Parabacteroides strain, culture, microbial inoculum and application of parabacteroides strain and culture

Abstract

The invention discloses a parabacteroides strain, a culture, a fungicide and application of the parabacteroides strain, the culture and the fungicide. In a first aspect of the present application, a strain of the genus Parabacteroides is provided, which can be a strain having a preservation number GDMCC No: 62664 or 62665, or a strain having at least 96.74% identity to the 16S rRNA of the strain, and / or having at least 87% average nucleotide identity to the genome of the strain. According to the embodiment of the invention, a new strain under parabacteroides is newly found, and the strain or a product based on the strain has good effects of preventing or treating tumors or cancers, metabolic diseases, inflammations or autoimmune diseases, infectious diseases and central nervous system diseases.

Description

Technical Field

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

[0002] The intestine is the largest microecological environment in the human body. It participates in many important physiological processes such as nutrient absorption, energy metabolism, tissue and organ development, immune defense, and endocrine regulation, and is closely related to human health. Intestinal microecology has made great progress in the past decade, and human understanding of it has continued to deepen. There are a large number of symbiotic microorganisms in the human intestine, and the total amount of genetic information they carry is 50 to 100 times that of the human genome, namely the "gut microbiome", also known as the "second genome" of humans. The intestinal microbiome is the largest and most direct external environment of the human body, and plays an indispensable role in maintaining human health.

[0003] Intestinal microorganisms include bacteria, chlamydia, rickettsia, mycoplasma, spirochetes, etc. Intestinal bacteria account for more than 95% of intestinal microorganisms, so intestinal microorganisms mainly refer to intestinal flora. With the continuous deepening of research, the impact and role of intestinal flora on intestinal and extra-intestinal diseases are becoming more and more clear. The intestinal flora is more like an organ with metabolic, immune and endocrine functions. It can affect the human digestive system, circulatory system and nervous system, and is closely related to the occurrence of various human chronic diseases (diabetes, hypertension, cardiovascular disease and brain disease, etc.) and tumors. Studying the relationship between intestinal flora and human health and disease is not only an important scientific research, but also has important significance and value in clinical diagnosis, treatment, and even transformation.

[0004] The two largest taxa in the intestinal flora of most adults are the Gram-positive Firmicutes and the Gram-negative Bacteroidetes, and others include a small number of Proteobacteria, Actinomycetes, Fusobacteria, and methanogens. Some bacteria in the Bacteroidetes phylum can benefit the host by preventing infection with potential pathogens, but there are also some conditional pathogens. At present, the most well-studied members of the Bacteroidetes phylum are Bacteroides, Prevotella, and Porphyromonas, but there are fewer studies on the bacteria of the genus Parabacteroides. Existing studies have shown that Parabacteroides goldsteinii and Parabacteroides distasonis have certain anti-inflammatory effects, and Parabacteroides distasonis is a potential probiotic for inhibiting colon cancer. Therefore, it is necessary to further explore intestinal Parabacteroides bacteria and discover strains with therapeutic effects. Summary of the invention

[0005] The present application aims to solve at least one of the technical problems existing in the prior art. To this end, the present application proposes a new strain of Parabacteroides, a culture, a bacterial agent and its application, which has a good therapeutic effect on tumors.

[0006] In a first aspect of the present application, a Parabacteroides strain is provided, wherein the Parabacteroides strain is any one of the following A1) to A5):

[0007] A1) strains having 16S rRNA that is at least 96.74%, 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) strains with a deposit number of GDMCC No: 62664 or GDMCC No: 62665;

[0009] A3) strains having at least 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity to the 16S rRNA of the strain described in A2);

[0010] A4) a strain having 16S rRNA with at least 97%, 98%, 98.65%, 99%, 99.5%, 99.9%, 100% identity to the sequence shown in SEQ ID NO: 2; preferably, the strain is selected from the species Parabacteroides hominis;

[0011] A5) a strain whose genome has an average nucleotide identity of at least 87%, 88%, 88.81%, 88.89%, 89%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% with the genome of any of the strains A1) to A4); and / or a strain whose genome has an alignment score of at least 70%, 73.54%, 74.55%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100 with the genome of any of the strains A1) to A4).

[0012] In some embodiments of the present application, A2) the sequence of 16S rRNA of the strain with the deposit number GDMCC No: 62664 is shown as SEQ ID NO: 1, and the sequence of 16S rRNA of the strain with the deposit number GDMCC No: 62665 is shown as SEQ ID NO: 2.

[0013] In some embodiments of the present application, the strain is derived from the human intestine.

[0014] In some embodiments of the present application, the strain is capable of high production of short-chain fatty acids.

[0015] In some embodiments of the present application, the strain can produce high amounts of acetic acid and / or propionic acid.

[0016] The strains according to the embodiments of the present application have at least the following beneficial effects:

[0017] The present application example newly discovered a new strain of the genus Parabacteroides, and the strain or a product based on the strain has a good effect in preventing or treating tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases.

[0018] The second aspect of the present application provides a culture of the aforementioned strain.

[0019] In some embodiments of the present application, the culture comprises any one of the following B1) to B5):

[0020] B1) fermentation medium of the strain; B2) supernatant of fermentation medium of the strain; B3) concentrated product of B1) or B2); B4) extract of B1) or B2); B5) dried product of any one of B1) to B4);

[0021] In some embodiments of the present application, the fermentation medium is a liquid medium.

[0022] In some embodiments of the present application, the culture is obtained using a liquid culture medium under anaerobic culture conditions.

[0023] In some embodiments of the present application, the fermentation medium is MM01 liquid medium, and / or the culture temperature is 35-42° C., and / or the culture time is 24-72 hours, and / or the pH of the culture medium is 6-10.

[0024] In a third aspect of the present application, a bacterial agent is further provided, which comprises the aforementioned Parabacteroides strain, or a metabolite thereof, or the aforementioned culture.

[0025] In a fourth aspect of the present application, a composition is further provided, wherein the composition or the raw material of the composition comprises the aforementioned Parabacteroides strain, or a metabolite thereof, or the aforementioned culture, or the aforementioned bacterial agent.

[0026] In some embodiments of the present application, the composition further comprises at least one of an excipient, a diluent, and a carrier.

[0027] In some embodiments of the present application, the composition further comprises an auxiliary material.

[0028] In some embodiments of the present application, the auxiliary material also includes at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a flavor.

[0029] In some embodiments of the present application, the composition is at least one of a medicine, a nutritional supplement, a food, a food additive, a feed, and a feed additive.

[0030] In some embodiments of the present application, the composition is a solid preparation or a liquid preparation.

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

[0032] In some embodiments of the present application, the composition further comprises a product instruction sheet.

[0033] In a fifth aspect of the present application, a pharmaceutical composition is provided, comprising the following C1) and C2):

[0034] C1) the aforementioned strain, or a metabolite of the aforementioned strain, or the aforementioned culture, or the aforementioned bacterial agent;

[0035] C2) Combination drugs.

[0036] In some embodiments of the present application, the combination drug of C2) includes at least one of tumor drugs, metabolic disease drugs, inflammatory or autoimmune disease drugs, infectious disease drugs, and central nervous system disease drugs.

[0037] In some embodiments of the present application, the tumor drug includes at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0038] In some embodiments of the present application, the chemotherapy drug includes at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin.

[0039] In some embodiments of the present application, the photosensitizer includes at least one of boron dipyrrole, dihydrochlorin, and rose bengal.

[0040] In some embodiments of the present application, the photothermal agent includes at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials.

[0041] In some embodiments of the present application, the immunotherapy drug includes at least one of an immune cell therapy drug and an immune checkpoint inhibitor.

[0042] In some embodiments of the present application, the immune cell therapy drug includes at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and a NK cell therapy drug.

[0043] In some embodiments of the present application, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, and KLRA.

[0044] In some embodiments of the present application, the pharmaceutical composition comprises 1×10 3 CFU / g~1×10 13 CFU / g or 1×10 3 CFU / mL~1×10 13 CFU / mL of Parabacteroides strains.

[0045] In some embodiments of the present application, each gram or each mL of 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 , 5×108 , 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 colony forming units (CFU) of the Parabacteroides spp.

[0046] In some embodiments of the present application, the Parabacteroides strain in the pharmaceutical composition is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium.

[0047] In some embodiments of the present application, the pharmaceutical composition or the raw material of the pharmaceutical composition includes about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80% of C1).

[0048] In some embodiments of the present application, the pharmaceutical composition or the raw materials of the pharmaceutical composition include C1) in a mass percentage of 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, 40-60%.

[0049] In some embodiments of the present application, the pharmaceutical composition further comprises at least one of an excipient, a diluent, and a carrier.

[0050] In some embodiments of the present application, the pharmaceutical composition further comprises an excipient.

[0051] In some embodiments of the present application, the auxiliary material also includes at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a flavor.

[0052] In some embodiments of the present application, the pharmaceutical composition is in a dosage form suitable for infants, children or adults.

[0053] In some embodiments of the present application, the pharmaceutical composition is in a dosage form for enteral administration or parenteral administration.

[0054] In some embodiments of the present application, the pharmaceutical composition is an oral preparation or an injectable preparation.

[0055] In some embodiments of the present application, the dosage form of the pharmaceutical composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and irrigations.

[0056] In some embodiments of the present application, the strain is capable of at least partially proliferating in the intestine of a subject.

[0057] In some embodiments of the present application, the pharmaceutical composition further comprises a product instruction sheet.

[0058] The sixth aspect of the present application provides a pharmaceutical combination product, comprising independently:

[0059] A first product, the first product comprising the aforementioned Parabacteroides strain, or a metabolite thereof, or the aforementioned culture, or the aforementioned bacterial agent;

[0060] The second product, the second product includes combination drugs.

[0061] In some embodiments of the present application, the combination drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory or autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug.

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

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

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

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

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

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

[0068] In some embodiments of the present application, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, and KLRA.

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

[0070] In some embodiments of the present application, each gram or each mL of the first product comprises at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×1010 , 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 colony forming units (CFU) of the Parabacteroides spp.

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

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

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

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

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

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

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

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

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

[0080] In some embodiments of the present application, the pharmaceutical combination product further comprises a product instruction sheet.

[0081] In a seventh aspect of the present application, there is provided use of the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product in the preparation of a medicament for preventing and / or treating a disease.

[0082] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0083] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, and a metastatic tumor.

[0084] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumors, chest cancer, bone tumors, soft tissue sarcomas, 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.

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

[0086] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma and leukemia.

[0087] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0088] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori and Fusobacterium nucleatum.

[0089] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia and metal ions.

[0090] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis and Sjögren's syndrome.

[0091] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses and fungi.

[0092] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder and depression.

[0093] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (l): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0094] In an eighth aspect of the present application, use of the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in preventing and / or treating diseases is provided.

[0095] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0096] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, and a metastatic tumor.

[0097] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumors, chest cancer, bone tumors, soft tissue sarcomas, 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.

[0098] In some embodiments of the present application, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow.

[0099] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma and leukemia.

[0100] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0101] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori and Fusobacterium nucleatum.

[0102] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia and metal ions.

[0103] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis and Sjögren's syndrome.

[0104] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses and fungi.

[0105] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder and depression.

[0106] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (k): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0107] 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 Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product;

[0108] In some embodiments of the present application, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0109] In some embodiments of the present application, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, and a metastatic tumor.

[0110] In some embodiments of the present application, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumors, chest cancer, bone tumors, soft tissue sarcomas, 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.

[0111] In some embodiments of the present application, the hematopoietic tumor includes at least one tumor of the blood, lymph and bone marrow.

[0112] In some embodiments of the present application, the tumor or cancer comprises oral cancer, salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, appendix cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampullary cancer, pancreatic cancer, gastric neuroendocrine tumor, duodenal and ampullary neuroendocrine tumor, jejunal - ileal neuroendocrine tumor, appendix neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma At least one of myoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, cutaneous fibrosarcoma protuberans, Merkel cell carcinoma, malignant melanoma of the skin, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, uterine carcinosarcoma, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of ocular adnexa, brain cancer, spinal cord tumor, differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical carcinoma, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma and leukemia.

[0113] In some embodiments of the present application, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses.

[0114] In some embodiments of the present application, the bacteria or virus includes at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori and Fusobacterium nucleatum.

[0115] In some embodiments of the present application, the metabolic disease includes a disease of abnormal metabolism of at least one of amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia and metal ions.

[0116] In some embodiments of the present application, the inflammatory or autoimmune disease comprises at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis and Sjögren's syndrome.

[0117] In some embodiments of the present application, the infectious disease includes a disease caused by at least one of bacteria, viruses and fungi.

[0118] In some embodiments of the present application, the central nervous system disease includes at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder and depression.

[0119] In some embodiments of the present application, the prevention and / or treatment of tumors includes being achieved by at least one of the following (a) to (k): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0120] In some embodiments of the present application, the Parabacteroides strain, culture, bacterial agent, composition, pharmaceutical composition or pharmaceutical combination product is administered at least once a day.

[0121] In some embodiments of the present application, when the administration frequency is twice or more per day, the same dose is administered each time.

[0122] In some embodiments of the present application, when the administration frequency is twice or more per day, each administration is at a different dose.

[0123] In some embodiments of the present application, the route of administration includes at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, intradermal injection, intramuscular injection, and intravenous injection.

[0124] In a tenth aspect of the present application, a method for preparing the aforementioned culture is provided, the method comprising the following steps:

[0125] The aforementioned Parabacteroides strain is taken and inoculated into a fermentation medium to obtain a culture.

[0126] In some embodiments of the present application, the method further comprises centrifuging the culture to collect the fermentation medium supernatant.

[0127] In some embodiments of the present application, the fermentation medium or the supernatant of the fermentation medium is concentrated by at least one of evaporation, freeze-drying, dialysis, extraction, and membrane separation to obtain a concentrated product.

[0128] In some embodiments of the present application, the method further comprises subjecting the fermentation medium or the supernatant of the fermentation medium to obtain an extract by at least one of extraction and solvent extraction.

[0129] In some embodiments of the present application, the method further comprises drying any one of the fermentation medium, the fermentation medium supernatant, the concentrated product, and the extract to obtain a dried product.

[0130] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0131] Figure 1 A and B in the figure are the colony morphologies of strains MNH 26621 and strain MNH 20645 after 48 hours of culture on anaerobic blood plates.

[0132] Figure 2 A and B in the figure are the microscopic morphologies of strain MNH 26621 and strain MNH 20645, respectively, with a scale bar of 5 μm.

[0133] Figure 3 A and B in the figure are the Gram-stained microscopic morphologies of strain MNH 26621 and strain MNH 20645, respectively.

[0134] Figure 4 A and B in the figure are the results of the tolerance of strains MNH 26621 and strain MNH 20645 to different pH values, respectively.

[0135] Figure 5 A and B in the figure are the results of the tolerance of strains MNH 26621 and strain MNH 20645 to different concentrations of NaCl, respectively.

[0136] Figure 6 A and B in the figure are the results of the tolerance of strains MNH 26621 and strain MNH 20645 to different concentrations of bile salts.

[0137] Figure 7 Phylogenetic tree based on 16S rRNA of strain MNH 26621 and other related strains.

[0138] Figure 8 Phylogenetic tree based on 16S rRNA of strain MNH 20645 and other related strains.

[0139] Fig. 9 Phylogenetic tree constructed based on 16S rRNA of strains MNH 26621 and MNH 20645 and other related strains.

[0140] Fig.10 This is the result of strain MNH 26621 regulating the immune activity of macrophages.

[0141] Fig.11 This is the result of strain MNH 20645 regulating the immune activity of macrophages.

[0142] Fig.12 This is the result of strain MNH 26621 regulating the immune activity of Primary PBMC.

[0143] Fig.13 A and B in the figure are the effects of strain MNH 26621 and strain MNH 20645 on IFNβ expression, respectively.

[0144] Fig.14 A to F in the figure are the weight change curves of the experimental group and the control group of the strain MNH 20645 in Example 5, the tumor volume change curves of the experimental group and the control group, the endpoint tumor volume of the experimental group and the control group, the endpoint tumor growth inhibition rate TGI of the experimental group and the control group, the response rate of the control group and the response rate of the experimental group. Among them, the response rate is presented as the tumor volume value, the dotted line represents the tumor volume with a 40% tumor inhibition rate relative to the control group, when the tumor volume of the mouse at the end of the test is greater than the tumor volume with a 40% tumor inhibition rate of the control group, it represents no response, and when the tumor volume of the mouse at the end of the test is less than or equal to the tumor volume with a 40% tumor inhibition rate of the control group, it represents a response.

[0145] Fig.15 A to F in the figure are respectively the tumor volume change curves of the experimental group and the control group mice of the strain MNH 26621 in Example 6, the endpoint tumor volume size of the experimental group and the control group mice, the endpoint tumor weight size of the experimental group and the control group mice, the endpoint tumor growth inhibition rate TGI of the experimental group and the control group mice, the response rate of the control group and the response rate of the experimental group. Among them, the response rate is presented as the tumor volume value, the dotted line represents the tumor volume with a 40% tumor inhibition rate relative to the control group, when the tumor volume of the mouse at the end of the test is greater than the tumor volume with a 40% tumor inhibition rate of the control group, it represents no response, and when the tumor volume of the mouse at the end of the test is less than or equal to the tumor volume with a 40% tumor inhibition rate of the control group, it represents a response.

