Strain, culture and application thereof

Abstract

The invention discloses a strain, a culture and application thereof. In a first aspect of the present application, a strain is provided, which may be a strain MNH19801 having a preservation number of GDMCC No: 61980, or a strain having at least 95% identity to the 16S rRNA of strain MNH19801, and / or having at least 87% average nucleotide identity to the genome of strain MNH19801. According to the embodiment of the invention, a new strain under the hispiraceae is newly found, and the strain or a product based on the strain has good effects of preventing or treating tumors, diabetes mellitus or other inflammatory diseases.

Description

Technical Field

[0001] The present application relates to the technical field of microbiology, and in particular to strains, cultures and applications thereof. Background Art

[0002] The intestine is the largest microecological environment in the human body. It participates in many important physiological processes such as nutrient absorption, energy metabolism, tissue and organ development, immune defense, and endocrine regulation, and is closely related to human health. Intestinal microecology has made great progress in the past decade, and human understanding of it has continued to deepen. 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, namely 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.

[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 intestinal flora on intestinal and extra-intestinal diseases have 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 and brain disease, etc.) and tumors in humans. Studying the relationship between 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 transformation.

[0004] Intestinal flora imbalance increases the incidence of colorectal cancer, and some intestinal microbial metabolites can directly slow down carcinogenesis or inhibit tumor occurrence. In addition to intestinal-related colon and rectal cancer, intestinal flora can also affect breast cancer, liver cancer, etc. More and more evidence shows that intestinal microorganisms can affect tumor formation, tumor development, and effectively cooperate with tumor treatment. How to regulate the immune and inflammatory responses caused by intestinal flora so that it will not induce tumors but can enhance the efficacy of anti-tumor drugs is still under discussion. If these problems can be solved, it will be of great significance to the innovation and change in the field of tumor treatment in the future. Therefore, the development of new methods to treat diseases requires the exploration of new bacterial species and strains and the characterization of new functional chemical entities. These will help accelerate the development of new microbiome-guided therapies based on new bacterial species and functional chemicals. Summary of the invention

[0005] The present application aims to solve at least one of the technical problems existing in the prior art. To this end, the present application proposes a new strain that can be used to treat tumors.

[0006] In the first aspect of the present application, a strain is provided, which is any one of the following A1) to A4):

[0007] A1) strains having 16S rRNA that is at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identical to the sequence shown in SEQ ID NO:1;

[0008] A2) strain MNH19801 with a deposit number of GDMCC No: 61980;

[0009] A3) strains having at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity to the 16S rRNA of the strain described in A2);

[0010] A4) a strain whose genome has an average nucleotide identity of at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% with the genome of any of the strains A1) to A3); and / or a strain whose genome has a alignment score of at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% with the genome of any of the strains A1) to A3).

[0011] In some embodiments of the present application, the sequence of 16S rRNA of the strain described in A2) is as shown in SEQ ID No.1.

[0012] In some embodiments of the present application, the strain is a strain of the Lachnospiraceae family.

[0013] In some embodiments of the present application, the strain is a strain of the genus Blautia in the family Lachnospiraceae.

[0014] In some embodiments of the present application, the strain is a new species (Blautia sp.) of the genus Blautia in the family Lachnospiraceae.

[0015] In some embodiments of the present application, the strain is a new species of the genus Oliverpabstia in the family Lachnospiraceae (Oliverpabstia sp.).

[0016] In some embodiments of the present application, the strain is highly acetic acid producing.

[0017] The strains according to the embodiments of the present application have at least the following beneficial effects:

[0018] The present application example newly discovered a new strain of Lachnospiraceae, and the strain or the product based on the strain has a good effect in preventing or treating tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases.

[0019] The second aspect of the present application provides a culture of the aforementioned strain.

[0020] In some embodiments of the present application, the culture comprises any one of the following C1) to C5):

[0021] C1) fermentation medium of the strain; C2) supernatant of fermentation medium of the strain; C3) concentrated product of C1) or C2); C4) extract of C1) or C2); C5) dried product of any one of C1) to C4);

[0022] In some embodiments of the present application, the fermentation medium is a liquid medium.

[0023] In some embodiments of the present application, the culture is obtained using a liquid culture medium under anaerobic culture conditions.

[0024] In some embodiments of the present application, the fermentation medium is MM01 liquid medium, and / or the culture temperature is 35-42° C., and / or the culture time is 24-72 hours, and / or the pH of the culture medium is 6-10.

[0025] The third aspect of the present application also provides a microbial agent, which includes the aforementioned strain, or its metabolites, or the aforementioned culture.

[0026] The fourth aspect of the present application further provides a composition, wherein the composition or the raw material of the composition includes the aforementioned strain, or the aforementioned culture, or the aforementioned microbial agent.

[0027] In some embodiments of the present application, the composition further comprises at least one of an excipient, a diluent, and a carrier.

[0028] In some embodiments of the present application, the composition further comprises an auxiliary material.

[0029] In some embodiments of the present application, the auxiliary material also includes at least one of a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor.

[0030] In some embodiments of the present application, the composition is at least one of a medicine, a nutritional supplement, a food, a food additive, a feed, and a feed additive.

[0031] In some embodiments of the present application, the dosage form of the composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and irrigations.

[0032] In a fifth aspect of the present application, a pharmaceutical composition is provided, comprising the following D1) and D2):

[0033] D1) the aforementioned strain, or a metabolite of the aforementioned strain, or the aforementioned culture, or the aforementioned microbial agent;

[0034] D2) Combination drugs.

[0035] In some embodiments of the present application, D2) includes at least one of tumor drugs, metabolic disease drugs, inflammatory disease drugs, autoimmune disease drugs, infectious disease drugs, and central nervous system disease drugs.

[0036] In some embodiments of the present application, the tumor drug includes at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0037] In some embodiments of the present application, the chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin.

[0038] In some embodiments of the present application, the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin, and rose bengal.

[0039] In some embodiments of the present application, the photothermal agent includes at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials.

[0040] In some embodiments of the present application, the immunotherapy drug includes at least one of an immune cell therapy drug and an immune checkpoint inhibitor.

[0041] In some embodiments of the present application, the immune cell therapy drug includes at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and a NK cell therapy drug.

[0042] In some embodiments of the present application, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, and BTLA.

[0043] In some embodiments of the present application, the pharmaceutical composition comprises 1×10 3 ~10 13 CFU / g strain.

[0044] In some embodiments of the present application, the pharmaceutical composition comprises at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×10 10 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 CFU / g strain.

[0045] In some embodiments of the present application, the strain in the pharmaceutical composition is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium.

[0046] In some embodiments of the present application, the pharmaceutical composition or the raw material of the pharmaceutical composition includes about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80% of D1).

[0047] In some embodiments of the present application, the pharmaceutical composition or the raw materials of the pharmaceutical composition include D1) in a mass percentage of 1-80%, 2-70%, 5-60%, 10-50%, or 20-40%.

[0048] In some embodiments of the present application, the pharmaceutical composition further comprises at least one of an excipient, a diluent, and a carrier.

[0049] In some embodiments of the present application, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0050] In some embodiments of the present application, the auxiliary material also includes at least one of a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor.

[0051] In some embodiments of the present application, the pharmaceutical composition is in a dosage form suitable for infants, children or adults.

[0052] In some embodiments of the present application, the pharmaceutical composition is in a dosage form for enteral administration or parenteral administration.

[0053] In some embodiments of the present application, the pharmaceutical composition is an oral preparation or an injectable preparation.

[0054] In some embodiments of the present application, the dosage form of the pharmaceutical composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and irrigations.

[0055] In some embodiments of the present application, the strain is capable of at least partially proliferating in the intestine of a subject.

[0056] In some embodiments of the present application, the pharmaceutical composition further comprises a product instruction sheet.

[0057] The sixth aspect of the present application provides a pharmaceutical combination product, comprising independently:

[0058] The first product comprises the aforementioned strain, or its metabolites, or the aforementioned culture, or the aforementioned microbial agent;

[0059] The second product, the second product includes combination drugs.

[0060] In some embodiments of the present application, the combination drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug.

[0061] In some embodiments of the present application, the pharmaceutical combination product further comprises a product instruction sheet.

[0062] In some embodiments of the present application, the tumor drug includes at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0063] In some embodiments of the present application, the chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin.

[0064] In some embodiments of the present application, the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin, and rose bengal.

[0065] In some embodiments of the present application, the photothermal agent includes at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials.

[0066] In some embodiments of the present application, the immunotherapy drug includes at least one of an immune cell therapy drug and an immune checkpoint inhibitor.

[0067] In some embodiments of the present application, the immune cell therapy drug includes at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and a NK cell therapy drug.

[0068] In some embodiments of the present application, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, and BTLA.

[0069] In some embodiments of the present application, the first product includes 1×10 3~10 13 CFU / g strain.

[0070] In some embodiments of the present application, the first product includes at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×10 10 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 CFU / g strain.

[0071] In some embodiments of the present application, the strain in the first product is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium.

[0072] In some embodiments of the present application, the first product and / or the second product further comprises at least one of an excipient, a diluent, and a carrier.

[0073] In some embodiments of the present application, the first product and the second product further independently include at least one pharmaceutically acceptable excipient.

[0074] In some embodiments of the present application, the auxiliary material also includes at least one of a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavor.

[0075] In some embodiments of the present application, the pharmaceutical combination product is an infant-suitable dosage form, a child-suitable dosage form, or an adult-suitable dosage form.

[0076] In some embodiments of the present application, the first product and the second product are independently in the form of a dosage form for enteral administration or a dosage form for parenteral administration.

[0077] In some embodiments of the present application, the first product and the second product are independently oral preparations or injectable preparations.

[0078] In some embodiments of the present application, the dosage forms of the first product and the second product are independently any one of powders, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and irrigations.

[0079] In some embodiments of the present application, the strain is capable of at least partially proliferating in the intestine of a subject.

[0080] The seventh aspect of the present application provides the use of the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in the preparation of a drug for preventing and / or treating a disease.

[0081] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0082] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor.

[0083] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer.

[0084] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of the following: myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0085] In some embodiments of the present application, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow.

[0086] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0087] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum.

[0088] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions.

[0089] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome.

[0090] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi.

[0091] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression.

[0092] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (l): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0093] The eighth aspect of the present application provides the use of the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in preventing and / or treating diseases.

[0094] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0095] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor.

[0096] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer.

[0097] In some embodiments of the present application, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow.

[0098] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of the following: myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0099] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0100] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum.

[0101] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions.

[0102] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome.

[0103] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi.

[0104] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression.

[0105] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (k): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0106] In a ninth aspect of the present application, a method for treating a disease is provided, the method comprising administering to a subject an effective dose of the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical combination product;

[0107] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0108] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor.

[0109] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer.

[0110] In some embodiments of the present application, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow.

[0111] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of the following: myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0112] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0113] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum.

[0114] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions.

[0115] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome.

[0116] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi.

[0117] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression.

[0118] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (k): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0119] In some embodiments of the present application, the strain, culture, microbial agent, composition, pharmaceutical composition or pharmaceutical combination product is administered at least once a day.

[0120] In some embodiments of the present application, when the administration frequency is twice or more per day, the same dose is administered each time.

[0121] In some embodiments of the present application, when the administration frequency is twice or more per day, each administration is at a different dose.

[0122] In some embodiments of the present application, the route of administration includes at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, and intramuscular injection.

[0123] In a tenth aspect of the present application, a method for preparing the aforementioned culture is provided, the method comprising the following steps:

[0124] The aforementioned strain is taken and inoculated into a fermentation medium to obtain a culture.

[0125] In some embodiments of the present application, the method further comprises centrifuging the culture to collect the fermentation medium supernatant.

[0126] In some embodiments of the present application, the fermentation medium or the supernatant of the fermentation medium is concentrated by at least one of evaporation, freeze-drying, dialysis, extraction, and membrane separation to obtain a concentrated product.

[0127] In some embodiments of the present application, the method further comprises subjecting the fermentation medium or the supernatant of the fermentation medium to obtain an extract by at least one of extraction and solvent extraction.

[0128] In some embodiments of the present application, the method further comprises drying any one of the fermentation medium, the fermentation medium supernatant, the concentrated product, and the extract to obtain a dried product.

[0129] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0130] Figure 1 This is the colony morphology of strain MNH 19801 after 48 hours of culture on anaerobic blood agar plate.

[0131] Figure 2 The microscopic morphology of strain MNH 19801 is shown with a scale bar of 2 μm.

[0132] Figure 3 The microscopic morphology of strain MNH 19801 is shown with a scale bar of 10 μm.

[0133] Figure 4 The figure shows the Gram-stained microscopic morphology of strain MNH 19801.

[0134] Figure 5 This is the microscopic morphology of the spores of strain MNH 19801.

[0135] Figure 6 The figure shows the tolerance of strain MNH 19801 to different pH values.

[0136] Figure 7 The figure shows the tolerance of strain MNH 19801 to different concentrations of NaCl.

[0137] Figure 8The figure shows the tolerance of strain MNH 19801 to different concentrations of bile salts.

