C18:0 10.92%); the remaining fatty acid types and their contents are detailed in Table 8 below.

[0237] Table 8. Fatty acid composition of strain MNH 46686

[0238] Example 6. Short-chain fatty acid (SCFA) determination of strain MNH 46686

[0239] 6.1 Bacteria Preparation

[0240] The strain MNH 46686 was inoculated into TSB liquid medium and cultured anaerobically at 37°C for 48 hours. The bacteria were collected by centrifugation and stored at -86°C until use.

[0241] 6.2 Preparation of Standards

[0242] Acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and hexanoic acid standards were weighed and prepared with ethyl acetate into eight mixed standard concentration gradients of 0.1 μg / mL, 0.5 μg / mL, 1 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, 50 μg / mL, and 100 μg / mL.

[0243] Take 600 μL of standard, add 25 μL of 4-methylvaleric acid with a final concentration of 500 μM as internal standard, mix well and add to the injection vial, enter GC-MS detection, injection volume 1 μL, split ratio 10:1, split injection.

[0244] 6.3 Metabolite extraction

[0245] Thaw the sample on ice, take 80 mg of sample into a 2 mL glass centrifuge tube, add 900 μL of 0.5% phosphoric acid to resuspend, shake and mix for 2 minutes, centrifuge at 14000g for 10 minutes, take 800 μL of supernatant, add an equal amount of ethyl acetate to extract, shake and mix for 2 minutes, centrifuge at 14000g for 10 minutes, take 600 μL of the upper organic phase, add 4-methylpentanoic acid with a final concentration of 500 μM as an internal standard, mix well and add to the injection vial, enter GC-MS detection, injection volume 1 μL, split ratio 10:1, split injection.

[0246] 6.4 Sample testing and analysis

[0247] The samples were separated using an Agilent DB-WAX capillary column (30 m × 0.25 mm ID × 0.25 μm) gas chromatography system. The temperature program was as follows: initial temperature 90°C, then increased at 10°C / min to 120°C, then at 5°C / min to 150°C, and finally at 25°C / min to 250°C, where it was maintained for 2 minutes. Helium carrier gas was used at a flow rate of 1.0 mL / min.

[0248] Mass spectrometry analysis was performed using an Agilent 7890A / 5975C gas chromatography-mass spectrometer (GC-MS). The inlet temperature was 250°C, the ion source temperature was 230°C, the transfer line temperature was 250°C, and the quadrupole temperature was 150°C. Electron impact ionization (EI) was used in full scan and SIM scanning modes, with an electron energy of 70 eV.

[0249] MSD ChemStation software was used to extract chromatographic peak areas and retention times, draw calibration curves, and calculate the content of short-chain fatty acids in the samples.

[0250] Table 9. Short-chain fatty acid (SCFA) production results of strain MNH 46686

[0251] The test results showed that strain MNH 46686 could synthesize a large amount of acetic acid, isobutyric acid, butyric acid and isovaleric acid during its growth.

[0252] Example 7. Regulation of macrophage immune activity by strain MNH 46686

[0253] The experimental method refers to Experimental Example 1 in the prior art CN112618576B.

[0254] THP-1 cells (Wuhan Pronocell Life Science Co., Ltd.) were treated with PMA at a final concentration of 5 ng / mL for 48 h to differentiate them into M0 macrophages.

[0255] The strain MNH46686 was added at an MOI (viable count: cell number) of 8-10:1. A control group of M1 macrophages (M0 macrophages induced with 20 ng / ml IFNγ and 10 pg / ml LPS for 24 hours) was also established. After 1 hour of culture, a cocktail of antibiotics (ampicillin, streptomycin, and colistin) was added to kill the bacteria. Culture was continued for another 23 hours, and the supernatant was collected.

[0256] Using BD TMThe Cytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit was used. After the supernatant samples were reacted with the antibody protein according to the instructions, the concentrations of cytokines such as IL-10, MIG, IL-6, MCP-1, RANTES, IL-1β, IL-8, and TNFα in the supernatant were detected by flow cytometry (results are shown in Figure 11).

[0257] The results showed that after co-culture of M0 macrophages with MNH46686, the macrophages clearly showed characteristics of differentiation into M1 macrophages. Among them, the levels of pro-inflammatory cytokines IL-6, IL-1β, IL-8, and IL-12 / IL-13P40 increased significantly, and TNFα showed an up-regulation trend; the levels of chemokines MIG, MCP-1, and RANTES increased significantly, and IP-10 showed an up-regulation trend; and the anti-inflammatory cytokine IL-10 showed a down-regulation trend.

[0258] Example 8. Regulation of Primary PBMC Immune Activity by Strain MNH 46686

[0259] The experimental method refers to Experimental Example 2 in the prior art CN112618576B.

[0260] The strain MNH46686 was co-cultured with primary PBMC cells at an MOI of 1 to 5 for 4 hours, and combined antibiotics (ampicillin, streptomycin, and colistin) were added to inhibit excessive proliferation of the strain. After 24 hours, the supernatant was collected and the cells were purified by BD Biosciences. TM The Cytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit was used to detect the concentrations of cytokines such as IFNγ, TNFα, IL-1β, IP-10 (CXCL-10), RANTES (CCL5), MIG (CXCL-9), IL-6, MCP-1 (CCL2), IL-10, IL-12 / IL-23P40, and IL-8 in the supernatant (see Figure 12 for the results).

[0261] The results showed that MNH46686 could induce a significant increase in the expression of inflammatory factors TNFα, IL-12 / IL-23P40, IL-6, IL-1β, IFNγ, IL-8, IP-10 (CXCL-10), MIG, and RANTES (CCL5); the expression of the inflammatory factor MCP-1 showed an increasing trend, but did not reach a significant change; the anti-inflammatory factor IL-10 was almost not expressed.

