Strain, culture, microbial agent, composition, pharmaceutical composition and application of strain, culture, microbial agent, composition and pharmaceutical composition

Abstract

The invention belongs to the technical field of biology, and discloses a strain, a culture, a microbial agent, a composition, a pharmaceutical composition and application thereof. In a first aspect of the present application, a strain is provided, which can be a strain having a preservation number of GDMCC No: 63077, a preservation name of MNH23031, or a strain having at least 95% identity with the 16S rRNA gene sequence of strain MNH23031, and / or having at least 95% average nucleotide consistency with the genome of strain MNH23031. 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, in particular to a strain, a culture, a microbial inoculum, a composition, a pharmaceutical composition and application thereof. BACKGROUND

[0002] The intestinal tract is the largest micro-ecological environment in the human body, participates in various 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. In the past decade, the field of intestinal microecology has developed by leaps and bounds, and human understanding of it has deepened. There are a large number of symbiotic microorganisms in the human intestinal tract, and the total amount of genetic information carried by them is 50-100 times that of the human genome, i.e. the “gut microbiome”, also known as the “second genome” of humans. The gut 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 influence 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, and 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. Research on the interrelationship between the intestinal flora and human health and disease not only has important scientific research significance, but also has important significance and value in clinical diagnosis, treatment and even transformation.

[0004] Intestinal flora imbalance can increase the incidence of colorectal cancer, and the metabolic products of some intestinal microorganisms can directly slow down the carcinogenic effect or inhibit tumor occurrence. Not only colorectal cancer related to the intestinal tract, but also breast cancer, liver cancer, etc. can be affected by the intestinal flora. 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 the intestinal flora so as to neither induce tumors nor enhance the efficacy of anti-tumor drugs is still under study. If these problems can be solved, it will have great significance for innovation and revolution in the field of tumor treatment in the future. Therefore, the development of new methods for treating diseases needs to explore new bacterial species and strains and the characterization of new functional chemical entities. These will help to accelerate the development of new microbiome-oriented therapies based on new bacterial species and functional chemical substances. SUMMARY

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

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

[0007] A1) a strain whose 16S rRNA gene sequence 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) a strain deposited with Guangdong Microbial Culture Collection Center, with the accession number of GDMCC No: 63077;

[0009] A3) a strain whose genome has at least 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 any one of the strains of A1) to A2);

[0010] A4) a strain whose genome has at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% alignment score to the genome of any one of the strains of A1) to A2).

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

[0012] In some embodiments of the present application, the strain is of the genus Lapidilactobacillus.

[0013] According to the strain of the embodiments of the present application, at least the following beneficial effects are achieved:

[0014] The strain of the embodiments of the present application, or the product based on the strain, has a good effect of preventing or treating tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases.

[0015] In a second aspect, the present application provides a culture of the aforementioned strain.

[0016] In some embodiments of the present application, the culture includes any one of the following C1) to C5):

[0017] C1) a fermentation culture of the aforementioned strain; C2) a supernatant of the fermentation culture of the aforementioned strain; C3) a concentrated product of the aforementioned C1) or C2); C4) an extract of C1) or C2); and C5) a dried product of any one of C1) to C4).

[0018] In some embodiments of the present application, the culture is obtained using a liquid medium under anaerobic culture conditions.

[0019] In some embodiments of the present application, the fermentation culture is obtained by inoculating the aforementioned strain into a fermentation medium, the fermentation medium is a liquid medium, the culture temperature is 35-42°C, the culture time is 24-72 hours, and the pH is 6-10.

[0020] In a third aspect of the present application, a microbial inoculant is provided, which comprises the aforementioned strain, or a metabolite thereof, or the aforementioned culture.

[0021] In a fourth aspect of the present application, a composition is provided, which comprises one or more of the aforementioned strain, the aforementioned culture, and the aforementioned microbial inoculant.

[0022] In some embodiments of the present application, the composition further comprises at least one of an excipient, a diluent, and a carrier.

[0023] In some embodiments of the present application, the composition further comprises an adjuvant, which comprises at least one of a lubricant, a wetting agent, an emulsifying agent, a suspension stabilizer, a preservative, a sweetener, and a flavoring agent.

[0024] In some embodiments of the present application, the composition is at least one of a drug, a nutritional supplement, a food, a food additive, a feed, and a feed additive.

[0025] In some embodiments of the present application, the composition is in any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a dripping pill, an injection, a suppository, an aerosol, an oral liquid, a paste, an emulsion, and an enema.

[0026] In a fifth aspect of the present application, a pharmaceutical composition is provided, which comprises D1) and D2) as follows:

[0027] D1) one or more of the aforementioned strain, a metabolite of the aforementioned strain, the aforementioned culture, and the aforementioned microbial inoculant;

[0028] D2) a combination drug.

[0029] In some embodiments of the present application, D2) comprises at least one of an oncology drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug.

[0030] In some embodiments of the present application, the oncology drug comprises at least one of a chemotherapeutic drug, a photosensitizer, a photothermal agent, and an immunotherapeutic drug.

[0031] In some embodiments of the application, the chemotherapeutic drug comprises at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, epirubicin.

[0032] In some embodiments of the application, the photosensitizer comprises at least one of boron dipyrromethene, chlorin, and Bengal red.

[0033] In some embodiments of the application, the photothermal agent comprises at least one of noble metal nanoparticles, organic polymer-based, carbon-based nanomaterials, magnetic nanomaterials, semiconductor nanomaterials.

[0034] In some embodiments of the application, the immunotherapeutic drug comprises at least one of an immune cell therapy drug, an immune checkpoint inhibitor.

[0035] In some embodiments of the application, the immune cell therapy drug comprises at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, an NK cell therapy drug.

[0036] In some embodiments of the application, the immune checkpoint inhibitor comprises an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA.

[0037] In some embodiments of the application, the pharmaceutical composition comprises 1 x 10 3 ~ 10 13 CFU per gram of strain.

[0038] In some embodiments of the application, the pharmaceutical composition comprises at least 1 x 10 3 , 2 x 10 3 , 5 x 10 3 , 1 x 10 4 , 2 x 10 4 , 5 x 10 4 , 1 x 10 5 , 2 x 10 5 , 5 x 10 5 , 1 x 10 6 , 2 x 10 6 , 5 x 10 6 , 1 x 10 7 , 2 x 10 7 , 5 x 10 7 , 1 x 10 8 , 2 x 10 8 , 5 x 10 8 , 1 x 10 9 , 2 x 10 9 , 5 x 109 1 x 10 10 2 x 10 10 5 x 10 10 1 x 10 11 2 x 10 11 5 x 10 11 1 x 10 12 2 x 10 12 5 x 10 12 1 x 10 13 CFU / g of the preceding strain.

[0039] In some embodiments of the present application, the strain in the pharmaceutical composition is a live bacteria, an attenuated bacteria, a killed bacteria, a lyophilized bacteria, or an irradiated bacteria.

[0040] In some embodiments of the present application, the pharmaceutical composition or the raw material of the pharmaceutical composition comprises D1) in a mass percentage of 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%.

[0041] In some embodiments of the present application, 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%.

[0042] In some embodiments of the present application, the pharmaceutical composition further comprises at least one of an excipient, a diluent, a carrier.

[0043] In some embodiments of the present application, the pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant.

[0044] In some embodiments of the present application, the adjuvant further comprises at least one of a lubricant, a wetting agent, an emulsifying agent, a suspension stabilizer, a preservative, a sweetener, a flavoring agent.

[0045] In some embodiments of the present application, the pharmaceutical composition is in a dosage form suitable for infants, children or adults.

[0046] In some embodiments of the present application, the pharmaceutical composition is in a dosage form for gastrointestinal administration or non-gastrointestinal administration.

[0047] In some embodiments of the present application, the pharmaceutical composition is in a dosage form for oral administration or injection.

[0048] In some embodiments of the present application, the pharmaceutical composition is in a dosage form of any one of powder, granules, tablets, capsules, gels, suspensions, drops, dripping pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, enemas.

[0049] In some embodiments of the present application, the strain is capable of at least partially proliferating in the intestinal tract of the subject.

[0050] In some embodiments of the present application, the pharmaceutical composition further comprises a product instruction.

[0051] In a sixth aspect of the present application, a pharmaceutical combination product is provided, comprising:

[0052] a first product, the first product comprising the aforementioned strain, or a metabolite thereof, or the aforementioned culture, or the aforementioned microbial inoculum;

[0053] a second product, the second product comprising a combination drug.

[0054] In some embodiments of the present application, the combination drug comprises at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, a central nervous system disease drug.

[0055] In some embodiments of the present application, the pharmaceutical combination product further comprises a product instruction.

[0056] In some embodiments of the present application, the tumor drug comprises at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, an immunotherapy drug.

[0057] In some embodiments of the present application, the chemotherapy drug comprises at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, epirubicin.

[0058] In some embodiments of the present application, the photosensitizer comprises at least one of boron dipyrromethene, chlorin, and Bengal red.

[0059] In some embodiments of the present application, the photothermal agent comprises at least one of noble metal nanoparticles, organic polymer-based, carbon-based nanomaterials, magnetic nanomaterials, semiconductor nanomaterials.

[0060] In some embodiments of the application, the immunotherapy drug comprises at least one of an immune cell therapy drug, an immune checkpoint inhibitor.

[0061] In some embodiments of the application, the immune cell therapy drug comprises at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, an NK cell therapy drug.

[0062] In some embodiments of the application, the immune checkpoint inhibitor comprises an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA.

[0063] In some embodiments of the application, the first product comprises 1 x 10 3 ~ 10 13 CFU / g of the strain.

[0064] In some embodiments of the application, the first product comprises at least 1 x 10 3 , 2 x 10 3 , 5 x 10 3 , 1 x 10 4 , 2 x 10 4 , 5 x 10 4 , 1 x 10 5 , 2 x 10 5 , 5 x 10 5 , 1 x 10 6 , 2 x 10 6 , 5 x 10 6 , 1 x 10 7 , 2 x 10 7 , 5 x 10 7 , 1 x 10 8 , 2 x 10 8 , 5 x 10 8 , 1 x 10 9 , 2 x 10 9 , 5 x 10 9 , 1 x 10 10 , 2 x 10 10 , 5 x 10 10 , 1 x 10 11 , 2 x 10 11 , 5 x 10 11 , 1 x 10 12 , 2 x 10 12 , 5 x 10 12 , 1 x 10 13 CFU / g of the strain.

[0065] In some embodiments of the present application, the strain MNH 23031 in the first product is live bacteria, attenuated bacteria, killed bacteria, lyophilized bacteria, or irradiated bacteria.

[0066] 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, a carrier.

[0067] In some embodiments of the present application, the first product and the second product further independently comprise at least one pharmaceutically acceptable excipient.

[0068] In some embodiments of the present application, the excipient further comprises at least one of a lubricant, a wetting agent, an emulsifying agent, a suspension stabilizer, a preservative, a sweetener, a flavoring agent.

[0069] In some embodiments of the present application, the pharmaceutical combination product is an infant-appropriate dosage form, a child-appropriate dosage form, or an adult-appropriate dosage form.

[0070] In some embodiments of the present application, the first product and the second product are respectively and independently a dosage form for gastrointestinal administration or a dosage form for non-gastrointestinal administration.

[0071] In some embodiments of the present application, the first product and the second product are respectively and independently an oral dosage or an injection dosage.

[0072] In some embodiments of the present application, the dosage form of the first product and the second product is respectively and independently any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a dripping pill, an injection, a suppository, an aerosol, an oral liquid, a paste, an emulsion, an enema.

[0073] In some embodiments of the present application, the strain is at least capable of partially proliferating in the intestinal tract of the subject.

[0074] In a seventh aspect of the present application, the use of the aforementioned strain, the aforementioned culture, the aforementioned microbial inoculum, the aforementioned composition, the aforementioned pharmaceutical composition, the aforementioned pharmaceutical combination product in the preparation of a medicament for preventing and / or treating a disease is provided.

[0075] In some embodiments of the present application, the disease comprises at least one of a tumor or a cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease.

[0076] In some embodiments of the present application, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, an adenoid tumor, or a metastatic tumor.

[0077] In some embodiments of the application, the solid tumor comprises 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, endocrine system cancer.

[0078] In some embodiments of the application, the tumor or cancer comprises at least one of oral cavity cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity 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, hepatic portal bile duct cancer, distal bile duct cancer, hepatopancreatic ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenum and ampulla neuroendocrine tumor, jejuno-ileum neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymus cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing's sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinosarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular adnexal lymphoma, brain cancer, spinal cord tumor, thyroid differentiated cancer, thyroid undifferentiated cancer, thyroid medullary cancer, parathyroid cancer, adrenal cortex cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0079] In some embodiments of the application, the hematopoietic tumor comprises a tumor of at least one of blood, lymph, and bone marrow.

[0080] In some embodiments of the application, the tumor or cancer comprises a tumor or cancer caused by bacteria or viruses.

[0081] In some embodiments of the application, the bacteria or viruses comprise at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, Fusobacterium nucleatum.

[0082] In some embodiments of the application, the metabolic disease comprises a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, metal ions.

[0083] 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, Sjogren's syndrome.

[0084] In some embodiments of the present application, the infectious disease comprises a disease caused by at least one of a bacterial, viral, or fungal infection.

[0085] In some embodiments of the present application, the central nervous system disease comprises at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, depression.

[0086] In some embodiments of the present application, the prevention and / or treatment of a tumor comprises at least one of (a) to (1) below: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) increasing tumor treatment response rate; (e) increasing efficacy of immunosuppressive drug treatment, for example, increasing 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 by 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) modulating the immune system of a subject to exert immunomodulatory effects; (k) promoting tumor cell senescence or apoptosis; (1) inhibiting angiogenesis in tumor tissue.

[0087] In an eighth aspect of the present application, there is provided use of the aforementioned strain, the aforementioned culture, the aforementioned microbial inoculant, the aforementioned composition, the aforementioned pharmaceutical composition, the aforementioned pharmaceutical combination product in preventing and / or treating a disease.

[0088] In some embodiments of the present application, the disease comprises at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease.

[0089] In some embodiments of the present application, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, an adenoid tumor, or a metastatic tumor.

[0090] In some embodiments of the present application, the solid tumor comprises 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, endocrine system cancer.

[0091] In some embodiments of the present application, the hematopoietic tumor comprises a tumor of at least one of blood, lymph, and bone marrow.

[0092] In some embodiments of the present application, the tumor or cancer comprises at least one of oral cavity cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity cancer, 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, hepatic portal bile duct cancer, distal bile duct cancer, hepatopancreatic ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenum and ampullary neuroendocrine tumor, jejunoileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymus 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, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinosarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular adnexal lymphoma, brain cancer, spinal cord tumor, thyroid differentiated cancer, thyroid undifferentiated cancer, thyroid medullary cancer, parathyroid cancer, adrenal cortex cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0093] In some embodiments of the present application, the tumor or cancer comprises a tumor or cancer caused by a bacterium or a virus.

[0094] In some embodiments of the present application, the bacterium or virus comprises at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, Fusobacterium nucleatum.

[0095] In some embodiments of the present application, the metabolic disease comprises a disease of abnormal metabolism of at least one of amino acid, organic acid, sugar, fat, purine, pigment, blood ammonia, metal ion.