[0138] Fig. 9 The phylogenetic tree was constructed based on the 16S rRNA of strain MNH 19801 and other strains with high similarity.

[0139] Fig.10 This is the result of strain MNH 19801 regulating the immune activity of macrophages.

[0140] Fig.11 The results of strain MNH 19801 regulating cytokine secretion by macrophages detected by CBA detection method. Among them, a~k represent the changes of macrophage secretion of IL-6, TNFα, RANTES, MIG, MCP-1, IL-2, IL-1β, IL-10, IFN-γ, IP-10, IL-12 / IL-23P40. In the figure, **: p<0.01, ***: p<0.001, ****: p<0.0001.

[0141] Fig.12 This is the promoting effect of strain MNH19801 on IFNβ transcription activity.

[0142] Fig.13 This is the inhibitory effect of strain MNH19801 on HDAC activity.

[0143] Fig.14 The results of the analysis of the immune activation of tumor mice by strain MNH 19801. A is the ratio of CD3 / CD45, B is the ratio of CD8 / CD45, A5 in A and B is the MNH 19801 experimental group, and CT is the control group.

[0144] Fig.15 The curve diagram of the tumor volume of mice in the control group and the experimental group. The data in the figure are displayed as mean ± standard deviation (Mean ± SD), and the statistical analysis is analyzed by two-way ANOVA with Sidak multiple comparison. The significant difference is indicated by *, *p < 0.05, **p < 0.01, no significant difference, no indication.

[0145] Fig.16 The figure shows the response rate of mice in the control group (A) and the experimental group (B) at the end of the experiment. The data are based on the tumor volume (mm 3 )show.

[0146] Fig.17 The change curve of tumor volume at different intervention time points in each group of mice; the mean value ± standard deviation of mouse tumor volume (mm 3) presents the data. The experimental endpoints were statistically analyzed by two-way ANOVA with Dunnett's multiple comparison analysis. Compared with the negative control group, the significant difference was as follows: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.

[0147] Fig.18 The tumor volume of each group of mice at the end point is shown in the figure, with the mean value of the mouse tumor volume ± standard deviation (mm 3 ) presents the data. The experimental endpoints were statistically analyzed by two-way ANOVA with Dunnett's multiple comparison analysis. Compared with the negative control group, the significant difference was as follows: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.

[0148] Fig.19 The figure is the tumor inhibition rate / tumor growth inhibition rate (TGI) of mice at the experimental end point. The mean ± standard deviation of mouse tumor volume (mm 3 ) presents the data. The experimental endpoints were statistically analyzed by two-way ANOVA with Dunnett's multiple comparison analysis. Compared with the Control group, the significant difference was as follows: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.

[0149] Fig. 20 The mouse response rate diagram of the negative control group (A), PD-1 group (B) and combination drug group (C) is the experimental endpoint. The data are based on the tumor volume of a single mouse (mm 3 )show.

[0150] Fig.21 This is the tumor volume change curve of mice in the negative control group, PD-1 group and combination drug group at different intervention time points in Example 12.

[0151] Fig. 22 This is the flow cytometry analysis of DC cell surface molecules MHCII, CD80 and CD86 in Example 13.

[0152] Fig.23 In Example 14, QPCR was used to detect the expression of M2 markers CD206 and Arg1 in RAW264.7 cells. DETAILED DESCRIPTION

[0153] The following will clearly and completely describe the concept of the present application and the technical effects produced in combination with the embodiments, so as to fully understand the purpose, features and effects of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of them. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present application.

[0154] The embodiments of the present application are described in detail below. The described embodiments are exemplary and are only used to explain the present application, and should not be understood as limiting the present application.

[0155] In the description of this application, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, "above", "below", "within", etc. are understood to include the number itself, and "about" means within the range of ±20%, 10%, 8%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, 0.1%, etc. of the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0156] In the description of the present application, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0157] The first aspect of the embodiments of the present application relates to a strain, which is named MNH 19801, deposited in the Guangdong Microbial Culture Collection Center (GDMCC), with the deposited name Lachnospiraceae sp.MNH19801, the deposited number GDMCC No: 61980, the deposited date October 27, 2021, and the deposited address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, and the proposed classification name is Lachnospiraceae sp.MNH19801.

[0158] Among them, the term "strain" refers to a member of a bacterial species with genetic characteristics, so that it can 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 presence of at least one recombinant gene, the presence of at least one mutant gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one non-natural plasmid, the presence of at least one antibiotic resistance box, or a combination thereof. The 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 case where a strain (compared to another strain of the same species) acquires or loses antibiotic resistance or acquires or loses biosynthetic ability (e.g., auxotrophic strains), strains or nutrients / metabolites can be distinguished by selection or counter-selection using antibiotics.

[0159] It is known in the art that the strains can be classified and identified by traditional classification methods and molecular biological methods. Traditional classification methods include, for example, cell morphology observation, Gram staining, flagella staining, various metabolic experiments, etc. Molecular biological methods include ribosomal RNA sequence determination methods, determination methods based on whole genome sequencing, etc. The strain involved in the embodiment of the present application is identified as a new genus under the Lachnospiraceae family based on the following taxonomic characteristics it has, including:

[0160] The bacteria have no spores, no flagella, are non-motile, are short rod-shaped, and are Gram-positive.

[0161] Inoculate in anaerobic blood plate culture medium, under anaerobic culture conditions (for example, anaerobic culture at 37°C for 48 hours), form visible colonies on the anaerobic blood plate culture medium, the colonies are round, with regular edges, a diameter of about 2mm, white, opaque, and no secretions are formed around the colonies.

[0162] The growth temperature range is 30-42°C, and the optimum growth temperature is about 37°C; it can grow in the pH range of 6-10, and the optimum growth pH is 7.0; it cannot grow under conditions where the NaCl concentration is higher than 2%; it can survive and grow in the bile salt concentration range of 0-0.25%, but the strain growth trend weakens with the increase of bile salt concentration; it cannot grow under aerobic conditions, but grows well under anaerobic conditions, and is an obligate anaerobic bacterium.

[0163] According to its obligate anaerobic characteristics, the API 20A test showed that it was sensitive to at least one of the antibiotics including gentamicin, erythromycin, chloramphenicol, tetracycline, penicillin, ampicillin and ceftriaxone; and resistant to at least one of the antibiotics including ciprofloxacin, co-trimoxazole and lincomycin.

[0164] In some embodiments, the strain or strain variant has the characteristic of high acetic acid production. Wherein, high acetic acid production refers to an acetic acid production of ≥200 μg / g in a short-chain fatty acid (SCFA) assay. Specifically, the acetic acid production is ≥300 μg / g, ≥400 μg / g, ≥500 μg / g, ≥600 μg / g, ≥700 μg / g, ≥800 μg / g, ≥900 μg / g, ≥1000 μg / g, ≥1100 μg / g, ≥1200 μg / g, ≥1300 μg / g, ≥1400 μg / g. For example, the above-mentioned strain or strain variant is inoculated in TSB liquid culture medium and anaerobically cultured at 37°C for 48 hours, and the bacteria are collected by centrifugation and the acetic acid production is detected to be ≥300μg / g, ≥400μg / g, ≥500μg / g, ≥600μg / g, ≥700μg / g, ≥800μg / g, ≥900μg / g, ≥1000μg / g, ≥1100μg / g, ≥1200μg / g, ≥1300μg / g, ≥1400μg / g.

[0165] In addition, the sequencing results of 16S rRNA of strain MNH19801 show that the sequence of its 16S rRNA is shown in SEQ ID No. 1. 16S rRNA is a ribosomal RNA of prokaryotes. The 16S rRNA gene consists of a variable region and a conserved region. The conserved region is common to all bacteria, while the variable region has different degrees of difference between different bacteria. By comparing the gene sequences of bacterial 16S rRNA, according to the sequence difference base and its evolutionary distance, a biological evolutionary tree can be drawn.

[0166] When the identity between the 16S rRNA gene sequences of two strains is less than 98.65%, it can be judged that they belong to different species (see, Kim, M., Oh, H.-S., Park, S.-C., & Chun, J. (2014). Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequences similarity for species demarcation of prokaryotes. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 64 (Pt 2), 346-351, and Liu, C., Du, M.-X., Abuduaini, R., Yu, H.-Y., Li, D.-H., Wang, Y.-J., Liu, S.-J. (2021). Enlightening the taxonomy darkness of human gut microbiomes with a culturedbiobank. Microbiome, 9(1), page P23; Pablo Yarza, Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNAgene sequences, Microbiology, volume 12, pages P635-P645, 2014).

[0167] When the identity between the 16S rRNA gene sequences of two strains is less than 95%, it can be judged that they belong to different genera (“Agenus definition for Bacteria and Archaea based on genome relatedness and taxonomic affiliation”, RA Barco, bioRxiv (doi: https: / / doi.org / 10.1101 / 392480); Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis, Jethro S. Johnson, Nature Communications 10, Article number: 5029 (2019)).

[0168] To this end, in some embodiments of the present application, variants of strain MNH19801 based on the above 16S rRNA sequence are provided, and these strain variants have at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity in 16S rRNA sequence compared with strain MNH19801 with a deposit number of GDMCC No: 61980, and thus are strains of the same species or different species within the same genus as strain MNH19801. It can also be reasonably inferred that these strains of the same species or different species of the same genus have similar biological activities. In other embodiments, the variant has at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% identity with the sequence shown in SEQ ID NO:1 in the 16S rRNA sequence.

[0169] The "identity" between the sequences of two nucleic acid molecules can be determined using a known computer algorithm, such as at least one of the GCG program packages, BLASTN or FASTA. Other commercial or publicly available programs may also be, for example, the DNAStar "MegAlign" program.

[0170] In addition, with the rapid development of second-generation and third-generation sequencing technologies, bacterial species identification based on whole genome sequencing has become possible, making the results of bacterial species identification more accurate. The average nucleotide identity (ANI) of bacterial genomes refers to the similarity of homologous genes between two bacterial genomes. In the field of bacterial taxonomy, it is generally believed that the ANI value needs to reach 98.65% or above to be identified as belonging to the same species (Kim, M., Oh, H.-S., Park, S.-C., & Chun, J. (2014). Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARYMICROBIOLOGY, 64(Pt 2), 346–351, and Liu, C., Du, M.-X., Abuduaini, R., Yu, H.-Y., Li, D.-H., Wang, Y.-J., Liu, S.-J. (2021). Enlightening the taxonomy darkness of human gutmicrobiomes with a cultured biobank. Microbiome, 9(1), P23; Pablo Yarza, Uniting the classification of cultured and uncultured bacteria and archaea using16SrRNAgene sequences, Microbiology, volume 12, pages P635-P645, 2014).

[0171] When the identity between the 16S rRNA gene sequences of two strains is less than 95%, it can be judged that they belong to different genera (“Agenus definition for Bacteria and Archaea based on genome relatedness and taxonomic affiliation”, RA Barco, bioRxiv (doi: https: / / doi.org / 10.1101 / 392480); Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis, Jethro S. Johnson, Nature Communications 10, Article number: 5029 (2019)).

[0172] Using the above method, a person skilled in the art can determine whether an isolated strain belongs to a new strain of the same genus but different species or the same species as strain MNH19801 in the Lachnospiraceae. For example, when the average nucleotide identity ANI value with the strain with the deposit number of GDMCC No: 61980 is at least 87%, such as at least 87%, 90%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100%, it can be determined to belong to the same genus or the same species.

[0173] At present, there are various mature ANI value calculation tools, such as local calculation software Jspecies ( / jspecies) and Gegenees ( / documentation.html), online calculation tools ANI caculator (http: / enveomics.gatech.edu / ), EzGenome ( / ezgenome / ani) and ANItools. Therefore, the ANI value of the isolated strain and MNH19801 can be calculated by any of the above local calculation software or online calculation tools or other tools or methods known in the art.

[0174] To this end, in some embodiments of the present application, variants of strain MNH19801 based on the above genome are provided, and these strain variants have at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity compared to strain MNH19801 with a deposit number of GDMCC No: 61980 or any of the above variants, and thus are strains of the same species or different species within the same genus as strain MNH19801. It can also be reasonably inferred that these strains of the same species or different species of the same genus have similar biological activities. In addition, it can also be judged by the alignment score of the corresponding genome. For example, compared with the strain MNH19801 with the deposit number GDMCCNo: 61980 or any other variant mentioned above, a variant with an alignment score of at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% also has the same or similar biological activity.

[0175] In some embodiments, based on the comparison results of 16S rRNA and ANI, the variant of the strain has at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity with the 16S rRNA of MNH19801, and at the same time, has at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity with the genome of MNH19801.

[0176] For example, a variant of the strain has at least 95% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0177] A variant of the strain has at least 96% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0178] A variant of the strain has at least 97% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0179] A variant of the strain has at least 98% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0180] A variant of the strain has at least 98.5% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0181] A variant of the strain has at least 98.65% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0182] A variant of the strain has at least 99% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0183] A variant of the strain has at least 99.5% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0184] A variant of the strain has at least 99.6% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0185] A variant of the strain has at least 99.7% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0186] A variant of the strain has at least 99.8% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0187] A variant of the strain has at least 99.9% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0188] A variant of the strain has at least 99.99% identity to the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity to the genome of MNH19801; or

[0189] The strain variants have 100% identity with the 16S rRNA of MNH19801 and at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity with the genome of MNH19801.