[0146] Fig.16A to F in the figure are the weight change curves of the mice in the experimental group and the control group of the strain MNH 20645 in Example 6, the tumor volume change curves of the mice in the experimental group and the control group, the endpoint tumor volume of the mice in the experimental group and the control group, the endpoint tumor growth inhibition rate TGI of the mice in the experimental group and the control group, the response rate of the control group and the response rate of the experimental group. Among them, the response rate is presented as the tumor volume value, the dotted line represents the tumor volume with a 40% tumor inhibition rate relative to the control group, when the tumor volume of the mouse at the end of the test is greater than the tumor volume with a 40% tumor inhibition rate of the control group, it represents no response, and when the tumor volume of the mouse at the end of the test is less than or equal to the tumor volume with a 40% tumor inhibition rate of the control group, it represents a response.

[0147] Fig.17 A to H in the figure respectively represent the tumor change curve of the A / B / C group in which strain MNH20645 was combined with Gemcitabine in Example 9, the endpoint tumor volume size of the A / B / C group, the endpoint tumor weight of the A / B / C group, the endpoint tumor growth inhibition rate TGI of the A / B / C group, the mouse weight change curve of the A / B / C group, the response rate of Group A, the response rate of Group B and the response rate of Group G.

[0148] Fig.18 These are the results of immune analysis of mice with liver cancer activated by strain MNH20645 in Example 10.

[0149] Fig.19 These are the results of immune analysis of mice with lung cancer activated by strain MNH20645 in Example 11.

[0150] Fig. 20 The results of immune analysis of mice bearing lung cancer tumors activated by strain MNH 26621 in Example 12.

[0151] Fig.21 This is the result of the correlation analysis between the relevant cells in the tumor tissue and the tumor endpoint volume in Example 12.

[0152] Fig. 22 These are the results of the correlation analysis between M2 macrophages, cytotoxic CD8+IFNγ+ cells and tumor weight in tumor tissue in Example 12.

[0153] In the above result figures, if significance analysis is involved, Unpaired t test, Two-Way ANOVA, Student's t test and other methods are used for significance analysis: no significant difference (ns), p value ≥ 0.05; *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001. DETAILED DESCRIPTION

[0154] The following will clearly and completely describe the concept of the present application and the technical effects produced in combination with the embodiments, so as to fully understand the purpose, features and effects of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of them. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present application.

[0155] The embodiments of the present application are described in detail below. The described embodiments are exemplary and are only used to explain the present application, and should not be understood as limiting the present application.

[0156] In the description of this application, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, "above", "below", "within", etc. are understood to include the number itself, and "about" means within the range of ±20%, 10%, 8%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, 0.1%, etc. of the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0157] In the description of the present application, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0158] The first aspect of the embodiments of the present application relates to Parabacteroides sp., including two strains named MNH 26621 and MNH20645. Among them, strain MNH 26621 is deposited in Guangdong Microbial Culture Collection Center (GDMCC), with the deposited name Parabacteroides sp. MNH26621, the deposited number GDMCC No: 62664, the deposited time July 28, 2022, the deposited address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, and the proposed classification name is Parabacteroides sp. The strain MNH20645 is deposited in the Guangdong Provincial Microbiological Culture Collection Center, with the deposited name Parabacteroides sp. MNH 20645, the deposited number GDMCC No: 62665, the deposited date July 28, 2022, and the deposited address is the 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences. The proposed taxonomic name is Parabacteroides sp.

[0159] Among them, the term "strain" refers to a member of a bacterial species with genetic characteristics, so that it can be distinguished from closely related members of the same bacterial species. The genetic characteristic can be the total or partial absence of at least one gene, the total or partial absence of at least one regulatory region (e.g., promoter, terminator, riboswitch, ribosome binding site), the presence of at least one recombinant gene, the presence of at least one mutant gene, the presence of at least one exogenous gene (a gene from another species), the presence of at least one non-natural plasmid, the presence of at least one antibiotic resistance box, or a combination thereof. The genetic characteristics between different strains can be identified by PCR amplification, optionally followed by DNA sequencing of the genomic region of interest or the entire genome. In the case where a strain (compared to another strain of the same species) acquires or loses antibiotic resistance or acquires or loses biosynthetic ability (e.g., auxotrophic strains), strains or nutrients / metabolites can be distinguished by selection or counter-selection using antibiotics.

[0160] It is known in the art that the strains 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 experiments, etc. Molecular biological methods include ribosomal RNA sequence determination, determination methods based on whole genome sequencing, etc. The strains MNH 26621 and strains MNH 20645 involved in the embodiments of the present application are identified as a new strain under the genus Parabacteroides based on the following traditional taxonomic characteristics and molecular biological methods, wherein the taxonomic characteristics include bacterial characteristics and colony characteristics, the bacteria have no spores, no flagella, no movement, rod-shaped or short rod-shaped, with a diameter of about (3-5) μm × (8-20) μm, Gram staining negative; under anaerobic culture conditions (for example, anaerobic culture at 37°C for 48h) visible colonies are formed, the colonies are round, the edges are regular and smooth, opaque, and no secretions are formed around the colonies.

[0161] In some embodiments, the strains are derived from the intestine, for example, any of the large intestine, small intestine, rectum, etc. of mammals, such as monotremes, marsupials, insectivores, shrews, scaphoids, dermoptera, chiroptera, primates, edentata, phlobothrium, lagomorpha, rodentia, carnivora, sirenians, hyraxes, tubulodonta, odd-toed ungulates, even-toed ungulates, cetaceans, etc. Common mammals include rodents (such as mice, rats, hamsters, guinea pigs), lagomorphs (such as rabbits), odd-toed ungulates (such as horses, donkeys), even-toed ungulates (such as sheep, goats, camels, cattle, pigs), primates (such as monkeys, gorillas, chimpanzees, humans), carnivores (such as dogs, cats), etc. It is understood that strains derived from the intestine can be isolated and extracted from feces, for example.

[0162] In some embodiments, the strain or strain variant has the characteristic of high production of short-chain fatty acids. Generally, short-chain fatty acids include saturated fatty acids with a chain length of 1 to 6 carbon atoms, such as formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, pivalic acid, caproic acid, etc. High production means that the production is above 100 μg / g according to the determination of short-chain fatty acids (SCFA).

[0163] In some embodiments, the strain or strain variant has the characteristics of high production of acetic acid and / or propionic acid. High production of acetic acid refers to the production of acetic acid in the short-chain fatty acid (SCFA) assay ≥ 100 μg / g, and high production of propionic acid refers to the production of propionic acid in the short-chain fatty acid (SCFA) assay ≥ 100 μg / g. Specifically, the acetic acid production is ≥200μg / g, ≥300μg / g, ≥400μg / g, ≥500μg / g, ≥600μg / g, ≥700μg / g, ≥800μg / g, ≥900μg / g, ≥1000μg / g, ≥1100μg / g, and the propionic acid production is ≥200μg / g, ≥300μg / g, ≥400μg / g, ≥500μg / g, ≥600μg / g, ≥700μg / g, ≥800μg / g, ≥900μg / g, ≥1000μg / g, ≥1100μg / g. For example, the above strain or strain variant is inoculated in TSB liquid culture medium and anaerobically cultured at 37°C for 48 hours, and the bacteria are collected by centrifugation to detect the acetic acid production of ≥100μg / g, ≥200μg / g, ≥300μg / g, ≥400μg / g, ≥500μg / g, ≥600μg / g, ≥700μg / g, ≥800μg / g, ≥900μg / g, ≥1000μg / g, and ≥1100μg / g; the propionic acid production is detected to be ≥100μg / g, ≥200μg / g, ≥300μg / g, ≥400μg / g, ≥500μg / g, ≥600μg / g, ≥700μg / g, ≥800μg / g, ≥900μg / g, ≥1000μg / g, and ≥1100μg / g.

[0164] In addition, the sequencing results of 16S rRNA of strain MNH 26621 show that the sequence of its 16S rRNA is shown in SEQID NO: 1, and the sequencing results of 16S rRNA of strain MNH 20645 show that the sequence of its 16S rRNA is shown in SEQID NO: 2. 16S rRNA is a ribosomal RNA of prokaryotes. The 16S rRNA gene consists of a variable region and a conserved region. The conserved region is common to all bacteria, while the variable region has different degrees of difference between different bacteria. By comparing the gene sequences of bacterial 16S rRNA, according to the sequence difference base and its evolutionary distance, a biological evolutionary tree can be drawn.

[0165] When the identity between the 16S rRNA gene sequences of two strains is less than 98.65%, it can be judged that they belong to different species (see, Kim, M., Oh, H.-S., Park, S.-C., & Chun, J. (2014). Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequences similarity for species demarcation of prokaryotes. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 64 (Pt 2), 346-351, and Liu, C., Du, M.-X., Abuduaini, R., Yu, H.-Y., Li, D.-H., Wang, Y.-J., Liu, S.-J. (2021). Enlightening the taxonomy darkness of human gut microbiomes with a cultured biobank.Microbiome, 9(1), page P23; Pablo Yarza, Uniting the classification of cultured and unculturedbacteria and archaea using 16S rRNAgene sequences, Microbiology, volume 12, pages P635-P645, 2014).

[0166] 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”, RA Barco, bioRxiv (doi: https: / / doi.org / 10.1101 / 392480); Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis, Jethro S. Johnson, Nature Communications 10, Article number: 5029 (2019)).

[0167] To this end, in some embodiments of the present application, variants of strain MNH 26621 based on the above 16S rRNA sequence are provided, and these strain variants have at least 95%, 96%, 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity in 16S rRNA sequence compared with strain MNH 26621 with a deposit number of GDMCC No: 62664, and thus are strains of the same species or different species within the same genus as strain MNH 26621. It can also be reasonably inferred that these strains of the same species or different species of the same genus have similar biological activities. In other embodiments, the variant has at least 95%, 96%, 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% identity with the sequence shown in SEQ ID NO:1 in the 16S rRNA sequence.

[0168] In some embodiments of the present application, variants of strain MNH 20645 based on the above 16S rRNA sequence are also provided, and these strain variants have at least 95%, 96%, 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity in 16S rRNA sequence compared with strain MNH 20645 with a deposit number of GDMCC No: 62665, and thus are strains of the same species or different species within the same genus as strain MNH 20645. It can also be reasonably inferred that these strains of the same species or different species of the same genus have similar biological activities. In yet other embodiments, the variant has at least 95%, 96%, 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100% identity with the sequence shown in SEQ ID NO:2 in the 16S rRNA sequence.

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

[0170] In addition, with the rapid development of second-generation and third-generation sequencing technologies, bacterial species identification based on whole genome sequencing has become possible, making the results of bacterial species identification more accurate. The average nucleotide identity (ANI) of bacterial genomes refers to the similarity of homologous genes between two bacterial genomes. In the field of bacterial taxonomy, it is generally believed that the ANI value needs to reach more than 95% to be identified as belonging to the same species (Jain C, Rodriguez-R LM, Phillippy A M, et al. High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries [J]. Nature Communications, 2018, 9 (1): 5114.).

[0171] Using the above method, a person skilled in the art can determine whether an isolated strain belongs to a new strain of the genus Parabacteroides that is the same species or a different species from strain MNH 26621 or MNH 20645. For example, when the average nucleotide identity ANI value with the strain with the deposit number GDMCC No: 62664 is at least 87%, such as at least 87%, 88%, 88.81%, 88.89%, 89%, 90%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100%, it can be determined that it belongs to a different species or the same species within the genus Parabacteroides; when the average nucleotide identity ANI value with the strain with the deposit number GDMCC No: 62664 is at least 87%, such as at least 87%, 88%, 88.81%, 88.89%, 89%, 90%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100%, it can be determined that it belongs to a different species or the same species within the genus Parabacteroides; When the average nucleotide identity ANI value of the strain No. 62665 is at least 87%, for example, at least 87%, 88%, 88.81%, 88.89%, 89%, 90%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or 100%, it can be determined that they belong to different species or the same species within the genus Parabacteroides.

[0172] At present, there are various mature ANI value calculation tools, such as local calculation software Jspecies ( / jspecies) and Gegenees ( / documentation.html), online calculation tools ANI caculator (http: / enveomics.gatech.edu / ), EzGenome ( / ezgenome / ani) and ANItools. Therefore, the ANI value of the isolated strain and MNH26621 or MNH 20645 can be calculated by any of the above local calculation software or online calculation tools or other tools or methods known in the art.

[0173] To this end, in some embodiments of the present application, variants of strains MNH 26621 or MNH20645 based on the above genome are provided, and these strain variants have at least 95%, 96%, 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% average nucleotide identity in the genome compared with the strain with the deposit number of GDMCC No: 62664 or GDMCC No: 62665 or any other variant described above, and thus are strains of the same species or different species within the genus Parabacteroides as strains MNH 26621 or MNH 20645. It can also be reasonably inferred that these strains of the same genus and species or different species of the same genus have similar biological activities. In addition, it can also be judged by the alignment score of the corresponding genome. For example, compared with the strain with the deposit number of GDMCC No: 62664 or GDMCC No: 62665 or any other variant mentioned above, variants with an alignment score of at least 60%, 70%, 73.54%, 74.55%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% also have the same or similar biological activity.

[0174] The term "variant" includes any bacteria produced from strain MNH 26621 or MNH 20645, such as bacteria produced by natural mutation or recombination (such as mutation caused by cell proliferation and cell division) or resistance selection, as well as mutation or recombination caused by any other mechanism such as radiation, virus, transposon or mutagenic chemicals. In addition, variants also include other strains isolated from the natural environment that have the same or similar taxonomic characteristics and properties as strains MNH 26621 or MNH 20645, generally including other strains of the genus Parabacteroides that are different from MNH 26621 or MNH 20645, or other strains with different phenotypic variations within the same species.

[0175] The second aspect of the embodiment of the present application relates to a culture of a strain. The term "culture" refers to a product obtained by culturing an isolated strain in a fermentation medium. The fermentation medium can be selected as a natural or artificial culture medium, for example, a solid culture medium or a liquid culture medium. In some embodiments, the fermentation medium includes peptone, such as at least one of meat peptone, casein peptone, whey peptone, soy peptone, etc. In addition, the fermentation medium may also include other inorganic salts or organic substances to promote bacterial growth.

[0176] In some embodiments, the culture may be a fermentation medium after strain culture, or a fermentation medium supernatant (e.g., obtained by centrifugation or the like), or a concentrated product of a fermentation medium or its supernatant (e.g., concentrated by at least one of evaporation, freeze drying, dialysis, extraction, membrane separation, etc.), or an extract of a fermentation medium or its supernatant (e.g., an extract obtained by at least one of extraction, extraction with a solvent such as water or an organic solvent, etc.), or a dried product obtained by drying any one of the fermentation medium or its supernatant or concentrated product or extract. In addition, in some specific embodiments, the extract may be an extraction for one or more specific components, such as an extraction for components with a specific molecular weight range, specific solubility, specific isoelectric point, etc.; or it may be an extraction not directed to one or several types of components. The specific extraction method includes extracting with at least one solvent such as methanol, ethanol, chloroform, water, acid, alkali, etc. in a specific process sequence under certain temperature conditions, for example, extracting in 75v / v% ethanol at 60-80°C for 1-10 minutes, or extracting in methanol at -30--10°C for 1-10 minutes, or extracting in methanol / chloroform / water solution at -40--20°C for 30-60 minutes, or extracting in deionized water at 95°C for 1-10 minutes, or extracting in perchloric acid, trichloroacetic acid, hydrochloric acid, sodium hydroxide solution at 0-4°C, etc., so as to extract the corresponding metabolites, such as short-chain fatty acids or short-chain fatty acid salts, such as at least one of acetic acid, butyric acid, valeric acid, acetate, propionate, and valerate.

[0177] In some embodiments, the culture is a culture obtained under anaerobic culture conditions using a liquid culture medium. For example, the liquid culture medium is an MM01 liquid culture medium (comprising about 5 g / L of peptone, about 5 g / L of trypticase, about 10 g / L of yeast powder, about 5 g / L of beef extract, about 5 g / L of glucose, and about 5 g / L of K 2 HPO 4 About 2g / L, sodium acetate about 2g / L, Tween 80 about 1mL / L, heme about 5mg / L, L-cysteine ​​hydrochloride about 0.5g / L, vitamin K1 about 1μL / L, inorganic salt solution about 8mL / L (calcium chloride about 0.25g / L, dipotassium hydrogen phosphate about 1g / L, potassium dihydrogen phosphate about 1g / L, magnesium sulfate about 0.5g / L, sodium bicarbonate about 10g / L, sodium chloride about 2g / L)). In some embodiments, during anaerobic culture, the culture temperature is 35-42°C, for example, 37°C. In some embodiments, during anaerobic culture, the culture time is 24-72 hours, for example, 36-60 hours, and can be optionally 48 hours. In some embodiments, during anaerobic culture, the pH of the culture medium is 6-10, and the pH can be 7.