[0096] 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, Sjogren's syndrome.

[0097] In some embodiments of the present application, the infectious disease comprises a disease caused by at least one of a bacterial, viral, or fungal infection.

[0098] In some embodiments of the present application, the central nervous system disease comprises at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, depression.

[0099] In some embodiments of the present application, the prevention and / or treatment of a tumor comprises at least one of (a) to (k): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) increasing tumor treatment response rate; (e) increasing efficacy of immunosuppressive drug treatment, for example, increasing 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 by 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) modulating the immune system of the subject to exert immunomodulatory effects; (k) promoting tumor cell senescence or apoptosis; (1) inhibiting angiogenesis in tumor tissue.

[0100] 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 inoculum, the aforementioned composition, the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical combination product.

[0101] In some embodiments of the present application, the disease comprises at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease.

[0102] In some embodiments of the present application, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, an adenoid tumor, or a metastatic tumor.

[0103] In some embodiments of the application, the solid tumor comprises 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, endocrine system cancer.

[0104] In some embodiments of the application, the hematopoietic tumor comprises a tumor of at least one of blood, lymph, and bone marrow.

[0105] In some embodiments of the application, the tumor or cancer comprises at least one of oral cavity cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity 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, hepatic portal bile duct cancer, distal bile duct cancer, hepatopancreatic ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenum and ampullary neuroendocrine tumor, jejuno-ileum neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymus cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing's sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinosarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular adnexal lymphoma, brain cancer, spinal cord tumor, thyroid differentiated cancer, thyroid undifferentiated cancer, thyroid medullary cancer, parathyroid cancer, adrenal cortex cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia.

[0106] In some embodiments of the application, the tumor or cancer comprises a tumor or cancer caused by a bacterium or a virus.

[0107] In some embodiments of the application, the bacterium or virus comprises at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, Fusobacterium nucleatum.

[0108] In some embodiments of the application, the metabolic disease comprises a disease of abnormal metabolism of at least one of amino acid, organic acid, sugar, fat, purine, pigment, blood ammonia, metal ion.

[0109] 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, Sjogren's syndrome.

[0110] In some embodiments of the present application, the infectious disease comprises a disease caused by at least one of a bacterial, viral, or fungal infection.

[0111] In some embodiments of the present application, the central nervous system disease comprises at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, depression.

[0112] In some embodiments of the present application, the prevention and / or treatment of a tumor comprises at least one of (a) to (k): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) increasing tumor treatment response rate; (e) increasing efficacy of immunosuppressive drug treatment, for example, increasing 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 by 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) modulating the immune system of the subject to exert immunomodulatory effects; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0113] In some embodiments of the present application, the number of administrations of the strain, culture, microbial inoculum, composition, pharmaceutical composition, or pharmaceutical combination product is at least once a day.

[0114] In some embodiments of the present application, when the number of administrations is more than twice a day, each administration is at the same dose.

[0115] In some embodiments of the present application, when the number of administrations is more than twice a day, each administration is at a different dose.

[0116] In some embodiments of the present application, the route of administration comprises at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, intramuscular injection.

[0117] In a tenth aspect of the present application, there is provided a method of preparing the culture as described above, the method comprising the steps of:

[0118] In some embodiments of the present application, the method further comprises inoculating the fermentation medium with the strain as described above.

[0119] In some embodiments of the present application, the method further comprises centrifuging the culture and collecting the supernatant of the fermentation medium.

[0120] In some embodiments of the present application, the method further comprises concentrating the fermentation medium or the supernatant of the fermentation medium by at least one of evaporation, lyophilization, dialysis, extraction, membrane separation to obtain a concentrated product.

[0121] In some embodiments of the present application, the method further comprises extracting the fermentation medium or the supernatant of the fermentation medium by at least one of extraction, solvent extraction to obtain an extract.

[0122] In some embodiments of the present application, the method further comprises drying any one of the fermentation medium, the supernatant of the fermentation medium, the concentrated product, the extract to obtain a dried product. BRIEF DESCRIPTION OF DRAWINGS

[0123] The above and other objects, features and advantages of the disclosed example embodiments will become more apparent from the following detailed description read in conjunction with the accompanying drawings. In the drawings, several embodiments of the disclosure are illustrated by way of example and not limitation in which like reference numerals represent similar, but not necessarily identical, elements or components, wherein:

[0124] Figure 1 A micro-morphology photograph of the strain MNH 23031 is shown.

[0125] Figure 2 A pH tolerance diagram of the strain MNH 23031 is shown.

[0126] Figure 3 A NaCl tolerance diagram of the strain MNH 23031 is shown.

[0127] Figure 4 A phylogenetic tree diagram of MNH 23031 is shown.

[0128] Figure 5 A diagram showing the results of the regulation of macrophage immune activity by the strain MNH 23031 is shown.

[0129] Figure 6 A diagram showing the results of the regulation of Primary PBMC immune activity by the strain MNH 23031 is shown.

[0130] Figure 7A schematic diagram showing the promotion of IFNβ reporter gene transcriptional activity by MNH23031 is shown.

[0131] Figure 8 A schematic diagram showing the tumor volume in mice in Example 6 over time is shown.

[0132] Figure 9 A schematic diagram showing the tumor volume in mice in Example 6 at the end of the experiment is shown.

[0133] Figure 10 A schematic diagram showing the tumor weight in mice in Example 6 at the end of the experiment is shown.

[0134] Figure 11 A schematic diagram showing the tumor growth inhibition rate in mice in Example 6 is shown.

[0135] Figure 12 A schematic diagram showing the response rate of the control group at the end of the experiment in Example 6 is shown.

[0136] Figure 13 A schematic diagram showing the response rate of the test group at the end of the experiment in Example 6 is shown.

[0137] Figure 14 A schematic diagram showing the tumor volume in mice in Example 8 over time is shown.

[0138] Figure 15 A schematic diagram showing the tumor volume in mice in Example 8 at the end of the experiment is shown.

[0139] Figure 16 A schematic diagram showing the tumor growth inhibition rate in mice in Example 8 is shown.

[0140] Figure 17 A schematic diagram showing the response rate of the control group at the end of the experiment in Example 8 is shown.

[0141] Figure 18 A schematic diagram showing the response rate of the test group at the end of the experiment in Example 8 is shown.

[0142] Deposit Information:

[0143] Deposit Unit: Guangdong Microbial Culture Collection Center (GDMCC).

[0144] Deposit Number: GDMCC No: 63077.

[0145] Deposit Time: December 23, 2022.

[0146] Address of deposit: 5th floor, No. 59, Building 59, Guangzhou Xianlie Middle Road 100 Courtyard, Guangdong Provincial Academy of Microbiology

[0147] The deposit name is MNH23031, and the proposed classification name is Lapidilactobacillus sp. DETAILED DESCRIPTION

[0148] The concept and the technical effects generated by the present application will be described below in conjunction 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 embodiments. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0149] The embodiments of the present application will be described in detail below. The described embodiments are exemplary and are used to explain the present application, and cannot be understood as limiting the present application.

[0150] In the description of the present application, if several meanings contain more than one, the meaning of multiple is more than two, greater than, less than, more than, etc. are understood as not including the number, above, below, within, etc. are understood as including the number, the meaning of about is within the number ±20%, 10%, 8%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, 0.1%, etc. If it is described that the first, second is only used to distinguish the purpose of the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the sequence of the indicated technical features.

[0151] In the description of the present application, the description of the terms "one embodiment", "some embodiments", "exemplary embodiment", "example", "specific example", 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 the present specification, the exemplary description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the described specific features, structures, materials or characteristics can be combined in any one or more embodiments or examples in a suitable manner.

[0152] A first aspect of embodiments of the present application relates to a strain, which is named MNH23031, deposited in the Guangdong Microbial Culture Collection Center (GDMCC), with the deposit name MNH23031, the deposit number GDMCC No: 63077, the deposit date December 23, 2022, and the deposit address 5th Floor, No. 59, Building, Guangzhou Institute of Microbiology, Chinese Academy of Sciences, 100 Moxie Middle Road, Guangzhou, China, and the proposed classification name Lapidilactobacillus sp.

[0153] The term "strain" refers to a member of a bacterial species that has specific genetic characteristics that allow it to be distinguished from closely related members belonging to the same bacterial species. The specific genetic characteristics can be the absence of all or part of at least one gene, the absence of all or part of at least one regulatory region (e.g., a promoter, a terminator, a riboswitch, a ribosome binding site), the presence of at least one recombinant gene, the presence of at least one mutated gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one non-native plasmid, the presence of at least one antibiotic resistance gene cassette, or a combination thereof. The genetic characteristics of a strain can be identified by PCR amplification, followed by DNA sequencing of the genomic region of interest or the entire genome. In the case where a strain gains or loses antibiotic resistance or gains or loses biosynthetic capabilities (e.g., an auxotrophic strain) compared to another strain of the same species, the strains or nutrients / metabolites can be distinguished by selection or counter-selection using antibiotics.

[0154] It is known in the art that bacterial species can be classified and identified by traditional classification methods and molecular biology methods. Traditional classification methods include, for example, cell morphology observation, Gram staining, flagella staining, various metabolic tests, etc. Molecular biology methods include ribosomal RNA sequence determination methods, whole genome sequencing-based determination methods, etc.

[0155] The strain MNH 23031 involved in embodiments of the present application has the following taxonomic characteristics: inoculated on MPYG solid plate medium, incubated anaerobically at 37°C for 48h, visible colonies were formed, the colonies were round, with regular and smooth edges, translucent, and no exudate was formed around the colonies. The strain MNH 23031 is gram-positive, microscopic morphological observation found that the strain MNH 23031 has no spores, no flagella, non-motile, spherical or short rod-shaped.

[0156] The strain MNH 23031 involved in the embodiments of the present application has a temperature growth range of 30-42°C, and an optimum growth temperature of 37°C. It can grow in a pH range of 5.0-10.0, and has an optimum growth pH of 6.0-8.0. It can tolerate a maximum of 4% NaCl. It is not tolerant to bile salts, and cannot grow when the bile salt concentration is greater than or equal to 0.10%. It can survive for 1h under aerobic conditions, with a survival rate of about 88%, and grows well under anaerobic conditions.

[0157] The antibiotic sensitivity test of the strain MNH 23031 was performed using the paper disc diffusion method. It was shown that the strain MNH 23031 is sensitive to antibiotics such as chloramphenicol, ciprofloxacin, ceftriaxone and penicillin, and is resistant to erythromycin, lincomycin, ampicillin, cotrimoxazole, gentamicin and tetracycline.

[0158] The 16S rRNA gene sequence of the strain MNH 23031 is shown as SEQ ID NO: 1.

[0159] 16S rRNA is a ribosomal RNA of prokaryotes, and the gene encoding 16S rRNA is composed of a variable region and a conserved region. The conserved region is common to all bacteria, while the variable region differs among different bacteria to varying degrees. By comparing the 16S rRNA gene sequences of bacteria, an evolutionary tree can be drawn according to the evolutionary distance based on the sequence difference basis.

[0160] When the identity between the 16S rRNA gene sequences of two strains is lower 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 cultured biobank. Microbiome, 9(1), p23; Pablo Yarza, Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNA gene sequences, Microbiology, volume 12, p635-p645, 2014).

[0161] When the identity between the 16S rRNA gene sequences of two strains is lower than 95%, it can be judged that they belong to different genera (“A genus definition for Bacteria and Archaea based on genome relatedness and taxonomic affiliation”, R. A. 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)).

[0162] To this end, in some embodiments of the present application, there are provided strain variants based on the above 16S rRNA gene sequences, which 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 the 16S rRNA sequence compared to strain MNH23031, and thus are strains of the same species or different species within the same genus as strain MNH23031. It can also be reasonably inferred that these strains of the same species or different species within the same genus have similar biological activities. In other embodiments, the 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 the 16S rRNA gene sequence with the sequence shown in SEQ ID NO: 1.

[0163] Among them, the "identity" between the sequences of two nucleic acid molecules can be determined using known computer algorithms, such as at least one of the GCG program package, BLASTN or FASTA, and other commercial or publicly available programs can also be, for example, DNAStar "MegAlign" program.

[0164] In addition, with the rapid development of second and third generation sequencing technology, bacterial identification based on whole genome sequencing becomes possible, which makes the bacterial identification results more accurate. Average nucleotide identity (ANI) of bacterial genome refers to the similarity of homologous genes between two bacterial genomes. In the field of bacterial taxonomy, it is generally believed that ANI value needs to reach 98.65% or above to be identified as belonging to the same bacterial 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 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 cultured biobank. Microbiome, 9(1), P23 pages; Pablo Yarza, Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNA gene sequences, Microbiology, volume 12, P635-P645 pages, 2014).

[0165] 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 ("A genus definition for Bacteria and Archaea based on genome relatedness and taxonomic affiliation", R. A. 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)).

[0166] Using the above method, a person skilled in the art can determine whether an isolated strain belongs to the same genus as strain MNH23031 or a new strain of the same species. For example, when the average nucleotide identity ANI value with the strain with accession number GDMCC No: 63077 is at least 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 or have similar biological activity.

[0167] There are various mature ANI value calculation tools at present, such as local operation 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 MNH23031 can be calculated by any of the above local operation software or online calculation tools or other tools or methods known in the art.

[0168] To this end, in some embodiments of the present application, provided are strain variants of the above-mentioned genome-based strain MNH23031, which 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 to the genome of strain MNH23031 or any of the above-mentioned other strain variants, and thus are strains of the same species or different species within the same genus as strain MNH23031 or any of the above-mentioned other strain variants. It can also be reasonably inferred that these strains of the same species or different species within the same genus have similar biological activities. In addition, it can also be judged by the alignment score of the corresponding genome, for example, variants with an alignment score of at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% compared to strain MNH23031 or any of the above-mentioned other strain variants also have the same or similar biological activities.

[0169] In some embodiments, in combination with the alignment results of 16S rRNA gene sequence and ANI, the variants of the strain 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 to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031.

[0170] For example, the variants of the strain have at least 95% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0171] The variants of the strain have at least 96% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0172] a variant of the strain has at least 97% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0173] a variant of the strain has at least 98% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0174] a variant of the strain has at least 98.5% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0175] a variant of the strain has at least 98.65% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0176] a variant of the strain has at least 99% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0177] a variant of the strain has at least 99.5% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0178] a variant of the strain has at least 99.6% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0179] a variant of the strain has at least 99.7% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0180] a variant of the strain has at least 99.8% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0181] a variant of the strain has at least 99.9% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0182] a variant of the strain has at least 99.99% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031; or,

[0183] a variant of the strain has 100% identity to the 16S rRNA gene sequence of MNH23031, while having at least 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 MNH23031.

[0184] The term "variant" includes any bacteria produced from strain MNH23031, such as mutations or recombination that occur through natural mutation or recombination (such as mutations caused by cell proliferation, cell division), or resistance screening, and any other mechanism such as radiation, viruses, transposons, or mutagenic chemicals. In addition, variants also include other strains isolated from natural environment that have the same or similar taxonomic characteristics and properties as strain MNH23031, and generally include other strains of the same genus but different species, or different phenotypic variations within the same species.