[0262] Example 9. Effect of strain MNH46686 on IFNβ expression

[0263] 1.1 Experimental methods

[0264] Interferon (IFN) receptor proteins are a type of cytokine secreted by host cells that regulate immune responses. Interferon is produced under stimulation by viruses, bacterial endotoxins, artificially synthesized double-stranded RNA, etc. Macrophages, lymphocytes, somatic cells, etc. in the human body can produce interferon. Among them, IFNβ belongs to type I interferon, which can promote the activity of NK cells, macrophages, and T lymphocytes. The experimental results in Example 12 of the present invention (Figures 16-18) show that MNH46686 increases NK cells during the anti-tumor process; further, the strain MNH46686 of the present invention was further verified by experiments to be able to promote the expression of IFNβ, thereby exerting antiviral, anti-tumor, and immunomodulatory effects. To verify whether MNH46686 can promote the expression of IFNβ, this study used THP-1 cells (Muen Company's self-built cell line) that have been constructed to carry the IFNβ gene promoter reporter gene (i.e., IFNβ-promoter reporter) to evaluate the effect of MNH46686 on IFNβ transcriptional activity.

[0265] Preparation of MNH46686 culture supernatant: strain MNH46686 was inoculated into MM01 liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the cells were removed by centrifugation. The culture supernatant was filtered through a 0.22 μm filter, aliquoted, and the collected material was stored at -86°C until use.

[0266] Positive drug group: MSA-2 (Sting agonist) (derived from TargetMol Cat No. T8798 (Target Molecule Corp.)) can promote the expression of IFNβ, and three gradients of 20 μM, 40 μM and 80 μM were set.

[0267] Untreated group: DMEM complete medium (10% FBS)

[0268] High group: containing 10% volume of MM01 bacterial culture

[0269] MNH46686 group: containing 10% volume of MNH46686 bacterial culture supernatant

[0270] THP-1-IFNβ-promoter reporter cells were seeded in 96-well plates at 1×10 cells per well. 5Cells were treated according to the designated groups. After 24 hours of culture, the cells were centrifuged at 300g for 5 minutes, the supernatant was removed, and 50 μL of 1× luciferase detection reagent was added. The reaction was allowed to proceed for 1 minute, and luminescence was measured using a microplate reader. Relative fluorescence values ​​were normalized to those of the untreated group and compared with those of the High group to evaluate the effect of MNH46686 on IFNβ transcriptional activity.

[0271] The results showed that the IFNβ gene reporter system constructed in the present invention (the gene reporter system mentioned in the present invention means: the reporter gene is inserted into the vector, the vector infects the cells, and the cell line expressing the reporter gene is screened) (see Document 1: Huashan Du, Tianmin Xu, Manhua Cui "cGAS-STING signaling in cancer immunity and immunotherapy" Biomedicine & Pharmacotherapy 133 (2021) 110972; Document 2: Jiang et al. "cGAS-STING, an important pathway in cancer immunotherapy" Journal of Hematology & Oncology (2020) 13: 81; Document 3: Khiem C. Lam et al. "Microbiota triggers STING-type I IFN-dependent monocyte reprogramming of the tumor microenvironment" Cell 184,5338–5356) showed a good response to positive drugs (Figure 13A). Further evaluation of samples using this system revealed that MNH46686 significantly promoted IFNβ transcriptional activity (Figure 13B). These results suggest that MNH46686 has potential for immunomodulatory, antiviral, and anti-tumor effects.

[0272] Example 10. Animal experiment on the prevention of liver cancer using strain MNH 46686

[0273] To verify the potential of strain MNH 46686 for tumor prevention and treatment, a mouse syngeneic tumor model was used to investigate the inhibition of liver cancer growth. This study has been ethically reviewed by the Muen Biotech Laboratory Animal Care and Use Committee.

[0274] Test strains: After thawing the glycerol cryopreserved tube of MNH 46686 at 37°C, the strain was inoculated on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain was inoculated into MM01 liquid medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured liquid was concentrated by centrifugation and resuspended in a solvent to obtain the purity and viable count (2.04×10 10 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0275] Tumor cells: H22 mouse hepatoma cells, CBP69230.

[0276] Experimental animals: The experimental mice were C57BL / 6J mice, 5 weeks old, a total of 20, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0277] Animal experiment: The mice were raised normally. After the quarantine period, they were randomly divided into two groups (control group and experimental group) according to their body weight, with 10 mice in each group and housed in separate cages. After grouping, H22 mouse liver cancer cells were injected intradermally to establish a mouse liver cancer model. The cell inoculation amount was 2×10 6 / mL, 0.1mL / piece.

[0278] Oral gavage administration began on the day of completion of inoculation (DAY 0). The control group was given culture medium, and the experimental group was given MNH 46686 bacteria. The gavage volume was 0.2 mL / mouse / time, and the administration frequency was 1 day / time. General observations were performed once a day during the quarantine period and once a day after administration during the administration period. The animals were weighed upon receipt, at the end of the quarantine period, and 3 times a week during the administration period. Starting 7 days after tumor cell inoculation (DAY 7), tumor measurements were performed 3 times a week, and tumor growth was recorded. The endpoint of this experiment was: 19 oral administrations were completed. At the end of the experiment, all mice were euthanized by cervical dislocation.

[0279] The experimental results are shown in Table 10 below.

[0280] Table 10. Overall statistics of mice at the end of the experiment

[0281] Figure 14 is a comparison of tumor tissue sizes after dissection of experimental mice. The left picture is the experimental group and the right picture is the control group.

[0282] The weight of the tumor tissues after mouse dissection was counted, and the results are shown in Table 11 below.

[0283] Table 11. Tumor tissue weight after mouse autopsy

[0284] The results showed that tumors in the control group grew rapidly, reaching an average weight of approximately 2.53g on day 19. Tumors in the experimental group averaged approximately 1.69g on day 19, showing a significantly slower growth rate than the control group. This experiment demonstrates that MNH 46686 can significantly reduce the growth rate of liver cancer. This suggests that the strain of the present invention can inhibit tumor growth and therefore has potential applications in tumor prevention and treatment.

[0285] Example 11. Experimental verification of the therapeutic effect of MNH 46686 on lung cancer

[0286] Experimental methods

[0287] To verify the therapeutic efficacy of strain MNH46686, we used a mouse syngeneic tumor model to investigate its ability to inhibit lung cancer growth. This experimental protocol has been ethically reviewed by the Muen Biotech Laboratory Animal Care and Use Committee.