[0190] The term "variant" includes any bacteria produced from strain MNH19801, such as bacteria produced by natural mutation or recombination (such as mutation caused by cell proliferation and cell division) or resistance selection, as well as mutation or recombination caused by any other mechanism such as radiation, virus, transposon or mutagenic chemicals. In addition, variants also include other strains isolated from the natural environment that have the same or similar taxonomic characteristics and properties as strain MNH19801, generally including other strains of the same genus but different species as MNH19801 under the Lachnospiraceae family, or other strains with different phenotypic variations within the same species.

[0191] The second aspect of the embodiment of the present application relates to a culture of a strain. The term "culture" refers to a product obtained by culturing an isolated strain in a fermentation medium. The fermentation medium can be selected as a natural or artificial culture medium, for example, a solid culture medium or a liquid culture medium. In some embodiments, the fermentation medium includes peptone, such as at least one of meat peptone, casein peptone, whey peptone, soy peptone, etc., and in addition, other inorganic salts or organic substances used to promote bacterial growth may also be included.

[0192] In some embodiments, the culture may be a fermentation medium after strain culture, or a supernatant obtained by centrifugation of the fermentation medium, or a product obtained by concentration of the fermentation medium or its supernatant by at least one of evaporation, freeze drying, dialysis, extraction, membrane separation, etc., or an extract obtained by extraction of the fermentation medium or its supernatant, solvent extraction (such as water or organic solvent), or a dried product obtained by drying any of the fermentation medium or its supernatant or concentrated product or extract. In addition, in some specific embodiments, the extract may be an extraction for one or more specific components, such as a component with a specific molecular weight range, specific solubility, specific isoelectric point, etc.; or it may be an extraction not directed to one or several types of components. The specific extraction method includes extracting with at least one solvent such as methanol, ethanol, chloroform, water, acid, alkali, etc. in a specific process sequence under certain temperature conditions, for example, extracting in 75v / v% ethanol at 60-80°C for 1-10 minutes, or extracting in methanol at -30--10°C for 1-10 minutes, or extracting in methanol / chloroform / water solution at -40--20°C for 30-60 minutes, or extracting in deionized water at 95°C for 1-10 minutes, or extracting in perchloric acid, trichloroacetic acid, hydrochloric acid, sodium hydroxide solution at 0-4°C, etc., so as to extract the corresponding metabolites, such as short-chain fatty acids or short-chain fatty acid salts, such as at least one of acetic acid, butyric acid, valeric acid, acetate, propionate, and valerate.

[0193] In some embodiments, the culture is a culture obtained under anaerobic culture conditions using a liquid culture medium. For example, the liquid culture medium is an MM01 liquid culture medium (including about 5 g / L of peptone, about 5 g / L of trypticase, about 10 g / L of yeast powder, about 5 g / L of beef extract, about 5 g / L of glucose, about 2 g / L of K2HPO4, about 2 g / L of sodium acetate, about 1 mL / L of Tween 80, about 5 mg / L of heme, about 0.5 g / L of L-cysteine ​​hydrochloride, about 1 μL / L of vitamin K1, and about 8 ml / L of inorganic salt solution (about 0.25 g / L of calcium chloride, about 1 g / L of dipotassium hydrogen phosphate, about 1 g / L of potassium dihydrogen phosphate, about 0.5 g / L of magnesium sulfate, about 10 g / L of sodium bicarbonate, and about 2 g / L of sodium chloride). In some embodiments, during anaerobic culture, the culture temperature is 35 to 42 ° C, for example, 37 ° C. In some embodiments, during anaerobic culture, the culture time is 24 to 72 hours, such as 36 to 60 hours, and can be 48 hours. In some embodiments, during anaerobic culture, the pH of the culture medium is 6 to 10, and can be 7.

[0194] It is understood that in the above-mentioned culture, compounds and / or cell fragments of the strain are contained, and / or metabolites secreted by the strain. Wherein, metabolites include small molecules (such as amino acids, nucleosides, nucleotides) and macromolecules (such as polypeptides, carbohydrates, nucleic acids, proteoglycans, lipids), which can be primary metabolites directly involved in normal cell functions, or secondary metabolites that are usually not required for basic cell functions, and metabolic intermediates produced in the synthesis of primary or secondary metabolites. Thus, biological functions similar to those of strains can be achieved by cultures of different types containing compounds and / or metabolites of the strain.

[0195] The third aspect of the embodiments of the present application relates to a microbial agent, which comprises one or more of the aforementioned strains or metabolites of the strains or the aforementioned cultures, for example, it may be a strain with a deposit number of GDMCC No: 61980 or its metabolites or the culture of the strain.

[0196] The fourth aspect of the embodiments of the present application relates to a pharmaceutical combination product, which comprises independently:

[0197] The first product includes the aforementioned strain, or the metabolite of the aforementioned strain, or the aforementioned culture, or the aforementioned microbial agent;

[0198] Second products, including combination drugs.

[0199] It is understood that the first product contains an effective amount of the aforementioned strain or the aforementioned culture. The first product and the second product can be administered at different time points and / or in different administration modes and / or at different administration intervals, respectively, in respective dosages.

[0200] In some embodiments, the combination drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug.

[0201] In some embodiments, the tumor drug may include at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0202] In some embodiments, the metabolic disease drugs include drugs used to treat abnormal amino acid metabolism (such as phenylketonuria), glycine metabolism, tyrosine metabolism), abnormal organic acid metabolism (methylmalonic acidemia, propionic acidemia), abnormal glucose metabolism (such as glycogen storage disease, galactosemia, diabetes), abnormal protein metabolism, abnormal fat metabolism (such as hypercholesterolemia, hyperlipidemia), abnormal purine metabolism (such as gout, self-destructive facies), abnormal pigment metabolism (such as porphyria), abnormal blood ammonia metabolism (such as hyperammonemia, citrin deficiency), and abnormal metal ion metabolism (such as hepatolenticular degeneration).

[0203] Taking diabetes as an example, diabetes drugs include at least one of insulin, metformin, pramide, sulfonylureas (such as glibenclamide, glipizide, glimepiride), methylcobalamin (such as repaglinide, nateglinide), thiazolidinediones (such as rosiglitazone, pioglitazone), DPP-4 inhibitors (such as sitagliptin, saxagliptin, linagliptin), GLP-1 receptor agonists (such as exenatide, liraglutide, semaglutide), SGLT2 inhibitors (such as canagliflozin, dapagliflozin, empagliflozin), entecavir, tenofovir, lamivudine, adefovir, telbivudine, simeprek, sofobuvir, interferon, ribavirin, etc.

[0204] In some embodiments, the pharmaceutical combination product is a dosage form suitable for infants, children or adults, a dosage form for gastrointestinal administration or parenteral administration, an oral preparation or an injectable preparation, and the strain in the pharmaceutical combination product is at least partially capable of proliferating in the intestine of the subject, and the strain can be any of live bacteria (such as bacteria, or attenuated bacteria, or freeze-dried bacteria, or irradiated bacteria) and dead bacteria.

[0205] The seventh aspect of the embodiments of the present application relates to the use of the aforementioned strain, the aforementioned culture, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in the preparation of a drug for preventing and / or treating a disease. In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0206] Wherein, term " tumor " or " tumor " refers to that cell division and proliferation in vivo exceed normal level, and a kind of abnormal mass or vegetation that grows out, and " cancer " generally refers to malignant tumor, and can be general in some cases in this application. Tumor or cancer generally include at least one of solid tumor, soft tissue tumor, hematopoietic tumor, glandular tumor or metastatic tumor. Solid tumor can be divided into epithelial cell derived tumor, fibroblast derived tumor etc. according to cell source characteristic. In addition, according to the difference of primary tumor lesion position, the type of solid tumor includes head and neck cancer, upper digestive tract cancer, lower digestive tract cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, osteoma, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, endocrine system cancer, hematological tumor includes at least one tumor in blood, lymph and bone marrow. Among them, head and neck cancer includes oral cancer (such as lip cancer, tongue cancer, mouth floor cancer, tonsil cancer, gum cancer, etc.), salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, etc. Upper digestive tract cancer includes esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, etc. Lower digestive tract cancer includes appendix cancer, colon cancer, rectal cancer, anal canal cancer, etc. Hepatobiliary system cancer includes liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampulla cancer, pancreatic cancer, etc. Neuroendocrine tumors include gastric neuroendocrine tumors, Duodenal and ampullary neuroendocrine tumors, jejunal-ileal neuroendocrine tumors, appendix neuroendocrine tumors, colorectal neuroendocrine tumors, pancreatic neuroendocrine tumors, etc. Chest cancer includes thymic carcinoma, lung cancer (such as small cell lung cancer, non-small cell lung cancer, etc.), malignant pleural mesothelioma, etc. Soft tissue tumors include angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma (formed by fibroblasts and collagen fibers), gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve Transvaginal carcinoma, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, etc. Skin cancer includes Merkel cell carcinoma, cutaneous malignant melanoma, etc. Female reproductive system cancer includes vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinoma sarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, etc. Male reproductive system cancer includes penile cancer, prostate cancer, testicular cancer, etc. Urinary system cancer includes kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethra cancer, etc. , eye cancers include eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular accessory organ lymphoma, etc., central nervous system cancers include brain cancer (such as glioma, meningioma, medulloblastoma, medullary pituitary adenoma, primary CNS lymphoma and central nervous system germ cell tumor, etc.), spinal cord tumors, etc., endocrine system cancers include differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, etc.Hematopoietic tumors include Hodgkin's lymphoma, non-Hodgkin's lymphoma (such as diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, Burkitt's lymphoma, lymph node marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, etc.), cutaneous lymphoma, plasma cell myeloma, leukemia, etc.

[0207] In addition, tumors or cancers also include tumors caused by bacteria or viruses, for example, tumors or cancers caused by at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum.

[0208] The term "metabolic disease" refers to the accumulation or lack of substances such as sugar, fat, and amino acids in the human body, which causes abnormal metabolic substance content in the body, and some diseases caused by it can be roughly divided into two categories: metabolic dysfunction and hypermetabolism. In some specific embodiments of the present application, metabolic diseases include abnormal amino acid metabolism (such as phenylketonuria, glycine metabolism, tyrosine metabolism), abnormal organic acid metabolism (methylmalonic acidemia, propionic acidemia), abnormal sugar metabolism (such as glycogen storage disease, galactosemia, diabetes), abnormal fat metabolism (such as hypercholesterolemia, hyperlipidemia), abnormal purine metabolism (such as gout, self-destructive face disease), abnormal pigment metabolism (such as porphyria), abnormal blood ammonia metabolism (such as hyperammonemia, citrin deficiency), and abnormal metal ion metabolism (such as hepatolenticular degeneration).

[0209] In some embodiments, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome;

[0210] In some embodiments, the infectious disease includes at least one of bacteria, viruses, and fungi. In some embodiments, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, and depression.

[0211] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product include preventing and / or treating tumors by at least one of the following (a) to (l): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0212] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product achieve the treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0213] Existing research results show that HDAC inhibitors have good anti-tumor effects on a variety of tumor cells, including tumor cells in the bladder, bone, breast, uterus, central nervous system, esophagus, lung, ovary, pancreas, bile duct cancer, prostate and other parts, and can cause these tumor cells to show obvious apoptosis, proliferation inhibition and cell cycle arrest, etc.; as an oncogene that regulates the onset of cancer, the activity reduction or inhibition of HDAC expression has been shown to have a variety of anti-cancer effects. In addition, TSA can also inhibit tumor angiogenesis by downregulating the expression of tumor angiogenesis-related genes caused by hypoxia and directly inhibiting the migration and proliferation of endothelial cells. Moreover, HDAC inhibitors can treat diabetes through a variety of mechanisms, including the inhibition of Pdxl (Park et al., 2008, J Clin Invest, 118, 2316-24), and enhancing the expression of transcription factor Ngn3 to increase the endocrine reservoir. 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 treatment method for late diabetic complications such as diabetic nephropathy and retinal ischemia (Christensen et al., 2011, Mol Med, 17 (5-6), 370-390). Therefore, the aforementioned strains, the aforementioned cultures, the aforementioned bacterial agents, the aforementioned compositions, the aforementioned pharmaceutical compositions, and the aforementioned pharmaceutical combination products achieve treatment or prevention of diseases (such as tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases) by inhibiting HDAC activity.

[0214] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product achieve the treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases) by upregulating proinflammatory factors and chemokines to stimulate immunity and / or by downregulating anti-inflammatory factors for immunomodulation. In some embodiments, the proinflammatory factor is selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0215] The eighth aspect of the embodiments of the present application relates to the use of the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in preventing and / or treating a disease. In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0216] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product include preventing and / or treating tumors by at least one of the following (a) to (l): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue. 。

[0217] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned drug combination product achieve the treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0218] The ninth aspect of the embodiments of the present application relates to a method for treating a disease, comprising administering to a subject an effective dose of the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical combination product.