[0185] A second aspect of the embodiments of the present application relates to a culture of the strain. The term "culture" refers to the product obtained by culturing the isolated strain in a fermentation medium. The fermentation medium can be a natural or artificial medium, such as a solid medium or a liquid medium. In some embodiments, the fermentation medium includes at least one of peptone, such as meat peptone, casein peptone, whey peptone, soybean peptone, and the like, and can further include other inorganic salts or organic substances for promoting bacterial growth.

[0186] In some embodiments, the culture can be the fermentation medium after the strain is cultured, or the supernatant obtained by centrifugation of the fermentation medium after the strain is cultured, or the product obtained by at least one of evaporation, freeze-drying, dialysis, extraction, membrane separation, and the like, of the fermentation medium or the supernatant after the strain is cultured, or the extract obtained by at least one of extraction, solvent extraction (such as water or organic solvent), and the like, of the fermentation medium or the supernatant after the strain is cultured, or the dried product obtained by drying any of the fermentation medium or the supernatant after the strain is cultured, the concentrated product, or the extract. In some specific embodiments, the extract can be an extract for a specific component or components, such as a component of a specific molecular weight range, a specific solubility, a specific isoelectric point, and the like, or can be an extract without targeting a specific component or components. Specific extraction methods include extraction with at least one of methanol, ethanol, chloroform, water, acid, base, and the like, under a specific process sequence and temperature conditions, such as extraction in 75 v / v% ethanol at 60-80°C for 1-10 minutes, or extraction in methanol at -30 to -10°C for 1-10 minutes, or extraction in a methanol / chloroform / water solution at -40 to -20°C for 30-60 minutes, or extraction in deionized water at 95°C for 1-10 minutes, or extraction in perchloric acid, trichloroacetic acid, hydrochloric acid, sodium hydroxide solution at 0-4°C, and the like, to extract corresponding metabolites, such as at least one of short-chain fatty acids or short-chain fatty acid salts, such as acetic acid, butyric acid, valeric acid, acetate, propionate, valerate.

[0187] In some embodiments, the culture is a culture obtained using a liquid medium under anaerobic culture conditions. For example, the liquid medium is a commercially available MM01 liquid medium (containing about 5 g / L of proteose peptone, about 5 g / L of trypticase, about 10 g / L of yeast extract, about 5 g / L of beef extract, about 5 g / L of glucose, about 2 g / L of K2HPO4, about 2 g / L of Na-acetate, about 1 mL / L of Tween 80, about 5 mg / L of hemin, about 0.5 g / L of L-cysteine hydrochloride, about 1 μL / L of vitamin K1, and about 8 mL / L of an inorganic salt solution (containing about 0.25 g / L of CaCl2, about 1 g / L of K2HPO4, about 1 g / L of KH2PO4, about 0.5 g / L of MgSO4, about 10 g / L of NaHCO3, and about 2 g / L of NaCl)). In some embodiments, the culture temperature is 35-42°C, for example, 37°C, when the culture is performed anaerobically. In some embodiments, the culture time is 24-72 hours, for example, 36-60 hours, for example, 48 hours, when the culture is performed anaerobically. In some embodiments, the pH of the culture medium is 6-10, for example, 7, when the culture is performed anaerobically.

[0188] It is understood that, in the above culture, compounds and / or cell fragments containing the strain, and / or metabolites secreted by the strain are included. The metabolites include small molecules (such as amino acids, nucleosides, nucleotides) and large molecules (such as polypeptides, carbohydrates, nucleic acids, proteoglycans, lipids), and can be primary metabolites directly involved in normal cell functions, or secondary metabolites not generally required for basic cell functions, as well as metabolic intermediates produced during the synthesis of primary or secondary metabolites. Thus, similar biological functions to the strain can be achieved by different types of cultures containing compounds and / or metabolites of the strain.

[0189] A third aspect of the embodiments of the present application relates to a microbial inoculant comprising one or more of the aforementioned strains or metabolites of the strains or the aforementioned cultures, for example, the strain MNH23031 or a metabolite thereof or a culture of the strain.

[0190] A fourth aspect of the embodiments of the present application relates to a composition comprising one or more of the aforementioned strains, cultures, microbial inoculants, or the composition being produced from one or more of the aforementioned strains, cultures, microbial inoculants as raw materials.

[0191] In some embodiments, the composition is used as at least one of a drug, a nutritional supplement, a food, a food additive, a feed, a feed additive. To this end, any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a dripping pill, an injection, a suppository, an aerosol, an oral solution, a paste, an emulsion, an enema can be used, taking into account the specific dosage form used.

[0192] In some embodiments, to form the above-mentioned compositions, at least one of excipients, diluents, carriers are further included. Generally, acceptable carriers or diluents are well known to those skilled in the art. For example, at least one of lactose, starch, glucose, sucrose, methyl cellulose, magnesium stearate, mannitol, sorbitol, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sugar syrup and methyl cellulose, etc. can be selected as carriers or excipients. At least one of ethanol, glycerol and water can be selected as diluents. In addition, it is understood that the specific carriers, excipients or diluents can be selected according to the intended application scenario of the composition and the administration mode, for example, different carriers, excipients or diluents can be selected for different uses such as bacterial agent, medicine, nutritional supplement, food, food additive, feed, feed additive, etc. and different dosage forms such as powder, granule, tablet, capsule, gel, suspension, drop, drop pill, injection, suppository, aerosol, oral liquid, ointment, emulsion, irrigation, etc. In addition, the composition can further include at least one of any suitable lubricants, wetting agents, binders, emulsifiers, suspension stabilizers, solubilizers, preservatives, sweeteners, flavors, etc. in addition to carriers, excipients or diluents. Lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, talc, mineral oil, etc. Binders include starch, gelatin, natural sugars and natural or synthetic gums. Natural sugars such as glucose, anhydrous lactose, free-flowing lactose, beta-lactose or corn sweetener. Natural or synthetic gums such as acacia gum, tragacanth gum, sodium alginate, carboxymethyl cellulose or polyethylene glycol. Preservatives include sodium benzoate, sorbic acid or p-hydroxybenzoic acid ester. By preparing the strain or strain culture into any of the above-mentioned compositions, the release of different requirements of effective active ingredients such as rapid release, sustained release, etc. can be achieved.

[0193] A fifth aspect of the embodiments of the present application relates to a pharmaceutical composition comprising D1) one or more of the aforementioned strains, metabolites thereof, cultures of the strains, the aforementioned microbial inoculants; and D2) a combination drug. Through the combination of drugs, the efficacy is improved. In some embodiments, the combination drug includes at least one of tumor drugs, metabolic disease drugs, inflammatory disease drugs, autoimmune disease drugs, infectious disease drugs, central nervous system disease drugs. In some embodiments, the tumor drug can be at least one of chemotherapy drugs, photosensitizers, photothermal agents, immunotherapy drugs.

[0194] The chemotherapeutic drug includes at least one of an alkylating agent (such as cyclophosphamide, ifosfamide, thiotepa, melphalan, carmustine, lomustine, semustine, nimustine, fotemustine, estramustine, methyl methanesulfonate, mechlorethamine, niuroura, phenylalanine mustard, chlorambucil, hexamethylmelamine, busulfan, temozolomide), a platinum (such as cisplatin, carboplatin, oxaliplatin, spiroplatin, lobaplatin, nedaplatin, dicycloplatin), a podophyllotoxin (such as podophyllotoxin, etoposide, teniposide, etoposide phosphate), a camptothecin (such as camptothecin, hydroxycamptothecin, irinotecan, topotecan), a taxane (such as paclitaxel, docetaxel), an anthracycline (such as daunorubicin, doxorubicin, epirubicin, pirarubicin, idarubicin, mitoxantrone, aclarubicin), a fluoropyrimidine (such as gemcitabine, capecitabine, 5-fluorouracil, difluorouracil, deoxyfluorouridine, tegafur, carmofur, trifluorouridine), an antibiotic (such as mitomycin, bleomycin, pirarubicin), and the like.

[0195] The photosensitizer includes at least one of a porphyrin (such as chlorin, phthalocyanine, bacteriochlorin), a boron dipyrromethene (such as fluoroboron dipyrromethene, azafuoroboron dipyrromethene), a Bengal red, and the like.

[0196] The photothermal agent includes at least one of a noble metal nanoparticle (such as a nanoparticle, nanorod, nanowire, nanosheet of gold, silver, platinum, palladium, and the like noble metal), an organic polymer (such as polypyrrole, polyaniline, polyethylene dioxythiophene, polystyrene sulfonate, indocyanine green, porphyrin), a carbon-based nanomaterial (such as graphene, carbon nanotube, cellulose carbonization product), a magnetic nanomaterial (such as Fe3O4), a semiconductor nanomaterial (such as copper sulfide, molybdenum sulfide, bismuth sulfide, antimony sulfide, gold sulfide, copper selenide, molybdenum selenide, bismuth selenide, antimony selenide, gold selenide).

[0197] The immunotherapeutic drug includes at least one of an immune cell therapy drug, an immune checkpoint inhibitor. The immune cell therapy drug includes at least one of a T cell therapy drug (such as CAR-T, TCR-T), a tumor infiltrating lymphocyte therapy drug, an NK cell therapy drug. 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, BTLA. In some specific embodiments, the inhibitor is an antibody, such as an anti-PD-1 antibody (such as pembrolizumab, nivolumab, camrelizumab), an anti-PD-L1 antibody (such as durvalumab, atezolizumab, avelumab), an anti-CTLA-4 antibody (such as ipilimumab), and the like.

[0198] In some embodiments, the metabolic disease drug comprises a drug for treating diseases of abnormal amino acid metabolism (e.g., phenylketonuria, glycine metabolism disease, tyrosine metabolism disease), diseases of abnormal organic acid metabolism (e.g., methylmalonic acidemia, propionic acidemia), diseases of abnormal sugar metabolism (e.g., glycogen storage disease, galactosemia, diabetes), diseases of abnormal fat metabolism (e.g., hypercholesterolemia, hyperlipidemia), diseases of abnormal purine metabolism (e.g., gout, self-mutilation), diseases of abnormal pigment metabolism (e.g., porphyria), diseases of abnormal ammonia metabolism (e.g., hyperammonemia, Hittlin protein deficiency), diseases of abnormal metal ion metabolism (e.g., liver degeneration).

[0199] For example, in the case of diabetes, the diabetes drug comprises at least one of insulin, metformin, pramlintide, sulfonylurea (e.g., glibenclamide, glipizide, glimepiride), mecobalamin (e.g., repaglinide, nateglinide), thiazolidinedione (e.g., rosiglitazone, pioglitazone), DPP-4 inhibitor (e.g., sitagliptin, saxagliptin, linagliptin), GLP-1 receptor agonist (e.g., exenatide, liraglutide, semaglutide), SGLT2 inhibitor (e.g., canagliflozin, dapagliflozin, empagliflozin), entecavir, tenofovir, lamivudine, adefovir, telbivudine, cobicistat, sofosbuvir, interferon, ribavirin, etc.

[0200] In some embodiments, the pharmaceutical composition further comprises a product insert.

[0201] In some embodiments, the pharmaceutical composition is in an infant-appropriate dosage form, a child-appropriate dosage form, or an adult-appropriate dosage form, is in a parenteral dosage form or a non-parenteral dosage form, is in an oral dosage or an injectable dosage, the strain in the pharmaceutical composition is capable of at least partially proliferating in the subject's intestinal tract, and the strain can be a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium.

[0202] In some embodiments, the pharmaceutical composition comprises 1 x 10 3 ~ 10 13 CFU / g of the strain, i.e., 1 x 10 3 ~ 1 x 10 13 colony-forming units (CFU) of the strain per g of the pharmaceutical composition, for example, at least 1 x 10 3 , 2 x 10 3 , 5 x 10 3 , 1 x 10 4 , 2 x 10 4 , 5 x 10 4 , 1 x 10 5 , 2 x 10 5 , 5 x 10 5 , 1 x 10 6 , 2 x 106 , 5 x 10 6 , 1 x 10 7 , 2 x 10 7 , 5 x 10 7 , 1 x 10 8 , 2 x 10 8 , 5 x 10 8 , 1 x 10 9 , 2 x 10 9 , 5 x 10 9 , 1 x 10 10 , 2 x 10 10 , 5 x 10 10 , 1 x 10 11 , 2 x 10 11 , 5 x 10 11 , 1 x 10 12 , 2 x 10 12 , 5 x 10 12 , 1 x 10 13 CFU / g strain.

[0203] In some embodiments, the pharmaceutical composition or the raw material of the pharmaceutical composition includes D1) in a mass percentage of 1% or more, for example, the pharmaceutical composition or the raw material of the pharmaceutical composition includes D1) in a mass percentage of 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%.

[0204] In some embodiments, the pharmaceutical composition or the raw material of the pharmaceutical composition includes D1) in a mass percentage of 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, 40-60%.

[0205] In some embodiments, the bacterial strain contained in the pharmaceutical composition described above is capable of at least partially colonizing the intestine of the subject, e.g., wherein at least 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% of the bacterial strain is capable of colonizing the intestine of the subject.

[0206] In some embodiments, the bacterial strain in the pharmaceutical composition is lyophilized. In some embodiments, the bacterial strain in the pharmaceutical composition is spray dried. In some embodiments, the bacterial strain in the pharmaceutical composition is lyophilized or spray dried and wherein it is viable. In some embodiments, the bacterial strain in the pharmaceutical composition is lyophilized or spray dried and wherein it is capable of partially or completely colonizing the intestine. In some embodiments, the lyophilized bacterial strain is reconstituted prior to administration. In some cases, reconstitution is by use of a diluent described herein.

[0207] In some embodiments, the pharmaceutical composition is administered orally. Oral administration can involve swallowing, so that the compound enters the gastrointestinal tract, and / or by buccal, lingual, or sublingual, administration by which the compound enters the blood stream directly from the mouth. Pharmaceutical formulations suitable for oral administration can include solid tablets, solid particles, semi-solid, and liquids (including multiple or dispersed systems), e.g., suspensions for syrups; soft or hard capsules; lozenges (including liquid-filled); chews; gels; fast dispersing dosage forms; liposomes; sprays; powders; and mouth washes, gums, and denture adhesive formulations.

[0208] In some embodiments, the pharmaceutical formulation is an enteric formulation, i.e., a gastric juice resistant formulation (e.g., resistant to the pH of the stomach) suitable for delivering the pharmaceutical composition to the intestine by oral administration. Enteric formulations can be particularly useful when the bacteria or another component of the composition is sensitive to acid, e.g., susceptible to degradation under stomach conditions.

[0209] In some embodiments, the enteric formulation comprises an enteric coating. In some embodiments, the formulation is an enteric coated dosage form. For example, the formulation can be an enteric tablet or enteric capsule, etc. The enteric coating can be a conventional enteric coating, such as a conventional coating for tablets, capsules, etc. for oral delivery. The formulation can comprise a film coating, such as a film layer of an enteric polymer, such as an acid insoluble polymer. In some embodiments, the enteric formulation is inherently enteric, such as gastroenteric, without the need for an enteric coating. In some embodiments, the formulation is an inherently enteric capsule (e.g., Oro-White® from Capsugel).