[0288] Test strains: After thawing the glycerol cryopreserved tube of MNH46686 at 37°C, inoculate it on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain is inoculated into MM01 liquid medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured liquid is centrifuged and concentrated, and the cells are resuspended in a solvent to obtain the purity and viable cell count (1.0-2.0×10 9 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0289] Tumor cells: LLC1 mouse lung cancer cells, CL-0140.

[0290] (1) Verification of the therapeutic effect of strain MNH46686 on tumors

[0291] Experimental animals: 120 5-week-old C57BL / 6J mice were purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0292] Animal experiment: The animals were raised normally. After the quarantine period, LLC1 lung cancer cells were subcutaneously inoculated to form an ectopic homologous tumor model. The cell inoculation volume was 2×10 6 / mL, 0.1mL / mouse. When the average tumor volume reaches 80-100mm 3The animals were randomly divided into two groups according to the tumor volume, group A (Control) and group F (MNH46686). Group BE was other test substances. Oral gavage was started on the day of grouping (D1). The control group was given the vehicle and group F was given MNH46686 bacteria. The oral gavage volume was 0.2 mL / animal / time, and the administration frequency was 1 day / time. General observation was performed once a day during the quarantine period and once a day after administration during the administration period. The animals were weighed at the time of receipt and at the end of the quarantine period. The animals were weighed twice a week after tumor inoculation. The day of tumor cell inoculation was D1. From D5 to grouping, the tumor diameter was measured once a day; after grouping, the tumor diameter was measured once every 2 days; when the average tumor volume was ≥1000 mm 3 The tumor diameter was measured once a day and the tumor growth was recorded. The endpoint of this experiment was the 20th day of administration. 3 ) were measured. At the endpoint of the study, all surviving mice were dissected and tumor weight and volume were measured. Statistical analysis and comparison were performed using the following parameters: tumor volume curve, endpoint tumor volume, endpoint tumor weight, and endpoint tumor inhibition rate (TGI). Tumor inhibition rate was calculated as (mean volume of control group minus mean volume of experimental group) / mean volume of control group × 100%. All data were expressed as mean ± SD and analyzed using GraphPad Prism 8.0.2 software. Pairwise comparisons were analyzed using the Student's t test, while two-way ANOVA with Sidak's multiple comparisons were used for two-way comparisons. Significant differences are indicated by *, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0293] At the end of the experiment (D21), the surviving mice were dissected and the tumor tissues were weighed. The results are summarized in the following table.

[0294] Table 12 Overall statistics of mice at the end of the experiment

[0295] The results showed that during the experiment, the statistical analysis of the tumor volume curve of group F (MNH46686) was significantly smaller than that of group A (Control) (Figure 15(A)). At the end of the experiment, the average tumor volume of mice in the control group was 1136.39 mm 3The average tumor tissue weight was about 1.33 g; the average tumor volume of mice in the MNH46686-treated group was 751.78 mm 3 The average tumor tissue weight was approximately 0.74 g. Tumor volume and weight in the MNH46686-treated group were significantly smaller than those in the control group (Table 12, Figures 15(B) and 15(C)). At the endpoint, the tumor growth inhibition rate (TGI) in Group F (MNO-863) was 36.3% (Figure 15(D)), significantly improving the tumor growth inhibition rate.

[0296] Example 12. Flow cytometry results of MNH 46686

[0297] This example explores the immunoassay of mice activated by MNH46686.

[0298] Experimental principle:

[0299] After preparing single-cell suspensions from tumor-bearing mouse tissue, cells are labeled with fluorescent markers. The proportion of cells expressing these markers can be measured using a flow cytometer. The distribution of these immune cells, to some extent, reflects the immune status of the mouse.

[0300] Experimental methods:

[0301] 1. Tumor tissue

[0302] 1. Preparation of tumor single cell suspension and cell counting

[0303] 0.3 g of tumor tissue was collected, rinsed with PBS, cut into small pieces, and pulverized. The pulverized tumor tissue was placed in 5 mL of collagenase IV digestion solution (1 mg / ml collagenase IV, 0.1 mg / ml DNase, 10% FBS) and digested at 37°C for 30 min. The digested tissue fluid was filtered through a 70 μm filter membrane to prepare a mouse tumor single-cell suspension, which was then analyzed and the total cell count was calculated.

[0304] 2. T cell / NK cell staining on tumor cell surface

[0305] Single-cell suspensions of mouse tumors were prepared and dyes (corresponding antibody combinations) T-Live-Tumor / NK-Live-Tumor and T-Tumor / NK-Tumor were added to stain T cells / NK cells on the tumor cell surface. The stained cells were resuspended in PBS and tested on an instrument.

[0306] 3. Staining of the intracellular transcription factor Foxp3 in tumor cells

[0307] Single-cell suspensions of mouse tumors were obtained, and dyes (corresponding antibody combinations) Treg-Live-Tumor and Treg-Surface-Tumor were added in sequence for staining. The stained cells were resuspended in PBS. Subsequently, fixative and permeabilization solutions were used in sequence according to the instructions of the detection kit, and Treg-Foxp3-Tumor nuclear stain was used for staining. The stained cells were resuspended in PBS and tested on the microscope.

[0308] 2. Peripheral blood

[0309] Peripheral blood was collected and lysed for 5 minutes using red blood cell lysis buffer. The cells were then centrifuged at 500 g and the supernatant removed. The cells were lysed again and resuspended in PBS to obtain a single-cell suspension. Single-cell suspensions of mouse peripheral blood were stained for T cells and NK cells by adding T-Live-Blood / NK-Live-Blood and T-Blood / NK-Blood dyes, respectively. The stained cells were then resuspended in PBS and analyzed.

[0310] 3. Spleen Tissue

[0311] 1. Preparation of Splenic Single Cell Suspension and Cell Counting

[0312] 0.1 g of spleen tissue was collected, the surface was rinsed with PBS and then the tissue was crushed. The crushed spleen tissue was suspended in PBS and filtered through a 70 μm filter membrane to prepare a mouse spleen single cell suspension for detection.