[0219] In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease. Among them, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, or a metastatic tumor. Solid tumors include at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumors, chest cancer, osteoma, soft tissue tumors, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer; hematopoietic tumors include at least one tumor in blood, lymph, and bone marrow. Tumors or cancers include tumors or cancers caused by bacteria or viruses (such as at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum). Metabolic diseases include at least one metabolic abnormality in amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions. Inflammatory or autoimmune diseases include at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome. Infectious diseases include diseases caused by at least one of bacteria, viruses, and fungi. Central nervous system diseases include at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, and depression.

[0220] In some embodiments, the prevention and / or treatment of tumors is achieved by at least one of the following (a) to (l): (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) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetate or acetate, propionate or propionate, butyrate or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the subject's immune system to exert an immunomodulatory effect; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0221] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned drug combination product achieve the treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0222] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product achieve the treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases) by upregulating proinflammatory factors and chemokines to stimulate immunity and / or by downregulating anti-inflammatory factors for immunomodulation. In some embodiments, the proinflammatory factor is selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0223] In some embodiments, the strain, culture, microbial agent, composition, pharmaceutical composition or pharmaceutical combination product is administered at least once a day. In some embodiments, when the administration frequency is more than twice a day, the same dose is administered each time. In some embodiments, when the administration frequency is more than twice a day, different doses are administered each time. In some embodiments, the route of administration includes at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, and intramuscular injection.

[0224] The term "subject", or patient, refers to any mammal. 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 alternative therapies, or reducing resistance to alternative therapies, or a combination thereof.

[0225] The term "prevention" is art-recognized and well-known in the art when used in connection with a condition such as local recurrence, and includes administration of a composition that reduces the occurrence of or delays the onset of symptoms of a medical condition in a subject relative to a subject not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable tumors in a population of patients 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.

[0226] The tenth aspect of the present application also relates to a method for preparing the aforementioned culture, which method comprises the following steps: taking the aforementioned strain, inoculating it into a fermentation medium, and obtaining a culture.

[0227] In some embodiments, the culture is centrifuged to collect the fermentation medium supernatant. In some embodiments, the fermentation medium or the fermentation medium supernatant is concentrated by at least one of evaporation, freeze drying, dialysis, extraction, and membrane separation to obtain a concentrated product. In some embodiments, the fermentation medium or the fermentation medium supernatant is extracted by at least one of extraction and solvent extraction to obtain an extract. In some embodiments, any one of the fermentation medium, the fermentation medium supernatant, the concentrated product, and the extract is dried to obtain a dried product.

[0228] The meanings of some abbreviations involved in the following embodiments are shown in Table 1.

[0229] Table 1. Meaning of some abbreviations

[0230] abbreviation Meaning / full name abbreviation Meaning / full name IND Indole experiment ESC Aesculin URE Urease GLY glycerin GLU glucose CEL Microscopic disaccharide MAN Mannitol MNE Mannose LAC lactose MLZ Melezitose SAC sucrose RAF Raffinose MAL maltose SOR Sorbitol SAL Salicylate RHA Rhamnose XY Xylose TRE Trehalose ARA Arabinose GEL gelatin

[0231] Some of the culture media involved in the following examples are as follows:

[0232] MM01 liquid culture medium, components: peptone 5g / L, tryptic casein 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K2HPO4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, heme 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1μL / L, inorganic salt solution 8ml / L (each 1L includes calcium chloride 0.25g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, sodium bicarbonate 10g, sodium chloride 2g).

[0233] Solid MM01 culture medium, composed of: peptone 5g / L, tryptic casein 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K2HPO4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, heme 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1μL / L, inorganic salt solution 8ml / L (each 1L 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.

[0234] AC liquid culture medium, composed of: peptone 20g / L, glucose 5g / L, yeast extract 3g / L, beef extract powder 3g / L, vitamin C 0.2g / L, pH 7.0.

[0235] Anaerobic blood agar plates (purchased from Huankai Microorganisms) were composed of 10 g / L casein pancreatic digest, 3 g / L heart pancreatic digest, 1 g / L corn starch, 5 g / L meat gastric digest, 5 g / L yeast extract powder, 5 g / L sodium chloride, 15 g / L agar, 50-100 mL / L sterile defibrinated sheep blood, 1000 mL distilled water, and a final pH of 7.3 ± 0.2.

[0236] Tryptic soy casein liquid medium TSB liquid medium, composition: tryptic soy casein 17g / L, soybean papain hydrolysate 3g / L, dipotassium hydrogen phosphate 2.5g / L, sodium chloride 5g / L, glucose 2.5g / L, pH 7.3±0.2.

[0237] The above culture medium can be prepared by conventional preparation methods and sterilization methods.

[0238] The above implementation is described below in conjunction with specific examples.

[0239] Example 1: Isolation and identification of strain MNH 19801

[0240] 1. Isolation of strains

[0241] The fecal sample was isolated from a healthy male volunteer in Guangzhou, Guangdong Province. The specific process is as follows:

[0242] The donor takes 2 to 5 grams of fresh feces and puts it into a sample collection and preservation tube. After shaking and homogenization, the processed feces sample is placed in an ice box and sent to the laboratory for strain isolation within 24 hours.

[0243] Physiological saline was dispensed in a biosafety cabinet, 9 ml / tube; anaerobic blood plates were prepared for strain isolation and transferred to an anaerobic workstation (Don Whitley Scientific H35) 24 h in advance, with sample information, culture medium type, isolation date, etc. marked.

[0244] Take a fresh stool sample and place it in the anaerobic operation station. Use a vortex shaker to shake for 1 min. Mix well. Pipette 1 mL of the sample into 9 mL of normal saline and mix well for 10 min. -1 dilution, and then serially diluted to 10 -6 Dilution solution for later use.

[0245] Take 10 respectively -6 The diluted solution was dropped onto the separation medium (anaerobic blood agar plate, Columbia blood agar plate and chocolate agar plate) at a drop volume of 100 μL / dish. The plate was spread evenly. After the surface of the plate was dry, the plate was inverted and cultured at 37°C for 3 to 5 days.

[0246] Observe the growth of the strain in the isolation medium and pick a single colony with a sterilized toothpick to purify the strain. The purified strain is placed at 37°C for anaerobically culture.

[0247] The pure culture strain numbered MNH 19801 was prepared into a 20% glycerol / water-bacteria solution and stored at -86°C.

[0248] 2. Identification of strains

[0249] 2.1 Morphological characteristics

[0250] The strain MNH 19801 was inoculated into anaerobic blood plate medium and cultured anaerobically at 37°C for 48 hours. Figure 1 As shown in the figure, it can be seen that visible colonies are formed on the anaerobic blood plate culture medium. The colonies are round, with regular edges, about 2 mm in diameter, white, opaque, and no secretions are formed around the colonies. Figures 2 to 5 Morphological observation under a microscope revealed that strain MNH 19801 had no spores, no flagella, no motility, and was Gram-positive.

[0251] 2.2 Physiological and biochemical characteristics

[0252] The growth temperature range of strain MNH 19801 is 30-42°C, and the optimal growth temperature is 37°C; Figure 6 , the strain can grow in the pH range of 6.0 to 10.0, and the optimal growth pH is 7.0; Figures 7-8 , strain MNH 19801 is difficult to grow under conditions where the NaCl concentration is higher than 2%, while it can survive and grow in the bile salt concentration range of 0-0.25%, but the growth trend of the strain weakens with the increase of bile salt concentration. Strain MNH 19801 cannot grow under aerobic conditions, but grows well under anaerobic conditions, and is an obligate anaerobic bacterium.

[0253] API 20A test

[0254] The test was performed using API 20A reagent strips (BioMérieux) according to the instructions. The culture conditions of the strain were 37°C and anaerobic. The results are shown in Table 2.

[0255] Table 2. API 20A test results for strain MNH 19801

[0256] Test items Test results Test items Test results IND Negative ESC Negative URE Negative GLY No acid production GLU No acid production CEL No acid production MAN No acid production MNE Acid production LAC No acid production MLZ No acid production SAC No acid production RAF Acid production MAL No acid production SOR No acid production SAL Acid production RHA Acid production XY Acid production TRE No acid production ARA No acid production GEL Negative

[0257] As can be seen from the table, strain MNH 19801 can utilize mannitol, raffinose, xylose, salicylate, and rhamnose as substrates to metabolize and produce acid, but other types of substrates cannot be used.

[0258] Antibiotic susceptibility testing

[0259] The antibiotic sensitivity test of strain MNH 19801 was carried out using the disk diffusion method. The test results are shown in Table 3.

[0260] Table 3. Antibiotic susceptibility test results of strain MNH 19801

[0261] antibiotic Diameter of inhibition zone (mm) Gentamycin (GEN) 10.74 Erythromycin (ERM) 111.41 Chloramphenicol (CLM) 30.39 Tetracycline (TET) 12.88 Penicillin (PEN) 18.79 Ciprofloxacin (CFX) 0 Co-trimoxazole (T / S) 0 Ampicillin (AMP) 11.17 Lincomycin (LIN) 0 Ceftriaxone (CTR) 33.18

[0262] The results refer to Table 3. The strain MNH 19801 is sensitive to antibiotics such as gentamicin, erythromycin, chloramphenicol, tetracycline, penicillin, ampicillin and ceftriaxone, but is resistant to ciprofloxacin, cotrimoxazole and lincomycin.

[0263] 2.3 16S rRNA

[0264] Fresh culture of strain MNH 19801 was taken, and the genomic DNA of the sample was extracted by SDS method, and then the purity and integrity of the DNA were detected by agarose gel electrophoresis.

[0265] First, the genomic DNA of the sample was extracted using the SDS method, and then the purity and integrity of the DNA was detected by agarose gel electrophoresis and quantified using Qubit. The extracted genome was sequenced using the Illumina platform and the Pacbio platform.

[0266] Illumina platform library construction and sequencing, the qualified DNA samples detected by electrophoresis were randomly broken into fragments of about 350 bp in length using Covaris ultrasonic crusher. Ultra TM DNA Library Prep Kit for Illumina (NEB, USA) was used to prepare the entire library through end repair, A-tailing, sequencing adapter addition, purification, PCR amplification and other steps. After the library was constructed, Qubit 2.0 was used for preliminary quantification, and the library was diluted to 2 ng / ul. Then, the insert fragment of the library was detected using Agilent 2100. After the insert size met expectations, the effective concentration of the library was accurately quantified using the Q-PCR method to ensure the quality of the library. After the library was qualified, PE150 sequencing was performed using Illumina NovaSeq.

[0267] PacBio platform library construction and sequencing, SMRT bell TM Template kit (version 2.0) was used to construct SMRT Bell library, and the qualified DNA samples detected by electrophoresis were sheared into target fragments of the size required for library construction using Covaris g-TUBE. After DNA damage repair and end repair, hairpin adapters were connected to the two ends of the DNA fragments using DNA ligase, and the DNA fragments were purified using AMpure PB magnetic beads. Blue Pippin fragments were used to screen fragments of a specific size, and the SMRT Bell library was screened for concentration using AMpure PB magnetic beads. Subsequently, DNA damage was repaired, and the SMRT Bell library was purified using AMpure PB magnetic beads again. The constructed library was quantified by Qubit concentration, and the size of the inserted fragment was detected using Agilent2100. Finally, the qualified library was sequenced using PacBio Sequel II.

[0268] The PacBio platform sequencing obtained 1.006Gbp of raw data, with an average sequence length of 82,146bp. The Illumina platform sequencing obtained 3.25Gbp of raw data, of which Q20 accounted for 96.19%.

[0269] Pacbio raw sequencing data was first processed by removing the connector to obtain valid data for further data analysis. Based on the valid data, Canu software was used for genome assembly to obtain a preliminary assembly result that can reflect the basic situation of the sample genome. Then, pbmm2 software was used to correct the splicing results based on the third-generation sequencing data, and then Pilon software was used to correct the results based on the Ilumina sequencing data to obtain the final assembly result. After obtaining the assembled genome, CheckM software was used to extract the full-length 16S sequence from the assembled genome. Sanger sequencing detected a 1,381bp 16S sequence, and a 1,540bp 16S sequence was extracted from the genome. Using blastn to compare the two sequences, the alignment results showed identity 100.0% and coverage 100.0%, so the 1,540bp 16S sequence extracted from the genome was selected for species identification.