[0210] In some embodiments, the formulation is a soft capsule. Soft capsules are capsules that have a certain elasticity and softness due to the addition of softening agents such as glycerol, sorbitol, maltitol and polyethylene glycol present in the capsule shell. Soft capsules can be produced, for example, on a gelatin or starch basis. Gelatin-based soft capsules are commercially available from various suppliers. Depending on the method of administration, such as oral or rectal administration, soft capsules can have various shapes, which can be, for example, round, oval, oblong or torpedo-shaped. Soft capsules can be produced by conventional processes, such as by the Scherer process, the Accogel process or the drop or blow molding process.

[0211] In some embodiments, the formulation is a microencapsulated capsule, which protects the composition from degradation until delivery to the target site, for example by rupture through chemical or physical stimuli such as pressure, enzymatic activity or physical disintegration, which can be triggered by a change in pH. Any suitable encapsulation method can be used. Exemplary encapsulation techniques include entrapment within a porous matrix, attachment or adsorption on a solid support surface, self-aggregation through flocculation or use of cross-linking agents, and mechanical containment behind a microporous membrane or microencapsulated capsule.

[0212] In some embodiments of the present application, the dosage form of the pharmaceutical composition is any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a drop pill, an injection, a suppository, an aerosol, an oral liquid, a paste, an emulsion, an enema. In some embodiments, the pharmaceutical composition further comprises at least one of an excipient, a diluent, a carrier. Generally, acceptable carriers or diluents are well known to those skilled in the art. For example, at least one of lactose, starch, glucose, sucrose, methyl cellulose, magnesium stearate, mannitol, sorbitol, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sugar syrup, and methyl cellulose, etc. can be selected as the carrier or excipient. At least one of ethanol, glycerol, and water can be selected as the diluent. In some embodiments, the pharmaceutical composition further comprises at least one of an auxiliary material, such as a lubricant, a wetting agent, an emulsifying agent, a suspension stabilizer, a preservative, a sweetener, a flavoring agent. The lubricant includes sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, talc, mineral oil, etc. The binder includes starch, gelatin, natural sugars, and natural or synthetic gum. Among them, the natural sugar is, for example, glucose, anhydrous lactose, free-flowing lactose, beta-lactose, or corn sweetener. The natural or synthetic gum is, for example, acacia gum, tragacanth gum, sodium alginate, carboxymethyl cellulose, or polyethylene glycol. The preservative includes sodium benzoate, sorbic acid, or p-hydroxy benzoic acid ester. By preparing the strain or the strain culture into any one of the above-mentioned compositions, the release of different requirements of the effective active ingredients therein, such as rapid release, sustained release, etc. can be achieved.

[0213] The sixth aspect of the embodiments of the present application relates to a pharmaceutical combination product, which comprises two separate products:

[0214] The first product comprises the aforementioned strain, or the aforementioned metabolite of the strain, or the aforementioned culture, or the aforementioned microbial inoculant;

[0215] The second product comprises a combination drug.

[0216] It can be 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 separately at different time points, and / or by different administration methods, and / or at different administration intervals, with respective dosages.

[0217] In some embodiments, the combination drug comprises at least one of a tumor drug, a metabolic disease drug, an inflammatory disease drug, an autoimmune disease drug, an infectious disease drug, a central nervous system disease drug.

[0218] In some embodiments, the tumor drug can be at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, an immunotherapy drug.

[0219] The chemotherapeutic drug includes at least one of an alkylating agent (such as cyclophosphamide, ifosfamide, thiotepa, melphalan, carmustine, lomustine, semustine, nimustine, fotemustine, estramustine, methyl methanesulfonate, mechlorethamine, niuroura, phenylalanine mustard, chlorambucil, hexamethylmelamine, busulfan, temozolomide), a platinum (such as cisplatin, carboplatin, oxaliplatin, spiroplatin, lobaplatin, nedaplatin, dicycloplatin), a podophyllotoxin (such as podophyllotoxin, etoposide, teniposide, etoposide phosphate), a camptothecin (such as camptothecin, hydroxycamptothecin, irinotecan, topotecan), a taxane (such as paclitaxel, docetaxel), an anthracycline (such as daunorubicin, doxorubicin, epirubicin, pirarubicin, idarubicin, mitoxantrone, aclarubicin), a fluoropyrimidine (such as gemcitabine, capecitabine, 5-fluorouracil, difluorouracil, deoxyfluorouridine, tegafur, carmofur, trifluorouridine), an antibiotic (such as mitomycin, bleomycin, pirarubicin), and the like.

[0220] The photosensitizer includes at least one of a porphyrin (such as chlorin, phthalocyanine, bacteriochlorin), a boron dipyrromethene (such as fluoroboron dipyrromethene, azaheterofluoroboron dipyrromethene), a Bengal red, and the like.

[0221] The photothermal agent includes at least one of a noble metal nanoparticle (such as a nanopoint, nanorod, nanowire, nanosheet of gold, silver, platinum, palladium, and the like noble metal), an organic polymer (such as polypyrrole, polyaniline, polyethylene dioxythiophene, polystyrene sulfonate, indocyanine green, porphyrin), a carbon-based nanomaterial (such as graphene, carbon nanotube, cellulose carbonization product), a magnetic nanomaterial (such as Fe3O4), a semiconductor nanomaterial (such as copper sulfide, molybdenum sulfide, bismuth sulfide, antimony sulfide, gold sulfide, copper selenide, molybdenum selenide, bismuth selenide, antimony selenide, gold selenide).

[0222] The immunotherapeutic drug includes at least one of an immune cell therapy drug, an immune checkpoint inhibitor. The immune cell therapy drug includes at least one of a T cell therapy drug (such as CAR-T, TCR-T), a tumor infiltrating lymphocyte therapy drug, an NK cell therapy drug. 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, BTLA. In some specific embodiments, the inhibitor is an antibody, such as an anti-PD-1 antibody (such as pembrolizumab, nivolumab, camrelizumab), an anti-PD-L1 antibody (such as durvalumab, atezolizumab, avelumab), an anti-CTLA-4 antibody (such as ipilimumab), and the like.

[0223] In some embodiments, the metabolic disease drugs include drugs for treating amino acid metabolism disorders (such as phenylketonuria), glycine metabolism disorders, tyrosine metabolism disorders), organic acid metabolism disorders (methylmalonic acidemia, propionic acidemia), glucose metabolism disorders (such as glycogen storage disease, galactosemia, diabetes), protein metabolism disorders, lipid metabolism disorders (such as hypercholesterolemia, hyperlipidemia), purine metabolism disorders (such as gout, disfigurement), pigment metabolism disorders (such as porphyria), blood ammonia metabolism disorders (such as hyperammonemia, citrinin deficiency), and metal ion metabolism disorders (such as Wilson's disease).

[0224] Taking diabetes as an example, diabetes medications include at least one of the following: insulin, metformin, pramidate, sulfonylureas (such as glibenclamide, glipizide, glimepiride), mecobalamin (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 canagliptin, dapagliptin, enpagliflozin), entecavir, tenofovir, lamivudine, adefovir, telbivudine, simexprevir, sofosbuvir, interferon, ribavirin, etc.

[0225] In some embodiments, the drug combination product is an infant-suitable, pediatric-suitable, or adult-suitable dosage form, is a gastrointestinal-administered or non-gastrointestinal-administered dosage form, is an oral or injectable dosage form, and the strains in the drug combination product are at least partially capable of proliferating in the intestine of the subject, and the strains can be any of live bacteria (such as bacteria, or attenuated bacteria, or lyophilized bacteria, or irradiated bacteria) or dead bacteria.

[0226] In some embodiments, the first product of the drug combination product includes 1×10 3 ~10 13 CFU / g strain, that is, 1×10⁻⁶ strains per gram of the first product. 3 Up to 1×10 13 A strain of colony-forming units (CFU), for example, the first product includes at least 1 × 10⁻⁶ CFU. 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 61 x 10 7 2 x 10 7 5 x 10 7 1 x 10 8 2 x 10 8 5 x 10 8 1 x 10 9 2 x 10 9 5 x 10 9 1 x 10 10 2 x 10 10 5 x 10 10 1 x 10 11 2 x 10 11 5 x 10 11 1 x 10 12 2 x 10 12 5 x 10 12 1 x 10 13 CFU / g strain.

[0227] In some embodiments, the pharmaceutical combination or the raw material of the pharmaceutical combination comprises D1) in an amount of 1% or more by mass, for example, the pharmaceutical combination or the raw material of the pharmaceutical combination comprises D1) in an amount of 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% by mass. In some embodiments, the pharmaceutical combination or the raw material of the pharmaceutical combination comprises D1) in an amount of 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, 40-60% by mass.

[0228] In some embodiments of the present application, the strain in the pharmaceutical combination is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium.

[0229] In some embodiments of the present application, the pharmaceutical combination product is in a dosage form suitable for infants, children or adults. In some of these embodiments, the first product and the second product are each independently a dosage form for administration via the gastrointestinal tract or a dosage form for administration other than via the gastrointestinal tract. In some of these embodiments, the first product and the second product are each independently an oral dosage or an injectable dosage. In some embodiments of the present application, the first product and the second product in the pharmaceutical combination product are each independently in any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a dripping pill, an injection, a suppository, an aerosol, an oral liquid, a paste, an emulsion, an enema. In some embodiments, the first product and the second product in the pharmaceutical combination product further independently comprise at least one of an excipient, a diluent, a carrier. Generally, acceptable carriers or diluents are well known to those skilled in the art. For example, at least one of lactose, starch, dextrose, sucrose, methyl cellulose, magnesium stearate, mannitol, sorbitol, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sugar syrup and methyl cellulose, etc. can be selected as the carrier or excipient. At least one of ethanol, glycerol and water can be selected as the diluent. In some embodiments, the first product and the second product in the pharmaceutical combination product further independently comprise at least one of an auxiliary material, such as a lubricant, a wetting agent, an emulsifying agent, a suspension stabilizer, a preservative, a sweetener, a flavoring agent. Lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, talc, mineral oil, etc. Binders include starch, gelatin, natural sugars such as dextrose, anhydrous lactose, free-flowing lactose, beta-lactose or corn sweetener. Natural or synthetic gum such as acacia, tragacanth, sodium alginate, carboxymethylcellulose or polyethylene glycol. Preservatives include sodium benzoate, sorbic acid or p-hydroxybenzoic esters. By preparing the strain or the culture of the strain into any one of the above compositions, the different requirements for the release of the active ingredient, such as fast release, sustained release, etc. can be achieved.

[0230] A 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, the aforementioned pharmaceutical combination product in the preparation of a medicament for preventing and / or treating a disease. In some of these embodiments, the disease comprises at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease.

[0231] 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, endocrine system cancer, blood tumor including at least one of blood, lymph and bone marrow tumor. The head and neck cancer includes oral cancer (such as lip cancer, tongue cancer, floor of mouth cancer, tonsil cancer, gum cancer, etc.), salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity cancer, sinus cancer, laryngeal cancer, etc., the upper digestive tract cancer includes esophageal cancer, esophagogastric junction cancer, gastric cancer, small intestinal cancer, etc., the lower digestive tract cancer includes appendix cancer, colon cancer, rectal cancer, anal canal cancer, etc., the liver and biliary 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., the neuroendocrine tumor includes gastric neuroendocrine tumor, duodenum and ampulla neuroendocrine tumor, jejunoileum neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, etc., the chest cancer includes thymus cancer, lung cancer (such as small cell lung cancer, non-small cell lung cancer, etc.), malignant pleural mesothelioma, etc., the soft tissue tumor includes angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma (formed by fibroblasts and collagen fibers), gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, etc., the skin cancer includes Merkel cell carcinoma, cutaneous malignant melanoma, etc., the female reproductive system cancer includes vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinosarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, etc., the male reproductive system cancer includes penile cancer, prostate cancer, testicular cancer, etc., the urinary system cancer includes renal cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, etc., the eye cancer includes eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular adnexal lymphoma, etc., the central nervous system cancer includes brain cancer (such as neuroglioma, meningioma, medulloblastoma, medulla pituitary adenoma, primary CNS lymphoma and central nervous system germ cell tumor, etc.), spinal cord tumor, etc., the endocrine system cancer includes thyroid differentiated cancer, thyroid undifferentiated cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortex cancer, 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, lymphoma of the marginal zone of lymph nodes, mucosa-associated lymphoid tissue lymphoma, etc.), cutaneous lymphoma, plasma cell myeloma, leukemia, etc.

[0232] In addition, the tumor or cancer also includes a tumor caused by at least one of a bacterium or a virus, for example, a tumor or cancer caused by at least one of a hepatitis B virus, a hepatitis C virus, a human papillomavirus, an Epstein-Barr virus, a Helicobacter pylori, and a Fusobacterium nucleatum.

[0233] The term "metabolic disease" refers to some diseases caused by accumulation or deficiency of substances such as sugar, fat, and amino acid in the human body, resulting in abnormal content of metabolic substances in the body, and can be roughly divided into two categories of metabolic dysfunction and metabolic hyperplasia. In some specific embodiments of the present application, the metabolic disease includes amino acid metabolic abnormality disease (such as phenylketonuria, glycine metabolic disease, tyrosine metabolic disease), organic acid metabolic abnormality disease (methylmalonic acidemia, propionic acidemia), sugar metabolic abnormality disease (such as glycogen accumulation disease, galactosemia, diabetes), fat metabolic abnormality disease (such as hypercholesterolemia, hyperlipidemia), purine metabolic abnormality disease (such as gout, self-mutilation syndrome), pigment metabolic abnormality disease (such as porphyria), blood ammonia metabolic abnormality disease (such as hyperammonemia, Hartnup disease), metal ion metabolic abnormality disease (such as hepatolenticular degeneration).

[0234] In some embodiments, the inflammatory or autoimmune disease includes 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, Sjogren's syndrome.

[0235] In some embodiments, the infectious disease includes a disease caused by at least one of a bacterial, viral, or fungal infection. In some embodiments, the disease caused by a bacterial infection includes a Gram-negative infection, including brucellosis, Campylobacter infection, cat scratch disease, cholera, E. coli infection, gonorrhea, Klebsiella infection, Enterobacter infection, Serratia infection, Legionella infection, meningococcal infection, pertussis, plague, Pseudomonas infection, Salmonella infection, Shigellosis, typhoid fever, tularemia, and the like, and a Gram-positive infection, including anthrax, C. difficile-induced colitis, diphtheria, Enterococcus infection, erysipelas, Listeriosis, Nocardiosis, Pneumococcal infection, Staphylococcal infection, Streptococcal infection, and the like. In some embodiments, the disease caused by a bacterial infection includes a spirochete infection, including syphilis, yaws, pinta, leptospirosis, Lyme disease, rat-bite fever, relapsing fever, and the like, and an anaerobic infection, including actinomycosis, Bacteroides infection, botulism, Clostridium infection, tetanus, and the like. In some embodiments, the disease caused by a fungal infection includes aspergillosis, blastomycosis, candidiasis, coccidioidomycosis, cryptococcosis, histoplasmosis, mucormycosis, paracoccidioidomycosis, sporotrichosis, and the like. In some embodiments, the infectious disease includes a viral infection, including dengue fever, Ebola infection, Marburg virus infection, Hantavirus infection, Lassa fever, South American hemorrhagic fever, West Nile virus infection, yellow fever, Zika virus infection, hand, foot, and mouth disease, polio, cytomegalovirus infection, herpes simplex virus infection, HIV infection, avian influenza, influenza, Middle East respiratory syndrome, SARS, SARS-CoV-2, and the like.