[0313] Take a small amount of the mouse spleen single cell suspension prepared above, add red blood cell lysis buffer, Spleen-Counting, pre-cooled PBS, etc. in sequence, complete the on-machine detection, and accurately count the mouse spleen single cell value.

[0314] The remaining mouse spleen single cell suspension prepared above was taken and red blood cell lysis buffer, PBS, etc. were added in sequence for cell treatment. The treated cell suspension was filtered through a 300-mesh filter and then tested on an instrument.

[0315] 2. Splenic Cell Surface T Cell / NK Cell Staining

[0316] Single cell suspensions of mouse spleen were obtained, and dyes T-Alexa Fluor647 anti-mouse FOXP3-SP / NK-Alexa Fluor647anti-mouse FOXP3-SP, T-Live-SP / NK-Live-SP, and T-SP / NK-SP were added to stain T cells / NK cells on the surface of spleen cells. The stained cells were resuspended in PBS and detected by the instrument.

[0317] 3. Spleen Cell Intracellular Transcription Factor Foxp3 Staining

[0318] Single cell suspension of mouse spleen was obtained, and dyes Treg-Fcblock-SP, Treg-Live-SP, and Treg-Surface-SP were added in sequence for staining. The stained cells were resuspended in PBS. Subsequently, according to the instructions of the detection kit, fixative, permeabilization solution, and Treg-SP nuclear staining were used for staining in sequence. The stained cells were resuspended in PBS and detected by the instrument.

[0319] Experimental results:

[0320] Combined with Figures 16 and 17, the results of the proportion of various immune cells in peripheral blood show that compared with the control group, the proportion of NK cells in the peripheral blood of mice was significantly increased, and immune T cells such as CD45+CD3+, CD45+CD4+, and CD45+CD8+ all showed an upward trend (see Figure 16G-J); Analysis of the correlation between various immune cells in peripheral blood and tumor endpoint volume (see Figure 17) shows that the tumor endpoint volume is significantly negatively correlated with the proportion of NK cells in peripheral blood, and there is no significant correlation with the proportion of other immune cells. This shows that MNH46686 can activate the mouse systemic immunity by increasing the proportion of NK cells in the peripheral blood of mice and exert an anti-tumor effect.

[0321] The proportion of M2 macrophages, immunosuppressive Treg cells, killer cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells in tumor tissues (see A to F in Figure 16) showed that compared with the control group, the proportion of killer cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells in mouse tumor tissues showed an upward trend, while M2 macrophages and immunosuppressive Treg cells showed a downward trend; analysis of the correlation between the above cells in tumor tissues and the tumor endpoint volume (see A to F in Figure 17) showed that the tumor endpoint volume was significantly positively correlated with the proportion of immunosuppressive Treg cells and M2 macrophages in tumor tissues, and was significantly positively correlated with the tumor killer cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells in tumor tissues. The results showed that MNH46686 can inhibit the proportion of M2 macrophages and immunosuppressive Treg cells in mouse tumors, increase the number of tumor-killing cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells, and activate the local immunity of mouse tumors, thereby exerting an anti-tumor effect. The strain MNH46686 can exert its anti-tumor activity by stimulating the activation of the mouse immune system.

[0322] Example 13. Experimental verification of the therapeutic effect of MNH 46686 combined with PD-1 antibody on lung cancer tumors:

[0323] 1. Experimental Methods

[0324] To validate the therapeutic efficacy of strain MNH46686 combined with a PD-1 antibody, a mouse syngeneic tumor model was used to investigate the inhibition of lung cancer growth. This experimental protocol has been reviewed by the Institutional Animal Care and Use Committee of Muen Biotech.

[0325] Test strains: After thawing the glycerol cryopreserved tube of MNH46686 at 37°C, the strain was inoculated on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain was inoculated into MM01 liquid medium and cultured anaerobically to obtain a sufficient number of viable bacteria. The cultured liquid was centrifuged and concentrated, and the cells were resuspended in a solvent to obtain the purity and viable count (9.23×10 8 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0326] Tumor cells: LLC1 mouse lung cancer cells, catalog number CL-0140.

[0327] Experimental animals: The experimental mice were C57BL / 6J mice, aged 5-6 weeks, a total of 108, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0328] Animal experiment: The animals were raised normally. After the quarantine period, LLC1 lung cancer cells were subcutaneously inoculated to form an ectopic homologous tumor model. The cell inoculation volume was 2×10 6 / mL, 0.1mL / mouse. When the average tumor volume reaches 60-100mm 3 At the time of the study, the animals were randomly divided into eight groups based on tumor volume: Group A (Negative Control), Group B (Positive Control, PD-1), Group E (MNH46686 & PD-1), and Groups C, D, and FH received other test substances. Dosing began on the day of grouping: Group A received vehicle and saline, Group B received vehicle and PD-1 antibody, and Group E received MNH46686 and PD-1 antibody. Vehicle and MNH46686 were administered by gavage at a volume of 0.2 mL per animal per day for a total of 20 doses. PD-1 antibody and saline were administered by intraperitoneal injection at a dose of 10 mg / kg every three days for a total of six doses. Animals were weighed upon receipt and at the end of the quarantine period. Animals were weighed twice weekly after tumor inoculation, including on the day of intraperitoneal injection. The day of tumor cell inoculation was designated Day 1. From D5 to grouping, the tumor diameter was measured once a day; after grouping, the tumor diameter was measured once every 3 days; and the tumor growth was recorded. The endpoint of the experiment was the 23rd day of administration. During the experiment, general clinical observation, body weight monitoring, tumor volume (mm 3) were measured. At the experimental endpoint, all surviving mice were dissected and tumor weight and volume were measured. Statistical analysis and comparison were performed using the following indicators: tumor volume curve, endpoint tumor volume, endpoint tumor weight, and endpoint tumor growth inhibition (TGI). TGI = 1 - (individual tumor volume - individual tumor volume at grouping) / (mean tumor volume of Group A - mean tumor volume of Group A at grouping) × 100%. All data are expressed as mean ± SD and analyzed using GraphPad Prism 8.0.2 software. For comparisons between three or more groups, one-way ANOVA with Tukey's multiple comparisons test was used. Two-way ANOVA with Sidak's multiple comparisons was used for two-factor analysis. Significant differences are indicated by *, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0329] 2. Experimental results of the combination of strain MNH46686 and PD-1 antibody for the treatment of lung cancer

[0330] At the end of the experiment (D24), the surviving mice were dissected and the tumor tissues were weighed. The results are summarized in Table 13.