[0270] The 16S rRNA gene sequence of strain MNH 19801 is as follows:

[0271] AACTTTATTAGAGAGTTTGATCCTGGCTCAGGATGAACGCTGGCGGCGTGCTTAAC

[0272] ACATGCAAGTCGAACGAAGCGCTTGAGAAGATTTCTTCGGAATGAAACTTAAGTGACTG

[0273] AGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGGGATAACAGTC

[0274] AGAAATGACTGCTAATACCGCATAAGCGCACAGTACCGCATGGTACGGTGTGAAAAAACT

[0275] CCGGTGGTATGGGATGGACCCGCGTCTGATTAGCTAGTTGGTGGGGTAACGGCCTACCA

[0276] AGGCGACGATCAGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGACACG

[0277] GCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGAAACCCTGA

[0278] TGCAGCGACGCCGCGTGAGGCGAAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGA

[0279] AGAAAATGACGGTACCTGACTAAGAAGCCCCGGCTAACTACGTGCCAGCCCGCGGT

[0280] AATACGTAGGGGGCAAGCGTTATCCGGGATTACTGGGTGTAAAGGGAGCGTAGACGGAC

[0281] TGGCAAGTCTGATGTGAAAACCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTCA

[0282] GTCTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATT

[0283] AGGAGGAACCACGATGGCGAAGGCGGCTTACTGGACGATAACTGACGTTGAGGCTCGA

[0284] AAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATA

[0285] CTAGGTGTCGGGAAGCAAAGCTTTTCGGTGCCGCAGCAAACGCAATAAGTATTCCACCT

[0286] GGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGG

[0287] TGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCGAT

[0288] CTGCCCGGACTGTAACGAGTCCTTTCCCTTCGGGGACAGAGAAGACAGGTGGTGCATG

[0289] GTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTT

[0290] ATCCTTAGTAGCCAGCAGGTAAAGCTGGGCACTCTGGGGAGACTGCCAGGGATAACCTG

[0291] GAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGATTTGGGCTACACACGTG

[0292] CTACAATGGCGTAAACAAAGGGAAGCAGCCCCGCGAGGGTGAGCAAATCTCAAAAATA

[0293] ACGTCCCAGTTCGGATTGTAGTCTGCAACTCGACTACATGAAGCTGGAATCGCTAGTAAT

[0294] CGCAGGTCAGCATACTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACA

[0295] CCATGGGAGTCGGATATGCCCGAAGCCAGTGACCCAACCTTAGGGAGGGAGCTGTCGA

[0296] AGGTGGAGCCGATAACTGGGGTGAAGTCGTAACAAGGTAGCCGTATCGGAAGGTGCGG

[0297] CTGGATCACCTCCTTTCT (SEQ ID NO: 1).

[0298] The measured sequence was analyzed with the data in GenBank by BLAST. The comparison results showed that strain MNH19801 belonged to the Lachnospiraceae family. The strains with higher similarity to MNH 19801 were Blautiabrookingsii (96.031%), Blautia faecicola (95.366%), Robinsoniella peoriensis (93.91%) and Oliverpabstia intestinalis (95.427%). The 16S rRNA gene sequence similarity between strain MNH 19801 and other strains of Lachnospiraceae was less than 95%. The low (<96.031%) 16S rRNA gene sequence similarity indicated that strain MNH 19801 might be a new genus or a new species of Lachnospiraceae.

[0299] The 16S rRNA gene sequences of MNH 19801 and related strains of Lachnospiraceae retrieved from GenBank and other databases were compared to construct a phylogenetic tree. The phylogenetic tree shows that strain MNH 19801 is closely related to Blautia faecicola KGMB01111 T (MK424331, superscript T indicates type strain) together formed a separate evolutionary branch (Bootstrap support value is 86).

[0300] The sequences of MNH 19801 and the model strains with high 16S rRNA gene sequence similarity in the NCBI database were aligned, and then the phylogenetic tree was constructed using the software MEGA5. The phylogenetic tree was constructed using the maximum likelihood method. The results are shown in the figure. Fig. 9The developmental tree nodes in the figure only show the values ​​with Bootstrap values ​​greater than 50%. It can be seen from the figure that strain MNH19801 and Blautia strain Blautia faecicola KGMB01111T (MK424331) and Blautia strain Blautia brookings ii strain SG772 form two different evolutionary branches (Bootstrap support value is 96).

[0301] 2.4 Genome analysis

[0302] According to the genome data assembled above, the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.6) was used to predict the genome genes. A total of 3292 CDS sequences were predicted, with an average CDS sequence length of 930bp. The model strain with the highest genome similarity was Blautia faecicola, with an average nucleotide similarity (ANI) of 80.29% and a gene coverage of 22.04%. Therefore, it can be identified as a new genus under the Lachnospiraceae family.

[0303] Potential antibiotic resistance genes in the genome were analyzed using the RGI pipeline (version: 4.2.2), with the antibiotic resistance gene database being CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi). No antibiotic resistance genes were identified.

[0304] The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.10.1+) to compare the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The detailed comparison results are shown in Table 4.

[0305] Table 4. Potential toxic genes of MNH19801

[0306] Strain Gene VFDB gene Gene name Comparison consistency (%) gene_00810 VFG000077 P 70.984 gene_00839 VFG013286 galE 64.583 gene_01307 VFG001967 glf 62.707

[0307] The analysis of potential secondary metabolism gene clusters in the genome was performed using antiSMASH5 (version: 5.1.1). The detailed alignment results are shown in Table 5.

[0308] Table 5. Potential secondary metabolism gene clusters of MNH19801

[0309]

[0310] 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.

[0311] Table 6. Potential primary metabolic gene clusters of MNH19801

[0312]

[0313] The butyrate production capacity of this strain was evaluated. The genes related to the butyrate production pathway in the existing technology (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: 2.10.1+). The detailed comparison results are shown in Table 7. The integrity of the butyrate production pathway was then calculated. It was found that the integrity of the butyrate production pathway of this strain was 10%.

[0314] Table 7. List of potential butyrate-producing genes in MNH19801

[0315] Strain Gene Butyrate production pathway name Gene name Comparison consistency (%) gene_01750 Butyrat_P3 HkDJ 73.315 gene_02233 Butyrat_P3 HkDJ 83.77

[0316] Fatty acid composition analysis

[0317] The strain MNH 19801 was inoculated on TSA plates and cultured anaerobically at 37°C for 48 hours. The bacteria were then collected and their fatty acids were extracted and methylated. The fatty acid composition of the strain MNH 19801 was analyzed using the fully automatic bacterial identification system (Microbial ID, Inc., Newark, Del.) (Kroppenstedt, 1985; Meier et al., 1993) of MIDI, USA.

[0318] The fatty acid composition of the experimental strain MNH 19801 is shown in Table 8. The main fatty acids (>10%) of MNH 19801 are 16-carbon saturated fatty acids (C16:0 28.92%) and iso-16-carbon saturated fatty acids (C16:0DMA 17.02%); the other types of fatty acids and their contents are detailed in Table 8 below.

[0319] Table 8. Fatty acid composition of strain MNH 19801

[0320]

[0321]

[0322] In the above table, “----” means no relevant data.

[0323] Based on the morphological, microscopic, physiological and biochemical characteristics, 16S rRNA gene sequence and genome information of strain MNH 19801, it was determined that strain MNH 19801 belongs to a new genus under the Lachnospiraceae.

[0324] Therefore, the strain MNH 19801 was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) with the deposit name Lachnospiraceae sp. MNH19801, the deposit number GDMCC No: 61980, the deposit date October 27, 2021, the deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, and the proposed taxonomic name is Lachnospiraceae sp. MNH19801. The deposited strain was identified as viable by the collection center.

[0325] Example 2: Short-chain fatty acid (SCFA) determination

[0326] Preparation of bacterial cells: The strain MNH 19801 was inoculated into TSB liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacterial cells were collected by centrifugation and stored at -86°C for later use.

[0327] Preparation of standard: Weigh acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid and hexanoic acid standards, and prepare eight mixed standard concentration gradients of 0.1μg / mL, 0.5μg / mL, 1μg / mL, 5μg / mL, 10μg / mL, 20μg / mL, 50μg / mL, and 100μg / mL with ethyl acetate. Take 600μL of standard, add 25μL of 4-methylvaleric acid with a final concentration of 500μM as internal standard, mix well and add to the injection bottle, enter GC-MS detection, injection volume 1μL, split ratio 10:1, split injection.

[0328] Metabolite extraction: Thaw the sample on ice, take 80 mg of the sample in a 2 mL glass centrifuge tube, add 900 μL of 0.5% phosphoric acid to resuspend, shake and mix for 2 min, centrifuge at 14000g for 10 min, take 800 μL of the supernatant, add an equal amount of ethyl acetate to extract, shake and mix for 2 min, centrifuge at 14000g for 10 min, 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 bottle, enter GC-MS detection, injection volume 1 μL, split ratio 10:1, split injection.

[0329] Sample detection and analysis: Agilent DB-WAX capillary column (30m×0.25mm ID×0.25μm) gas chromatography system was used to separate the samples. Program temperature rise: initial temperature 90℃, temperature rise to 120℃ at 10℃ / min, then temperature rise to 150℃ at 5℃ / min, and finally temperature rise to 250℃ at 25℃ / min and maintain for 2min. The carrier gas was helium with a flow rate of 1.0mL / min.

[0330] Agilent 7890A / 5975C gas chromatograph was used for mass spectrometry analysis. The injection port 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) source, full scan and SIM scanning mode, electron energy 70 eV.

[0331] The chromatographic peak area and retention time were extracted using MSD ChemStation software. A calibration curve was drawn to calculate the content of short-chain fatty acids in the sample. The results are shown in Table 9 below.

[0332] Table 9. Short chain fatty acid (SCFA) production results of strain MNH 19801

[0333] SCFA Types Yield (μg / g) Acetic acid 1429.94 Propionic acid 3.78 Isobutyric acid 0.08 Butyric acid 9.04 Isovalerate 1.26 Valeric acid 0.07 Hexanoic acid 0.34

[0334] The results showed that strain MNH 19801 was able to synthesize a large amount of acetic acid and a small amount of propionic acid, isovaleric acid, butyric acid, isobutyric acid, hexanoic acid and valeric acid during its growth.

[0335] Example 3: MNH 19801 bacterial cell preparation method

[0336] Use a sterile toothpick to pick up the MNH 19801 monoclone and transfer it to 10ml AC liquid culture medium for anaerobic culture at 37℃ for 1 day. Take 1ml of the culture medium for mass spectrometry identification. After the identification result is correct, take 2ml of the bacterial suspension and transfer it to 80ml AC liquid culture medium and culture it at 37℃ on the anaerobic operation table. After 24 hours of culture, take 30ml of the bacterial suspension and transfer it to 400ml AC liquid culture medium and culture it at 37℃ on the anaerobic operation table. After 24 hours of culture, take 1ml of the bacterial liquid for mass spectrometry identification. After the identification result is correct, transfer all the bacterial liquid to a 1L centrifuge bottle and centrifuge it at 6000rpm and 4℃ for 30min. Take the precipitate and resuspend it with AC liquid culture medium: sterile glycerol (4:1).

[0337] Pipette 0.1 ml of the MNH 19801 bacterial solution and glycerol mixture into 0.9 ml of saline for gradient dilution to 10 -8 Gradient, concentration is 10 -6 , 10 -7 , 10 -8 100 μl of the gradient dilutions were added to the anaerobic blood plate culture medium, glass beads were added and shaken, and the viable bacterial concentration (CFU) was determined by the plate count method.

[0338] Dispense the prepared bacterial solution into sterile cryopreservation tubes, with 0.2 ml of bacterial solution in each tube and 10 tubes for each bacterial strain. Freeze and store at -80°C. After complete freezing (24 hours), take out a tube of frozen bacteria, thaw to room temperature, and measure CFU by dilution coating method for later use.

[0339] Example 4: Regulation of macrophage immune activity by strain MNH 19801

[0340] After incubating strain MNH 19801 with macrophages (Wuhan Pronocell Life Science Co., Ltd.) for 24 hours, the supernatant was collected and the concentration of the secreted cytokine IL-1β was detected by ELISA. For the specific method, please refer to Experimental Example 1 in CN112618576A.

[0341] Results Fig.10 After co-culture of macrophages and MNH 19801, the macrophages clearly showed the characteristics of differentiation into M1 macrophages, and their IL-1β secretion was significantly increased.

[0342] After co-incubating strain MNH19801 with macrophages for 24 hours, the supernatant was collected and the concentrations of secreted proinflammatory cytokines IL-6, TNFα, RANTES (CCL5), MIG (CXCL-9), MCP-1 (CCL2), IL-2, IL-1β, IL-10, IFN-γ, IP-10, and IL-12 / IL-23P40 were detected by CBA method. The results showed that after co-culture of macrophages with MNH20237, macrophages clearly showed the characteristics of differentiation into M1 macrophages. Specifically, the secretion of IL-6, RANTES, MIG, MCP-1, IL-1β, IP-10, and IL-12 / IL-23P40 increased significantly.

[0343] Example 5: MNH 19801 regulates the immune activity of Primary PBMC

[0344] After incubating strain MNH 19801 with Primary PBMC cells (purchased from TPCS, batch number A19Z289100) for 24 hours, the supernatant was collected and the concentrations of secreted cytokines IL-1β and IFNγ were detected by ELISA. For specific methods, please refer to Experimental Example 2 in CN112618576A.

[0345] After strain MNH 19801 was co-incubated with primary PBMC cells for 24 hours, the supernatant was collected and the concentrations of secreted cytokines TNFα, RANTES (CCL5), MIG (CXCL-9), IL-12 / IL-23P40, IL-1β, IL-6, IL-8, MCP-1 (CCL2), and IP-10 were detected by CBA method.