[0236] In some embodiments, the central nervous system disease includes at least one of a stroke, schizophrenia, drug addiction, Alzheimer’s disease, Parkinson’s disease, anxiety, depression, and the like.

[0237] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial agent, the aforementioned composition, the aforementioned pharmaceutical composition, the aforementioned pharmaceutical combination product prevents and / or treats a tumor by at least one of (a) to (1) as follows: (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) increasing tumor treatment response rate; (e) increasing efficacy of immunosuppressive drug treatment, for example, increasing 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 by 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) modulating the immune system of the subject to exert immunomodulatory effects; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0238] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned composition, the aforementioned pharmaceutical composition, the aforementioned pharmaceutical combination product treats or prevents a disease (such as a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0239] Current research results show that HDAC inhibitors exhibit good anti-tumor effects on various tumor cells, including bladder, bone, breast, uterine, central nervous system, esophagus, lung, ovary, pancreas, cholangiocarcinoma, prostate, etc. These tumor cells exhibit obvious cell apoptosis, proliferation inhibition, and cell cycle arrest, etc. As a cancer gene that regulates the onset of cancer, the reduced or inhibited expression of HDAC activity has shown various anti-cancer effects. In addition, TSA can also inhibit tumor angiogenesis by down-regulating 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 various mechanisms, including the inhibition of Pdxl (Park et al., 2008, J Clin Invest, 118, 2316-24), the enhancement of the expression of transcription factor Ngn3 to increase endocrine library progenitor cells (Haumaitre et al., 2008, Mol Cell Biol, 28, 6373-83) and the enhancement of insulin expression (Molsey et al., 2003, J Biol Chem, 278, 19660-6), etc. HDAC inhibition is also a promising treatment for advanced diabetic complications such as diabetic nephropathy and retinal ischemia (Christensen et al., 2011, Mol Med, 17(5-6), 370-390). Therefore, the aforementioned strain, the aforementioned culture, the aforementioned microbial inoculum, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product achieve the treatment or prevention of diseases (such as tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases) by inhibiting the activity of HDAC.

[0240] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned microbial inoculum, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product achieve the treatment or prevention of diseases (such as tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, central nervous system diseases) by up-regulating pro-inflammatory factors, chemotactic factors to stimulate immunity and / or by down-regulating anti-inflammatory factors to achieve immune regulation. In some embodiments, the pro-inflammatory factors are selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0241] The eighth aspect of this application relates to the use of the aforementioned strains, cultures, microbial agents, compositions, pharmaceutical compositions, and pharmaceutical combination products in the prevention and / or treatment of diseases. In some embodiments, the disease includes at least one of tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases.

[0242] In some embodiments, the aforementioned strains, cultures, microbial agents, compositions, pharmaceutical compositions, and pharmaceutical combination products include the ability to prevent and / or treat 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 drugs, such as improving the efficacy of PD-1 drugs; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing antitumor drug resistance; (h) inhibiting HDAC activity by at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, or 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 senescence or apoptosis; and (l) inhibiting angiogenesis in tumor tissue.

[0243] In some embodiments, the aforementioned strains, cultures, microbial agents, compositions, pharmaceutical compositions, and pharmaceutical combination products achieve the treatment or prevention of diseases (such as tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0244] In some embodiments, the aforementioned strains, cultures, compositions, pharmaceutical compositions, and pharmaceutical combination products achieve the treatment or prevention of diseases (such as tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases) by stimulating immunity through upregulation of pro-inflammatory factors and chemokines and / or by immunomodulation through downregulation of anti-inflammatory factors. In some embodiments, the pro-inflammatory factors are selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0245] A ninth aspect of the embodiments of this application relates to a method of 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, and the aforementioned pharmaceutical combination product.

[0246] In some embodiments, the disease comprises at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease. In some embodiments, the tumor or cancer comprises at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, an adenoid tumor, or a metastatic tumor. In some embodiments, the solid tumor comprises at least one of a head and neck cancer, an upper gastrointestinal cancer, a lower gastrointestinal cancer, a hepatobiliary system cancer, a neuroendocrine tumor, a thoracic cancer, a bone tumor, a soft tissue tumor, a skin cancer, a breast cancer, a female reproductive system cancer, a male reproductive system cancer, a urinary system cancer, an ocular cancer, a central nervous system cancer, an endocrine system cancer. In some embodiments, the hematopoietic tumor comprises a tumor of at least one of blood, lymph, and bone marrow. In some embodiments, the tumor or cancer comprises a tumor or cancer caused by at least one of a bacterium or a virus (e.g., at least one of a hepatitis B virus, a hepatitis C virus, a human papillomavirus, an Epstein-Barr virus, a Helicobacter pylori, a Fusobacterium nucleatum). In some embodiments, the metabolic disease comprises a disease of at least one of an amino acid, an organic acid, a sugar, a fat, a purine, a pigment, blood ammonia, a metal ion. 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, Sjogren's syndrome. In some embodiments, the infectious disease comprises a disease caused by at least one of a bacterium, a virus, a fungus. In some embodiments, the central nervous system disease comprises at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, depression.

[0247] In some embodiments, the prevention and / or treatment of the tumor is achieved by at least one of (a) to (l): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) increasing tumor treatment response rate; (e) increasing efficacy of immunosuppressive drug treatment, e.g., increasing 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 by 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) modulating immune system of the subject to exert immunomodulatory effect; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0248] In some embodiments, the foregoing strain, the foregoing culture, the foregoing microbial inoculant, the foregoing composition, the foregoing pharmaceutical composition, the foregoing pharmaceutical combination product achieves treatment or prevention of a disease (e.g., a neoplastic or cancerous disease, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0249] In some embodiments, the foregoing strain, the foregoing culture, the foregoing microbial inoculant, the foregoing composition, the foregoing pharmaceutical composition, the foregoing pharmaceutical combination product achieves treatment or prevention of a disease (e.g., a neoplastic or cancerous disease, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease) by upregulating proinflammatory factors, chemokine stimulation of immunity and / or by downregulating antiinflammatory factors. In some embodiments, the proinflammatory factors are selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, TNFα.

[0250] In some embodiments, the strain, the culture, the microbial inoculant, the composition, the pharmaceutical composition, or the pharmaceutical combination product is administered at least once daily. In some embodiments, the strain, the culture, the microbial inoculant, the composition, the pharmaceutical composition, or the pharmaceutical combination product is administered at least twice daily, each time at the same dose. In some embodiments, the strain, the culture, the microbial inoculant, the composition, the pharmaceutical composition, or the pharmaceutical combination product is administered at least twice daily, each time at a different dose. In some embodiments, the route of administration comprises at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, intramuscular injection.

[0251] The term "subject," or "patient," refers to any mammal. A subject or patient described as "in need of" treatment (or prevention) of a disease refers to a mammal that would benefit from treatment (or prevention) of a disease. Examples include any one or more animals classified as a mammal, such as, for example, Monotremata, Marsupialia, Notoryctemorphia, Golden-mole, Scandentia, Dermoptera, Pteropodidae, Primates, Solenodon, Sciuromorpha, Lagomorpha, Rodentia, Carnivora, Sirenia, Lagomorpha, Odd-toed ungulates, Even-toed ungulates, Cetacea, etc. Common mammals include Rodentia (e.g., mice, rats, hamsters, guinea pigs), Lagomorpha (e.g., rabbits), Odd-toed ungulates (e.g., horses, donkeys), Even-toed ungulates (e.g., sheep, goats, camels, cattle, pigs), Primates (e.g., monkeys, gorillas, chimpanzees, humans), Carnivora (e.g., dogs, cats), etc. It is understood that the subject can be healthy, or can have a disease at any stage of development, such as at least one of a neoplastic or cancerous disease, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, a central nervous system disease, etc.

[0252] The term "treating" a disease in a subject or "treating" a subject having or suspected of having a disease refers to administering a drug treatment, e.g., administering one or more agents, to the subject, so as to reduce or prevent at least one symptom of the disease from worsening. Thus, in one embodiment, "treating" refers to, inter alia, delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing efficacy of alternative therapy, or decreasing resistance to alternative therapy, or combinations thereof.

[0253] The term "preventing" is art-recognized and, as used in connection with a condition such as local recurrence, is well-understood in the art and includes administering a composition that reduces or delays the onset of a symptom of a medical condition in a subject relative to a subject that does not receive the composition. Thus, prevention of cancer includes, e.g., reducing the number of detectable tumors in a patient population that receives 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.

[0254] A tenth aspect of the present application also relates to a method of making the culture of the foregoing, the method comprising the steps of: taking the strain of the foregoing, inoculating into a fermentation medium, and obtaining the culture.

[0255] In some embodiments, the method further comprises centrifuging the culture and collecting the fermentation medium supernatant. In some embodiments, the method further comprises concentrating the fermentation medium or the fermentation medium supernatant by at least one of evaporation, lyophilization, dialysis, extraction, membrane separation, to obtain a concentrated product. In some embodiments, the method further comprises extracting the fermentation medium or the fermentation medium supernatant by at least one of extraction, solvent extraction, to obtain an extract. In some embodiments, the method further comprises drying any of the fermentation medium, the fermentation medium supernatant, the concentrated product, the extract, to obtain a dried product.

[0256] The meanings of the abbreviations for the moieties referred to in the following examples are as shown in Table 1 below.

[0257] Table 1.

[0258] Abbreviation Meaning / Full name Abbreviation Meaning / Full name IND indomethacin ESC esculetin URE urease GLY glycerol GLU glucose CEL cellobiose MAN mannitol MNE mannose LAC lactose MLZ meltobiose SAC sucrose RAF raffinose MAL maltose SOR sorbitol SAL salicin RHA rhamnose XYL xylose TRE trehalose ARA arabinose GEL gelatin

[0259] The meanings of the abbreviations for the moieties referred to in the following examples are as shown in Table 1 below.

[0260] MM01 liquid medium, consisting of: peptone 5 g / L, trypticase 5 g / L, yeast extract 10 g / L, beef extract 5 g / L, glucose 5 g / L, K2HPO4 2 g / L, Na-acetate 2 g / L, Tween 80 1 mL / L, hemin 5 mg / L, L-cysteine hydrochloride 0.5 g / L, vitamin K1 1 μL / L, inorganic salt solution 8 ml / L (0.25 g CaCl2, 1 g K2HPO4, 1 g KH2PO4, 0.5 g MgSO4, 10 g NaHCO3, 2 g NaCl per 1 L), distilled water 1000 mL, final pH 7.3 ± 0.2.

[0261] MM01 solid medium, consisting of: peptone 5 g / L, trypticase 5 g / L, yeast extract 10 g / L, beef extract 5 g / L, glucose 5 g / L, K2HPO4 2 g / L, Na-acetate 2 g / L, Tween 80 1 mL / L, hemin 5 mg / L, L-cysteine hydrochloride 0.5 g / L, vitamin K1 1 μL / L, inorganic salt solution 8 ml / L (0.25 g CaCl2, 1 g K2HPO4, 1 g KH2PO4, 0.5 g MgSO4, 10 g NaHCO3, 2 g NaCl per 1 L), agar 15 g / L, distilled water 1000 mL, final pH 7.3 ± 0.2.

[0262] AC liquid medium, consisting of: peptone 20 g / L, glucose 5 g / L, yeast extract 3 g / L, beef extract 3 g / L, vitamin C 0.2 g / L, pH 7.0.

[0263] Anaerobic blood agar plate (purchased from Hangzhou Microorganism), consisting of: trypticase 10 g / L, heart infusion 3 g / L, corn starch 1 g / L, meat infusion 5 g / L, yeast extract 5 g / L, NaCl 5 g / L, agar 15 g / L, sterile defibrinated sheep blood 50-100 mL / L, distilled water 1000 mL, final pH 7.3 ± 0.2.

[0264] Trypticase soy broth (TSB) liquid medium, consisting of: trypticase 17 g / L, soy papain hydrolysate 3 g / L, K2HPO4 2.5 g / L, NaCl 5 g / L, glucose 2.5 g / L, pH 7.3 ± 0.2.

[0265] The MPYG medium involved in the embodiments of the present application adopts the PYG MEDIUM (modified) formula of Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures ([https: / / www.dsmz.de / microorganisms / medium / pdf / DSMZ_Medium104.pdf]).

[0266] The solvent (also referred to as PBS-Cys (Gly)) involved in the embodiments of the present application is prepared by uniformly mixing PBS-Cys (phosphate buffer containing 0.05% cysteine hydrochloride) and 100% glycerol at a ratio of 3:1.

[0267] The above-mentioned culture medium can be prepared by using conventional preparation methods and sterilization methods.

[0268] The above-mentioned embodiments are described below in combination with specific examples.

[0269] Example 1:

[0270] Isolation and identification of strain MNH23031.

[0271] 1. Isolation of the strain

[0272] The strain MNH 23031 was isolated from a stool sample of a healthy volunteer in Dali City, Yunnan Province. The specific process is as follows:

[0273] The donor took 2-5 grams of fresh stool, put it into a sample collection and preservation tube, and then homogenized the treated stool sample in an ice box within 24 hours and sent it to the laboratory for strain isolation.

[0274] Physiological saline was divided in a biological safety cabinet, 9 ml / tube; strain isolation medium was prepared, including anaerobic blood agar plate, Columbia blood agar plate, chocolate agar plate, etc., and was transferred into an anaerobic workstation 24 hours in advance, and the sample information, medium type, isolation date, etc. were marked.

[0275] The fresh stool sample was placed in an anaerobic operation station (Don Whitley Scientific A95) and was shaken for 1 min using a vortex shaker, mixed uniformly, 1 mL of the sample was taken and mixed in 9 mL of physiological saline, and then gradient dilution was performed to 10 -1 dilution, and then gradient dilution was performed to 10 -6 dilution, and then gradient dilution was performed to 10

[0276] Take 10 -6 Add the diluted solution dropwise to the separation medium (anaerobic blood agar plate, Columbia blood agar plate, chocolate agar plate) at a rate of 100 μL / plate. Spread the solution evenly and allow the plate surface to dry. Then, invert the plate and incubate at 37°C for 3-5 days.

[0277] The growth of the strains in the isolation medium was observed, and single colonies were picked with sterile toothpicks for strain purification. The purified strains were anaerobically cultured at 37°C for 24–48 h. The pure culture strain numbered MNH 23031 was prepared into a 20% glycerol / water-bacterial suspension and stored at -86°C.

[0278] 2. Identification of strains

[0279] 2.1 Morphological characteristics

[0280] Figure 1 A microscopic morphological photograph of strain MNH 23031 is shown.

[0281] Strain MNH 23031 was inoculated on MPYG solid agar plates and anaerobically cultured at 37°C for 48 hours. Visible colonies formed on the plates. The colonies were round, with regular and smooth edges, and were translucent. No secretions formed around the colonies. The strain was Gram-positive. Microscopic morphological observation revealed that the strain was non-spore-forming, non-flagellated, non-motile, and spherical or short rod-shaped.

[0282] 2.2 Physiological and Biochemical Characteristics

[0283] Figure 2 A schematic diagram of the pH tolerance of strain MNH 23031 is shown.