[0331] Table 13 Overall statistics of mice at the end of the experiment

[0332] The results showed that during the experiment, the statistical analysis of the tumor volume curve of Group E (MNH46686 & PD-1) was significantly smaller than that of Group A (Negative Control) and Group B (Positive Control) (Figure 19). At the end of the experiment, the average tumor volume of mice in the negative control group was 1279.80 mm 3 The average tumor tissue weight was about 1.28g; the average tumor volume of the positive control group (Positive Control) mice was 710.29mm 3 The average tumor tissue weight was about 0.92g; the average tumor volume of mice in the MNH46686 & PD-1 antibody treatment group was 346.99mm 3The average tumor tissue weight reached approximately 0.47g. Compared to the negative control group, the endpoint tumor volume and weight of mice treated with MNH46686 and PD-1 antibody were significantly reduced. Compared to the positive control group, both endpoint tumor volume and weight showed a downward trend (Figures 20 and 21). At the endpoint, the tumor growth inhibition rate (TGI) in Group B (Positive Control) was 47.3%, and in Group E (MNH46686 and PD-1) was 77.2%, significantly higher than that in the positive control group (Figure 22). At the endpoint, the response rate in Group A (Negative Control), Group B (Positive Control), and Group E (MNH46686 and PD-1) was 20.0%, 60.0%, and 87.5%. The combination of MNH46686 and PD-1 antibody significantly improved the response rate of tumors (Figure 22). The data in the figures are presented as mean ± standard deviation (Mean ± SD). Statistical analysis was performed using two-way ANOVA with Sidak's multiple comparisons. Significant differences are indicated by *, *p < 0.05.

[0333] Example 14. Inhibitory effect of strain MNH46686 on histone deacetylase (HDAC) activity

[0334] To verify whether MNH46686 has an inhibitory effect on histone deacetylase activity, this study used the HDAC Inhibitor Drug Screening Kit (Fluorometric) purchased from Abcam to perform in vitro HDAC activity inhibition detection.

[0335] Preparation of MNH46686 culture supernatant: strain MNH46686 was inoculated into liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the cells were removed by centrifugation. The culture supernatant was filtered through a 0.22 μm filter, aliquoted, and the collected material was frozen at -80°C for later use.

[0336] Assay sample preparation: 1) Control group: MM01 culture medium was diluted 10-fold with PBS to obtain a 10% MM01 content; 2) Positive control group: TSA, an HDAC inhibitor, was diluted with PBS to a final concentration of 40 μM; 3) MNH46686 group: MNH46686 culture supernatant was diluted 10-fold with PBS to obtain an experimental sample containing 10% bacterial supernatant.

[0337] Preparation of HDACs detection reaction reagents: Prepare an appropriate amount of detection reaction system according to the kit instructions. Each reaction requires 50 μL of reaction reagent.

[0338] HDACs activity assay: Add 50 μL of test sample to a 96-well white plate, followed by 50 μL of each reaction reagent, mix thoroughly, and incubate at 37°C for 30 minutes. Add 10 μL of Lysine Developer to each well and mix thoroughly to terminate the reaction. Incubate the plate at 37°C for 30 minutes. Finally, measure the fluorescence intensity of the samples using a microplate reader. The reader settings are Ex. = 350-380 nm, Em. = 440-460 nm. For HDACs inhibitory activity analysis, set the fluorescence intensity of the control to 100%. Divide the fluorescence intensity of the positive control and MNH46686 by the control and multiply by 100% to obtain relative HDACs activity.

[0339] The results (see Figure 23) show that compared to the HDAC activity of the control group, the supernatant of MNH46686 significantly inhibited HDAC activity, similar to the effect of TSA, the HDAC inhibitor control group. These results suggest that MNH46686 can inhibit HDAC activity and, by inhibiting HDAC activity, can be used to prevent or treat diseases mediated by HDAC activity.

[0340] The function of HDACs in cancer is associated with the abnormal expression or function of genes that promote cell proliferation and tumorigenic phenotypes. In certain cancers, HDACs primarily regulate the onset of cancer and have been described as oncogenes. Reduced or inhibited HDAC expression has been shown to have a variety of anti-cancer effects, such as cell cycle arrest and inhibition of proliferation, apoptosis, differentiation, and senescence, as well as disruption of angiogenesis. Therefore, the strain MNH46686 of the present invention has the ability to inhibit HDAC activity, suggesting that the drugs or compositions of the present invention can be used to prevent or treat tumors or cancers mediated by HDAC activity. Because genes regulated by HDACs play an important role in angiogenesis, the drugs or compositions of the present invention can be used to prevent tumor metastasis.

[0341] Diabetes is a group of diseases in which low levels of insulin and / or peripheral insulin resistance lead to hyperglycemia. It has been proposed that inhibition of HDAC is used to treat diabetes by various mechanisms, including the inhibition of Pdxl (Park et al., 2008, J Clin Invest, 118, 2316-24), enhancing the expression of transcription factor Ngn3 to increase endocrine reservoirs. Progenitor cells (Haumaitre et al., 2008, Mol Cell Biol, 28, 6373-83) and enhanced insulin expression (Molsey et al., 2003, J Biol Chem, 278, 19660-6) etc. HDAC inhibition is also a promising method for treating complications of late diabetes such as diabetic nephropathy and retinal ischemia (Christensen et al., 2011, Mol Med, 17 (5-6), 370-390). Therefore, the compositions of the present invention can be used to treat or prevent diabetes mediated by HDAC activity.

[0342] The compositions of the present invention are used to treat or prevent diabetes. In a preferred embodiment, the compositions of the present invention are used to treat or prevent type I diabetes. In a preferred embodiment, the compositions of the present invention are used to treat or prevent type II diabetes. In certain embodiments, the compositions of the present invention are used to treat or prevent diabetes, wherein the treatment or prevention is achieved by reducing or preventing HDAC activation.