[0346] The results are as follows Fig.11 As shown in a~11k, MNH19801 can induce increased expression of inflammatory factors TNFα, IL-1β, IL-6, and MCP-1.

[0347] Example 6: Effect of strain MNH19801 on IFNβ expression

[0348] To verify whether MNH19801 can promote the expression of IFNβ, the effect of MNH19801 on the transcriptional activity of IFNβ was evaluated using THP-1 cells (cell line constructed by Muen) carrying the IFNβ gene promoter reporter gene (i.e., IFNβ-promoter reporter).

[0349] The construction steps of THP-1 cells carrying the IFNβ gene promoter reporter gene (THP-1-IFNβ-promoter reporter) include: inserting the reporter gene into the vector, infecting the cells with the vector, and screening to obtain the cell line expressing the reporter gene (specifically Du H, Xu T, Cui M. cGAS-STING signaling in cancer immunity and immunotherapy. Biomed Pharmacother. 2021; 133: 110972.; Jiang M, Chen P, Wang L, et al. cGAS-STING, an important pathway in cancer immunotherapy. J Hematol Oncol. 2020; 13(1): 81.; Lam KC, Araya RE, Huang A, et al. Microbiota triggers STING-type I IFN-dependent monocyte reprogramming of the tumor microenvironment. Cell. 2021; 184(21): 5338-5356. e21.).

[0350] Preparation of MNH19801 culture supernatant: The strain MNH19801 was inoculated into MM01 liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacteria were removed by centrifugation. The culture supernatant was filtered with a 0.22 μm filter, aliquoted, and the collected material was stored at -80°C for later use.

[0351] The samples to be tested include:

[0352] 1) Control group: PBS diluted with 10% volume of MM01 bacterial culture medium;

[0353] 2) Positive control MSA-2 group: MSA-2 (Sting agonist, TargetMol Cat No. T8798) was diluted with PBS to a final concentration of 40 μM;

[0354] 3) MNH19801 group: PBS diluted with 10% volume of MNH19801 bacterial culture supernatant.

[0355] THP-1-IFNβ-promoter reporter cells were seeded in 96-well plates at 1×10 cells per well. 5cells. Treat the cells according to the set groups. After culturing for 24 hours, centrifuge at 300g for 5 minutes, remove the culture supernatant, add 50μL 1×Luciferase detection reagent, react for 1 minute, and detect the luminescence chemiluminescence value with an enzyme reader. The relative fluorescence value (Relative Luminescence) was normalized to the Control group to evaluate the effect of MNH19801 on IFNβ transcription activity.

[0356] The results are as follows Fig.12 As shown, MNH19801 can significantly promote IFNβ transcription activity, and the effect is higher than that of MSA-2 as a positive control.

[0357] Interferon (IFN) is a type of cytokine secreted by host cells that can regulate immune responses. Viruses, bacterial endotoxins, artificially synthesized double-stranded RNA, etc. can stimulate the production of interferon. 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, thereby playing antiviral, antitumor, and immunomodulatory effects. Therefore, the results of this embodiment show that MNH19801 can play an immunomodulatory, antiviral, and antitumor effect by promoting the transcriptional activity of IFNβ.

[0358] Example 7: Inhibition of histone deacetylase activity by strain MNH19801

[0359] In order to verify whether MNH19801 has the function of inhibiting the activity of histone deacetylases (HDAC), this example uses the HDAC Inhibitor Drug Screening Kit (Fluorometric) purchased from Abcam to detect the in vitro HDAC activity inhibition effect.

[0360] Preparation of MNH19801 culture supernatant: strain MNH19801 was inoculated into liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacteria were removed by centrifugation. The culture supernatant was filtered with a 0.22 μm filter, aliquoted, and the collected material was frozen and stored at -80°C for later use.

[0361] Preparation of samples to be tested:

[0362] 1) Control group: PBS diluted with 10% volume of MM01 bacterial culture medium;

[0363] 2) Positive control TSA group: TSA (HDAC inhibitor) was diluted with PBS to a final concentration of 10 μM;

[0364] 3) MNH19801 group: PBS diluted with 10% volume of MNH19801 bacterial culture supernatant.

[0365] HDACs activity detection: Add 50μL of the sample to be tested to a 96-well white plate, then add 50μL of the reaction reagent prepared according to the kit instructions, mix well, incubate at 37℃ for 30min, add 10μL Lysine Developer to the reaction well and mix well to terminate the reaction. Incubate at 37℃ for 30min. Finally, use an ELISA reader to detect the fluorescence intensity of the sample. The ELISA reader settings are Ex. = 350~380nm, Em. = 440~460nm. HDACs inhibition activity analysis takes the fluorescence intensity value of the Control group as 100%, and converts the relative fluorescence intensity of the positive control group and MNH19801 into the relative HDACs activity.

[0366] The results are as follows Fig.13 As shown, MNH19801 can significantly inhibit HDAC activity, which is similar to the effect of HDACs inhibitor TSA (10 μM), so it can be used as a substitute for existing HDAC inhibitors.

[0367] Existing research results show that HDAC inhibitors have good anti-tumor effects on a variety of tumor cells, including tumor cells in the bladder, bone, breast, uterus, central nervous system, esophagus, lung, ovary, pancreas, bile duct cancer, prostate and other parts, and can cause these tumor cells to show obvious apoptosis, proliferation inhibition and cell cycle arrest. As an oncogene that regulates the onset of cancer, the reduction or inhibition of HDAC expression has been shown to have a variety of anti-cancer effects. For example, TSA (Trichostatin A), as an HDAC inhibitor, can upregulate the expression of cell cycle protein kinase inhibitors p21 and p27, downregulate the content of cyclinA, cyclinD1, and cyclinD2, and inhibit the activation of CDK, thereby causing tumor cells to stagnate in the G1 or G2 phase, blocking the continued growth of tumor cells (such as MCF-7 breast cancer cells and Hela cervical cancer cells) and promoting their apoptosis (see the role and mechanism of Trichostatin A in tumor immunotherapy, Zhou Yi, China Pharmaceutical Biotechnology). In addition, TSA can also inhibit tumor angiogenesis by downregulating the expression of tumor angiogenesis-related genes caused by hypoxia and directly inhibiting the migration and proliferation of endothelial cells. In human osteosarcoma, TSA was also found to inhibit the expression of HIF-1α and VEGF, induce the degradation of their receptors, and inhibit tumor angiogenesis by upregulating the transcription of p53 and VHL (Yang QC, Zeng BF, Shi ZM, et al. Inhibition of hypoxia-induced angiogenesis by trichostatin Aviasuppression of HIF-1a activity in human osteosarcoma. J Exp Clin Cancer Res, 2006, 25 (4): 593-599.).

[0368] HDAC inhibitors also have anti-fibrotic effects, and 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 treats diabetes by multiple mechanisms, including inhibition of Pdxl (Park et al., 2008, J Clin Invest, 118, 2316-24), enhancing the expression of transcription factor Ngn3 to increase the endocrine reservoir. 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 composition of the present invention can be used to treat or prevent diabetes mediated by HDAC activity.

[0369] In addition, studies have shown that HDACs are essential for mitochondrial adaptation to IL-1β production in macrophages through non-histone deacetylation, and can promote lipopolysaccharide-induced acute inflammation and high-fat diet-induced chronic inflammation by enhancing NLRP3-dependent activation of caspase 1. This suggests that inhibition of HDACs can also be used to prevent or treat inflammatory diseases.

[0370] Therefore, the strains provided in the examples of the present application can be used to prevent or treat HDAC activity-mediated tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases, etc., by inhibiting HDAC activity. Moreover, since the genes regulated by HDAC play an important role in angiogenesis, they can also be used to prevent tumor metastasis.

[0371] Example 8: Animal experiment on the treatment of liver cancer using strain MNH 19801

[0372] To verify whether strain MNH 19801 can be used for tumor prevention and treatment, a mouse homologous tumor model was used to perform an experiment to inhibit liver cancer growth. This study has been reviewed by the Muen Biotechnology Management and Use Committee.

[0373] Test strains: The MNH 19801 strain in Example 3 was thawed in a glycerol cryopreservation tube at 37°C and inoculated on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain was inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in a solvent (PBS-Cys) to obtain the purity and viable count (2.04×1010 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0374] Tumor cells: H22 mouse hepatoma cells, CBP69230.

[0375] Experimental animals: The experimental mice were C57BL / 6J mice, 5 weeks old, a total of 20 mice, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0376] Animal experiment: The animals were raised normally. After the quarantine period, they were randomly divided into a control group and an experimental group according to their body weight, with 10 mice in each group and kept in separate cages. After grouping, the mouse liver cancer model was established by intradermal injection of H22 mouse liver cancer cells. The cell inoculation amount was 2×10 6 / mL, 0.1mL / mouse. Oral gavage began on the day of completion of inoculation (DAY 0). The control group was given the vehicle, and the experimental group was given MNH 19801 bacteria. The gavage volume was 0.2mL / mouse / time, and the frequency of administration was 1 day / time. General observation was carried out once a day during the quarantine period and once a day after administration during the administration period. The animals were weighed 3 times a week when they were received, at the end of the quarantine period, and during the administration period. Starting from 7 days after tumor cell inoculation (DAY7), 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. The experimental results are shown in Tables 10 and 11.

[0377] Table 10. Overall statistics of mice at the end of the experiment

[0378]

[0379] Table 11. Tumor tissue weight after mouse autopsy

[0380]

[0381]

[0382] The results showed that the tumors in the control group grew rapidly, with the average weight of tumor tissue reaching about 2.53g on D19, while the average weight of tumor tissue in the experimental group was about 1.8g on day 19, and the growth rate was significantly slower than that of the control group, indicating that MNH 19801 can significantly reduce the growth rate of liver cancer tumor tissue.

[0383] Example 9: Immunoassay of tumor-activated mice by strain MNH19801

[0384] 1. Preparation of spleen single cell suspension and cell counting

[0385] 0.1 g of spleen tissue from the experimental endpoint mice (tumor-bearing mice) in Example 8 was taken, the surface was washed with PBS and then the tissue was pulverized. The pulverized spleen tissue was suspended in PBS and filtered through a 70 μm filter membrane to prepare a mouse spleen single cell suspension.

[0386] Take a small amount of the mouse spleen single cell suspension prepared above, add red blood cell lysis solution, Spleen-Counting, pre-cooled PBS, etc. in sequence, complete the on-machine detection in the flow cytometer, and accurately count the mouse spleen single cell value.

[0387] The remaining mouse spleen single cell suspension prepared above was taken, and red blood cell lysis solution, PBS, etc. were added in sequence for cell treatment. The treated cell suspension was filtered through a 300-mesh filter and then tested on a flow cytometer.

[0388] 2. Spleen cell surface T cell / NK cell staining

[0389] A single cell suspension of mouse spleen was obtained, and dyes T-Alexa Fluor647 anti-mouse FOXP3-SP / NK-Alexa Fluor647 anti-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 tested on a flow cytometer.

[0390] Statistical analysis was performed using one-way ANOVA with Dunnet's multiple comparison, *p<0.05, **p<0.01, ***p<0.001.

[0391] The results are as follows Fig.14 As shown in A and B, compared with the control group, the ratio of CD3 and CD8 in mouse spleen cells was significantly upregulated, and the ratio of T cells and NK cells increased. Therefore, MNH19801 can activate the systemic immunity of mice by increasing the ratio of T cells / NK cells in mouse spleen, thereby exerting an anti-tumor effect.

[0392] Example 10: Animal experiment on the use of strain MNH 19801 for lung cancer treatment

[0393] To verify whether strain MNH 19801 can be used for the prevention and treatment of lung cancer, a mouse syngeneic tumor model was used to perform an experiment to inhibit lung cancer growth. This study has been reviewed by the Muen Biotechnology Management and Use Committee.

[0394] Test strains: The MNH 19801 strain in Example 3 was thawed in a glycerol cryopreservation tube at 37°C and inoculated on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain was inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in a solvent (PBS-Cys) to obtain the purity and viable count (1.75×10 8 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0395] Tumor cells: LLC1 mouse lung cancer cells, CL-0140.