[0284] like Figure 2 As shown, strain MNH 23031 has a temperature range of 30–42℃, with an optimal growth temperature of 37℃; it can grow in a pH range of 5.0 to 10.0, with an optimal growth pH of 6.0–8.0.

[0285] Figure 3 A schematic diagram of the NaCl tolerance of strain MNH 23031 is shown.

[0286] like Figure 3 As shown, it can tolerate up to 4% NaCl.

[0287] Table 2 shows the results of antibiotic susceptibility testing of strain MNH 23031 using the disc diffusion method.

[0288] Table 2.

[0289]

[0290]

[0291] As shown in Table 2, the strain MNH 23031 is sensitive to antibiotics such as chloramphenicol, ciprofloxacin, ceftriaxone and penicillin; the strain MNH 23031 is resistant to erythromycin, lincomycin, ampicillin, cotrimoxazole, gentamicin and tetracycline.

[0292] 2.3 Determination of 16S rRNA gene of the strain MNH 23031

[0293] Fresh culture of the strain MNH 23031 was taken, and the genomic DNA of the sample was extracted by the SDS method. The extracted genomic DNA template was used to amplify the 16S rRNA gene sequence.

[0294] The amplification primers were 1492R (5'-AGAGTTTGATCATGGCTCAG-3') (SEQ ID NO. 2) and 27F (5'-TAGGGTTACCTTGTTACGACTT-3') (SEQ ID NO. 3). The amplified PCR product was purified, and then sequenced by ABI3730XL analyzer (commercially available), thereby obtaining a 16S rRNA gene sequence of 1429 bp, the sequence of which is shown in SEQ ID NO. 1:

[0295]

[0296] The sequence was aligned with the NCBI Nucleotide collection (nr / nt) database, and the closest species was Lapidilactobacillus wuchangensis, with a similarity of 99.72%. The species classification information of the strain was preliminarily determined, i.e., the strain MNH23031 was preliminarily determined to be Lapidilactobacillus wuchangensis.

[0297] The sequence was aligned with the 16S rRNA gene sequences of related genera, and a phylogenetic tree was constructed using MEGA5, Figure 4 The phylogenetic tree of MNH23031 is shown in the schematic diagram.

[0298] As shown in Figure 4 Strain MNH23031 and Lapidilactobacillus wuchangensis, Lapidilactobacillus gannanensis form a separate evolutionary branch.

[0299] 2.4 Genome analysis

[0300] The genome of strain MNH23031 was fragmented by ultrasonic method, and the fragmentation length was ~ 350 bp, and then an Illumina sequencing library was constructed using a standard DNA library kit (commercially available NEB Ultra TM ). The constructed sequencing library was sequenced by NovaSeq (Illumina) gene sequencer (commercially available) with double-end 150 bp. The sequencing obtained 1.26 Gbp data, and the Q20 accounted for 98.225%.

[0301] The raw sequencing data of the genome was filtered using fastp (version: 0.20.0), and the filtered raw data was used for genome assembly using SPAdes (version: v3.14.0). The genome assembly obtained a total length of 2.30 Mbp, N50 length of 57.5 kbp, and GC content of 40.62%.

[0302] The genome gene was analyzed by genome gene prediction using prokka (version: 1.14.5), a prokaryotic analysis software. A total of 2124 CDS sequences were predicted, with an average CDS sequence length of 881 bp.

[0303] The average nucleotide identity (ANI) and alignment fraction (AF) between the whole genome sequence of MNH23031 and reference organisms were determined to evaluate the degree of genome correlation using GTDB-Tk (version: 1.7.0).

[0304] Table 3 shows the ANI and AF alignment results between the whole genome sequence of MNH23031 and reference organisms.

[0305] Table 3.

[0306]

[0307]

[0308] The strain with the highest average nucleotide identity to the genome of strain MNH 23031 is Lapidilactobacillus gannanensis (GCA_003946675.1), which is shown as a model strain in NCBI, with an average nucleotide similarity (ANI) of 98.73% and an alignment fraction (AF) of 85.48%.

[0309] The 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). For detailed comparison information, refer to Table 4.

[0310] Table 4.

[0311] Strain gene Resistance gene Gene name Alignment identity (%) MNH23031_01097 ARO:3000375 ErmB 97.96

[0312] The analysis of potential virulence factors and related genes in the genome was performed using NCBI blastp (version: 2.7.1+) to align the virulence factor database VFDB (http: / / www.mgc.ac.cn / cgibin / VFs / v5 / main.cgi, updated on September 19, 2019). ErmB is resistant to streptomycin, macrolide antibiotics, lincomycin antibiotics, streptomycin B antibiotics, and streptomycin A antibiotics, which is consistent with the results of the antibiotic sensitivity test.

[0313] Table 5 shows the alignment results of potential virulence genes of strain NH23031.

[0314] Table 5.

[0315] Strain gene VFDB gene Gene name Alignment identity (%) MNH23031_00113 VFG002190 cpsA 64.557 MNH23031_00239 VFG000077 clpP 70.000 MNH23031_00277 VFG006717 lap 60.141 MNH23031_00331 VFG048830 gnd 66.525 MNH23031_00649 VFG002182 cpsI 62.602 MNH23031_00871 VFG002165 efaA 60.215 MNH23031_00913 VFG002182 cpsI 62.670 MNH23031_01248 VFG000080 clpE 60.822 MNH23031_01263 VFG000964 hasC 70.748 MNH23031_01920 VFG002182 cpsI 61.892 MNH23031_01923 VFG000964 hasC 70.742 MNH23031_01941 VFG037100 pilB 64.234

[0316] Table 6 shows the results of the comparison of potential primary metabolic gene clusters of strain MNH23031.

[0317] The analysis of potential primary metabolic gene clusters in the genome was performed using gutSMASH5 (version 1.0.0). The "Pyruvate to acetate-formate E. coli" process breaks down pyruvate to produce acetic acid, which is then converted into butanol and acetone. This process requires the participation of acetyl-CoA and is presumably a step in the tricarboxylic acid cycle.

[0318] Table 6.

[0319]

[0320] Table 7 shows the results of the comparison of potential secondary metabolic gene clusters of strain MNH23031.

[0321] Analysis of potential secondary metabolic gene clusters in the genome was performed using antiSMASH6 (version 6.0.1). "RiPP-like" ribosomes are synthesized and post-translational modified peptides. "T3PKS" type III polyketide synthases are involved in the synthesis of polyketides.

[0322] Table 7.

[0323] Gene cluster range Type From To Most similar known gene cluster Similarity Region 5.1 RiPP-like 57337 67636 - - Region 17.1 T3PKS 13138 53272 - -

[0324] 2.5 Determination of Short-Chain Fatty Acids (SCFA) in Strain MNH23031

[0325] SCFA testing method:

[0326] Sample pretreatment: Take bacterial samples of strain MNH23031 into a 2mL EP tube, add 1mL of pure water, vortex for 10s, add steel beads, grind at 40Hz for 4min, sonicate for 5min (ice-water bath), repeat 3 times; centrifuge the sample at 4℃, 5000rpm for 20min, transfer 0.8mL of supernatant into a 2mL EP tube; add 0.1mL of 50% H2SO4 to the EP tube, vortex for 10s, shake for 10min, sonicate for 10min (ice-water bath); then centrifuge the sample at 4℃, 10000rpm for 15min, and incubate at -20℃ for 30min;

[0327] The supernatant was collected into a sample vial and analyzed by GC-MS. The Shimadzu GC2030-QP2020 NX gas chromatograph-mass spectrometer was equipped with an Agilent HP-FFAP capillary tube (30m×250μm×0.25μm, J&W Scientific, Folsom, CA, USA).

[0328] Table 8 shows the results of short-chain fatty acid (SCFA) assay of strain MNH23031.

[0329] Table 8.

[0330]

[0331]

[0332] Based on the morphological characteristics, microscopic characteristics, physiological and biochemical characteristics, 16S rRNA gene sequence and genomic information of strain MNH 23031, it is determined that strain MNH 23031 belongs to Lapidilactobacillus gannanensis.

[0333] Example 2:

[0334] Strain MNH23031 modulates the immune activity of macrophages.

[0335] The test method refers to the experimental example 1 in the prior art CN112618576B.

[0336] Test strain: After the glycerol stock tube of MNH23031 strain was thawed at 37℃, it was inoculated on anaerobic blood agar plates under anaerobic conditions for activation. The activated strain was inoculated in MM01 liquid medium, centrifuged after anaerobic culture, and the bacterial sediment was resuspended with appropriate PBS. According to the instructions of LIVE / DEAD BacLight Bacterial Viability Kit kit, MNH23031 bacterial flow counting was performed to obtain a strain sample that met the test requirements.

[0337] THP-1 (Wuhan Punsai Life Science Co., Ltd.) was treated with PMA at a final concentration of 5 ng / mL for 48 hours to differentiate into M0 macrophages.

[0338] Strain MNH23031 was added at a ratio of MOI (viable bacteria number: cell number) = 10:1. At the same time, M1 macrophage control group (M0 macrophages induced by 20 ng / ml IFNγ+10 pg / ml LPS for 24 hours) was set. After 1 hour of culture, mixed antibiotics (1 mg / mL Ampicillin; 5 mg / mL Streptomycin; 1 mg / mL Colistin) were added to kill bacteria, and the culture was continued for 23 hours. The supernatant was collected.

[0339] BD TMCytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit kit, according to the instructions, the supernatant sample is reacted with the detection protein, and then the concentrations of cytokines IL-10, MIG, IP-10, IL-12 / IL-13P40, IL-6, MCP-1, RANTES, IL-1β, IL-8 and TNF in the supernatant are detected by flow cytometry as shown in Figure 5

[0340] Figure 5 The results of the strain MNH23031 regulating macrophage immune activity are shown in the schematic diagram.

[0341] The results show that after co-culturing macrophages and MNH23031, the macrophages obviously exhibit the characteristics of M1 type macrophage differentiation. Among them, the contents of pro-inflammatory factors IL-6, IL-1β, IL-8 and IL-12 / IL-13P40 are significantly increased; the contents of chemotactic factors MCP-1, RANTES, MIG and IP-10 are significantly increased.

[0342] Example 3:

[0343] Strain MNH23031 regulates the immune activity of Primary PBMC.

[0344] The test method refers to the experimental example 2 in the prior art CN112618576B.

[0345] Test strain: After thawing the glycerol stock of MNH23031 strain at 37℃, inoculate it on anaerobic blood agar plates under anaerobic conditions for activation, inoculate the activated strain into MM01 liquid medium, centrifuge after anaerobic culture, resuspend the bacterial precipitate with appropriate PBS, according to the instructions of LIVE / DEAD BacLight Bacterial Viability Kit kit, perform MNH23031 bacterial flow counting, and obtain a strain sample that meets the test requirements.

[0346] Commercially available Primary PBMC (purchased from TPCS, batch number A19Z289100) was used, and after recovery, it was cultured in PRMI1640 complete culture medium (10% heat-inactivated FBS, containing 1% L-glutamine, 0.1% ps (penicillin-streptomycin mixture), 10 mg / mL DNAse, and the function of DNAse is to avoid agglutination).

[0347] ​Primary PBMCs were co-incubated with MNH 23031 for 24 hours: bacteria were added at an MOI (live bacteria:cells) ratio of 1:1, and a PBS control group was set up (PBS was co-cultured with Primary PBMCs for 24 hours). After anaerobic culture for 1 hour, the bacteria were killed. The culture was continued for 23 hours, and the supernatant was collected.

[0348] Using BD TM The Cytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit was used to detect the concentrations of cytokines IL-10, MIG, IP-10, IL-12 / IL-13P40, IL-6, MCP-1, RANTES, IL-1β, IFN-γ, IL-8, and TNF in the supernatant using flow cytometry. No results were obtained for IL-10 and IFN-γ. MIG decreased, while IL-12 / IL-13P40, RANTES, and TNF showed no significant changes. Results are as follows. Figure 6 As shown.

[0349] Figure 6 A schematic diagram showing the results of the regulation of the immunomodulatory activity of strain MNH23031 on primary PBMCs is presented.

[0350] The results showed that MNH23031 could induce a significant increase in inflammatory factors MCP-1, IL-1β, IP-10 (CXCL-10), IL-6, and IL-8.

[0351] Example 4:

[0352] Effects of strain MNH23031 on IFNβ expression.

[0353] Interferon (IFN) receptor proteins are a class of cytokines secreted by host cells that regulate immune responses. Viruses, bacterial endotoxins, and synthetic double-stranded RNA can stimulate interferon production. IFNβ belongs to type I interferon and can promote the activity of NK cells, macrophages, and T lymphocytes, thereby exerting antiviral, antitumor, and immunomodulatory effects. To verify whether MNH23031 can promote IFNβ expression, this study used a pre-constructed THP-1 cell line (a cell line built by Muen Biotech) carrying the IFNβ gene promoter reporter gene (i.e., IFNβ-promoter reporter) to evaluate the effect of MNH23031 on IFNβ transcriptional activity.

[0354] The construction steps of THP-1 cells carrying the IFNβ gene promoter reporter gene include: reporter gene insertion into a vector, vector infection of cells, and screening of a cell line expressing the reporter gene (for details, see the literature: Huashan Du, Tianmin Xu, Manhua Cui“cGAS-STING signaling in cancer immunity and immunotherapy”Biomedicine & Pharmacotherapy 133 (2021) 110972; Jiang et al.“cGAS-STING, an important pathway in cancer immunotherapy”Journal of Hematology & Oncology (2020) 13:81; Khiem C. Lam et al.“Microbiota triggers STING-type I IFN-dependent monocyte reprogramming of the tumor microenvironment”Cell 184, 5338-5356).

[0355] Preparation of MNH23031 culture supernatant: the strain MNH23031 was inoculated in MM01 liquid medium, and cultured anaerobically at 37°C for 48 hours. The bacterial bodies were removed by centrifugation, and the culture supernatant was filtered with a 0.22 μm filter, then divided and stored at -80°C for later use.

[0356] Control group: DMEM complete medium (10% FBS) containing 10% volume of MM01 bacterial culture medium;

[0357] MSA-2 group: positive control of IFNβ activation (40 μM)

[0358] MNH23031 group: containing 10% volume of MNH23031 bacterial culture supernatant.

[0359] THP-1-IFNβ-promoter reporter cells were inoculated in a 96-well plate at 1×10 5Cells were treated according to the set group. After 24 hours of continuous culture, the cells were centrifuged at 300g for 5 minutes, the culture supernatant was removed, 50 μL of 1x luciferase detection reagent was added, and the reaction was allowed to proceed for 1 minute. The luminescence chemiluminescence value was detected by an enzyme-labeled instrument. The relative fluorescence value (Relative Luminescence) was normalized by the Control group to evaluate the effect of MNH23031 on the transcriptional activity of IFNβ.

[0360] Figure 7 A schematic diagram showing that MNH23031 promotes the transcriptional activity of the IFNβ reporter gene is shown.

[0361] The results of the experiment are shown in Table 1. Figure 7 As shown in Table 1, strain MNH23031 can significantly promote the transcriptional activity of IFNβ. Type I interferon IFNβ has been shown to be able to achieve anti-tumor through immune regulation, and has potential functional effects on anti-viral tumors. Therefore, these results show that strain MNH23031 can achieve anti-tumor through immune regulation, and has potential functional effects on anti-viral tumors.