[0343] The strain MNH46686 of the present invention can inhibit HDAC activity, activate T cells, and achieve anti-tumor purposes.

[0344] The compositions of the present invention have HDAC inhibitory activity, and thus they can be used to treat inflammatory bowel diseases mediated by HDAC activity.

[0345] Example 15 MNH46686 combined with PD-1 to activate mouse immune analysis

[0346] After preparing single-cell suspensions from tumor-bearing mouse tissue, cells are labeled with fluorescent markers. The proportion of cells expressing these markers can be measured using a flow cytometer. The distribution of these immune cells, to some extent, reflects the immune status of the mouse.

[0347] Experimental methods:

[0348] Tumor tissue was removed, the surface rinsed with PBS, cut into small pieces, and pulverized. The pulverized tumor tissue was placed in 5 mL of collagenase IV digestion solution (1 mg / ml collagenase IV, 0.1 mg / ml DNase, 10% FBS) and digested at 37°C for 30 min. The digested tissue fluid was filtered through a 70 μm filter membrane to prepare a mouse tumor single-cell suspension, which was then analyzed and the total number of cells in the tissue was counted. The mouse tumor single-cell suspension was stimulated with stimulant for 4 hours. After washing and resuspending the cells, the dyes (corresponding antibody combination) Live-Tumor and IFN / TNF-Surface-Tumor were added sequentially for staining. The stained cells were resuspended in PBS. Subsequently, the cells were treated with fixative and permeabilization solution according to the detection kit instructions, and stained with IFN / TNF-Tumor nuclear stain. The stained cells were resuspended in PBS and analyzed.

[0349] Flow cytometry data were processed using CyExpert and statistical analysis was performed using Graphpad Prism V9. Statistical analysis was performed using one-way ANOVA with Dunnet's multiple comparison, ns not significant*p<0.05,**p<0.01,***p<0.001.

[0350] Experimental results:

[0351] The proportions of immune cells with tumor-killing effects in tumor tissues, including CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells, were found to be upregulated in the tumor tissues of mice in the MNH46686+PD-1 group compared with those in the positive control group (PD-1 group). The proportions of immune cells with tumor-killing effects in the MNH46686+PD-1 group were found to be upregulated in the tumor tissues of mice in the positive control group (PD-1 group). The correlation between the above cells in tumor tissue and the tumor endpoint weight (see Figures 25A to 25D) shows that tumor weight is significantly negatively correlated with the proportion of tumor-killing cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells in tumor tissue; thus, it can be seen that MNH46686 can activate local tumor immunity in mice by increasing the proportion of tumor-killing cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells, thereby enhancing the anti-tumor effect of PD-1 antibodies.

[0352] Example 16 Experimental study on the efficacy of strain MNH46686 in lung cancer tumors

[0353] 1.2.1 Experimental methods

[0354] To investigate the effect of MNH46686 dosage on tumor treatment, we used a mouse syngeneic tumor model to investigate the inhibition of lung cancer growth. This experimental protocol has been reviewed by the Institutional Animal Care and Use Committee of Muen Biotech.

[0355] Test strains: After thawing the glycerol cryopreserved tube of MNH46686 at 37°C, the strain was inoculated on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain was inoculated into MM01 liquid medium and cultured anaerobically to obtain a sufficient number of viable bacteria. The cultured bacterial solution was centrifuged and concentrated, and the cells were resuspended in a solvent to obtain the purity and viable count (4.1×10 10 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0356] Tumor cells: LLC1 mouse lung cancer cells, catalog number CL-0140.

[0357] Experimental animals: The experimental mice were C57BL / 6J mice, aged 5-6 weeks, a total of 60, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0358] Animal experiment: The animals were raised normally. After the quarantine period, LLC1 lung cancer cells were subcutaneously inoculated to form an ectopic homologous tumor model. The cell inoculation volume was 2×10 6 / mL, 0.1mL / mouse. When the average tumor volume reaches 60-100mm 3 The patients were randomly divided into three groups according to tumor volume: group A (Negative Control), group B (High-MNH46686), and group C (Low-MNH46686). The drugs were given on the day of grouping. Group A (Negative Control) was given the negative control substance PBS-Cys (glycan), and group B (high-MNH46686) was given 10 -1 Dilution (8.2×10 8 CFU / mouse), group C (Low-MNH46686) was given 10 -2 Dilution (8.2×10 7 CFU / animal), administered by oral gavage, with a gavage volume of 0.2 mL / animal / time, and a dosing frequency of 1 day / time, for a total of 21 doses. General clinical observations were performed once a day during the trial. Animals were weighed upon receipt and at the end of the quarantine period; animals were weighed twice a week after tumor inoculation. Tumor diameter was measured once a day from D5 to grouping; and once every 2 days after grouping. When the average tumor volume in any group reached or exceeded 1000 mm 3The experiment can be terminated if the average tumor volume of any group from Group B to Group C is significantly smaller than that of Group A or the average tumor growth inhibition rate (TGI) of the group reaches or exceeds 30%; or if the average tumor volume of any group of mice reaches or exceeds 2000mm 3 The experiment was terminated if the patient reached the endpoint. All surviving mice were dissected or euthanized at the endpoint, and tumor weight and volume were measured. Statistical analysis and comparison were performed using tumor volume curves, endpoint tumor volume, endpoint tumor weight, and endpoint tumor growth inhibition (TGI). TGI = 1 - (individual tumor volume - individual tumor volume at grouping) / (mean tumor volume of group A - mean tumor volume of group A at grouping) × 100%. All data are expressed as mean ± SD and analyzed using GraphPad Prism 8.0.2 software. Comparisons between three or more groups were performed using one-way ANOVA with Dunnett's multiple comparisons test. Two-way ANOVA with Sidak's multiple comparisons was used for two-factor analysis. Significant differences are indicated by *, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0359] 1.2.2 Results of the experimental study on the efficacy of strain MNH46686 in lung cancer tumors

[0360] At the end of the experiment (D27), the surviving mice were dissected and the tumor tissues were weighed. The results are summarized in Table 14.