[0178] It is understandable that, in the above-mentioned culture, the compound and / or cell debris of the bacterial strain are contained, and / or the metabolites secreted by the bacterial strain. Wherein, metabolites include small molecules (such as amino acids, nucleosides, nucleotides) and macromolecules (such as polypeptides, carbohydrates, nucleic acids, proteoglycans, lipids). These metabolites can be primary metabolites directly involved in normal cell functions, or secondary metabolites that are usually not needed by basic cell functions, and metabolic intermediates produced in primary or secondary metabolite building-up processes. Thus, biological functions similar to bacterial strains can be realized by different types of cultures containing compounds and / or metabolites of bacterial strains.

[0179] A third aspect of an embodiment of the present application relates to a bacterial agent, which comprises one or more of the aforementioned strains or metabolites of the strains or the aforementioned cultures, for example, it may be a strain with a deposit number of GDMCC No: 62664 or its metabolites or the culture of the strain, and / or a strain with a deposit number of GDMCC No: 62665 or its metabolites or the culture of the strain.

[0180] The fourth aspect of the embodiment of the present application relates to a composition, the composition includes the aforementioned strain or its metabolite or the aforementioned culture or the aforementioned microbial agent, or the raw material for preparing the composition includes the aforementioned strain or its metabolite or the aforementioned culture or the aforementioned microbial agent. In some embodiments, the composition is used as at least one of a medicine, a nutritional supplement, a food, a food additive, a feed, and a feed additive. For this reason, considering the specific dosage form used, it can be any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop, a pill, an injection, a suppository, an aerosol, an oral liquid, an ointment, an emulsion, and an irrigant. In some embodiments, in order to form the above-mentioned composition, the composition also includes at least one component in an excipient, a diluent, and a carrier. Generally, an acceptable carrier or diluent is well known to those skilled in the art. For example, at least one of lactose, starch, glucose, sucrose, methylcellulose, magnesium stearate, mannitol, sorbitol, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, syrup and methylcellulose can be used as a carrier or excipient. At least one of ethanol, glycerol and water can also be used as a diluent. In addition, it is understood that the carrier, excipient or diluent can be specifically selected according to the expected application scenario and mode of administration of the composition, for example, for different uses such as bacterial agents, drugs, nutritional supplements, foods, food additives, feeds, feed additives, and powders, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, lavage agents, etc., and different dosage forms can be targeted to select different carriers, excipients or diluents. In addition, the composition may also include excipients other than carriers, excipients or diluents, and the excipients may specifically be at least one of any suitable lubricants, wetting agents, binders, emulsifiers, suspension stabilizers, solubilizers, preservatives, sweeteners, flavors, etc. The specific selection of these excipients is conventional, so only some of the ingredients are described by way of example: 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, wherein natural sugars such as glucose, anhydrous lactose, free-flowing lactose, β-lactose or corn sweeteners, natural or synthetic gums such as gum arabic, tragacanth, sodium alginate, carboxymethyl cellulose or polyethylene glycol; preservatives include sodium benzoate, sorbic acid or parabens. By making the strain or strain culture into any of the above-mentioned compositions, the release of different requirements of the effective active ingredients therein is achieved, such as rapid release, sustained release, etc. In some embodiments, the composition also includes a product instruction sheet.

[0181] The fifth aspect of the embodiments of the present application relates to a pharmaceutical composition, which includes C1) the aforementioned Parabacteroides strain or its metabolite, or the aforementioned culture, or the aforementioned bacterial agent; and C2) a combined drug. The improvement of drug efficacy is achieved by combining drugs. In some embodiments, the combined drug includes at least one of a tumor drug, a metabolic disease drug, an inflammatory or autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug. In some embodiments, the tumor drug may include at least one of a chemotherapeutic drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0182] Among them, chemotherapy drugs include alkylating agents (such as cyclophosphamide, ifosfamide, thiotepa, melphalan, carmustine, lomustine, semustine, nimustine, fotemustine, estramustine, nitrogen methyl, nitrogen mustard, nitrosamic acid mustard, chlorambucil, hexamethylmelamine, busulfan, temozolomide), platinums (such as cisplatin, carboplatin, oxaliplatin, suplatin, lobaplatin, nedaplatin, dicycloplatin), podophyllotoxins (such as podophyllotoxin, etoposide, teniposide, etoposide phosphate), camptothecin At least one of alkaloids (such as camptothecin, hydroxycamptothecin, irinotecan, topotecan), taxanes (such as paclitaxel, docetaxel), anthracyclines (such as daunorubicin, doxorubicin, epirubicin, pirarubicin, idarubicin, mitoxantrone, aclarubicin), fluoropyrimidines (such as gemcitabine, capecitabine, 5-fluorouracil, bifuroxidine, deoxyfluridine, tegafur, carmofur, trifluridine), antibiotics (such as mitomycin, bleomycin, bleomycin), etc.

[0183] The photosensitizer includes at least one of porphyrins (such as dihydrochlorins, phthalocyanines, bacteriochlorins), boron dipyrroles (such as fluoroboron dipyrroles, azafluoroboron dipyrroles), and rose bengal.

[0184] Photothermal agents include precious metal nanoparticles (such as nanodots, nanorods, nanowires, nanosheets, etc. of precious metals such as gold, silver, platinum, and palladium), organic polymers (such as polypyrrole, polyaniline, polyethylene dioxythiophene, polystyrene sulfonate, indocyanine green, porphyrin), carbon-based nanomaterials (such as graphene, carbon nanotubes, cellulose carbonization products), magnetic nanomaterials (such as Fe 3 O 4 ), at least one of semiconductor nanomaterials (such as copper sulfide, molybdenum sulfide, bismuth sulfide, antimony sulfide, gold sulfide, copper selenide, molybdenum selenide, bismuth selenide, antimony selenide, and gold selenide).

[0185] Immunotherapy drugs include at least one of immune cell therapy drugs and immune checkpoint inhibitors. Among them, immune cell therapy drugs include at least one of T cell therapy drugs (such as CAR-T, TCR-T), tumor infiltrating lymphocyte therapy drugs, and NK cell therapy drugs. Immune checkpoint inhibitors include inhibitors targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, and KLRA.

[0186] The term "immune checkpoint inhibitor" is an antagonist that targets the immune checkpoint protein, which can enhance proteins that stimulate immune responses or block proteins that inhibit immune responses, thereby showing anti-cancer effects through immune responses. In some embodiments, the immune checkpoint inhibitor can be a protein or peptide, such as a soluble fusion protein; it can be an antibody or its antigen-binding fragment that binds to the immune checkpoint protein to be inhibited; it can be an inhibitory nucleic acid (such as a siRNA molecule, a shRNA molecule, an antisense RNA) that specifically binds to the mRNA encoding the immune checkpoint protein.

[0187] In some specific embodiments, the inhibitor is an antibody, such as anti-PD-1 antibodies (such as Camrelizumab, Cemiplimab (cemiplimab-rwlc), Dostarlimab, Enlonstobart, Nivolumab, Pembrolizumab, Penpulimab, Prolgolimab, Pucotenlimab, Retifanlimab, Serplulimab, Sintilimab, Tislelizumab, Toripalimab, Zimberelimab), anti-PD-L1 antibodies (such as Adebrelimab, Atezolizumab, Avelumab, Cosibelimab, Durvalumab, Envafolimab, Socazolimab, Sugemalimab, Tagitanlimab), anti-CTLA-4 antibodies (such as Ipilimumab, Tremelimumab), etc.

[0188] In some embodiments, the metabolic disease drugs include drugs used to treat abnormal amino acid metabolism (such as phenylketonuria, glycine metabolism disease, tyrosine metabolism disease), abnormal organic acid metabolism (methylmalonic acidemia, propionic acidemia), abnormal glucose metabolism (such as glycogen storage disease, galactosemia, diabetes), abnormal fat metabolism (such as hypercholesterolemia, hyperlipidemia), abnormal purine metabolism (such as gout, self-destructive facies), abnormal pigment metabolism (such as porphyria), abnormal blood ammonia metabolism (such as hyperammonemia, citrin deficiency), and abnormal metal ion metabolism (such as hepatolenticular degeneration).

[0189] Taking diabetes as an example, diabetes drugs include at least one of insulin, metformin, pramide, sulfonylureas (such as glibenclamide, glipizide, glimepiride), methylcobalamin (such as repaglinide, nateglinide), thiazolidinediones (such as rosiglitazone, pioglitazone), DPP-4 inhibitors (such as sitagliptin, saxagliptin, linagliptin), GLP-1 receptor agonists (such as exenatide, liraglutide, semaglutide), SGLT2 inhibitors (such as canagliflozin, dapagliflozin, empagliflozin), entecavir, tenofovir, lamivudine, adefovir, telbivudine, simeprek, sofobuvir, interferon, ribavirin, etc.

[0190] In some embodiments, the pharmaceutical composition further comprises a product instruction sheet.

[0191] In some embodiments, the pharmaceutical composition is a dosage form suitable for infants, children or adults, a dosage form for gastrointestinal administration or parenteral administration, an oral preparation or an injection, the strain in the pharmaceutical composition is at least partially capable of proliferating in the intestine of the subject, and the Parabacteroides strain can be a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium or an irradiated bacterium.

[0192] In some embodiments, the pharmaceutical composition comprises 1×10 3 CFU / g~1×10 13 CFU / g of Parabacteroides strain, that is, each gram of the pharmaceutical composition contains 1×10 3 Up to 1×10 13 Colony forming units (CFU) of the Parabacteroides strain, for example, each gram of the pharmaceutical composition includes at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5, 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×10 10 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 In some other embodiments, the pharmaceutical composition comprises 1×10 3 CFU / mL~1×10 13 CFU / mL of the Parabacteroides strain, i.e., each mL of the pharmaceutical composition contains 1×10 3 Up to 1×10 13 Colony forming units (CFU) of the Parabacteroides strain, for example, each mL of the pharmaceutical composition includes at least 1×10 3 , 2×10 3 , 5×10 3 , 1×10 4 , 2×10 4 , 5×10 4 , 1×10 5 , 2×10 5 , 5×10 5 , 1×10 6 , 2×10 6 , 5×10 6 , 1×10 7 , 2×10 7 , 5×10 7 , 1×10 8 , 2×10 8 , 5×10 8 , 1×10 9 , 2×10 9 , 5×10 9 , 1×10 10 , 2×10 10 , 5×1010 , 1×10 11 , 2×10 11 , 5×10 11 , 1×10 12 , 2×10 12 , 5×10 12 , 1×10 13 The colony forming units (CFU) of the Parabacteroides spp.

[0193] In some embodiments, the pharmaceutical composition or the raw material of the pharmaceutical composition includes C1) at a mass percentage of more than 1%, for example, the pharmaceutical composition or the raw material of the pharmaceutical composition includes about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 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 C1). In some embodiments, the pharmaceutical composition or the raw material of the pharmaceutical composition includes C1) in an amount of 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, or 40-60% by mass.

[0194] In some embodiments, the Parabacteroides strain contained in the above-mentioned pharmaceutical composition is capable of at least partially proliferating in the intestine of the subject, for example, 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%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 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 Parabacteroides strains were able to proliferate in the intestines of the subjects.

[0195] In some embodiments, the bacterial strain is freeze-dried in the pharmaceutical composition. In some embodiments, the bacterial strain is spray-dried in the pharmaceutical composition. In some embodiments, the bacterial strain is freeze-dried or spray-dried in the pharmaceutical composition and wherein it is alive. In some embodiments, the bacterial strain is freeze-dried or spray-dried in the pharmaceutical composition and wherein it is able to partially or completely colonize the intestine. In some embodiments, the freeze-dried bacterial strain is reconstituted prior to administration. In some cases, reconstitution is performed by using a diluent.

[0196] In some embodiments, the pharmaceutical composition is administered orally. Oral administration may involve swallowing, whereby the compound enters the gastrointestinal tract, and / or oral, lingual, or sublingual administration, whereby the compound enters the bloodstream directly from the oral cavity. Pharmaceutical dosage forms suitable for oral administration include solid suppositories, solid particulates, semisolids, and liquids (including multiphase or dispersed systems) such as tablets; soft or hard capsules containing multi-particles or nanoparticles, liquids (e.g., aqueous solutions), emulsions, or powders; lozenges (including liquid fillers); chewable gels; rapidly dispersible dosage forms; ovules; sprays; and buccal / mucosal adhesive patches.

[0197] In some embodiments, the pharmaceutical formulation is an enteric formulation, i.e., a gastro-resistant formulation (e.g., resistant to gastric pH) suitable for delivering the pharmaceutical composition to the intestine by oral administration. Enteric formulations may be particularly useful when the Parabacteroides strain or another component of the pharmaceutical composition is acid-sensitive, e.g., susceptible to degradation under gastric conditions.

[0198] In some embodiments, the enteric preparation comprises an enteric coating. In some embodiments, the preparation is an enteric-coated dosage form. For example, the preparation can be an enteric-coated tablet or an enteric-coated capsule, etc. The enteric coating can be a conventional enteric coating, such as a conventional coating for tablets, capsules, etc. for oral delivery. The preparation can include a film coating, such as a film layer of an enteric polymer, such as an acid-insoluble polymer. In some embodiments, the enteric preparation is enteric in nature, such as gastrointestinal and does not require an enteric coating. In some embodiments, the preparation is an inherent enteric capsule (e.g., from Capsugel).

[0199] In some embodiments, the preparation is a soft capsule. A soft capsule is a capsule with certain elasticity and softness due to the addition of a softener such as glycerol, sorbitol, maltitol and polyethylene glycol present in the capsule shell. Soft capsules can be produced, for example, on the basis of gelatin or starch. Soft capsules based on gelatin are commercially available from various suppliers. According to the method of administration, for example, oral or rectal administration, soft capsules can have various shapes, and they can be, for example, circular, oval, elliptical or torpedo-shaped. Soft capsules can be produced by conventional processes, for example, by Scherer process, Accogel process or droplet or blow molding process.

[0200] In some embodiments, the formulation is a microencapsulated capsule that protects the composition from degradation by encapsulation until it is delivered to the target location, for example, by chemical or physical stimulation such as pressure, enzymatic activity or physical decomposition and rupture, which may 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 by flocculation or the use of a cross-linking agent, and mechanical containment behind a microporous membrane or microencapsulated capsule.

[0201] In some embodiments of the present application, the dosage form of the pharmaceutical composition is any one of powder, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and lavage agents. In some embodiments, the pharmaceutical composition also includes at least one of an excipient, a diluent, and 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, methylcellulose, magnesium stearate, mannitol, sorbitol, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, syrup, and methylcellulose can be used as a carrier or excipient. And at least one of ethanol, glycerol, and water can also be used as a diluent. In some embodiments, the pharmaceutical composition also includes excipients, such as at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a flavor. For example, 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, β-lactose or corn sweeteners, natural or synthetic gums such as gum arabic, tragacanth, sodium alginate, carboxymethyl cellulose or polyethylene glycol; preservatives include sodium benzoate, sorbic acid or parabens. By making the strain or strain culture into any of the above-mentioned pharmaceutical compositions, the release of the effective active ingredients therein according to different requirements, such as rapid release, sustained release, etc., can be achieved.

[0202] The sixth aspect of the embodiments of the present application relates to a pharmaceutical combination product, which comprises independently:

[0203] A first product, comprising the aforementioned Parabacteroides strain, or a metabolite of the aforementioned Parabacteroides strain, or the aforementioned culture, or the aforementioned bacterial agent;

[0204] Second products, including combination drugs.

[0205] It is understood that the first product contains an effective amount of the aforementioned strain or the aforementioned culture. The first product and the second product can be administered at different time points, and / or in different administration modes, and / or at different administration intervals, in respective doses.

[0206] In some embodiments, the combination drug in the second product includes at least one of a tumor drug, a metabolic disease drug, an inflammatory or autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug. In some embodiments, the tumor drug may include at least one of a chemotherapeutic drug, a photosensitizer, a photothermal agent, and an immunotherapy drug.

[0207] Among them, chemotherapy drugs include alkylating agents (such as cyclophosphamide, ifosfamide, thiotepa, melphalan, carmustine, lomustine, semustine, nimustine, fotemustine, estramustine, nitrogen methyl, nitrogen mustard, nitrosamic acid mustard, chlorambucil, hexamethylmelamine, busulfan, temozolomide), platinums (such as cisplatin, carboplatin, oxaliplatin, suplatin, lobaplatin, nedaplatin, dicycloplatin), podophyllotoxins (such as podophyllotoxin, etoposide, teniposide, etoposide phosphate), camptothecin At least one of alkaloids (such as camptothecin, hydroxycamptothecin, irinotecan, topotecan), taxanes (such as paclitaxel, docetaxel), anthracyclines (such as daunorubicin, doxorubicin, epirubicin, pirarubicin, idarubicin, mitoxantrone, aclarubicin), fluoropyrimidines (such as gemcitabine, capecitabine, 5-fluorouracil, bifuroxidine, deoxyfluridine, tegafur, carmofur, trifluridine), antibiotics (such as mitomycin, bleomycin, bleomycin), etc.