[0362] The drugs prepared from the Lapidilactobacillus sp. provided by the present application can significantly promote the transcriptional activity of IFNβ. Type I interferon IFNβ has been shown to be able to reestablish the synergy between innate immunity and acquired immunity in the tumor microenvironment, and is used for the treatment of refractory drug-resistant cancer (see "Targeting the tumor Microenvironment with interferon-β Bridges innate and adaptive immune responses, Yang X1, cancer cell, 2014, doi: 10.1016 / j.ccr.2013.12.004."). Therefore, these results show that MNH23031 or the drugs prepared from the strain of Lapidilactobacillus sp. provided by the present application can achieve anti-tumor through immune regulation, and have potential functional effects on anti-viral tumors.

[0363] Example 5:

[0364] Inhibition of histone deacetylase (HDAC) activity by strain MNH23031.

[0365] To verify whether the strain MNH23031 has inhibitory effect on histone deacetylase activity, the HDAC Inhibitor Drug Screening Kit (Fluorometric) purchased from Abeam Company was used to detect the inhibitory effect of the strain MNH23031 on the histone deacetylase activity in vitro.

[0366] Preparation of the culture supernatant of the strain MNH23031: the strain MNH23031 was inoculated in a liquid medium and cultured anaerobically at 37°C for 48 hours, and then centrifuged to remove the bacterial bodies. The culture supernatant was filtered with a 0.22 μM filter, aliquoted, and stored at -80°C until use.

[0367] Preparation of the test samples:

[0368] 1) Control group (Control), the MM01 medium was diluted 10 times with PBS to obtain 10% MM01;

[0369] 2) Positive control group (TSA group), the HDAC inhibitor Trichostatin A (TSA) at a concentration of 10 μM;

[0370] 3) MNH23031 group, the culture supernatant of the strain MNH23031 was diluted 10 times with PBS to obtain the test sample containing 10% bacterial supernatant.

[0371] Preparation of the reagents for the histone deacetylase detection reaction: the detection reaction system was prepared according to the instructions of the kit, and 50 μL of the reaction reagent was required for each reaction.

[0372] Detection of the histone deacetylase activity: 50 μL of the test sample was added to a 96-well white plate, and then 50 μL of the reaction reagent was added, mixed thoroughly, incubated at 37°C for 30 minutes, 10 μL of Lysine Developer was added to the reaction well and mixed thoroughly to terminate the reaction. The plate was incubated at 37°C for 30 minutes. Finally, the fluorescence intensity of the sample was detected using an enzyme marker. The enzyme marker was set as Ex. = 350-380 nM and EM. = 440-460 nM. The fluorescence intensity value of the Control was set as 100%, and the fluorescence intensity of the positive control group and the MNH23031 group was divided by the control and multiplied by 100% to obtain the relative histone deacetylase activity.

[0373] The experiment showed that, compared with the histone deacetylase activity of the Control group, the supernatant of the strain MNH23031 had inhibitory effect on the HDAC activity. These results suggest that the strain MNH23031 has potential effects on T cell activation and anti-tumor.

[0374] Example 6:

[0375] Verification test of tumor treatment effect of strain MNH23031.

[0376] Inhibition of lung cancer growth test was carried out using mouse homologous tumor model to verify whether strain MNH23031 could be used for prevention and treatment of lung cancer. This study has been reviewed by the Muen Biological Animal Management and Use Committee.

[0377] Test strain: After the strain MNH23031 glycerol stock tube was thawed at 37°C, it was inoculated on anaerobic blood agar plates in an anaerobic workstation for activation. The activated strain was inoculated in MM01 liquid medium and anaerobically cultured to obtain sufficient amount of culture. The cultured bacterial liquid was centrifuged and concentrated, and then the bacterial bodies were resuspended with solvent to obtain the test substance with a purity and viable bacterial count (8.7 x 10 9 CFU / mL) meeting the requirements of animal test.

[0378] Tumor cells: LLC1 mouse lung cancer cells were purchased from Wuhan Punsai Life Science and Technology Co., Ltd.

[0379] Test animals: C57BL / 6J mice were used for the test, and the mice were 5 weeks old, purchased from Guangdong Yekang Biological Technology Co., Ltd.

[0380] Animal test: normal feeding, after the quarantine period, LLC1 lung cancer cells were subcutaneously inoculated to form an ectopic homologous tumor model, and the cell inoculation amount was 2 x 10 6 / mL, 0.1 mL per mouse. When the average tumor volume reached 60-100 mm 3 , the mice were randomly divided into 10 groups according to the tumor volume, including groups A to J, the control group-A group (Control), the test group-B group (MNH23031), and other groups for other test substances. Gavage was started on the same day of grouping (D1). The control group was given solvent, and the B group was given MNH23031 bacteria. The gavage volume was 0.2 mL per mouse per time, and the drug administration frequency was 1 day per time. General observation was performed once a day during the quarantine period and once a day after drug administration during the drug administration period. The animal body weight was measured 3 times a week at the end of the quarantine period and during the drug administration period. The tumor diameter was measured once a day from the fifth day after tumor inoculation to the day before grouping; the tumor diameter was measured once every 2 days after grouping; when the average tumor volume was ≥1000 mm 3 , the tumor diameter was measured once a day, and the tumor growth was recorded.

[0381] The end point of this test was: when the average tumor volume of any group in groups B to J was ≥1000 mm 3 , and was significantly smaller than the control group A or the average tumor growth inhibition rate (TGI) was ≥30%; or the average tumor volume of any group of mice was greater than 2000 mm3 .

[0382] The trial included routine clinical observation, weight monitoring, and tumor volume (mm). 3 After reaching the experimental endpoint, all surviving mice were dissected and their tumors were collected. The tumor weight and volume were measured and calculated. Finally, statistical analysis and comparisons were performed on the changes in tumor volume curve, the tumor volume at the endpoint, the tumor weight at the endpoint, and the tumor growth inhibition rate at the endpoint (TGI).

[0383] Tumor growth inhibition rate (TGI) = 1 - (individual tumor volume - individual tumor volume at grouping) / (mean volume of tumors in group A - mean volume of tumors in group A at grouping) × 100%. All data are expressed as mean ± SD. GraphPadPrism 8.0.2 software was used for plotting and statistical analysis. For pairwise comparisons, Student's t-test was used. For two-way analysis, two-way ANOVA combined with Sidak multiple comparisons was used. Significance was indicated by *: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0384] The experimental results of strain MNH23031 in tumor treatment are as follows: Figure 8 to Figure 13 And as shown in Table 9.

[0385] Table 9 shows the overall statistics of the mice at the experimental endpoint.

[0386] Table 9.

[0387]

[0388] Figure 8 A schematic diagram of the change in mouse tumor volume over time is shown.

[0389] Figure 9 A schematic diagram showing the control of mouse tumor volume at the end of the experiment is shown.

[0390] Figure 10 A schematic diagram showing the comparison of mouse tumor weight at the end of the experiment is shown.

[0391] Figure 11 A schematic diagram showing the control of tumor growth inhibition rate in mice is presented.

[0392] Figure 12The response rate of the control group at the end of the experiment is shown.

[0393] Figure 13 The response rate of the test group at the end of the experiment is shown.

[0394] As shown in Table 9, Figure 8

[0395] As shown in Table 9, Figure 9 and Figure 10 The data is shown as mean ± SD, and the statistical analysis is performed by Student’s t test. Significant differences are indicated by *, p < 0.05, and **, p < 0.01. No significant differences are not indicated. At the end of the experiment, the average tumor volume of the control group was 1962.17 mm 3 , and the average tumor weight was about 2.475 g. The average tumor volume of the test group, i.e., the MNH23031 treatment group, was 1192.69 mm 3 , and the average tumor weight was about 1.627 g. The tumor volume and tumor weight of the MNH23031 treatment group were significantly smaller than those of the control group.

[0396] As shown in Table 9, Figure 11 The data is shown as mean ± SD, and the statistical analysis is performed by Student’s t test. Significant differences are indicated by *, p < 0.05, and **, p < 0.01. No significant differences are not indicated. At the end of the experiment, the tumor growth inhibition rate (TGI) of group B (MNH23031) was 40.3%, which was significantly higher than that of the control group, indicating a significant inhibitory effect on tumor growth.

[0397] Figure 12 As shown in Table 9, 3 The response rate of group A (Negative Control) at the end of the experiment was 14.29%.

[0398] As shown in Table 9, Figure 13 The response rate of group B (MNH23031) was 57.14%, indicating that MNH23031 significantly improved the response rate of tumor treatment.

[0399] Example 7: ​​

[0400] Strain MNH23031 activates immune analysis of tumor mice.

[0401] After the tissues of tumor-bearing mice in Example 6 were prepared into single cell suspensions, specific molecules labeled with fluorescent molecules were labeled, and the proportion of cells expressing specific molecules could be detected in a flow cytometer. The proportion distribution of immune cells to some extent reflects the immune status of the mice. The experimental steps are as follows:

[0402] The tumor tissue was rinsed with PBS, cut into small pieces and pulverized, and the pulverized tumor tissue was placed in a 5 mL collagenase IV digestion system (1 mg / mL collagenase IV, 0.1 mg / mL DNAase, 10% FBS) and digested at 37°C for 30 minutes. The digested tissue liquid was filtered through a 70 μM filter membrane to prepare a mouse tumor single cell suspension, which was detected by a machine to count the total number of cells in the tissue.

[0403] The mouse tumor single cell suspension was taken, and dyes (corresponding antibody combination) Live / Fc, T-TAM-Surface were added in sequence for staining, and the stained cells were resuspended with PBS. Subsequently, according to the instructions of the detection kit (Cytofix Cytoperm Fixation Permeabilization Kit, purchased from BD), the fixed liquid and membrane-breaking liquid were used in sequence, and intranuclear staining was performed for staining. The stained cells were resuspended with PBS, and the machine was detected. TM Fixation Permeabilization Kit, purchased from BD) instructions in sequence using fixation liquid, membrane-breaking liquid processing, nuclear staining for staining; stained cells using PBS resuspension, machine detection.

[0404] The mouse tumor single cell suspension was taken, and SP-Live and Treg Surface were added for staining, and the stained cells were resuspended with PBS. Subsequently, according to the instructions of the detection kit (Foxp3 / Transcription Factor Staining Buffer Set, purchased from Invitrogen), the fixed liquid and membrane-breaking liquid were used in sequence, and the antibody Foxp3 was added for staining. The stained cells were resuspended with PBS, and the machine was detected.

[0405] The mouse tumor single cell suspension was taken, and dyes (corresponding antibody combination) Live / Fc, T-Tumor Stain were added for tumor cell surface T cell staining, and the stained cells were resuspended with PBS. The mouse tumor single cell suspension was taken, and dyes (corresponding antibody combination) Live / Fc, MDSC Stain were added in sequence for staining, and the stained cells were resuspended with PBS. The machine was detected.

[0406] Flow cytometry data were processed by CyExpert and analyzed by Graphpad Prism V9. Statistical analysis was performed by one-way ANOVA with Dunnet’s Multiple Comparison, ns not significant *p<0.05, **p<0.01, ***p<0.001.

[0407] Results showed that MNH23031 can activate the local immunity of mouse tumor and play an anti-tumor effect.

[0408] Example 8:

[0409] Verification test of the therapeutic effect of strain MNH23031 on liver cancer tumor.

[0410] To verify whether strain MNH23031 has the effect of treating tumors, a mouse homologous tumor model was used to perform a liver cancer growth inhibition test. The test protocol has been reviewed by the Muen Biological Animal Ethics Committee.

[0411] Test strain: After thawing the glycerol stock of MNH23031 strain at 37°C, inoculate it on anaerobic blood agar plates in an anaerobic workstation for activation. Inoculate the activated strain into MM01 liquid medium and cultivate anaerobically to obtain a sufficient number of live bacteria. Centrifuge the cultured bacterial solution to concentrate it, then resuspend the bacterial bodies with solvent to obtain the test substance with a purity and viable bacterial count (2.0 x 10 9 CFU / mL) meeting the requirements of animal tests.

[0412] Tumor cells: H22 mouse hepatoma cells were purchased from Nanjing Kebai Biological Technology Co., Ltd.

[0413] Test animals: BALB / C mice were used for the test, aged 5-6 weeks, purchased from Guangdong Sijia Jiada Biological Technology Co., Ltd.

[0414] Animal test: normal feeding, after the quarantine period, subcutaneously inoculate H22 hepatoma cells to form an ectopic homologous tumor model, the cell inoculation amount is 2 x 10 5 / mL, 0.1 mL per mouse. When the average tumor volume reaches 80-100 mm 3At the time of tumor volume stratification, the control group-A group (Control) and the test group-B group (MNH23031) were randomly divided, and the C, D, E, and F groups were other test substances. The animals were administered intragastrically on the day of grouping. The control group was given the vehicle, and the B group was given MNH23031 bacteria. The intragastric volume was 0.2 mL per animal per time, and the administration frequency was once a day. During the quarantine period and after administration each day during the administration period, general observations were made once a day. The animals were weighed at the end of the quarantine period, and the animals were weighed each time the tumor diameter was measured after tumor inoculation. The animals were weighed before dissection at the end of the experiment. Day D1 was the day of tumor cell inoculation. During the experiment, general clinical observations were made once a day. The animals were weighed at the end of the quarantine period. The animals were weighed twice a week after tumor inoculation. From the fifth day after tumor inoculation to the day of grouping, the tumor diameter was measured once a day. After grouping, the tumor diameter was measured once every two days. When the average tumor volume of any of the B, C, D, E, or F groups reached or exceeded 1000 mm 3 The tumor diameter was measured once a day.

[0415] When the average tumor volume of any of the B, C, D, E, or F groups reached or exceeded 1000 mm 3 , the average tumor volume of any group was significantly smaller than that of the control group, or the average tumor growth inhibition rate (TGI) of the group reached or exceeded 30%, the experiment was terminated. If the average tumor volume of any group of mice was greater than 2000 mm 3 , the experiment was terminated. After reaching the endpoint of the experiment, all surviving mice were dissected and euthanized.

[0416] After reaching the endpoint of the experiment, all surviving mice were dissected and euthanized, and the tumor weight and tumor volume were measured. Finally, statistical analysis and comparison of tumor volume curve changes (Tumor volume), endpoint tumor volume (Tumor volume at endpoint), endpoint tumor weight (Tumor weight at endpoint), and endpoint tumor growth inhibition rate (Tumor inhibition rate at endpoint, TGI) were performed. The tumor inhibition rate was calculated as follows: (average volume of the control group - volume of the test group) / average volume of the control group x 100%. All data were expressed as mean ± SD, and were plotted and statistically analyzed using GraphPad Prism 8.0.2 software. For pairwise comparisons, the Student's t test was used for analysis. For two-factor investigation, the 2-way ANOVA method was used in combination with Sidak's multiple comparison for analysis. Significant differences were indicated by *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

[0417] The results of the experiment of strain MNH23031 on liver cancer tumor treatment are shown in Table 10. Figure 14 to Figure 17

[0418] Table 10 shows the statistics of the overall situation of mice at the end of the experiment.