[0396] Experimental animals: The experimental mice were C57BL / 6J mice, 5 weeks old, a total of 24 mice, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0397] 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 amount was 2×10 6 / mL, 0.1mL / mouse. When the average tumor volume reaches 60-100mm 3 At 14:00 onset, the animals were randomly divided into a control group and an experimental group according to the tumor volume. On the day of grouping (D1), intragastric administration was started. The control group was given a solvent (PBS-Cys), and the experimental group was given MNH19801 bacteria. The intragastric volume was 0.2 mL / mouse / 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, at the end of the quarantine period, and 3 times a week during the administration period. From D5 to before 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 ≥1000mm 3 The tumor diameter was measured once a day and the tumor growth was recorded. The end point of this experiment was when the average tumor volume of the experimental group was ≥1000mm 3 , and significantly less than the control group or the average tumor growth inhibition rate (TGI) ≥ 30%; or the average tumor volume of any group of mice is greater than 2000mm 3 During the experiment, general clinical observation, body weight monitoring, tumor volume (mm 3) measurement, after reaching the end point of the experiment, all surviving mice were dissected and sampled, and the tumor weight and tumor volume were measured. Finally, the changes in the curve of tumor volume (Tumor volume), the end point tumor volume (Tumor volume at endpoint), the end point tumor weight (Tumor weight at endpoint) and the end point tumor growth inhibition rate (Tumor Growth inhibition rate at endpoint, TGI) were statistically analyzed and compared. Tumor growth inhibition rate (TGI) = 1-(individual tumor volume-individual tumor volume at grouping) / (average tumor volume of the control group-average tumor volume of the control group at grouping) × 100%. All data were expressed in the form of mean ± SD, and GraphPad Prism 8.0.2 software was used for drawing and statistical analysis. For pairwise comparisons, the t test (Student's t test) was used for analysis. For two-factor investigations, two-way ANOVA with Sidak's multiple comparisons was used for analysis. The significant differences were indicated by *, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0398] The experimental results are shown in Table 12:

[0399] Table 12: Overall statistics of mice at the end of the experiment

[0400]

[0401] refer to Fig.15 During the experiment, the statistical analysis results of the tumor volume curve of the experimental group were significantly smaller than that of the control group. Combined with Table 12, at the end of the experiment, the average tumor volume of the mice in the control group was 1962.17 mm 3 The average tumor tissue weight was about 2.475 g; the average tumor volume of the experimental group mice was 1275.27 mm 3 The average weight of tumor tissue was about 1.769 g. The tumor volume and weight of mice in the experimental group were significantly smaller than those in the control group.

[0402] At the end of the experiment, the tumor growth inhibition rate (TGI) of the experimental group was 36.1%, which was significantly higher than that of the control group, and had a significant effect in inhibiting tumor growth.

[0403] refer to Fig.16 At the end of the trial, the response rate of the control group was 14.29%, and the response rate of the experimental group was 37.5%. Therefore, MNH19801 can significantly improve the response rate of tumor treatment.

[0404] Example 11: Experimental verification of the therapeutic effect of strain MNH 19801 on PD-1 combined with lung cancer

[0405] The present invention orders several male C57BL / 6J mice, and tumor cells are inoculated after quarantine to construct an ectopic homologous tumor model of lung cancer. The day of tumor cell inoculation is D1. At D9, mice with abnormal tumor growth are eliminated and randomly stratified and grouped according to the tumor volume of the day. There are 3 groups, 8 mice / group, divided into a negative control group, a positive control group (PD-1 group), and a combined medication group (PD-1+MNH19801). The negative control group was given a solvent and saline, and the positive control group was given a solvent and PD-1 antibody; the combined medication group was given MNH19801 and PD-1 antibody; the solvent and MNH19801 were administered by gavage, the gavage volume was 0.2mL / mouse / time, and the frequency of administration was 1 day / time; PD-1 antibody and saline were administered by intraperitoneal injection, the dosage was 10mg / kg, and the frequency of administration was once every 3 days. The medication intervention was carried out according to the grouping on the day of grouping. During the experiment, animal characteristics, tumor changes, tumor size, and mouse weight were regularly monitored. At the end of the experiment, weight changes, tumor volume changes, response rates, and tumor inhibition rates were analyzed. Throughout the experiment, the weight of all mice increased steadily.

[0406] At the end of the experiment, compared with the negative control group, the tumor volume of the PD-1 group and the combination group showed a significant downward trend, and the tumor volume of the combination group was significantly reduced. Compared with the PD-1 monotherapy group, the tumor volume of the combination group also showed a downward trend.

[0407] like Fig.17 As shown, at D23 of medication, the tumor volume of mice in the negative control group, PD-1 group and combination group began to show significant differences (p < 0.001, ***), and the difference gradually increased with the extension of medication time. In addition, from D17 of medication until the end of the experiment at D28, the tumor volume of mice in the PD-1 group was higher than that in the PD-1 and MNH19801 combination group, and at D25 of medication, the tumor volume of mice in the PD-1 group and the PD-1 and MNH19801 combination group showed significant differences (p < 0.05, *).

[0408] Depend on Fig.18 It can be seen that at the end of the experiment, compared with the negative control group, the tumor volume of the PD-1 group and the combination group showed a significant downward trend, among which the tumor volume of the mice in the combination group was significantly reduced. And compared with the PD-1 monotherapy group, the tumor volume of the combination group showed a more significant downward trend.

[0409] Depend on Fig.19It can be seen that at the end point of the experiment, compared with the negative control group, both the PD-1 group and the combination drug group showed significant tumor inhibition rate (TGI), with a tumor inhibition rate ≥ 50%, and the tumor inhibition rate of the combination drug group was significantly better than that of the PD-1 monotherapy group.

[0410] Fig. 20 A-20C reflects the response rate of mice in each group at the experimental endpoint. Compared with the negative control group, the PD-1 response rates of the PD-1 group and the combination drug group were significantly increased, and the PD-1 response rate of the combination drug group was higher than that of the PD-1 monotherapy group.

[0411] In summary, the combination of MNH19801 and PD-1 showed better efficacy in treating lung cancer, reducing the growth of lung cancer in mice and increasing the PD-1 response rate.

[0412] Example 12: Experimental verification of the therapeutic effect of strain MNH 19801 on PD-1 combined with pancreatic cancer

[0413] The present invention ordered several male C57BL / 6J mice, and after the quarantine, the mice were inoculated with pancreatic cancer KPC cells to construct a pancreatic cancer ectopic homologous tumor model. The day of tumor cell inoculation was D1, and when the tumor grew to an average volume of 60 mm 3 ~100mm 3 At the same time, mice with abnormal tumor growth were removed and randomly stratified and grouped according to tumor volume. A total of 3 groups, 8 mice / group, were set up, divided into negative control group, positive control group (PD-1 group), and combined drug group (PD-1+MNH19801). The negative control group was given vehicle and saline, and the positive control group was given vehicle and PD-1 antibody; the combined drug group was given MNH19801 and PD-1 antibody; the vehicle and MNH19801 were administered by gavage, with a gavage volume of 0.2mL / mouse / time, and a dosing frequency of 1 day / time; PD-1 antibody and saline were administered by intraperitoneal injection, with a dose of 10mg / kg, and a dosing frequency of once every 3 days. On the day of grouping, drug intervention was carried out according to the grouping. During the experiment, animal characteristics, tumor changes, tumor size, and mouse weight were regularly monitored. At the end of the experiment, weight changes and tumor volume changes were analyzed. The weight of all mice increased steadily throughout the experiment.

[0414] At the end of the experiment, compared with the negative control group, the tumor volume of the PD-1 group and the combination group showed a significant downward trend, and the tumor volume of the combination group was significantly reduced. Compared with the PD-1 monotherapy group, the tumor volume of the combination group also showed a downward trend.

[0415] like Fig.21As shown in the tumor growth curve, from D17 after medication until the experimental endpoint D33, the tumor volume of mice in the PD-1 medication group was higher than that in the PD-1 and MNH19801 combination medication group, and on D33, there was a significant difference in the tumor volume between the PD-1 medication group and the PD-1 and MNH19801 combination medication group (the experimental endpoint was statistically analyzed using two-way ANOVA with Dunnett's multiple comparison analysis. *, p < 0.05).

[0416] Therefore, the combination of MNH19801 and PD-1 showed better efficacy in treating pancreatic cancer than PD-1 alone, reducing the growth of lung cancer in mice and increasing the PD-1 response rate.

[0417] Example 13: Evaluation of the effect of strain MNH19801 on differentiation and maturation of mouse bone marrow-derived DC cells

[0418] This embodiment is based on the principle that dendritic cells (DC) play an important role in immune activation. It has been confirmed that DC is the only antigen presenting cell (APC) that can significantly stimulate the proliferation of naive T cells, while other types of APC (such as mononuclear macrophages, B cells, etc.) can only stimulate activated or memory T cells. Therefore, DC is the initiator of the adaptive T cell immune response of the body and plays an extremely important role in tumor immunity. For mice, the most common method is to induce DC from bone marrow cells, that is, bone marrow-derived DC (Bone Marrow-Derived Dendritic Cells, BMDC). Therefore, this study uses this model and uses flow cytometry to detect biomarkers related to differentiation and maturation of DC cell surfaces (CD11c, MHCII, CD80, CD86, etc.) to evaluate the effect of strains on DC differentiation and maturation phenotypes.

[0419] Experimental preparation: The DMEM complete medium of this example is: DMEM medium containing 10% fetal bovine serum (FBS), 1% Penicillin-Streptomycin, 1% L-glutamine, and 1% HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer, pH buffer range of 6.8-8.2).

[0420] BMDC induction medium: DMEM complete medium containing 25 ng / mL GM-CSF (granulocyte-macrophage colony-stimulating factor) and 10 ng / mL IL-4 (interleukin-4).

[0421] Preparation of MNH19801 culture supernatant: The strain MNH19801 was inoculated into MM01 liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacteria were removed by centrifugation. The culture supernatant was filtered with a 0.22 μm filter, aliquoted, and the collected material was stored at -80°C for later use.

[0422] Control group: DMEM complete medium;

[0423] MM01 group: BMDC induction medium containing 10% MM01 bacterial culture medium.

[0424] MNH19801 group: BMDC induction medium containing 10% MNH19801 bacterial culture supernatant.

[0425] BMDC surface marker staining solution: CD45-AF700, CD11C-PC5.5, MHC-II-APC, CD86-PE-Cy7, and CD80-FITC are prepared at a ratio of 1:100, with 100 μL for each reaction system.

[0426] Experimental steps: Extraction of mouse bone marrow cells: Take the femur and tibia of healthy wild-type mice (C57BL / 6), cut both ends, use a 1mL syringe to absorb pre-cooled PBS to blow out the bone marrow cells from the femur and tibia, and disperse them by blowing. After grinding, centrifuge at 1000rpm for 5 minutes. The cells are lysed and finally resuspended in PBS to complete the preparation of single cell suspension.

[0427] Induction and culture of BMDCs: centrifuge and precipitate bone marrow cells, resuspend the cells in 10 mL BMDC induction medium, transfer to a 10 cm dish, and culture in a 37°C constant temperature incubator for 6 days for differentiation. Half of the BMDC induction medium is replaced every two days. After 6 days of culture, relatively mature BMDC cells can be obtained.

[0428] Treatment of BMDC cells with strain supernatant: BMDC cells were centrifuged, resuspended, counted, and inoculated in 12-well plates at 1×106 / well. Cells were treated according to the conditions of the Control group, MM01 group, and MNH19801 group. After 24 hours of culture, cells were collected by centrifugation, washed with PBS, and stained with BMDC surface marker staining solution for 30 minutes. Finally, BMDC cells were detected by flow cytometry to analyze the expression of DC cell MHCII, CD80, CD86 and molecules related to differentiation and maturation.

[0429] Experimental results: Fig. 22Flow cytometry analysis of DC cell surface molecules MHCII, CD80 and CD86. The results showed that compared with the MM01 group, MNH19801 significantly upregulated the expression of MHCII molecules in DC cells; it also significantly promoted the upregulation of CD80 and CD86 molecules. These results suggest that MNH19801 has the effect of promoting DC differentiation and maturation, and therefore may promote the activation of anti-tumor immunity.

[0430] Fig. 22 Data were presented as mean ± standard deviation (Mean ± SD), and one-way ANOVA and Tukey's multiple comparisons test were used for statistical analysis. Significant differences were indicated by *. Significant significance analysis: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.

[0431] Example 14 Evaluation of the effect of strain MNH19801 in inhibiting the M2 TAM phenotype

[0432] This embodiment is based on the principle that tumor-associated macrophages (TAM) play an important role in the malignant progression of tumors. TAM can be roughly divided into M1 and M2 types. M1 plays a pro-inflammatory role and inhibits tumor growth, while M2 plays an anti-inflammatory role, inhibits anti-tumor immune activation, and promotes the malignant progression of tumors. Therefore, for M2 macrophages, this study established an in vitro method of inducing mouse mononuclear macrophage leukemia cells (RAW264.7) with IL-4 (interleukin-4) in vitro to construct an M2 TAM model, and detected the changes in the expression levels of M2 macrophage biomarkers CD206 and Arg1 by real-time fluorescence quantitative PCR technology, which was used to evaluate the effect of strains on the phenotype of M2 macrophages.

[0433] Experimental preparation: Preparation of MNH19801 culture supernatant: The strain MNH19801 was inoculated into MM01 liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacteria were removed by centrifugation. The culture supernatant was filtered with a 0.22 μm filter, aliquoted, and the collected material was stored at -80°C for later use.

[0434] The DMEM complete medium of this embodiment is: DMEM medium: fetal bovine serum (FBS): P / S (Penicillin-Streptomycin) = 9:1:0.1; for example, 45 mL DMEM medium + 5 mL fetal bovine serum (FBS) + 500 μL P / S

[0435] Control group: DMEM complete medium containing 5% volume of MM01 bacterial culture medium;

[0436] TAM group: DMEM complete medium containing 20 ng / mL IL4 and 5% MM01 bacterial culture medium

[0437] MNH19801 group: DMEM complete medium containing 20 ng / mL IL4 and 5% MNH19801 bacterial culture supernatant.