[0208] The photosensitizer includes at least one of porphyrins (such as dihydrochlorins, phthalocyanines, bacteriochlorins), boron dipyrroles (such as fluoroboron dipyrroles, azafluoroboron dipyrroles), and rose bengal.

[0209] Photothermal agents include precious metal nanoparticles (such as nanodots, nanorods, nanowires, nanosheets, etc. of precious metals such as gold, silver, platinum, and palladium), organic polymers (such as polypyrrole, polyaniline, polyethylene dioxythiophene, polystyrene sulfonate, indocyanine green, porphyrin), carbon-based nanomaterials (such as graphene, carbon nanotubes, cellulose carbonization products), magnetic nanomaterials (such as Fe 3 O 4 ), at least one of semiconductor nanomaterials (such as copper sulfide, molybdenum sulfide, bismuth sulfide, antimony sulfide, gold sulfide, copper selenide, molybdenum selenide, bismuth selenide, antimony selenide, and gold selenide).

[0210] Immunotherapy drugs include at least one of immune cell therapy drugs and immune checkpoint inhibitors. Among them, immune cell therapy drugs include at least one of T cell therapy drugs (such as CAR-T, TCR-T), tumor infiltrating lymphocyte therapy drugs, and NK cell therapy drugs. Immune checkpoint inhibitors include inhibitors targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, and KLRA.

[0211] In some specific embodiments, the inhibitor is an antibody, such as anti-PD-1 antibodies (such as Camrelizumab, Cemiplimab (cemiplimab-rwlc), Dostarlimab, Enlonstobart, Nivolumab, Pembrolizumab, Penpulimab, Prolgolimab, Pucotenlimab, Retifanlimab, Serplulimab, Sintilimab, Tislelizumab, Toripalimab, Zimberelimab), anti-PD-L1 antibodies (such as Adebrelimab, Atezolizumab, Avelumab, Cosibelimab, Durvalumab, Envafolimab, Socazolimab, Sugemalimab, Tagitanlimab), anti-CTLA-4 antibodies (such as Ipilimumab, Tremelimumab), etc.

[0212] In some embodiments, the metabolic disease drugs include drugs used to treat abnormal amino acid metabolism (such as phenylketonuria, glycine metabolism disease, tyrosine metabolism disease), abnormal organic acid metabolism (methylmalonic acidemia, propionic acidemia), abnormal glucose metabolism (such as glycogen storage disease, galactosemia, diabetes), abnormal fat metabolism (such as hypercholesterolemia, hyperlipidemia), abnormal purine metabolism (such as gout, self-destructive facies), abnormal pigment metabolism (such as porphyria), abnormal blood ammonia metabolism (such as hyperammonemia, citrin deficiency), and abnormal metal ion metabolism (such as hepatolenticular degeneration).

[0213] Taking diabetes as an example, diabetes drugs include at least one of insulin, metformin, pramide, sulfonylureas (such as glibenclamide, glipizide, glimepiride), methylcobalamin (such as repaglinide, nateglinide), thiazolidinediones (such as rosiglitazone, pioglitazone), DPP-4 inhibitors (such as sitagliptin, saxagliptin, linagliptin), GLP-1 receptor agonists (such as exenatide, liraglutide, semaglutide), SGLT2 inhibitors (such as canagliflozin, dapagliflozin, empagliflozin), entecavir, tenofovir, lamivudine, adefovir, telbivudine, simeprek, sofobuvir, interferon, ribavirin, etc.

[0214] In some embodiments, the first product of the pharmaceutical combination product comprises 1×10 3 CFU / g~1×10 13 CFU / g of the Parabacteroides strain, i.e., each gram of the first product contains 1×10 3 Up to 1×1013 Colony forming units (CFU) of the Parabacteroides strain, for example, 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 In some other embodiments, the first product includes 1×10 3 CFU / mL~1×10 13 CFU / mL of the Parabacteroides strain, i.e., each mL of the first product contains 1×10 3 Up to 1×10 13 colony forming units (CFU) of the Parabacteroides strain, for example, 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 colony forming units (CFU) of the Parabacteroides spp.

[0215] In some embodiments, the pharmaceutical combination product or the raw material of the pharmaceutical combination product includes 1% or more of the first product by weight, for example, the pharmaceutical combination product or the raw material of the pharmaceutical combination product includes about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, %, 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 first product. In some embodiments, the pharmaceutical combination product or the raw material of the pharmaceutical combination product includes 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, 40-60% by mass of the first product.

[0216] In some embodiments, the Parabacteroides strain in the pharmaceutical combination product is at least partially capable of proliferating in the intestinal tract of the subject, and the strain can be any of living bacteria (such as bacteria, attenuated bacteria, freeze-dried bacteria, irradiated bacteria) and dead bacteria.

[0217] In some embodiments, the first product and the second product in the drug combination product are infant-suitable dosage forms, children-suitable dosage forms or adult-suitable dosage forms. In addition, the first product and the second product can be independently a dosage form for administration through the gastrointestinal tract or a dosage form for administration beyond the gastrointestinal tract, and can be an oral agent or an injection. In some embodiments of the present application, the dosage forms of the first product and the second product in the drug combination product are independently any one of powders, granules, tablets, capsules, gels, suspensions, drops, pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, and lavage agents. In some embodiments, the first product and the second product in the drug combination product also independently include at least one of an excipient, a diluent, and a carrier. By making a parabacteroides strain or a metabolite of a parabacteroides strain, or a culture, or a bacterial agent into any of the above-mentioned drug combination products, the release of different requirements of the effective active ingredients therein, such as rapid release, sustained release, etc., is achieved.

[0218] The seventh aspect of the embodiments of the present application relates to the use of the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product in the preparation of a medicament for preventing and / or treating a disease. In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0219] Wherein, the term "tumor" or "tumor" refers to a kind of abnormal mass or vegetation that cells in the body divide and proliferate beyond normal levels, and "cancer" generally refers to malignant tumors, which can be used in some cases in this application. Tumor or cancer generally includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, glandular tumors and metastatic tumors. Solid tumors can be divided into epithelial cell-derived tumors, fibroblast-derived tumors and the like according to cell-derived characteristics. In addition, according to the difference in the location of the primary tumor lesion, the type of solid tumor includes head and neck cancer, upper digestive tract cancer, lower digestive tract cancer, hepatobiliary system cancer, neuroendocrine tumor, chest cancer, osteoma, soft tissue sarcoma, 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, and hematological tumors include at least one tumor in blood, lymph and bone marrow. Among them, head and neck cancer includes oral cancer (such as lip cancer, tongue cancer, mouth floor cancer, tonsil cancer, gum cancer, etc.), salivary gland cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, nasal cancer, paranasal sinus cancer, laryngeal cancer, etc. Upper digestive tract cancer includes esophageal cancer, gastroesophageal junction cancer, gastric cancer, small intestine cancer, etc. Lower digestive tract cancer includes appendix cancer, colon cancer, rectal cancer, anal canal cancer, etc. Hepatobiliary system cancer includes liver cancer, intrahepatic bile duct cancer, gallbladder cancer, hilar bile duct cancer, distal bile duct cancer, hepatopancreatic ampulla cancer, pancreatic cancer, etc. Neuroendocrine tumors include gastric neuroendocrine tumors, Duodenal and ampullary neuroendocrine tumors, jejunal-ileal neuroendocrine tumors, appendix neuroendocrine tumors, colorectal neuroendocrine tumors, pancreatic neuroendocrine tumors, etc. Chest cancer includes thymic carcinoma, lung cancer (such as small cell lung cancer, non-small cell lung cancer, etc.), malignant pleural mesothelioma, etc. Soft tissue sarcomas include angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma (formed by fibroblasts and collagen fibers), gastrointestinal stromal tumors, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral blood cancer Schwannoma, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, skin fibrosarcoma protuberans, etc. Skin cancer includes Merkel cell carcinoma, cutaneous malignant melanoma, etc. Female reproductive system cancer includes vulvar cancer, vaginal cancer, cervical cancer, uterine body cancer, uterine carcinoma sarcoma, uterine body sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, etc. Male reproductive system cancer includes penile cancer, prostate cancer, testicular cancer, etc. Urinary system cancer includes kidney cancer, renal pelvis cancer, ureter cancer, bladder cancer, urethral cancer etc. Eye cancers include eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, ocular accessory organ lymphoma, etc. Central nervous system cancers include brain cancer (such as glioma, meningioma, medulloblastoma, medullary pituitary adenoma, primary CNS lymphoma and central nervous system germ cell tumor, etc.), spinal cord tumors, etc. Endocrine system cancers include differentiated thyroid cancer, anaplastic thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenal cortical carcinoma, etc.Hematopoietic tumors include Hodgkin's lymphoma, non-Hodgkin's lymphoma (such as diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, Burkitt's lymphoma, lymph node marginal zone lymphoma, mucosa-associated lymphoid tissue lymphoma, etc.), cutaneous lymphoma, plasma cell myeloma and leukemia.

[0220] In addition, tumors or cancers also include tumors caused by bacteria or viruses, for example, tumors or cancers caused by at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori and Fusobacterium nucleatum.

[0221] The term "metabolic disease" refers to the accumulation or lack of substances such as sugar, fat, and amino acids in the human body, which causes abnormal metabolic substance content in the body, and some diseases caused by it can be roughly divided into two categories: metabolic dysfunction and hypermetabolism. In some specific embodiments of the present application, metabolic diseases include abnormal amino acid metabolism (such as phenylketonuria, glycine metabolism, tyrosine metabolism), abnormal organic acid metabolism (methylmalonic acidemia, propionic acidemia), abnormal sugar metabolism (such as glycogen storage disease, galactosemia, diabetes), abnormal fat metabolism (such as hypercholesterolemia, hyperlipidemia), abnormal purine metabolism (such as gout, self-destructive face disease), abnormal pigment metabolism (such as porphyria), abnormal blood ammonia metabolism (such as hyperammonemia, citrin deficiency), and abnormal metal ion metabolism (such as hepatolenticular degeneration).

[0222] The term "inflammation" refers to a pathological process dominated by defense response caused by damage to inflammatory factors in living tissues with a vascular system, which is manifested as degeneration, exudation and proliferation of local tissues, wherein inflammatory factors include physical factors, chemical factors, biological factors and immune responses. The term "autoimmune disease" refers to a disease in which the body's immune system function abnormally, causing the body to attack its own tissues. 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 and Sjögren's syndrome.

[0223] In some embodiments, infectious diseases include diseases caused by at least one of bacteria, viruses and fungi. Among them, diseases caused by bacterial infection can be divided into Gram-negative infection and Gram-positive infection according to the type of bacterial Gram staining, Gram-negative infection includes: brucellosis, Campylobacter infection, cat scratch disease, cholera, Escherichia coli infection, gonorrhea, Klebsiella infection, Enterobacter infection, Serratia infection, Legionella infection, meningococcal infection, whooping cough, plague, Pseudomonas infection, Salmonella infection, Shigellosis, typhoid fever, tularemia, etc., Gram-positive infection includes: anthrax, Clostridium difficile-induced colitis, diphtheria, enterococcal infection, erysipelothrixosis, listeriosis, nocardiosis, pneumococcal infection, staphylococcal infection, streptococcal infection, etc. Other classification methods also include spirochete infections classified by bacterial morphology: syphilis, yaws, pinta, leptospirosis, Lyme disease, rat-bite fever, relapsing fever, etc., and anaerobic infections that determine whether the pathogens need oxygen include: actinomycosis, bacteroid infection, botulism, clostridium infection, tetanus, etc. Diseases caused by fungal infections include: aspergillosis, dermatitis blastomycosis, candidiasis, coccidioidomycosis, cryptococcosis, histoplasmosis, mucormycosis, paracoccidioidomycosis, sporotrichosis, etc. Infectious diseases caused by viruses include dengue fever, Ebola virus 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, poliomyelitis, cytomegalovirus infection, herpes simplex virus infection, HIV infection, avian influenza, influenza, Middle East respiratory syndrome, SARS, SARS-CoV-2, etc.

[0224] In some embodiments, the central nervous system disease comprises at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety disorder and depression.

[0225] In some embodiments, the aforementioned strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition, and the aforementioned pharmaceutical combination product include preventing and / or treating tumors by at least one of the following (a) to (l): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcription activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell aging or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0226] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0227] Existing research results show that HDAC inhibitors have good anti-tumor effects on a variety of tumor cells, including tumor cells in the bladder, bone, breast, uterus, central nervous system, esophagus, lung, ovary, pancreas, bile duct cancer, prostate and other parts, and can cause these tumor cells to show obvious apoptosis, proliferation inhibition and cell cycle arrest, etc.; as an oncogene that regulates the onset of cancer, the activity reduction or inhibition of HDAC expression has been shown to have a variety of anti-cancer effects. In addition, TSA can also inhibit tumor angiogenesis by downregulating the expression of tumor angiogenesis-related genes caused by hypoxia and directly inhibiting the migration and proliferation of endothelial cells. Moreover, HDAC inhibitors can treat diabetes through a variety of mechanisms, including the inhibition of Pdxl (Park et al., 2008, J Clin Invest, 118, 2316-24), and enhancing the expression of transcription factor Ngn3 to increase the endocrine reservoir. Progenitor cells (Haumaitre et al., 2008, Mol Cell Biol, 28, 6373-83) and enhanced insulin expression (Molsey et al., 2003, J Biol Chem, 278, 19660-6), etc. HDAC inhibition is also a promising treatment method for late diabetic complications such as diabetic nephropathy and retinal ischemia (Christensen et al., 2011, Mol Med, 17 (5-6), 370-390). Therefore, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves 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.

[0228] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by upregulating proinflammatory factors and chemokines to stimulate immunity and / or by downregulating anti-inflammatory factors for immunomodulation. In some embodiments, the proinflammatory factor is selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0229] The eighth aspect of the embodiments of the present application relates to the use of the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product in preventing and / or treating a disease. In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease and a central nervous system disease.

[0230] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product comprises preventing and / or treating tumors by at least one of the following (a) to (l): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, valeric acid or valerate in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0231] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0232] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by upregulating proinflammatory factors and chemokines to stimulate immunity and / or by downregulating anti-inflammatory factors for immunomodulation. In some embodiments, the proinflammatory factor is selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0233] A ninth aspect of the embodiments of the present application relates to a method for treating a disease, comprising administering to a subject an effective dose of the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical combination product.

[0234] In some embodiments, the disease includes at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease. Among them, the tumor or cancer includes at least one of a solid tumor, a soft tissue tumor, a hematopoietic tumor, a glandular tumor, and a metastatic tumor. Solid tumors include at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary cancer, neuroendocrine tumors, chest cancer, osteoma, soft tissue sarcoma, skin cancer, breast cancer, female reproductive system cancer, male reproductive system cancer, urinary system cancer, eye cancer, central nervous system cancer, and endocrine system cancer; hematopoietic tumors include at least one tumor in blood, lymph, and bone marrow. Tumors or cancers include tumors or cancers caused by bacteria or viruses (such as at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori, and Fusobacterium nucleatum). Metabolic diseases include at least one metabolic abnormality in amino acids, organic acids, sugars, fats, purines, pigments, blood ammonia, and metal ions. Inflammatory or autoimmune diseases include at least one of psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjögren's syndrome. Infectious diseases include diseases caused by at least one of bacteria, viruses, and fungi. Central nervous system diseases include at least one of stroke, schizophrenia, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, and depression.

[0235] In some embodiments, the prevention and / or treatment of tumors is achieved by at least one of the following (a) to (l): (a) inhibiting tumor volume growth; (b) inhibiting tumor weight increase; (c) inhibiting tumor cell growth; (d) improving tumor treatment response rate; (e) improving the efficacy of immunosuppressive drug treatment, such as improving the efficacy of anti-PD-1 antibody treatment; (f) preventing and / or inhibiting tumor cell spread or metastasis; (g) reducing anti-tumor drug resistance; (h) inhibiting HDAC activity through at least one of acetate or acetate, propionate or propionate, butyrate or butyrate, valeric acid or valeric acid in SCFA; (i) inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) regulating or activating the subject's immune system; (k) promoting tumor cell senescence or apoptosis; (l) inhibiting angiogenesis in tumor tissue.

[0236] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by inhibiting HDAC activity and / or promoting IFNβ transcriptional activity.

[0237] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product achieves treatment or prevention of diseases (such as tumors or cancer, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases) by upregulating proinflammatory factors and chemokines to stimulate immunity and / or by downregulating anti-inflammatory factors for immunomodulation. In some embodiments, the proinflammatory factor is selected from at least one of IFNγ, IFNβ, IL-1β, IL-6, IL-1β, MCP-1, MIG, RANTES, and TNFα.

[0238] In some embodiments, the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition or the aforementioned pharmaceutical combination product is administered at least once a day. In some embodiments, when the administration frequency is more than twice a day, the same dose is administered each time. In some embodiments, when the administration frequency is more than twice a day, different doses are administered each time. In some embodiments, the route of administration includes at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, intradermal injection, intramuscular injection, and intravenous injection.