[0361] Table 14 Overall statistics of mice at the end of the experiment

[0362] During the experiment, except for erythema, scabs, tumor cavitation or low body temperature caused by the tumor, no other abnormalities were observed in all mice. At the end of the experiment, the statistical analysis results of the tumor volume curve of Group C (Low-MNH46686) were significantly smaller than those of Group A (Negative Control), and there were no significant differences between the other groups and Group A (as shown in Figure 26). At the end of the experiment (D27), the average tumor volume of Groups A to C was 1067.54 mm 3 、838.01mm3 and 561.50mm 3 , with average tumor weights of 1.39g, 1.18g, and 0.72g, respectively. At the endpoint, the tumor volumes and weights of Groups B through C compared with those of Group A are shown in Figures 27 and 28 . At the endpoint, the TGIs of Groups B through C were 23.00% and 50.30%, respectively. At the endpoint, the TGIs of Groups B through C compared with those of Group A are shown in Figure 30 . The response rates in Group A, Group B, and Group C were 22.22%, 33.33%, and 66.67%, respectively (Figure 30 ).

[0363] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of a Lachnospiraceae microbial strain, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation culture broth of the Lachnospiraceae microbial strain, or a supernatant of the fermentation culture broth, in the preparation of a drug for preventing and / or treating tumors, characterized in that: The Lachnospiraceae microbial strain belongs to a new genus and species of the Lachnospiraceae family (Lachnospiraceae sp.), and the Lachnospiraceae microbial strain has a 16S rDNA of the nucleotide sequence shown in SEQ ID No. 1 or a 16S rDNA with at least 90% identity thereto, such as at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.65%, 98.7%, 99% identity, such as 97%, 98.7% or 99% identity; The tumor includes a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by viral infection or a tumor caused by bacterial infection, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Epstein-Barr virus infection, and Helicobacter pylori (HP) infection and Fusobacterium nucleatum (F. nucleatum) infection.

2. The use according to claim 1, characterized in that The Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in Guangdong Microbial Culture Collection Center with the deposited name of Lachnospiraceae sp. MNH 46686 and the deposited number of the strain is GDMCC No: 62002; Alternatively, the Lachnospira microbial strain has an average nucleotide identity ANI value of at least 95% with the Lachnospira microbial strain MNH 46686, preferably, the average nucleotide identity ANI value is 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

3. A composition comprising a Lachnospiraceae microbial strain belonging to a new genus and species of the Lachnospiraceae family, wherein the Lachnospiraceae microbial strain has a 16S rDNA sequence of the nucleotide sequence shown in SEQ ID No. 1 or a 16S rDNA sequence having at least 90% identity thereto, such as at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.65%, 98.7%, 99% identity, such as 97%, 98.7% or 99% identity; preferably, the composition is an oral composition; Preferably, the composition further comprises a pharmaceutically acceptable carrier; Preferably, the Lachnospiraceae microbial strain and the pharmaceutically acceptable carrier are present in an enteric coating.

4. The composition according to claim 3, characterized in that The Lachnospiraceae microbial strain includes the Lachnospiraceae microbial strain MNH 46686, which is deposited in the Guangdong Provincial Microbial Culture Collection Center with the deposited name of Lachnospiraceae sp. MNH 46686 and the deposited number of the strain is GDMCC No: 62002; Alternatively, the Lachnospira microbial strain has an average nucleotide identity ANI value of at least 95% with the Lachnospira microbial strain MNH 46686, preferably, the average nucleotide identity ANI value is 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or 100%.

5. The composition according to any one of claims 3 to 4, characterized in that The invention comprises living cells or living bacteria or attenuated bacteria or irradiated bacteria or inactivated bacteria of the Lachnospiraceae microbial strain or a culture of the Lachnospiraceae microbial strain or a metabolite of the Lachnospiraceae microbial strain or a fermentation broth of the Lachnospiraceae microbial strain or a supernatant of the fermentation broth.

6. The composition according to claim 5, wherein The composition further comprises a combined drug; the combined drug is selected from at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, an immunotherapy drug, and an enhanced cell therapy drug; Preferably, the chemotherapy drugs include one or more of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin or epirubicin; Preferably, the photosensitizer comprises one or more of boron dipyrrole, chlorin or red bengal; Preferably, the photothermal agent is one or more of indocyanine green, new indocyanine green or gold nanoparticle rods; Preferably, the immunotherapy drug comprises a PD-1 antibody, a CTLA-4 antibody, a PD-L1 antibody, and a PD-L1 inhibitor; more preferably, the PD-L1 inhibitor is selected from durvalumab, atezolizumab, or avelumab; the PD-1 antibody or PD-L1 antibody is selected from pembrolizumab or nivolumab; the CTLA-4 antibody is selected from ipilimumab; more preferably, the effective dose of the PD-1 inhibitor is 1 to 11 mg / kg; preferably, the effective dose of PD-1 includes: 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg; Preferably, the enhanced cell therapy comprises CAR-T; Preferably, the content of microbial strains in the composition per unit dose includes 2×10 7 ~9×10 10 CFU / mL or 1×10 6 ~2×10 10 CFU / mg of Lachnospiraceae microbial strains; the unit dose is preferably 4×10 7 CFU / mL, 5×10 7 CFU / mL, 6×10 7 CFU / mL, 7×10 7 CFU / mL, 8×10 7 CFU / mL, 1×10 8 CFU / mL, 2×10 8 CFU / mL, 3×10 8 CFU / mL, 4×10 8 CFU / mL, 5×10 8 CFU / mL, 6×10 8 CFU / mL, 7×10 8 CFU / mL, 8×10 8 CFU / mL, 9×10 8 CFU / mL, 1×10 9 CFU / mL, 2×10 9 CFU / mL, 3×10 9 CFU / mL, 4×10 9 CFU / mL, 5×10 9 CFU / mL, 6×10 9 CFU / mL, 7×10 9 CFU / mL, 8×10 9 CFU / mL, 9×10 9 CFU / mL, 2×10 10 or 1.2×10 6 CFU / mg, 4×10 6 CFU / mg, 5×10 7 CFU / mg, 6×10 7 CFU / mg, 7×10 7 CFU / mg, 8×10 7 CFU / mg, 9×10 7 CFU / mg, 1×10 8 CFU / mg, 2×10 8 CFU / mg, 3×10 8 CFU / mg, 4×10 8 CFU / mg, 5×10 8 CFU / mg, 6×10 8 CFU / mg, 7×10 8 CFU / mg, 8×10 8 CFU / mg, 9×10 8 at least one of CFU / mg.