[0438] After the RAW264.7 cells were counted, they were plated in a 6-well plate, with 5*10 cells per well. 5 / 2mL, cultured in a 5% CO2 incubator at 37°C, and the cells adhered for 24 hours. On the second day, the old culture medium was removed with a pipette, and 2mL of the corresponding culture medium of Control, TAM, and MNH19801 were added, mixed gently, and placed in a 5% CO2, 37°C incubator for 2 days. After the culture, the cells were washed with PBS, and RNA was extracted (TAKARA MiniBEST Universal RNA Extraction Kit, TAKARA), reverse transcribed (PrimeScriptTM RT reagent Kit with gDNA Eraser, TAKARA), and qPCR was used to detect the expression of CD206 and Arg1, and the expression of internal reference GAPDH (TB Premix Ex Taq TM II, TAKARA). All operations were performed according to the instructions of the kit.

[0439] CD206 primers: F-CTCTGTTCAGCTATTGGACGC (SEQ ID NO: 2); R-CGGAATTTCTGGGATTCAGCTTC (SEQ ID NO: 3);

[0440] Arg1 primers: F-CATTGGCTTGCGAGACGTAGAC (SEQ ID NO: 4); R-GCTGAAGGTCTCTTCCATCACC (SEQ ID NO: 5);

[0441] GAPDH primers: F-GTGTTCCTACCCCCAATGGTT (SEQ ID NO: 6); R-ATTGTCATACCAGGAAATGAGCTT (SEQ ID NO: 7).

[0442] Experimental results: Fig.23The expression of M2 markers CD206 and Arg1 in RAW264.7 cells was detected by QPCR. The results showed that IL-4 significantly promoted the significant upregulation of the expression of macrophage M2 biomarkers CD206 and Arg1, indicating that IL-4 can successfully induce the M2 transformation of RAW265.7 cells. The culture supernatant of strain MNH19801 significantly inhibited the significant expression of CD206 and Arg1 caused by IL-4, indicating that MNH19801 has the ability to inhibit the M2 transformation of RAW264.7 cells induced by IL-4. These results suggest that MNH26621 has the ability to relieve the immunosuppression of M2 macrophages in the tumor microenvironment, thereby promoting the activation of anti-tumor immunity.

[0443] Fig.23 Data were presented as mean ± standard deviation (Mean ± SD), and one-way ANOVA and Tukey's multiple comparisons test were used for statistical analysis. Significant differences were indicated by *. Significant significance analysis: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.

[0444] The present application is described in detail above in conjunction with the embodiments, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present application. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

Claims

1. A strain characterized in that Any one of the following A1) to A4): A1) strains having 16S rRNA that is at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identical to the sequence shown in SEQ ID NO:1; A2) strain MNH19801 with a deposit number of GDMCC No: 61980; A3) strains having at least 95%, 96%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity to the 16S rRNA of the strain described in A2); A4) a strain whose genome has an average nucleotide identity of at least 87%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% with the genome of any of the strains A1) to A3); and / or a strain whose genome has an alignment score of at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% with the genome of any of the strains A1) to A3); Preferably, the sequence of 16S rRNA of the strain A2) is as shown in SEQ ID No.1; Preferably, the strain is a strain of the Lachnospiraceae family; Preferably, the strain is a strain of the genus Blautia in the family Lachnospiraceae; Preferably, the strain is a new species of the genus Blautia in the family Lachnospiraceae (Blautia sp.); Preferably, the strain is a new strain of the genus Oliverpabstia in the family Lachnospiraceae (Oliverpabsti asp.); Preferably, the strain is capable of producing high acetic acid.

2. A culture of the strain according to claim 1; Preferably, the culture comprises any one of the following C1) to C5): C1) fermentation medium of the strain; C2) the supernatant of the fermentation medium of the strain; C3) the concentrated product of C1) or C2); C4) an extract of C1) or C2); C5) a dried product of any one of C1) to C4); Preferably, the fermentation medium is a liquid medium; Preferably, the culture is obtained using a liquid culture medium under anaerobic culture conditions; Preferably, the fermentation medium is MM01 liquid medium, and / or the culture temperature is 35-42° C., and / or the culture time is 24-72 hours, and / or the pH of the culture medium is 6-10.

3. A microbial agent, characterized in that: The microbial agent comprises the strain according to claim 1, or a metabolite of the strain according to claim 1, or a culture according to claim 2.

4. A composition, characterized in that The composition or the raw material of the composition comprises the strain according to claim 1, or comprises the culture according to claim 2, or comprises the microbial agent according to claim 3; Preferably, the composition further comprises at least one of an excipient, a diluent, and a carrier; Preferably, the composition further comprises an auxiliary material; Preferably, the auxiliary material also includes at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a flavoring agent; Preferably, the composition is at least one of a medicine, a nutritional supplement, a food, a food additive, a feed, and a feed additive; Preferably, the dosage form of the composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and irrigation solutions.

5. A pharmaceutical composition, characterized in that Including the following D1) and D2): D1) The strain according to claim 1, or the metabolite of the strain according to claim 1, or the culture according to claim 2, or the microbial agent according to claim 3; D2) combined medication; Preferably, the combination drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug; Preferably, the pharmaceutical composition further comprises a product instruction sheet; Preferably, the tumor drug includes at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug; Preferably, the chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin; Preferably, the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin, and red bengal; Preferably, the photothermal agent includes at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials; Preferably, the immunotherapy drug includes at least one of an immune cell therapy drug and an immune checkpoint inhibitor; Preferably, the immune cell therapy drug includes at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and a NK cell therapy drug; Preferably, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, and BTLA; Preferably, the pharmaceutical composition comprises 1×10 3 ~10 13 CFU / g of the strain; Preferably, the pharmaceutical composition comprises at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×10 10 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 CFU / g of the strain; Preferably, the strain in the pharmaceutical composition is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium or an irradiated bacterium; Preferably, the pharmaceutical composition or the raw material of the pharmaceutical composition comprises 1-80%, 40-60% by mass of D1); Preferably, the pharmaceutical composition or the raw material of the pharmaceutical composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, %, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80% of D1); Preferably, the pharmaceutical composition or the raw material of the pharmaceutical composition comprises D1) in a mass percentage of 1-80%, 2-70%, 5-60%, 10-50%, 20-40%; Preferably, the pharmaceutical composition further comprises at least one of an excipient, a diluent, and a carrier; Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient; Preferably, the auxiliary material includes at least one of a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavoring agent; Preferably, the pharmaceutical composition is in a dosage form suitable for infants, children or adults; Preferably, the pharmaceutical composition is a dosage form for gastrointestinal administration or parenteral administration; Preferably, the pharmaceutical composition is an oral preparation or an injection; Preferably, the dosage form of the pharmaceutical composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and lavage agents; Preferably, the strain is capable of at least partially proliferating in the intestine of a subject.

6. A pharmaceutical combination product, characterized in that Includes standalone: A first product, wherein the first product comprises the strain according to claim 1, or comprises a metabolite of the strain according to claim 1, or comprises the culture according to claim 2, or comprises the microbial agent according to claim 3; a second product, wherein the second product comprises a combination drug; Preferably, the combination drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug; Preferably, the pharmaceutical combination product further comprises a product instruction sheet; Preferably, the tumor drug includes at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug; Preferably, the chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin; Preferably, the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin, and red bengal; Preferably, the photothermal agent includes at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials; Preferably, the immunotherapy drug includes at least one of an immune cell therapy drug and an immune checkpoint inhibitor; Preferably, the immune cell therapy drug includes at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and a NK cell therapy drug; Preferably, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, and BTLA; Preferably, the first product comprises 1×10 3 ~10 13 CFU / g of the strain; Preferably, the first product comprises at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×10 10 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 CFU / g of the strain; Preferably, the strain in the first product is live bacteria, attenuated bacteria, killed bacteria, freeze-dried bacteria or irradiated bacteria; Preferably, the first product and / or the second product further comprises at least one of an excipient, a diluent, and a carrier; Preferably, the first product and the second product further independently include at least one pharmaceutically acceptable excipient; Preferably, the auxiliary material includes at least one of a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavoring agent; Preferably, the pharmaceutical combination product is a dosage form suitable for infants, children or adults; Preferably, the first product and the second product are independently in the form of a dosage form for gastrointestinal administration or a dosage form for parenteral administration; Preferably, the first product and the second product are independently oral preparations or injections; Preferably, the first product and the second product are independently any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and lavage agents; Preferably, the strain is capable of at least partially proliferating in the intestine of a subject.

7. Use of the strain according to claim 1, the culture according to claim 2, the microbial agent according to claim 3, the composition according to claim 4, the pharmaceutical composition according to claim 5, or the pharmaceutical combination product according to claim 6 in the preparation of a drug for preventing and / or treating a disease; Preferably, the disease includes at least one of tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; Preferably, the solid tumor comprises at least one of head and neck cancer, upper digestive tract cancer, lower digestive tract cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer; Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow; Preferably, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, esophageal-gastric junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma , synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum; Preferably, the metabolic disease includes at least one metabolic abnormality of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome; Preferably, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi; Preferably, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression; Preferably, the prevention and / or treatment of tumors comprises at least one of the following (a) to (l): (a) Inhibit tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) Improve the response rate of tumor treatment; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) preventing and / or inhibiting the spread or metastasis of tumor cells; (g) reduce anti-tumor drug resistance; (h) inhibiting HDAC activity by at least one of acetate or acetate, propionate or propionate, butyrate or butyrate, valeric acid or valerate among SCFAs; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the immune system of the subject to exert an immunomodulatory effect; (k) Promote tumor cell senescence or apoptosis; (l) Inhibit angiogenesis in tumor tissue.

8. Use of the strain according to claim 1, the culture according to claim 2, the microbial agent according to claim 3, the composition according to claim 4, the pharmaceutical composition according to claim 5, or the pharmaceutical combination product according to claim 6 in preventing and / or treating a disease; Preferably, the disease includes at least one of tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; Preferably, the solid tumor comprises at least one of head and neck cancer, upper digestive tract cancer, lower digestive tract cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer; Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow; Preferably, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, esophageal-gastric junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma , synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum; Preferably, the metabolic disease includes at least one metabolic abnormality of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome; Preferably, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi; Preferably, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression; Preferably, the prevention and / or treatment of tumors comprises at least one of the following (a) to (l): (a) Inhibit tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) Improve the response rate of tumor treatment; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) preventing and / or inhibiting the spread or metastasis of tumor cells; (g) reduce anti-tumor drug resistance; (h) inhibiting HDAC activity by at least one of acetate or acetate, propionate or propionate, butyrate or butyrate, valeric acid or valerate among SCFAs; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the immune system of the subject to exert an immunomodulatory effect; (k) promoting tumor cell senescence or apoptosis; (l) Inhibit angiogenesis in tumor tissue.

9. A method for treating a disease, comprising administering to a subject an effective dose of the strain of claim 1, the culture of claim 2, the microbial agent of claim 3, the composition of claim 4, the pharmaceutical composition of claim 5, or the pharmaceutical combination product of claim 6; Preferably, the disease includes at least one of tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor or a metastatic tumor; Preferably, the solid tumor comprises at least one of head and neck cancer, upper digestive tract cancer, lower digestive tract cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, bone tumor, soft tissue tumor, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer; Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow; Preferably, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, esophageal-gastric junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma , synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum; Preferably, the metabolic disease includes at least one metabolic abnormality of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome; Preferably, the infectious disease includes a disease caused by at least one of bacteria, viruses, and fungi; Preferably, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder, and depression; Preferably, the prevention and / or treatment of tumors comprises at least one of the following (a) to (l): (a) Inhibit tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) Improve the response rate of tumor treatment; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of PD-1 drug treatment; (f) preventing and / or inhibiting the spread or metastasis of tumor cells; (g) reduce anti-tumor drug resistance; (h) inhibiting HDAC activity by at least one of acetate or acetate, propionate or propionate, butyrate or butyrate, valeric acid or valerate among SCFAs; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating the immune system of the subject to exert an immunomodulatory effect; (k) Promote tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue; Preferably, the strain, the culture, the microbial agent, the composition, the pharmaceutical composition or the pharmaceutical combination product is administered at least once a day; Preferably, when the administration frequency is twice or more per day, the same dose is administered each time; Preferably, when the administration frequency is twice or more per day, each administration is at a different dose; Preferably, the administration route includes at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, and intramuscular injection.

10. A method for preparing the culture according to claim 2, characterized in that The following steps are involved: Taking the strain according to claim 1, inoculating it into a fermentation medium to obtain a culture; Preferably, the method further comprises centrifuging the culture to collect the fermentation medium supernatant; Preferably, the method further comprises concentrating the fermentation medium or the supernatant of the fermentation medium by at least one of evaporation, freeze-drying, dialysis, extraction, and membrane separation to obtain a concentrated product; Preferably, the method further comprises subjecting the fermentation medium or the supernatant of the fermentation medium to obtain an extract by at least one of extraction and solvent extraction; Preferably, the method further comprises drying any one of the fermentation medium, the fermentation medium supernatant, the concentrated product and the extract to obtain a dry product.

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