[0239] The term "subject", or patient, refers to any mammal. A subject or patient described as "in need" refers to a mammal in need of treatment (or prevention) of a disease. For example, it includes any one or more animals under the class Mammalia, such as Monotremes, Marsupials, Insectivora, Homarus, Climbing, Dermoptera, Chiroptera, Primates, Edentata, Pholidota, Lagomorpha, Rodentia, Carnivora, Sirenians, Hyraxes, Tubulodonta, Perissodactyla, Artiodactyla, Cetacea and other animals. Common mammals include Rodentia (such as mice, rats, hamsters, guinea pigs), Lagomorpha (such as rabbits), Perissodactyla (such as horses, donkeys), Artiodactyla (such as sheep, goats, camels, cattle, pigs), Primates (such as monkeys, gorillas, chimpanzees, humans), Carnivora (such as dogs, cats), etc. It is understood that the subject may be healthy, or may suffer from a disease at any stage of development, such as at least one of a tumor or cancer, a metabolic disease, an inflammatory or autoimmune disease, an infectious disease, and a central nervous system disease.

[0240] The term "treating" a disease in a subject or "treating" a subject having or suspected of having a disease refers to administering a drug therapy to the subject, such as administering one or more agents (e.g., including the aforementioned Parabacteroides strain, the aforementioned culture, the aforementioned bacterial agent, the aforementioned composition, the aforementioned pharmaceutical composition, or the aforementioned pharmaceutical combination product), thereby reducing or preventing at least one symptom of the disease from worsening. Therefore, in one embodiment, "treating" refers in particular to delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing the efficacy of alternative therapies, or reducing resistance to alternative therapies, or a combination thereof.

[0241] The term "prevention" is art-recognized and well-known in the art when used in connection with a condition such as local recurrence, and includes administration of a composition that reduces the occurrence of or delays the onset of symptoms of a medical condition in a subject relative to a subject not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable tumors in a population of patients receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable tumors in a treated population relative to an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0242] The tenth aspect of the present application also relates to a method for preparing the aforementioned culture, which method comprises the following steps: taking the aforementioned strain, inoculating it into a fermentation medium, and obtaining a culture.

[0243] In some embodiments, the culture is centrifuged to collect the fermentation medium supernatant. In some embodiments, the fermentation medium or the fermentation medium supernatant is concentrated by at least one of evaporation, freeze drying, dialysis, extraction, and membrane separation to obtain a concentrated product. In some embodiments, the fermentation medium or the fermentation medium supernatant is extracted by at least one of extraction and solvent extraction to obtain an extract. In some embodiments, any one of the fermentation medium, the fermentation medium supernatant, the concentrated product, and the extract is dried to obtain a dried product.

[0244] The meanings of some abbreviations involved in the following embodiments are shown in Table 1.

[0245] Table 1. Meaning of some abbreviations

[0246]

[0247]

[0248] Some of the culture media involved in the following examples are as follows:

[0249] MM01 liquid culture medium, components: peptone 5g / L, trypticase 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K 2 HPO 4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, heme 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1μL / L, inorganic salt solution 8mL / L (each 1L includes calcium chloride 0.25g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, sodium bicarbonate 10g, sodium chloride 2g).

[0250] Solid MM01 medium, composed of: peptone 5g / L, trypticase 5g / L, yeast powder 10g / L, beef extract 5g / L, glucose 5g / L, K 2 HPO 4 2g / L, sodium acetate 2g / L, Tween 80 1mL / L, heme 5mg / L, L-cysteine ​​hydrochloride 0.5g / L, vitamin K1 1μL / L, inorganic salt solution 8mL / L (each 1L includes calcium chloride 0.25g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, sodium bicarbonate 10g, sodium chloride 2g), agar 15g / L.

[0251] AC liquid culture medium, composed of: peptone 20g / L, glucose 5g / L, yeast extract 3g / L, beef extract powder 3g / L, vitamin C 0.2g / L, pH 7.0.

[0252] Anaerobic blood agar plates (purchased from Huankai Microorganisms) were composed of: 10 g / L casein pancreatic digest, 3 g / L heart pancreatic digest, 1 g / L corn starch, 5 g / L meat gastric digest, 5 g / L yeast extract powder, 5 g / L sodium chloride, 15 g / L agar, 50-100 mL / L sterile defibrinated sheep blood, pH 7.3±0.2.

[0253] Columbia blood agar plates (purchased from Huankai Microorganisms) were composed of: tryptic peptone 12 g / L, animal tissue protein digest 5 g / L, corn starch 1 g / L, beef extract 3 g / L, yeast extract 3 g / L, sodium chloride 5 g / L, agar 13.5 g / L, polymyxin B 0.01 g / L, nalidixic acid 0.01 g / L, sterile defibrinated sheep blood or 50-100 mL / L, pH 7.3±0.2.

[0254] Chocolate agar plate (purchased from Huankai Microorganisms), composed of: polyvalent peptone 18 g / L, starch 1 g / L, sodium chloride 5 g / L, agar 15 g / L, defibrinated sheep blood 40 g / L, pH 7.4±0.2.

[0255] Tryptic soy casein liquid medium TSB liquid medium, composition: tryptic soy casein 17g / L, soybean papain hydrolysate 3g / L, dipotassium hydrogen phosphate 2.5g / L, sodium chloride 5g / L, glucose 2.5g / L, pH 7.3±0.2.

[0256] MYPG solid plate culture medium, composed of: peptone 5g / L, malt extract powder 3g / L, yeast extract powder 3g / L, glucose 10g / L, agar 20g / L, pH 6.2±0.2.

[0257] The above culture medium can be prepared by conventional preparation methods and sterilization methods.

[0258] The above implementation is described below in conjunction with specific examples.

[0259] Example 1: Isolation and identification of strains MNH 26621 and MNH 20645

[0260] 1. Isolation of strains

[0261] The strain MNH 26621 was isolated from a stool sample of a healthy volunteer in Dali City, Yunnan Province, and the strain MNH20645 was isolated from a stool sample of a male healthy volunteer in Guangzhou City, Guangdong Province. The specific process is as follows:

[0262] (1) Volunteers collect 2 to 5 grams of fresh feces and put them into a sample collection and storage tube. After shaking and homogenization, the processed fecal sample is placed in an ice box and sent to the laboratory for strain isolation within 24 hours.

[0263] (2) Dispense physiological saline in a biosafety cabinet, 9 mL / tube; prepare strain isolation culture medium and transfer it to an anaerobic workstation (Don Whitley Scientific H35) 24 h in advance, labeling the sample information, culture medium type, isolation date, etc.

[0264] (3) Place a fresh stool sample in an anaerobic operating station and vortex for 1 min to mix. Pipette 1 mL of the sample into 9 mL of saline and mix for 10 min. -1 dilution, and then serially diluted to 10 -6 Dilution solution for later use.

[0265] (4) Take 10 respectively -6 The diluted solution was dropped onto the separation medium (anaerobic blood agar plate, Columbia blood agar plate and chocolate agar plate) at a drop volume of 100 μL / dish. The plate was spread evenly. After the surface of the plate was dry, the plate was inverted and cultured at 37°C for 3 to 5 days.

[0266] (5) Observe the growth of the strain in the isolation medium and pick a single colony with a sterilized toothpick to purify the strain. The purified strain is placed at 37°C and cultured anaerobically.

[0267] (6) The pure culture strain was prepared into a 20% glycerol / water bacterial solution and stored at -86°C.

[0268] 2. Identification of strains

[0269] 2.1 Morphological characteristics

[0270] The strain MNH 26621 was inoculated on MYPG solid plate medium and cultured anaerobically at 37°C for 48 hours. Figure 1 As shown in A, visible colonies are formed on the plate culture medium. The colonies are round, with regular and smooth edges, about 2 mm in diameter, opaque, and no secretions are formed around the colonies. Figure 2 A and Figure 3 A. Microscopic morphological observation revealed that the strain had no spores, no flagella, was non-motile, was short rod-shaped, about 3-5 μm × 8-15 μm in diameter, and was Gram-negative.

[0271] The strain MNH 20645 was inoculated on MYPG solid plate medium and cultured anaerobically at 37°C for 48 hours. Figure 1 As shown in B, visible colonies are formed on the plate culture medium. The colonies are round, with regular and smooth edges, and are opaque. No secretions are formed around the colonies. Figure 2 B and Figure 3 B. Microscopic morphological observation revealed that the strain had no spores, no flagella, was non-motile, rod-shaped, with a diameter of about 5μm×10~20μm; Gram staining was negative.

[0272] 2.2 Physiological and biochemical characteristics

[0273] The temperature growth range of strain MNH 26621 is 30-42°C, and the optimal growth temperature is 37°C. Figures 4 to 6 A, the strain can grow in the pH range of 7.0-10.0, with the optimal growth pH of 7.0-9.0; it can tolerate up to 2% NaCl; it can survive and grow in the bile salt concentration range of 0-0.10%, and cannot grow when the bile salt concentration is greater than or equal to 0.15%. In addition, the strain cannot grow under aerobic conditions, but grows well under anaerobic conditions, and is an obligate anaerobic bacterium.

[0274] The temperature growth range of strain MNH 20645 is 30-42℃, and the optimal growth temperature is 37℃. Figures 4 to 6 B. This strain is sensitive to pH, and its optimal growth pH is about 7.2 ( Figures 5-6 The medium pH is about 7.2 when the concentration of bile salt is 0%); it can tolerate up to 2% NaCl; it is intolerant to bile salts and cannot grow when the bile salt concentration is greater than or equal to 0.1%. In addition, the strain can survive for 4 hours under aerobic conditions with a survival rate of about 80%, and grows well under anaerobic conditions.

[0275] API 20A test

[0276] The test was performed using API 20A reagent strips (BioMérieux) according to the instructions. The culture conditions of the strain were 37°C and anaerobic. The results are shown in Table 2.

[0277] Table 2. API 20A test results for strains MNH 26621 and MNH 20645

[0278]

[0279] As can be seen from the table, strains MNH 26621 and MNH 20645 can utilize glucose, lactose, sucrose, xylose, arabinose, mannose, raffinose, and rhamnose as substrates for metabolism and acid production, while other types of substrates cannot be used.

[0280] Antibiotic susceptibility testing

[0281] The antibiotic sensitivity test of strain MNH 26621 was carried out using the disk diffusion method. The test results are shown in Table 3.

[0282] Table 3. Antibiotic susceptibility test results of strains MNH 26621 and MNH 20645

[0283]

[0284]

[0285] The results refer to Table 3. The strain MNH 26621 is sensitive to antibiotics such as ampicillin, erythromycin, ciprofloxacin, lincomycin, chloramphenicol, penicillin, gentamicin, tetracycline and ceftriaxone, but is resistant to cotrimoxazole. The strain MNH20645 is sensitive to antibiotics such as erythromycin, chloramphenicol, tetracycline, penicillin, ampicillin, lincomycin and ceftriaxone, but is resistant to cotrimoxazole, ciprofloxacin and gentamicin.

[0286] 2.316S rRNA

[0287] Fresh cultures of strains MNH 26621 and MNH 20645 were taken, and the genomic DNA of the samples was extracted using the SDS method. The purity and integrity of the DNA were then detected by agarose gel electrophoresis and quantified using Qubit.

[0288] The extracted genomic DNA template was used to amplify 16S rRNA, and the amplification primers were:

[0289] 1492R: AGAGTTTGATCATGGCTCAG (SEQ ID NO: 3)

[0290] 27F: TAGGGTTACCTTGTTACGACTT (SEQ ID NO: 4).

[0291] The amplified PCR product was purified and then sequenced using ABI3730XL. The 1422 bp 16S rRNA sequence of strain MNH 26621 was obtained as follows:

[0292] CATGGAGGCTTACACATGCAAGTCGAGGGGCATCGGGGTGTAGCAATACATTGCCGGCGACCGGCGCACGGGTGAGTAACGCGTATGCAACTTACCTATCAGAGGGGAATAGCCCGGCGAAAGTCGGATTAATGCCCCATAAAACAGGGGTCCCGCATGGGACTATTTGTTAAAGATTCATCGCTGATAGATAGGCATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAACCGACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACACGGTCCAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGACGTGAGTCTGAACCAGCCAAGTCGCGTGAAGGAAGAAGGTTCTATGGATTGTAAACTTCTTTTATACGGGAATAAAGTGCACCACGTGTGGTGTTTTGTATGTAcCGTATGAATAAGCATCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTTGTTTTTTAAGTCAGCGGTGAAAGTTTGTGGCTCAACCATAAAATTGCCGTTGAAACTGGAAGACTTGAGTGTGTTTGAGGTAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCTTACTAAACCATAACTGACACTGAAGCACGAAAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCAGTAAACGATGATTACTAGGAGTTTGCGATAGACAGTAAGCTCTACAGCGAAAGCGTTAAGTAATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGG

[0293] GGGCCCGCACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACC

[0294] CGGGTTTGAACGTAGTCGGACCGGCCTTGAAAGAGGCCTTCTAGCAaTAGCTGATTACG

[0295] AGGTGCTGCATGGTTGTCGTCAGCTCGTGCCGTGAGGTGTCGGCTTAAGTGCCATAACG

[0296] AGCGCAACCCTTATCACTAGTTACTAACAGGTAAAGCTGAGGACTCTGGTGAGACTGCC

[0297] AGCGTAAGCTGTGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACATCCGG

[0298] GGCGACACACGTGTTACAATGGCATGGACAAAGGGCAGCTACACAGCGATGTGATGCTA

[0299] ATCTCCAAACCATGTCTCAGTTCGGATCGGAGTCTGCAACTCGACTCCGTGAAGCTGGA

[0300] TTCGCTAGTAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACACA

[0301] CCGCCCGTCAAGCCATGGGAGCCGGGGGTACCTGAAGTCCGTAACCGCGAGGATCGGC

[0302] CTAGGGGAAAA (SEQ ID NO: 1).

[0303] This sequence was compared with the NCBI Nucleotide collection (nr / nt) database, and the closest species was Parabacteroides merdae with a similarity of 96.34%. The species classification information of this strain was preliminarily determined, that is, it was preliminarily determined that MNH 26621 belonged to the genus Parabacteroides and was a new species.

[0304] The 16S rRNA sequence of 1406 bp obtained for strain MNH 20645 is as follows:

[0305] GGCTTACACAGTGCAAGTCTGAGGGGCAGCGGGGGTAGCAATACCTGCCGGCGAC

[0306] CGGCGCACGGGTGAGTAACGCGTATGCAACTTGCCTATCAGAGGGGAATAGCCCGGCGA

[0307] AAGTCGGATTAATGCTCCATAAAACAGGGGTCCCGCATGGGAATATTTGTTAAAGATTCA

[0308] TCGCTGATAGATAGGCATGCGTTCCATTAGGCAGTTGGCGGGGTAACGGCCCACCAAAC

[0309] CGACGATGGATAGGGGTTCTGAGAGGAAGGTCCCCCACATTGGAACTGAGACACGGTC

[0310] CAAACTCCTACGGGAGGCAGCAGTGAGGAATATTGGTCAATGGCCGGGAGGCTGAACC

[0311] AGCCAAGTCGCGTGAAGGATGAAGGATCTATGGTTTGTAAACTTCTTTTATACGGGAATA

[0312] AAGTGTGGGACGTGTCCCATTTTGTATGTACCGTATGAATAAGCATCGGCTAACTCCGTG

[0313] CCAGCAGCCGCGGTAATACGGAGGATGCGAGCGTTATCCGGATTTATTGGGTTTAAAGG

[0314] GTGCGTAGGTGGTTGTTTAAGTCAGCGGTGAAAGTTTGTGGCTCAACCATAAAATTGCC

[0315] GTTGAAACTGGATGACTTGAGTATATTTGAGGTAGGCGGAATGCGTGGTGTAGCGGTGA

[0316] AATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCTTACTAAACTATAACTGAC

[0317] ACTGAAGCACGAAAGCGTGGGGATCAAACAGGATTAGATACCCTGGTAGTCCACGCAGT

[0318] AAACGATGATTACTAGGAGTTTGCGATAGACAGTAAGCTCTACAGCGAAAGCGTTAAGT

[0319] AATCCACCTGGGGAGTACGCCGGCAACGGTGAAACTCAAAGGAATTGACGGGGGCCCG

[0320] CACAAGCGGAGGAACATGTGGTTTAATTCGATGATACGCGAGGAACCTTACCCGGGTTT

[0321] GAACGCATTCGGACCGGAGTGGAAaCGCTCCTTTCAGCAATGACCGTTTGCGAGgTGCT

[0322] GCATGgtTGTCGTCAGCTCGTGCCGTGAGgTGTCGGCTTAAGTGCCATAaCGAGCGCAAC

[0323] CCTTATCACTAGTTACTAACAGGTCAAGCTGAGGACTCTGGTGAGACTGCCAGCGTAAG

[0324] CTGTGAGGAAGGTGGGGATGACGTCAAATCAGCACGGCCCTTACATCCGGGGCGACAC

[0325] ACGTGTTACAATGGCATGGACAAAGGGCAGCTACACAGCGATGTGATGCAAATCTCCAA

[0326] ACCATGTCTCAGTTCGGATCGGAGTCTGCAACTCGACTCCGTGAAGCTGGATTCGCTAG

[0327] TAATCGCGCATCAGCCATGGCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGT

[0328] CAAGCCATGGGAGCCGGGGGTACCTGAAGTCCGTAACCGCAAGGATCGGCC(SEQ ID NO:2)。

[0329] This sequence was compared with the NCBI Nucleotide collection (nr / nt) database, and the closest species was Parabacteroides hominis with a similarity of 99.85%. It was preliminarily determined that strain MNH 20645 was Parabacteroides hominis.