7. The composition according to claim 5, wherein The composition further comprises a pharmaceutically acceptable carrier; preferably, the carrier is selected from one or more of a diluent, a dispersant, an excipient, a stabilizer, a lubricant, and a disintegrant; Preferably, the drug is in the form of a liquid preparation, a solid preparation, a capsule preparation, a sustained-release preparation, and a nanoformulation.

8. The composition of claim 5, wherein at least 50% of the bacteria are viable, for example, at least 90% are viable.

9. The composition of claim 5, wherein the composition is in the form of a powder, microencapsulated powder, capsule, tablet, lozenge, granule, emulsion, suspension, suppository, beverage, food, medicine or nutraceutical, food additive, dietary supplement or dairy product.

10. Use of the composition according to any one of claims 3 to 9 in the preparation of a medicament for preventing and / or treating at least one of metabolic diseases, diabetes, autoimmune diseases, infectious diseases, central nervous system diseases, and inflammatory bowel diseases.

11. Use of the composition according to any one of claims 3 to 9 in the preparation of a drug for anti-tumor or tumor growth inhibition. Preferably, inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid, propionic acid, butyric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one of acetic acid, propionic acid, butyric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect; The tumor includes a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from at least one of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, kidney cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by a virus or a tumor caused by bacteria, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Helicobacter pylori infection (HP), Epstein-Barr virus infection, and tumors caused by Helicobacter pylori (HP) infection and Fusobacterium nucleatum (F.nucleatum) infection.

12. Use of the composition according to any one of claims 3 to 9 in the preparation of a medicament for treating and / or preventing a disease mediated by HDAC activity, preferably, the disease mediated by HDAC activity comprises at least one of a tumor, a metabolic disease, diabetes, an autoimmune disease, an infectious disease, a central nervous system disease, and an inflammatory bowel disease; preferably, the tumor comprises a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor; preferably, the tumor is selected from the group consisting of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, and ovarian cancer. , pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a tumor caused by viral infection or a tumor caused by bacterial infection, such as human papillomavirus infection (HPV), hepatitis B or C infection (HBV, HCV), human immunodeficiency virus infection (HIV), Epstein-Barr virus infection, and Helicobacter pylori (HP) infection, Fusobacterium nucleatum (F.nucleatum) infection.

13. A Lachnospiraceae microbial strain, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation broth of the Lachnospiraceae microbial strain, or a supernatant of the fermentation broth, characterized in that: The Lachnospiraceae microbial strain belongs to a new genus and species (Lachnospiraceae sp.) of the Lachnospiraceae family. The nucleotide sequence of 16S rDNA of the Lachnospiraceae microbial strain is shown in SEQ ID No.

1.

14. The Lachnospiraceae microbial strain, or the culture of the Lachnospiraceae microbial strain, or the metabolite of the Lachnospiraceae microbial strain, or the fermentation broth of the Lachnospiraceae microbial strain, or the supernatant of the fermentation broth according to claim 13, characterized in that: The Lachnospiraceae microbial strain includes Lachnospiraceae microbial strain MNH 46686, which is preserved in Guangdong Microbial Culture Collection Center with the preservation name of Lachnospiraceae sp. MNH 46686 and the preservation number of the strain being GDMCC No: 62002.

15. A method for treating or preventing tumors or diabetes, comprising the step of administering to a subject in need thereof an effective amount of the Lachnospiraceae microbial strain of claim 13 or 14, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation broth of the Lachnospiraceae microbial strain, or a supernatant of the fermentation broth, or the composition of any one of claims 3 to 9; Preferably, the tumor comprises a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from one or more of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a viral tumor or a bacterial tumor, such as a tumor caused by human papillomavirus infection, hepatitis B infection or hepatitis C infection, human immunodeficiency virus infection, Helicobacter pylori infection, Epstein-Barr virus infection or Fusobacterium nucleatum infection; Preferably, the strain prevents and / or treats tumors by inhibiting tumor growth, and inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one short-chain fatty acid or short-chain fatty acid salt of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect.

16. The Lachnospiraceae microbial strain according to claim 13 or 14, or a culture of the Lachnospiraceae microbial strain, or a metabolite of the Lachnospiraceae microbial strain, or a fermentation broth or supernatant of the fermentation broth of the Lachnospiraceae microbial strain, or the composition according to any one of claims 3 to 9, for use in preventing and / or treating tumors or diabetes; Preferably, the tumor comprises a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; preferably, the tumor is selected from one or more of liver cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, breast cancer, cervical cancer, ovarian cancer, pancreatic cancer, bile duct cancer, renal cancer, glioma, liposarcoma, melanoma, lymphoma, small cell lung cancer, squamous cell carcinoma, chondrosarcoma and fibrosarcoma; or the tumor is a viral tumor or a bacterial tumor, such as a tumor caused by human papillomavirus infection, hepatitis B infection or hepatitis C infection, human immunodeficiency virus infection, Helicobacter pylori infection, Epstein-Barr virus infection or Fusobacterium nucleatum infection; Preferably, the strain prevents and / or treats tumors by inhibiting tumor growth, and inhibiting tumor growth includes at least one of the following: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) inhibiting tumor cell metastasis; (g) reducing PD-1 resistance; (h) increasing the NK ratio in peripheral blood to activate the subject's systemic immunity; (i) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (j) inhibiting tumors through at least one short-chain fatty acid or short-chain fatty acid salt of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valeric acid in SCFA; (k) inhibiting tumors by regulating the subject's immune system to exert an immunomodulatory effect.

Images