[0419] Table 10.

[0420]

[0421] Figure 14 The schematic diagram of the change of tumor volume of mice over time is shown.

[0422] Figure 15 The schematic diagram of the control of tumor volume of mice at the end of the experiment is shown.

[0423] Figure 16 The schematic diagram of the tumor growth inhibition rate of mice is shown.

[0424] Figure 17 The schematic diagram of the response rate of the control group at the end of the experiment is shown.

[0425] Figure 18 The schematic diagram of the response rate of the experimental group at the end of the experiment is shown.

[0426] As shown in Table 10, at the end of the experiment, the surviving mice were dissected, and the tumor tissues were weighed. Figure 14 The data is shown as mean ± standard deviation (Mean ± SD), and the statistical analysis is analyzed by two-way ANOVA (2way ANOVA) with Sidak multiple comparison, and the significant difference is indicated by *, p < 0.05. During the experiment, the statistical analysis result of the tumor volume curve of group B (MNH23031) was significantly smaller than that of group A (Control).

[0427] As shown in Table 10, at the end of the experiment, the surviving mice were dissected, and the tumor tissues were weighed.

[0428] As shown in Table 10, at the end of the experiment, the surviving mice were dissected, and the tumor tissues were weighed. Figure 15 The data is shown as mean ± standard deviation (Mean ± SD), and the statistical analysis is analyzed by two-way ANOVA (2way ANOVA) with Sidak multiple comparison, and the significant difference is indicated by *, p < 0.05. During the experiment, the statistical analysis result of the tumor volume curve of group B (MNH23031) was significantly smaller than that of group A (Control). 3 3 The average tumor volume of mice in the control group at the end of the experiment was 1180.09 mm

[0429] As shown in Table 10, at the end of the experiment, the surviving mice were dissected, and the tumor tissues were weighed. Figure 16 ​​The data is shown as mean ± SD, the statistical analysis is analyzed by Student's t test, the significant difference is represented by *, *p<0.05, **p<0.01. At the end of the experiment, the tumor growth inhibition rate (TGI) of group B (MNH23031) was 33.2%, higher than that of the control group;

[0430] As shown in Figure 17 At the end of the experiment, the response rate of group A was 0%, as shown in Figure 18 The response rate of group B was 44.44%, and the response rate was significantly improved.

[0431] Example 9:

[0432] Strain MNH23031 activates immune analysis of tumor mice.

[0433] After the tissues of tumor-bearing mice in Example 8 were prepared into single cell suspension, specific molecules labeled with fluorescent molecule markers were labeled, and the proportion of cells expressing specific molecules could be detected in flow cytometry. The proportion distribution of immune cells reflects the immune status of mice to some extent. The experimental steps are as follows:

[0434] The tumor tissue was washed with PBS, cut into small pieces and crushed, and the crushed tumor tissue was placed in a 5mL collagenase IV digestion system (1mg / mL collagenase IV, 0.1mg / mL DNAase, 10% FBS) 37℃ digestion treatment for 30min; the digestion treatment of tissue liquid was filtered by 70μM filter membrane, and mouse tumor single cell suspension was prepared, and the total cell number in the tissue was detected by machine.

[0435] The mouse tumor single cell suspension was taken, and the dyes (corresponding antibody combination) Live / Fc, T-TAM-Surface were added in turn for staining, and the stained cells were resuspended with PBS; then, according to the instructions of the detection kit (Cytofix Cytoperm TM Fixation Permeabilization Kit, purchased from BD) in turn using fixing solution, membrane breaking solution treatment, intranuclear staining for staining; the stained cells were resuspended with PBS, and the machine was detected.

[0436] Take mouse tumor single cell suspension, add SP-Live, Treg Surface respectively for staining, resuspend the cells after staining with PBS, resuspend the cells after staining with PBS, then, according to the instructions of the detection kit (Foxp3 / Transcription Factor Staining Buffer Set, purchased from Invitrogen) in turn using fixing fluid, membrane breaker, add antibody Foxp3 staining for staining; resuspend the cells after staining with PBS, and detect on the machine.

[0437] Take mouse tumor single cell suspension, add dye (corresponding antibody combination) Live / Fc, T-TumorStain for tumor cell surface T cell staining, resuspend the cells after staining with PBS, and detect on the machine; take mouse tumor single cell suspension, add dye (corresponding antibody combination) Live / Fc, MDSC Stain for staining, resuspend the cells after staining with PBS, and detect on the machine.

[0438] Flow analysis data are processed by CyExpert, and statistical processing is performed by Graphpad Prism V9. Statistical analysis uses one-way ANOVA with Dunnet's Multiple Comparison, ns not significant, p<0.05, **p<0.01, ***p<0.001.

[0439] The results show that MNH23031 can activate the local immunity of mouse tumor and play an anti-tumor role.

[0440] Although a number of embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that many changes, modifications, and alterations to the embodiments described herein can be made. It should be understood that various alternatives to the embodiments of the present disclosure described herein can be employed in practicing the present disclosure. The appended claims are intended to cover all such alternatives and equivalents.

Claims

1. A bacterial strain, characterized in that, It is any one of the following A1) to A4): A1) Strains whose 16S rRNA gene sequence is 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% identical to the sequence shown in SEQ ID NO: 1; A2) The strain deposited at Guangdong Provincial Microbial Culture Collection Center, with accession number GDMCC No: 63077; A3) Strains whose genomes share at least 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% average nucleotide similarity with the genomes of any of the strains described in A1) to A2); A4) A strain whose genome is aligned with the genome of any of the strains described in A1) to A2) at a score of at least 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99%.

2. A culture, characterized in that, The culture comprises any one of the following C1) to C5): C1): Fermentation culture of the strain described in claim 1; C2): The supernatant of the fermentation culture of the strain described in claim 1; C3): A concentrated product of C1) or C2); C4): Extracts of C1 or C2); C5): The dried product of any one of C1) to C4); Preferably, the fermentation culture is obtained by the following method: the strain described in claim 1 is inoculated into a liquid culture medium and cultured for 24 to 72 hours at a pH of 6 to 10 and a culture temperature of 35 to 42°C to obtain the fermentation culture.

3. A microbial inoculant, characterized in that, The microbial agent comprises the strain described in claim 1; Or the metabolites of the strain described in claim 1; Or the culture as described in claim 2.

4. A composition, characterized in that, The composition or the raw materials of the composition include one or more of the strains of claim 1, the cultures of claim 2, and the microbial agents of claim 3; Preferably, the composition further includes at least one of an excipient, a diluent, and a carrier; Preferably, the composition further includes excipients, said excipients including at least one selected from lubricants, wetting agents, binders, emulsifiers, suspending stabilizers, solubilizers, preservatives, sweeteners, and flavorings. Preferably, the composition is at least one of a drug, nutritional supplement, food, food additive, feed, and 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 irrigants.

5. A pharmaceutical composition, characterized in that, Including the following D1) and D2): D1) One or more of the strain of claim 1, the metabolites of the strain of claim 1, the culture of claim 2, and the microbial agent of claim 3; D2) Combined medications; Preferably, the combination drugs include at least one of the following: oncology drugs, metabolic disease drugs, inflammatory disease drugs, autoimmune disease drugs, infectious disease drugs, and central nervous system disease drugs; Preferably, the pharmaceutical composition further includes a product instruction manual; Preferably, the tumor drug includes at least one of chemotherapy drugs, photosensitizers, photothermal agents, and immunotherapy drugs; 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 borodipyrrole, dihydroporphyrin, and Bengal red; 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 immune cell therapy drugs and immune checkpoint inhibitors; Preferably, the immunotherapy drug includes at least one of T-cell therapy drugs, tumor-infiltrating lymphocyte therapy drugs, and NK-cell therapy drugs; Preferably, the immune checkpoint inhibitor includes an inhibitor that targets 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 The strain described in CFU / g; 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 The strain described in CFU / g; Preferably, the bacterial strain in the pharmaceutical composition is a live bacterium, attenuated bacterium, killed bacterium, freeze-dried bacterium, or irradiated bacterium; Preferably, the pharmaceutical composition or the raw materials of the pharmaceutical composition include D1 in a mass percentage of 1-80% or 40-60%; Preferably, the pharmaceutical composition or the raw materials of the pharmaceutical composition comprise, by weight percentages of approximately 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% of D1); Preferably, 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%. Preferably, the pharmaceutical composition further includes at least one of an excipient, a diluent, and a carrier; Preferably, the pharmaceutical composition further includes pharmaceutically acceptable excipients; Preferably, the excipients include at least one of lubricant, wetting agent, emulsifier, suspending stabilizer, preservative, sweetener, and flavoring; Preferably, the pharmaceutical composition is a dosage form suitable for infants, children, or adults; Preferably, the pharmaceutical composition is a gastrointestinal dosage form or a non-gastrointestinal dosage form; Preferably, the pharmaceutical composition is an oral dosage form or an injectable dosage form; Preferably, the dosage form of the pharmaceutical composition is any one of the following: powder, granules, tablets, capsules, gel, suspension, drops, pills, injection, suppository, aerosol, oral liquid, ointment, emulsion, and irrigation solution; Preferably, the strain is at least partially capable of proliferating in the intestine of the subject.

6. The use 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, in the preparation of medicaments for the prevention and / or treatment of diseases; Preferably, the disease includes at least one of the following: tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, adenomas, or metastatic tumors; Preferably, the solid tumor includes at least one of the following: head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic 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, and endocrine system cancer. Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph, and bone marrow; Preferably, the tumors or cancers include oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity cancer, sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestinal 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 ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampulla neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoidoma, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, and rhabdomyosarcoma. Synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, endometrial cancer, uterine carcinosarcoma, endometrial sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastoma, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureteral cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of the ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid carcinoma, undifferentiated thyroid carcinoma, medullary thyroid carcinoma, parathyroid carcinoma, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia (at least one of the following). Preferably, the tumor or cancer includes tumors or cancers 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 a disease involving abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease includes at least one of the following: psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye syndrome, 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 diseases caused by infection with 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, prevention and / or treatment of tumors includes achieving this through at least one of the following (a) to (l): (a) Inhibits tumor growth; (b) Inhibits tumor weight gain; (c) Inhibits tumor cell growth; (d) Improve the response rate to tumor treatment; (e) Enhance the efficacy of immunosuppressive drugs, such as enhancing the efficacy of PD-1 drugs; (f) To prevent and / or inhibit the spread or metastasis of tumor cells; (g) Reduce resistance to antitumor drugs; (h) HDAC activity is inhibited by at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyate, or valeric acid or valerate in SCFA; (i) Inhibit HDAC activity and / or promote IFNβ transcriptional activity; (j) It exerts an immunomodulatory effect by regulating the subject's immune system; (k) Promotes tumor cell senescence or apoptosis; (l) Inhibits angiogenesis in tumor tissue.

7. The use 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, in the prevention and / or treatment of diseases; Preferably, the disease includes at least one of the following: tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, adenomas, or metastatic tumors; Preferably, the solid tumor includes at least one of the following: head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic 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, and endocrine system cancer. Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph, and bone marrow; Preferably, the tumors or cancers include oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity cancer, sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestinal 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 ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampulla neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoidoma, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, and rhabdomyosarcoma. Synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, endometrial cancer, uterine carcinosarcoma, endometrial sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastoma, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureteral cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of the ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid carcinoma, undifferentiated thyroid carcinoma, medullary thyroid carcinoma, parathyroid carcinoma, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia (at least one of the following). Preferably, the tumor or cancer includes tumors or cancers 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 a disease involving abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease includes at least one of the following: psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye syndrome, 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 diseases caused by infection with 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, prevention and / or treatment of tumors includes achieving this through at least one of the following (a) to (l): (a) Inhibits tumor growth; (b) Inhibits tumor weight gain; (c) Inhibits tumor cell growth; (d) Improve the response rate to tumor treatment; (e) Enhance the efficacy of immunosuppressive drugs, such as enhancing the efficacy of PD-1 drugs; (f) To prevent and / or inhibit the spread or metastasis of tumor cells; (g) Reduce resistance to antitumor drugs; (h) HDAC activity is inhibited by at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyate, or valeric acid or valerate in SCFA; (i) Inhibit HDAC activity and / or promote IFNβ transcriptional activity; (j) It exerts an immunomodulatory effect by regulating the subject's immune system; (k) Promotes tumor cell senescence or apoptosis; (l) Inhibits angiogenesis in tumor tissue.

8. A method of 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, or the pharmaceutical composition of claim 5; Preferably, the disease includes at least one of the following: tumor or cancer, metabolic disease, inflammatory or autoimmune disease, infectious disease, and central nervous system disease; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, adenomas, or metastatic tumors; Preferably, the solid tumor includes at least one of the following: head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic 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, and endocrine system cancer. Preferably, the hematopoietic tumor includes at least one tumor of the blood, lymph, and bone marrow; Preferably, the tumors or cancers include oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cavity cancer, sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestinal 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 ampulla cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampulla neuroendocrine tumor, jejunal-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoidoma, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, and rhabdomyosarcoma. Synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, endometrial cancer, uterine carcinosarcoma, endometrial sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastoma, penile cancer, prostate cancer, testicular cancer, renal cancer, renal pelvis cancer, ureteral cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of the ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid carcinoma, undifferentiated thyroid carcinoma, medullary thyroid carcinoma, parathyroid carcinoma, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, leukemia (at least one of the following). Preferably, the tumor or cancer includes tumors or cancers 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 a disease involving abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions; Preferably, the inflammatory or autoimmune disease includes at least one of the following: psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye syndrome, 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 diseases caused by infection with 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, prevention and / or treatment of tumors includes achieving this through at least one of the following (a) to (l): (a) Inhibits tumor growth; (b) Inhibits tumor weight gain; (c) Inhibits tumor cell growth; (d) Improve the response rate to tumor treatment; (e) Enhance the efficacy of immunosuppressive drugs, such as enhancing the efficacy of PD-1 drugs; (f) To prevent and / or inhibit the spread or metastasis of tumor cells; (g) Reduce resistance to antitumor drugs; (h) HDAC activity is inhibited by at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyate, or valeric acid or valerate in SCFA; (i) Inhibit HDAC activity and / or promote IFNβ transcriptional activity; (j) It exerts an immunomodulatory effect by regulating the subject's immune system; (k) Promotes tumor cell senescence or apoptosis; (l) Inhibits 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 daily; Preferably, when the application is performed more than twice a day, the same dose is used each time; Preferably, when the application is performed more than twice a day, a different dose is used each time; Preferably, 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.

9. A method for preparing the culture according to claim 2, characterized in that, Includes the following steps: The strain described in claim 1 was inoculated into a fermentation medium to obtain a culture. Preferably, the process further includes centrifuging the culture and collecting the supernatant from the fermentation culture; Preferably, the method further includes concentrating the culture or the supernatant of the fermentation culture by at least one of evaporation, freeze drying, dialysis, extraction, and membrane separation to obtain a concentrated product; Preferably, the method further includes treating the fermentation culture or the supernatant of the fermentation culture by at least one of extraction and solvent extraction to obtain an extract; Preferably, the method further includes drying any one of the fermentation culture medium, the fermentation culture supernatant, the concentrated product, and the extract to obtain a dried product.

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