[0330] The identity of the 16S rRNA of strain MNH 26621 (SEQ ID NO: 1) and the 16S rRNA of MNH 20645 (SEQ ID NO: 2) was 96.74%.

[0331] The 16S rRNA sequences of MNH 26621 and MNH 20645 measured above and the 16S rRNA sequences of related genera were aligned using the MUSCLE method, and finally the maximum likelihood method was used to perform 1000 similarity calculations, and a phylogenetic tree was constructed using MEGA X. Figure 7 , Figure 8 and Fig. 9 In the evolutionary tree, MNH 20645 and Parabacteroideshominis NSJ 79 are in the same branch with a short evolutionary distance, while MNH 26621 forms a separate evolutionary branch with a long evolutionary distance from other species.

[0332] In order to further determine the average nucleotide identity of the genomes of strains MNH 26621, MNH 20645 and other members of the genus Parabacteroides, the genomes of the isolated original strains of MNH 26621 and MNH 20645 were further prepared, sequenced, assembled and analyzed. The genomic DNA samples of the original strains of MNH 26621 and MNH 20645 that passed the electrophoresis test were fragmented by ultrasound, with a fragment length of about 350bp, and then the Illumina sequencing library was constructed using a standard DNA library construction kit (NEB UltraTM). The constructed sequencing library was sequenced with Illumina NovaSeq for double-end 150bp sequencing. MNH 26621 sequencing obtained 1.27Gbp of data, of which Q20 accounted for: 97.17%. MNH 20645 sequencing obtained 1.30Gbp of data, of which Q20 accounted for: 97.04%.

[0333] The raw genome sequencing data was filtered using fastp (version: 0.20.0) with filtering parameters: "--poly_g_min_len10--poly_x_min_len 10-q 15-u 40-n 5-l 50". The filtered raw data was assembled using SPAdes (version: v3.14.0) with assembly parameters "--isolate--cov-cutoff 10". The MNH26621 genome was assembled to obtain a total gene length of 5.06 Mbp, an N50 length of 145.2 kbp, and a GC content of 46.17%. The MNH20645 genome was assembled to obtain a total gene length of 5.19 Mbp, an N50 length of 105.3 kbp, and a GC content of 46.27%.

[0334] The genomic genes were predicted and analyzed using the prokaryotic analysis software genome annotation pipeline prokka (version: 1.14.5), with the parameter "--gcode 11--evalue 1e-09". A total of 1833 CDS sequences were predicted for MNH 26621, with an average CDS sequence length of 1055 bp. A total of 4192 CDS sequences were predicted for MNH 20645, with an average CDS sequence length of 1103 bp. The genome information of all model strains under the genus Parabacteroides was downloaded from NCBI, and the degree of genome relevance was determined by evaluating the average nucleotide identity (ANI) and alignment score (AF) of the whole genome sequences of MNH 26621 and MNH 20645 with these reference organisms. The alignment results are shown in Table 4 below:

[0335] Table 4. Comparison of the whole genome sequences of strains MNH 26621 and MNH 20645 with other strains of the genus Parabacteroides

[0336]

[0337] The model strain with the highest average nucleotide identity with the genome of strain MNH 26621 was Parabacteroideshominis, with an average nucleotide similarity (ANI) of 88.87% and an alignment score (AF) of 75.27%. The full genome information comparison between MNH 26621 and known species showed that strain MNH 26621 and known species Parabacteroidesjohnsonii, Parabacteroides merdae, Parabacteroides massiliensis, Parabacteroidesacidifaciens, Parabacteroides gordonii, Parabacteroides faecis, Parabacteroidesbouchesdurhonensis, Parabacteroides goldsteinii, Parabacteroides timonensis, Parabacteroides provencensis, Parabacteroides chinchillae, and Parabacteroidesdistasonis can be clearly distinguished from genotypic characteristics.

[0338] The model strain with the highest average nucleotide identity to the genome of strain MNH 20645 is Parabacteroides hominis, with an average nucleotide identity (ANI) of 99.92% and an alignment score (AF) of 97.14%. Therefore, MNH20645 can be identified as Parabacteroides hominis.

[0339] The average nucleotide identity between the MNH 26621 genome and the MNH 20645 genome was 88.89%, and the alignment score was 74.55% (with the MNH 20645 genome as the reference sequence).

[0340] The average nucleotide identity between the MNH 20645 genome and the MNH 26621 genome was 88.81%, and the alignment score was 73.54% (with the MNH 26621 genome as the reference sequence).

[0341] 2.4 Genome analysis

[0342] Based on the previously assembled genome data, the RGI pipeline (version: 4.2.2) was used to analyze potential antibiotic resistance genes, and the antibiotic resistance gene database was CARD (version: 3.0.0, https: / / card.mcmaster.ca / analyze / rgi).

[0343] The comparison results are shown in Table 5.

[0344] Table 5. Drug resistance gene information of strains MNH 26621 and MNH 20645

[0345] Strain Gene Resistance genes Gene name Comparison consistency (%) MNH 26621_01561 ARO:3000191 Q 96.41 MNH 20645_00602 ARO:3001302 ikB 100 MNH 20645_02466 ARO:3000191 Q 99.84

[0346] NCBI blastp (version 2.7.1+) was used to analyze the potential virulence factors and related genes in the genome and compared with the virulence factor database VFDB (virulence factor database, http: / / www.mgc.ac.cn / cgi-bin / VFs / v5 / main.cgi, updated on September 19, 2019). The comparison results are shown in Table 6.

[0347] Table 6. Potential toxic genes of strains MNH 26621 and MNH 20645

[0348]

[0349]

[0350] gutSMASH5 (version 1.0.0) was used to analyze the potential primary metabolic gene clusters in the genome, and the alignment results are shown in Table 7.

[0351] Table 7. Potential primary metabolic gene clusters of strain MNH 26621 and strain MNH 20645

[0352]

[0353] 2.5 Short-chain fatty acids (SCFA) determination

[0354] (1) Preparation of bacterial cells

[0355] The strain MNH 26621 was inoculated into a liquid culture medium and cultured anaerobically at 37°C for 48 hours. The bacterial cells were collected by centrifugation and stored at -86°C for later use.

[0356] (2) Preparation of standard products

[0357] Acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid and hexanoic acid standards were weighed and prepared with ethyl acetate into eight mixed standard concentration gradients of 0.1 μg / mL, 0.5 μg / mL, 1 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, 50 μg / mL and 100 μg / mL.

[0358] Take 600 μL of standard, add 25 μL of 4-methylvaleric acid with a final concentration of 500 μM as internal standard, mix well and add to the injection bottle, enter GC-MS detection, injection volume 1 μL, split ratio 10:1, split injection.

[0359] (3) Metabolite extraction

[0360] The collected bacterial samples were thawed on ice, 80 mg of the sample was taken into a 2 mL glass centrifuge tube, and 900 μL of 0.5% phosphoric acid was added to resuspend, shaken and mixed for 2 min, centrifuged at 14000 g for 10 min, 800 μL of the supernatant was taken, and an equal amount of ethyl acetate was added for extraction, shaken and mixed for 2 min, centrifuged at 14000 g for 10 min, 600 μL of the upper organic phase was taken, 4-methylpentanoic acid with a final concentration of 500 μM was added as an internal standard, mixed and added to the injection bottle, and entered the GC-MS detection, the injection volume was 1 μL, the split ratio was 10:1, and the split injection was performed.

[0361] (4) Sample testing and analysis

[0362] The samples were separated using an Agilent DB-WAX capillary column (30m×0.25mm ID×0.25μm) gas chromatography system. The temperature was programmed as follows: the initial temperature was 90°C, then the temperature was raised to 120°C at 10°C / min, then to 150°C at 5°C / min, and finally to 250°C at 25°C / min and maintained for 2min. The carrier gas was helium with a flow rate of 1.0mL / min.

[0363] Agilent 7890A / 5975C gas chromatograph was used for mass spectrometry analysis. The injection port temperature was 250°C, the ion source temperature was 230°C, the transfer line temperature was 250°C, and the quadrupole temperature was 150°C. Electron impact ionization (EI) source, full scan and SIM scanning mode, electron energy 70 eV.

[0364] The chromatographic peak area and retention time were extracted using MSD ChemStation software. The calibration curve was drawn and the content of short-chain fatty acids in the samples was calculated. The results are shown in Table 8.

[0365] Table 8. Short chain fatty acid (SCFA) production results of strain MNH 26621

[0366] Acetic acid Butyric acid Isovalerate Valeric acid Hexanoic acid Propionic acid Isobutyric acid SCFA production (μg / g) 1111.8 0.85 85.01 0.11 0.189 658.35 29.1

[0367] It can be seen from the results in the table that the strain MNH 26621 provided in this example produces high amounts of acetic acid and propionic acid, and low amounts of butyric acid, valeric acid and caproic acid.

[0368] The above method was used to detect the short-chain fatty acid (SCFA) production of strain MNH 20645. The results are shown in Table 9.

[0369] Table 9. Short chain fatty acid (SCFA) production results of strain MNH 20645

[0370] Acetic acid Butyric acid Isovalerate Valeric acid Hexanoic acid Propionic acid Isobutyric acid SCFA production (μg / g) 985.06 0.56 81.19 0.078 0.106 1118.9 38.69

[0371] It can be seen from the results in the table that the strain MNH 20645 provided in this example produces high amounts of acetic acid and propionic acid, and low amounts of butyric acid, valeric acid and caproic acid.

[0372] Combining the morphological, physiological, biochemical, 16S and whole genome identification results, strain MNH 26621 has significant differences from other existing strains of the genus Parabacteroides and can be identified as a new species of the genus Parabacteroides.

[0373] The strain MNH 26621 was deposited in Guangdong Microbial Culture Collection Center (GDMCC) with the deposit name Parabacteroides sp. MNH 26621, the deposit number GDMCC No: 62664, the deposit date July 28, 2022, the deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, and the proposed taxonomic name is Parabacteroides sp. The deposited strain was identified as viable by the collection center.

[0374] Combining the morphological, physiological, biochemical, 16S and whole genome identification results, strain MNH 20645 can be identified as Parabacteroides hominis.

[0375] The strain MNH 20645 was deposited in Guangdong Microbial Culture Collection Center (GDMCC) with the deposit name Parabacteroides sp. MNH 20645, the deposit number GDMCC No: 62665, the deposit date July 28, 2022, and the deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Institute of Microbiology, Guangdong Academy of Sciences, and the proposed taxonomic name is Parabacteroides sp. The deposited strain was identified as viable by the collection center.

[0376] Example 2: Regulation of the immune activity of macrophages by strains

[0377] THP-1 (Wuhan Pronosai Life Science Co., Ltd.) was treated with PMA at a final concentration of 5 ng / mL for 48 hours to differentiate into M0 macrophages. M0 macrophages were co-incubated with strains (MNH 26621 or MNH 20645) for 24 hours. The strains were added at a ratio of MOI (viable count: cell count) = 10:1. At the same time, an M1 macrophage control group (M0 macrophages were induced with 20 ng / mL IFNγ + 10 pg / mL LPS for 24 hours) was set up. After anaerobic culture for 1 hour, mixed antibiotics (1 mg / mL Ampicillin, 5 mg / mL Streptomycin, 1 mg / mL Colistin) were added to kill the bacteria, and the culture was continued for 23 hours, and the supernatant was collected.

[0378] The BD Cytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit was used to react the supernatant sample with the detection protein according to the instructions, and the concentrations of cytokines such as IL-10, IL-8, IL-1β, MCP-1, RANTES and TNFα in the supernatant were detected by flow cytometry.

[0379] The results for strain MNH 26621 are as follows Fig.10 As shown in the figure, after co-culture of macrophages with strain MNH 26621, macrophages clearly showed the characteristics of differentiation into M1 macrophages. Among them, the content of anti-inflammatory factor IL-10 was significantly downregulated, the content of pro-inflammatory factor IL-1β was significantly upregulated, and the contents of IL-6, TNFα, IL-12 / IL-13P40 showed an upward trend; the contents of chemokines RANTES and MCP-1 were significantly upregulated, and the contents of MIG and IP-10 showed an upward trend.

[0380] The results for strain MNH 20645 are as follows Fig.11 As shown in the figure, after co-culture of macrophages with strain MNH 20645, macrophages also showed obvious characteristics of differentiation into M1 macrophages. Among them, the contents of chemokines MCP-1 and RANTES were significantly upregulated, the contents of MIG and IP-10 showed an upregulation trend, the contents of pro-inflammatory factors IL-1β, IL-6, IL-12 / IL-13P40, and IL-8 were significantly upregulated, and the content of anti-inflammatory factor IL-10 showed an upregulation trend.

[0381] Example 3: Regulation of Primary PBMC Immune Activity by Strain

[0382] After the strain (MNH 26621 or MNH 20645) was co-incubated with Primary PBMC cells for 24 hours, the supernatant was collected, and a control group in which the strain MNH 26621 was replaced with PBS was set up. The supernatant samples were reacted with the detection proteins according to the instructions using the BD Cytometric Bead Array (CBA) Human Soluble Protein Master Buffer Kit, and the concentrations of cytokines IFNγ, TNFα, IL-1β, IP-10 (CXCL-10), RANTES (CCL5), MIG (CXCL-9), IL-6, IL-10, MCP-1 (CCL2), IL-8, and IL-12 / IL-23P40 in the supernatant were detected by flow cytometry.

[0383] The results are as follows Fig.12 As shown, strain MNH 26621 can significantly induce a significant increase in inflammatory factors IL-1β, IL-6, IL-8 and chemokines MCP-1 (CCL2) and RANTES (CCL5); the expression of induced inflammatory factors TNFα and IP-10 (CXCL-10) tends to increase.

[0384] In addition, the results of strain MNH 20645 showed that the expression of inflammatory factors TNFα, IL-1β, and IL-6 tended to increase.

[0385] Example 4: Effect of strains on IFNβ expression

[0386] Interferon (IFN) receptor proteins are a type of cytokine secreted by host cells that can regulate immune responses. Viruses, bacterial endotoxins, and synthetic double-stranded RNA can stimulate the production of interferons. IFNβ belongs to type I interferon, which can promote the activity of NK cells, macrophages, and T lymphocytes, thereby playing antiviral, anti-tumor, and immunomodulatory roles. In order to verify whether the strain can promote the expression of IFNβ, this study used THP-1 cells (a cell line built by Muen) that had been constructed to carry the IFNβ gene promoter reporter gene (i.e., IFNβ-promoter reporter) to evaluate the effect of the strain on IFNβ transcriptional activity.

[0387] Preparation of strain culture supernatant: The strain (MNH 26621 or MNH 20645) was inoculated into MM01 liquid culture medium, cultured anaerobically at 37°C for 48 hours, and the bacteria were removed by centrifugation. The culture supernatant was filtered through a 0.22 μm filter, aliquoted, and the collected material was stored at -80°C for later use.

[0388] Control group: DMEM complete medium (10% FBS) containing 10% volume of MM01 bacterial culture medium.

[0389] MSA-2 group: IFNβ activation positive control (40 μM).

[0390] Strain group: containing 10% volume of the culture supernatant of the strain (MNH 26621 or MNH 20645).

[0391] THP-1-IFNβ-promoter reporter cells were seeded in 96-well plates at 1×10 cells per well. 5 cells. Treat the cells according to the set groups. After continuing to culture for 24 hours, the cells were centrifuged at 300g for 5 minutes, the culture supernatant was removed, 50μL 1×luciferase detection reagent was added, the reaction was carried out for 1 minute, and the luminescence chemiluminescence value was detected by an enzyme reader. The relative fluorescence value (Relative Luminescence) was normalized with the Control group to evaluate the effect of the strain on IFNβ transcription activity.

[0392] The results are as follows Fig.13 As shown, the results show that strains MNH 26621 and strains MNH 20645 can significantly promote IFNβ transcription activity. These results show that the strains provided in the examples of this application have potential functional effects in immunomodulation, anti-virus, and anti-tumor.

[0393] Example 5: Animal experiment on the use of strains for liver cancer treatment

[0394] To verify whether strains MNH 26621 or MNH 20645 can be used for tumor prevention and treatment, a mouse syngeneic tumor model was used to perform an experiment to inhibit liver cancer growth. This study has been reviewed by the Munn Institutional Animal Care and Use Committee.

[0395] Test strains: After the glycerol cryopreservation tubes of the strains in Example 1 were thawed at 37°C, they were inoculated on anaerobic blood plates in an anaerobic workstation for activation. The activated strains were inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in a solvent (PBS-Cys: phosphate buffer containing 0.05% cysteine ​​hydrochloride) to obtain the purity and viable count (3.5×10 8 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0396] Tumor cells: H22 mouse hepatoma cells, CBP69230.

[0397] Experimental animals: The experimental mice were BALB / C mice, aged 5-6 weeks, a total of 140 mice, purchased from Guangdong Sijiajingda Biotechnology Co., Ltd.

[0398] Animal experiment: The animals were raised normally. After the quarantine period, the mouse liver cancer model was established by intradermal injection of H22 mouse liver cancer cells. The cell inoculation amount was 2×10 6 / mL, 0.1mL / mouse. The day of inoculation was recorded as D1. At D7, mice with abnormal tumor growth were eliminated and randomly stratified and grouped according to tumor volume, 12 mice / group. The negative control group A (Control) was given culture medium, the experimental group F was given strain MNH 26621, and the experimental group G was given strain MNH 20645. The endpoint of this experiment was: 12 oral administrations were completed. During the experiment, animal characteristics, tumor changes, tumor size, and mouse weight were regularly monitored. At the end of the experiment, weight changes, tumor volume changes, response rate, and tumor inhibition rate were analyzed. At the end of the experiment, all mice were euthanized by cervical dislocation. The results obtained are shown in Tables 10 to 11 below, and can be used for reference. Fig.14 .

[0399] Table 10. Overall statistics of mice at the end of the experiment with strain MNH 20645

[0400]

[0401] Table 11. Tumor tissue weight after dissection of experimental mice of strain MNH 20645

[0402]

[0403]

[0404] The results in the figure show that the weight of the mice increased steadily during the administration period, and there was no significant difference between the experimental group (Group G) and the control group (Group A) of the strain; compared with the mouse tumors in the control group, the mouse tumors in the experimental group were significantly smaller, and the TGI increased significantly, showing a significant inhibitory effect on tumor growth. Relative to the low response rate of the control group, the response rate of the experimental group was significantly improved. For example, the experimental group of strain MNH 20645 had a response rate of up to 45.45%, while the response rate of the control group was only 8.33%. In summary, the strain MNH 20645 of the embodiment shows good efficacy in anti-liver cancer and can significantly reduce the growth rate of liver cancer in mice.

[0405] At the same time, the experimental results also showed that strain MNH 26621 exhibited certain efficacy in anti-liver cancer and could reduce the growth rate of liver cancer in mice.

[0406] Example 6: Animal experiment on the use of strains for lung cancer treatment

[0407] To verify whether the strain can be used for the prevention and treatment of lung cancer, a mouse homologous tumor model was used to perform an experiment to inhibit lung cancer growth. This study has been reviewed by the Experimental Animal Care and Use Committee of Muen Biotechnology.

[0408] Test strains: The glycerol cryopreserved tubes of the strains in Example 1 were thawed at 37°C and inoculated on anaerobic blood plates in an anaerobic workstation for activation. The activated strains were inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in a solvent (PBS-Cys) to obtain the purity and viable count (3.5×10 8 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0409] Tumor cells: LLC1 mouse lung cancer cells, CL-0140.

[0410] Experimental animals: The experimental mice were C57BL / 6J mice, 5 weeks old, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0411] Animal Experimentation:

[0412] The mice were raised normally. After the quarantine period, the mouse lung cancer model was established by intradermal injection of LLC1 mouse lung cancer cells. The cell inoculation amount was 2×10 7 / mL, 0.1mL / mouse. The day of inoculation was recorded as D1. At D7, mice with abnormal tumor growth were eliminated and randomly stratified and grouped according to tumor volume, 12 mice / group. The negative control group (Control) was given culture medium, and the experimental group was given the corresponding strain. Drug administration began on the day of grouping, the gavage volume was 0.2mL / mouse / time, and the administration frequency was 1 day / time. The endpoint of this experiment was D21: 14 gavage administrations were completed. During the experiment, animal characteristics, tumor changes, tumor size, and mouse weight were regularly monitored. At the end of the experiment, weight changes, tumor volume changes, response rate, and tumor inhibition rate were analyzed. At the end of the experiment, all mice were euthanized by cervical dislocation. The results obtained are shown in the table below and Figure 15-16 .

[0413] Table 12. Overall statistics of mice at the end of the experiment

[0414]

[0415] Table 13. Tumor tissue weight after mouse autopsy

[0416]

[0417]

[0418] Fig.15 and Fig.16 The results showed that the weight of the mice increased steadily during the administration period, and there was no significant difference between the experimental group and the respective control group; compared with the mouse tumors in the respective control groups, the tumor volume and tumor weight of the mice in the experimental group were significantly reduced, and the TGI increased significantly, showing a significant inhibitory effect on tumor growth. Relative to the low response rate of the respective control groups, the response rate of the experimental group was significantly improved. For example, the experimental group of strain MNH 26621 had a response rate of up to 72.73%, while the control group had only 36.36%; the implementation group of strain MNH 20645 had a response rate of up to 50%, while the control group had only 16.67%. In summary, the strains of the embodiment show good efficacy in anti-lung cancer and can significantly reduce the growth rate of lung cancer in mice.

[0419] Example 7: Experimental verification of the effect of strains on PD-1 combined with liver cancer treatment

[0420] To verify the effect of the combination of strains and anti-PD-1 antibodies on tumor treatment, a mouse homologous tumor model was used to perform an experiment to inhibit liver cancer growth. This experimental protocol has been approved by the Animal Ethics Committee of Muen Biotechnology.

[0421] The experimental method refers to Example 5. For the strains in Example 1, negative control groups 1 and 2, positive control groups (PD-1) 1 and 2, and combination groups (strain + PD-1) 1 and 2 were set up respectively. General clinical observation, body weight monitoring, tumor volume (mm 3 ) measurements, and when the experiment endpoint was reached, all surviving mice were dissected to measure tumor weight and calculate tumor volume. Finally, statistical analysis and comparison were performed on the changes in tumor volume curve (Tumor volume), endpoint tumor volume (Tumor volume at endpoint), endpoint tumor weight (Tumor weight at endpoint), and endpoint tumor growth inhibition rate (Tumor Growth inhibition rate at endpoint, TGI).

[0422] Experimental results show that the combination of the strain and anti-PD-1 antibodies can inhibit the growth of liver cancer tumors, reduce the growth rate of liver cancer in mice, and have a certain effect on the treatment of liver cancer.

[0423] Example 8: Experimental verification of the effect of strains on PD-1 combined with lung cancer treatment

[0424] To verify the effect of combining strains with PD-1 antibodies on tumor treatment, a mouse homologous tumor model was used to perform an experiment to inhibit lung cancer growth. This experimental protocol has been approved by the Animal Ethics Committee of Muen Biotechnology.

[0425] Test strains: The glycerol cryopreserved tubes of the strains in Example 1 were thawed at 37°C and inoculated on anaerobic blood plates in an anaerobic workstation for activation. The activated strains were inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient amount of culture. The cultured bacterial solution was concentrated by centrifugation and resuspended in a solvent (PBS-Cys) to obtain the purity and viable count (1.0×10 8 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0426] Tumor cells: LLC1 mouse lung cancer cells, CL-0140.

[0427] Experimental animals: The experimental mice were C57BL / 6J mice, 5 weeks old, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0428] PD-1 antibody: anti-mPD-1 antibody (RMP1-14), prepared into 2 mg / mL solution with normal saline, sourced from BioXcell.

[0429] Animal Experimentation:

[0430] The mice were raised normally. After the quarantine period, the mouse lung cancer model was established by intradermal injection of LLC1 mouse lung cancer cells. The cell inoculation amount was 2×10 7 / mL, 0.1mL / mouse. The day of inoculation was recorded as D1. On D5, mice with abnormal tumor growth were removed and randomly divided into groups according to tumor volume, 8 mice / group, and the grouping was as follows:

[0431] Table 14. Grouping of combined experiments

[0432]

[0433] The PBS-Cys (glycerol) is prepared by uniformly mixing PBS-Cys (phosphate buffer containing 0.05% cysteine ​​hydrochloride) and 100% glycerol in a ratio of 3:1.

[0434] The negative control group A (Negative Control) was given negative control PBS-Cys (Gly) (ig) and negative control saline (ip), the positive control group B (PD-1) was given negative control PBS-Cys (Gly) and positive control anti-PD-1 antibody (ip), and the F and G groups were given test strains (MNH 26621 (ig), MNH 20645 (ig)) and anti-PD-1 antibodies, respectively. The administration began on the day of grouping. The intragastric volume of PBS-Cys (Gly) and each test strain was 0.2 mL / mouse, the injection volume of saline was 5 mL / kg, and the injection volume of anti-PD-1 antibody was 10 mg / kg. The administration frequency of PBS-Cys (Gly) and each test strain was once a day, and the administration was given until the end of the experiment (20 times in total); saline and anti-PD-1 antibodies were administered once every 3 days, for a total of 6 times.

[0435] After reaching the endpoint of the experiment, all surviving mice were dissected, the tumor weight was measured and the tumor volume was calculated. Finally, statistical analysis and comparison were performed on the changes in the tumor volume curve (Tumor volume), endpoint tumor volume (Tumor volume at endpoint), endpoint tumor weight (Tumor weight at endpoint) and endpoint tumor growth inhibition rate (Tumor Growth inhibition rate at endpoint, TGI).

[0436] The results showed that the combination of strains and PD-1 antibodies has a certain efficacy in treating lung cancer. The negative control of the endpoint tumor volume was 1279.80 mm 3 , positive control group (PD-1 antibody): 710.29mm 3, MNH 20645 combined with PD-1 antibody: 627.71 mm 3 , MNH 26621 combined with PD-1 antibody: 675.53 mm 3 Compared with the positive control group, the tumor volume of the MNH 20645 combination group and the MNH 26621 combination group showed a downward trend. The tumor growth inhibition rate (TGI) was statistically analyzed. The positive control group (PD-1) TGI was 47.3%; the strain MNH 20645 combined with PD-1 antibody TGI: 53.9%; the strain MNH 26621 combined with PD-1 antibody TGI: 50.0%. Compared with the positive control group, the TGI of the MNH 20645 combination group and the MNH 26621 combination group was increased to a certain extent. Therefore, the MNH 20645 combined with PD-1 antibody group and the MNH 26621 combined with PD-1 antibody group had a certain inhibitory effect on tumor growth.

[0437] Example 9: Experimental results of the combination of strain MNH20645 and Gemcitabine for the treatment of pancreatic cancer

[0438] To verify the effect of combining strain MNH20645 with gemcitabine on tumor treatment, a mouse homologous tumor model was used to perform an experiment to inhibit lung cancer growth. This experimental protocol has been approved by the Animal Ethics Committee of Muen Biotechnology.

[0439] Test strains: After thawing the glycerol cryopreserved tube of MNH20645 strain at 37°C, inoculate it on an anaerobic blood plate in an anaerobic workstation for activation. The activated strain is inoculated into MM01 liquid culture medium and cultured anaerobically to obtain a sufficient number of viable bacteria. The cultured bacterial solution is concentrated by centrifugation and resuspended in a solvent to obtain the purity and viable count (1.30×10 10 CFU / mL) of the test substances that meet the requirements of animal experiments.

[0440] Tumor cells: KPC mouse pancreatic cancer cells.

[0441] Experimental animals: The experimental mice were C57BL / 6J female mice, 7-8 weeks old, purchased from Guangdong Yaokang Biotechnology Co., Ltd.

[0442] Animal experiment: The animals were raised normally. After the quarantine period, KPC mouse pancreatic cancer cells were subcutaneously inoculated to form an ectopic homologous tumor model. The inoculation volume was 3×10 6 cells / mouse, 0.1mL / mouse. When the average tumor volume reaches 60-100mm 3 The patients were randomly divided into 10 groups according to the tumor volume. The grouping is as follows:

[0443] Table 15. Grouping of combined experiments

[0444]

[0445] The PBS-Cys (glycerol) is prepared by uniformly mixing PBS-Cys (phosphate buffer containing 0.05% cysteine ​​hydrochloride) and 100% glycerol in a ratio of 3:1.

[0446] On the day of grouping, oral administration began. Groups A and B were given PBS-Cys (glycan), and group G was given MNH20645. The oral administration volume was 0.2 mL / mouse. When the average tumor volume of the negative control group A grew to 120 mm 3 ~150mm 3 At 14:00 onset, groups A, B, and G began to receive drugs via intraperitoneal injection. Group A was given normal saline at a dose of 5 mg / kg, and groups B and G were given Gemcitabine at a dose of 50 mg / kg. The frequency of administration was once every 3 days, for a total of 6 times. The animals were weighed upon receipt and at the end of the quarantine period; the animals were weighed twice a week after tumor inoculation, and the day of intraperitoneal injection was used; the day of tumor cell inoculation was D1. From Day 7 to before grouping, the tumor diameter was measured once a day; after grouping, the tumor diameter was measured once every 3 days; when the average tumor volume of any group reached or exceeded 600mm 3 The tumor diameter was measured every 2 days, and the tumor growth was recorded. The experiment was terminated on D35. General clinical observation, body weight monitoring, tumor volume (mm 3 ) measurements, and when the experiment endpoint was reached, all surviving mice were dissected to measure tumor weight and calculate tumor volume. Finally, statistical analysis and comparison were performed on the changes in tumor volume curve (Tumor volume), endpoint tumor volume (Tumor volume at endpoint), endpoint tumor weight (Tumor weight at endpoint), and endpoint tumor inhibition rate (Tumor inhibition rate at endpoint, TGI).

[0447] Tumor inhibition rate: (average volume of control group minus volume of experimental group) / average volume of control group × 100%, all data are expressed in mean ± SD form, and GraphPad Prism 8.0.2 software is used for plotting and statistical analysis. For comparisons between more than three groups, one-way ANOVA method with Dunnett's multiple comparisons test was used for analysis, and two-way ANOVA with Sidak multiple comparisons was used for analysis of two-factor investigations. Significant differences are indicated by *, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0448] The results are shown in the following table and Fig.17 shown.

[0449] Table 16 Statistical table of overall status of mice at the end of the experiment

[0450]

[0451] Table 17 Tumor tissue weight after mouse dissection

[0452]

[0453]

[0454] The results showed that the average tumor endpoint volume in the negative control group was 961.75 mm 3 The average tumor volume at the endpoint of the positive control group (Gemcitabine) was 605.41 mm 3 The mean tumor volume at the end point of mice treated with MNH20645 combined with Gemcitabine was significantly reduced to 429.52 mm 3 ; On D35, the average weight of tumor tissue in the negative control group mice was about 0.92g, the average weight of tumor tissue in the positive control group mice was about 0.68g, and the average weight of tumor tissue in the experimental group was about 0.48g on the 35th day. MNH20645 combined with Gemcitabine treatment can inhibit tumor growth.

[0455] Fig.17 A shows the change curve of tumor volume at different intervention time points in each group of mice; Fig.17 B shows the tumor volume size at the end point of each group of mice. Fig.17 The mean ± standard deviation of mouse tumor volume (mm 3 ) to present the data. Fig.17Statistical analysis using two-way ANOVA showed that compared with the tumors in the negative control group, the tumor volumes of the mice in the positive control group and the MNH20645 combined with Gemcitabine experimental group were significantly reduced. Compared with the positive control group, the tumor volume of the mice in the MNH20645 combined with Gemcitabine experimental group was significantly reduced. Fig.17 Statistical analysis using one-way ANOVA with Dunnett's multiple comparison analysis showed that at the end point, the tumor volumes of mice in the positive control group and the MNH20645 combined with Gemcitabine experimental group were significantly smaller than those in the negative control group. Fig.17 Statistical analysis using one-way ANOVA with Dunnett's multiple comparison analysis showed that at the endpoint, the tumor volume of the positive control group showed a decreasing trend compared with the tumor of the negative control group, but it did not reach significance, while the tumor volume of the mice in the MNH20645 combined with Gemcitabine experimental group was significantly reduced. Fig.17 D is the mouse endpoint TGI graph. One-Way ANOVA analysis showed that at the endpoint, the endpoint TGI of mice in the positive control group and the MNH20645 combined with Gemcitabine experimental group were significantly greater than that in the negative control group. Fig.17 Figure E shows the weight of mice during the drug administration period. The results show that the weight of mice increased during the drug administration period. Two-Way ANOVA showed that there was no significant difference between the MNH20645 combined drug and the negative control and positive control groups. Fig.17 It can be clearly seen from A to D that MNH 20645 combined with Gemcitabine can significantly inhibit the growth of pancreatic cancer tumors.

[0456] Fig.17 In F to H, the changes in tumor volume of a single mouse are presented as tumor volume values. The dotted line represents the tumor volume at a 40% tumor inhibition rate relative to the negative control group. When the mouse volume is larger than the tumor volume at a 40% tumor inhibition rate in the negative control group, it means that the tumor inhibition rate is <40%, which is not responsive to anti-tumor efficacy. When the mouse volume is smaller than the tumor volume at a 40% tumor inhibition rate in the negative control group, it means that the tumor inhibition rate is ≥40%, which is responsive to anti-tumor efficacy. Compared with the 0% response rate in the negative control group, the response rate in the positive control group was 50%. The MNH20645 combined with Gemcitabine experimental group significantly increased the response rate, up to 62.5%.

[0457] In summary, MNH20645 combined with Gemcitabine can inhibit pancreatic cancer tumor growth and significantly reduce the growth rate of pancreatic cancer in mice.

[0458] Example 10: Immunoassay of MNH20645-activated liver cancer tumor mice

[0459] This embodiment is based on the following principle: after taking the tissue of tumor-bearing mice and preparing it into a single cell suspension, specific molecules with fluorescent molecule markers are labeled, and the proportion of cells expressing the specific molecules can be detected in a flow cytometer; and the proportion distribution of immune cells reflects the immune status of the mice to a certain extent.

[0460] Take an appropriate amount of tumor tissue from the experimental endpoint mice (tumor-bearing mice) in Example 5, rinse the surface with PBS, cut into small pieces and crush, place the crushed tumor tissue in 5mL collagenase IV digestion solution system (1mg / mL collagenase IV, 0.1mg / mL DNA enzyme, 10% FBS) at 37°C for digestion for 30min; the digested tissue fluid is filtered through a 70μm filter membrane to prepare a mouse tumor single cell suspension, which is tested on the machine to count the total number of cells in the tissue. Take the mouse tumor single cell suspension, add dyes (corresponding antibody combinations) Live / Fc and T-Tumor respectively for tumor cell surface T cells, and the stained cells are resuspended with PBS and tested on the machine; take the mouse tumor single cell suspension, add dyes (corresponding antibody combinations) Live / Fc and MDSC Stain in sequence for staining, and the stained cells are resuspended with PBS and tested on the machine. Single cell suspension of mouse tumor was obtained, and SP-Live and TAM Surface were added for staining respectively. The stained cells were resuspended in PBS, and then, according to the instructions of the detection kit, the fixative and permeabilization solution were used in sequence, and the antibody CD206 was added for staining; the stained cells were resuspended in PBS and tested on the machine.

[0461] The flow cytometry data were processed by CyExpert and statistically analyzed by Graphpad Prism V9. Statistical analysis was performed using one-way ANOVA with Dunnet's multiple comparison, ns not significant*p<0.05,**p<0.01,***p<0.001.

[0462] The results of the proportion of various immune cells in tumor tissue are as follows Fig.18 As shown, compared with the control group, the proportion of immune cells CD4+% / CD45+ and M1 macrophages in the MNH20645-treated group showed an upward trend, while the proportion of MDSC cells showed a downward trend.

[0463] Example 11: Immunoassay of MNH20645-activated lung cancer tumor mice

[0464] This embodiment is based on the following principle: after taking the tissue of tumor-bearing mice and preparing it into a single cell suspension, specific molecules with fluorescent molecule markers are labeled, and the proportion of cells expressing the specific molecules can be detected in a flow cytometer; and the proportion distribution of immune cells reflects the immune status of the mice to a certain extent.

[0465] Take an appropriate amount of tumor tissue from the experimental endpoint mice (tumor-bearing mice) in Example 6, take the tumor tissue, rinse the surface with PBS, cut it into small pieces and crush it, place the crushed tumor tissue in 5mL collagenase IV digestion solution system (1mg / mL collagenase IV, 0.1mg / mL DNase, 10% FBS) and digest it at 37°C for 30min; the digested tissue fluid is filtered through a 70um filter membrane to prepare a mouse tumor single cell suspension, which is tested on the machine to count the total number of cells in the tissue. Take the mouse tumor single cell suspension, add SP-Live and TAM Surface respectively for staining, resuspend the stained cells with PBS, resuspend the stained cells with PBS, then, use the fixative and membrane permeabilization solution in sequence according to the instructions of the detection kit, add the antibody CD206 for staining; the stained cells are resuspended with PBS and tested on the machine. Single cell suspension of mouse tumor was obtained, and SP-Live and Treg Surface were added for staining respectively. The stained cells were resuspended with PBS, and then treated with fixative and permeabilization solution in sequence according to the instructions of the detection kit, and antibody Foxp3 was added for staining; the stained cells were resuspended with PBS and tested on the machine.

[0466] Take an appropriate amount of spleen tissue from the experimental endpoint mouse (tumor-bearing mouse) in Example 6, rinse the surface with PBS and grind it, and then enter the red blood cell lysis step: add red blood cell lysis solution for lysis for 5 minutes, add PBS to terminate the lysis, centrifuge at 500g to remove the supernatant; after washing with PBS once, resuspend in PBS to obtain a spleen single cell suspension. Take the mouse spleen single cell suspension, add stimulant to stimulate for 4 hours, centrifuge at 500g to obtain the supernatant, add SP-Live and T-IFN / TNF Surface respectively for staining, resuspend the stained cells with PBS, resuspend the stained cells with PBS, then, use the fixative and membrane permeabilization solution in sequence according to the instructions of the detection kit, add antibody IFN / TNF-α for staining; resuspend the stained cells with PBS and detect them on the machine.

[0467] The flow cytometry data were processed by CyExpert and statistically analyzed by Graphpad Prism V9. Unpaired t test was used for statistical analysis, ns not significant*p<0.05,**p<0.01,***p<0.001.

[0468] Spleen tissue results Fig.19 As shown in A to D, compared with the control group, cytotoxic immune T cells such as TNF% / CD4+, TNF+% / CD8+, IFN+% / CD4+, and IFN+% / CD8+ in the spleen of mice showed an up-regulated trend.

[0469] Tumor tissue results Fig.19 As shown in E to G, compared with the control group, the proportion of M1 macrophages in the MNH20645-treated group showed an upward trend, while the proportion of M2 cells and Treg cells showed a downward trend.

[0470] Therefore, strain MNH20645 can exert anti-tumor activity by stimulating the activation of the mouse immune system.

[0471] Therefore, the strain MNH 20645 provided in the examples of the present application can exert anti-tumor activity by stimulating the activation of the mouse immune system.

[0472] Example 12: Immunoassay of MNH26621-activated lung cancer tumor mice

[0473] This embodiment is based on the following principle: after taking the tissue of tumor-bearing mice and preparing it into a single cell suspension, a specific molecule with a fluorescent molecule marker is labeled, and the proportion of cells expressing the specific molecule can be detected in a flow cytometer. The proportion distribution of immune cells reflects the immune status of mice to a certain extent.

[0474] Take an appropriate amount of tumor tissue from the experimental endpoint mice (tumor-bearing mice) in Example 6, take the tumor tissue, rinse the surface with PBS, cut it into small pieces and crush it, place the crushed tumor tissue in 5mL collagenase IV digestion solution system (1mg / mL collagenase IV, 0.1mg / mL DNase, 10% FBS) and digest it at 37°C for 30min; the digested tissue fluid is filtered through a 70um filter membrane to prepare a mouse tumor single cell suspension, which is tested on the machine to count the total number of cells in the tissue. Take the mouse tumor single cell suspension, add dyes (corresponding antibody combinations) T-Live-Tumor / NK-Live-Tumor, T-Tumor / NK-Tumor to stain the surface of tumor cells T cells / NK cells, resuspend the stained cells with PBS, and detect them on the machine. Take the mouse tumor single cell suspension, add dyes (corresponding antibody combinations) MDSC-Live-Tumor, MDSC-Tumor to stain tumor cells MDSC cells, resuspend the stained cells with PBS, and detect them on the machine. Take the mouse tumor single cell suspension, add the dye (corresponding antibody combination) TAM-Live-Tumor and TAM-Tumor-Surface to stain the tumor cells TAM cells, resuspend the stained cells with PBS and detect on the machine. Take the mouse tumor single cell suspension, add the stimulant to stimulate for 4h, add the dye (corresponding antibody combination) Live-Tumor and Tumor-IFN-Surface to stain the tumor cells, resuspend the stained cells with PBS; then, according to the instructions of the detection kit, use the fixative and membrane permeabilization solution in sequence, use T-IFN / TNF for staining, resuspend the stained cells with PBS and detect on the machine. Take the mouse tumor single cell suspension, add the dye (corresponding antibody combination) Treg-Live-Tumor and Treg-Surface-Tumor in sequence to stain, resuspend the stained cells with PBS; then, according to the instructions of the detection kit, use the fixative and membrane permeabilization solution in sequence, use Treg-Foxp3-Tumor nuclear staining for staining; resuspend the stained cells with PBS and detect on the machine.

[0475] Take peripheral blood and perform red blood cell lysis steps: add red blood cell lysis solution to lyse for 5 minutes, add PBS to stop lysis, centrifuge at 500g to remove supernatant; repeat red blood cell lysis once more, resuspend in PBS to obtain peripheral blood single cell suspension. Take mouse peripheral blood single cell suspension, add dyes T-Live-Blood / NK-Live-Blood and T-Blood / NK-Blood to stain T cells / NK cells on the surface of peripheral blood cells, resuspend the stained cells in PBS and detect on the machine.

[0476] The flow cytometry data were processed by CyExpert and statistically analyzed by Graphpad Prism V9. Statistical analysis was performed using one-way ANOVA with Dunnet's multiple comparison, ns not significant*p<0.05,**p<0.01,***p<0.001.

[0477] The results of the proportion of various immune cells in peripheral blood showed that compared with the control group, NK cells and CD45+CD19+ cells in the peripheral blood of mice showed an up-regulation trend (see Fig. 20 M~N). It can be seen that MNH26621 can activate the systemic immunity of mice by increasing the proportion of NK cells and CD45+CD19+ cells in the peripheral blood of mice and exerting anti-tumor effects.

[0478] The results of the respective proportions of CD45+ cells, CD45+CD3+ cells, CD45+CD8+ cells, NK cells, M1 macrophages, M2 macrophages, immunosuppressive Treg cells, MDSC cells, cytotoxic cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD8+TNFα+ cells in tumor tissues can be found in Fig. 20 A~L, compared with the control group, the CD45+CD3+ cells in the mouse tumor tissue ( Fig. 20 B), cytotoxic cells CD4+IFNγ+ cells ( Fig. 20 D) was significantly upregulated, while the proportions of CD45+ cells, CD45+CD8+ cells, NK cells, M1 macrophages, cytotoxic cells CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD8+TNFα+ cells showed an upward trend; the proportion of immunosuppressive Treg cells was significantly downregulated, while the proportions of M2 macrophages and MDSC cells showed a downward trend.

[0479] Analyze the correlation between the above cells in tumor tissue and tumor endpoint volume (see Fig.21 A~D), it can be seen that the tumor endpoint volume is significantly positively correlated with the proportion of immunosuppressive Treg cells and M2 macrophages in tumor tissue, and is significantly negatively correlated with the proportion of tumor-killing cells CD8+IFNγ+ cells and CD8+TNFα+ cells in tumor tissue.

[0480] Analyze the correlation between M2 macrophages, cytotoxic CD8+IFNγ+ cells and tumor weight in tumor tissue (see Fig. 22 A and B), it can be seen that tumor weight is significantly positively correlated with the M2 macrophages in tumor tissue, and significantly negatively correlated with the proportion of tumor-killing cells CD8+IFNγ+ cells in tumor tissue.

[0481] It can be seen that the strain provided in the examples of the present application can activate the local immunity of mouse tumors and exert an anti-tumor effect by inhibiting the proportion of MDSC cells, M2 macrophages, and immunosuppressive Treg cells in mouse tumors, and increasing the proportion of tumor CD45+CD3+ cells, CD45+ cells, CD45+CD8+ cells, NK cells, M1 macrophages, cytotoxic cells CD4+IFNγ+ cells, CD8+IFNγ+ cells, CD4+TNFα+ cells, and CD4+TNFα+ cells.

[0482] Therefore, the strain MNH26621 provided in the examples of the present application can exert anti-tumor activity by stimulating the activation of the mouse immune system.

[0483] The present application is described in detail above in conjunction with the embodiments, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present application. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

Claims

1. A strain of the genus Parabacteroides, characterized in that it is any one of the following A1) to A5): A1) A strain whose 16S rRNA has at least 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity with the sequence shown in SEQ ID NO: 1; A2) A strain with the deposit number GDMCC No: 62664 or GDMCC No: 62665; A3) A strain whose 16S rRNA has at least 96.74%, 97%, 98%, 98.5%, 98.65%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% identity with the 16S rRNA of the strain described in A2); A4) A strain whose 16S rRNA has at least 97%, 98%, 98.65%, 99%, 99.5%, 99.9%, 100% identity with the sequence shown in SEQ ID NO: 2; Preferably, the strain is selected from the species Parabacteroides hominis; A5) A strain whose genome has an average nucleotide identity of at least 87%, 88%, 88.81%, 88.89%, 89%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, 100% with the genome of the strain described in any one of A1) to A4); and / or, a strain whose genome has a alignment score of at least 70%, 73.54%, 74.55%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% with the genome of the strain described in any one of A1) to A4); Preferably, the sequence of the 16S rRNA of the strain with the deposit number GDMCC No: 62664 is as shown in SEQ ID NO: 1, and the sequence of the 16S rRNA of the strain with the deposit number GDMCC No: 62665 is as shown in SEQ ID NO: 2; Preferably, the strain is derived from the human intestine; Preferably, the strain can produce high yields of short-chain fatty acids; Preferably, the strain can produce high yields of acetic acid and / or propionic acid.

2. A culture of the Parabacteroides strain according to claim 1; Preferably, the culture includes any one of the following B1) to B5): B1) The fermentation medium of the strain; B2) The supernatant of the fermentation medium of the strain; B3) A concentrated product of B1) or B2); B4) An extract of B1) or B2); B5) A dried product of any one of B1) to B4); Preferably, the fermentation medium is a liquid medium; Preferably, the culture is obtained using a liquid medium under anaerobic culture conditions; Preferably, the fermentation medium is MM01 liquid medium, and / or the culture temperature is 35-42°C, and / or the culture time is 24-72 hours, and / or the pH of the medium is 6-10.

3. Bacterial agent, characterized in that, the bacterial agent comprises the Parabacteroides strain described in claim 1, or the metabolite of the Parabacteroides strain described in claim 1, or the culture described in claim 2.

4. Composition, characterized in that, the composition or the raw material of the composition comprises the Parabacteroides strain described in claim 1, or the metabolite of the Parabacteroides strain described in claim 1, or the culture described in claim 2, or the bacterial agent described in claim 3; Preferably, the composition further comprises at least one of excipients, diluents, carriers; Preferably, the composition further comprises adjuvants; Preferably, the adjuvants further comprise at least one of lubricants, wetting agents, binders, emulsifiers, suspension stabilizers, solubilizers, preservatives, sweeteners, fragrances; Preferably, the composition is at least one of drugs, nutritional supplements, foods, food additives, feeds, feed additives; Preferably, the composition is a solid preparation or a liquid preparation; Preferably, the dosage form of the composition is any one of powders, granules, tablets, capsules, gels, suspensions, drops, dripping pills, injections, suppositories, aerosols, oral liquids, ointments, emulsions, lavage solutions; Preferably, the composition further comprises a product instruction manual.

5. Pharmaceutical composition, characterized in that, it comprises the following C1) and C2): C1) the Parabacteroides strain described in claim 1, or the metabolite of the Parabacteroides strain described in claim 1, or the culture described in claim 2, or the bacterial agent described in claim 3; C2) a combined drug; Preferably, the combined drug comprises at least one of anti-tumor drugs, drugs for metabolic diseases, drugs for inflammatory or autoimmune diseases, drugs for infectious diseases, drugs for central nervous system diseases; Preferably, the anti-tumor drugs comprise at least one of chemotherapy drugs, photosensitizers, photothermal agents, immunotherapy drugs; Preferably, the chemotherapy drugs comprise at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, epirubicin; Preferably, the photosensitizers comprise at least one of boron dipyrrole, chlorin, erythrosin B; Preferably, the photothermal agents comprise at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, semiconductor nanomaterials; Preferably, the immunotherapy drugs comprise at least one of immunocyte therapy drugs, immune checkpoint inhibitors; Preferably, the immunocyte therapy drugs comprise at least one of T cell therapy drugs, tumor-infiltrating lymphocyte therapy drugs, NK cell therapy drugs; Preferably, the immune checkpoint inhibitor includes an inhibitor targeting at least one of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, TIGIT, VISTA, BTLA, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, KLRA; Preferably, the pharmaceutical composition comprises 1×10 3 CFU / g~1×10 13 CFU / g or 1×10 3 CFU / mL~1×10 13 CFU / mL of the Parabacteroides strain; Preferably, each gram or milliliter of 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 colony forming units (CFU) of the Parabacteroides strain; Preferably, the Bacteroides strain in the pharmaceutical composition is live bacteria, attenuated bacteria, killed bacteria, freeze-dried bacteria or irradiated bacteria; Preferably, the pharmaceutical composition or the raw materials of the pharmaceutical composition include 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% of C1); Preferably, the pharmaceutical composition or the raw materials of the pharmaceutical composition include 1-80%, 2-70%, 5-60%, 10-50%, 20-40%, 40-60% of C1); Preferably, the pharmaceutical composition further includes at least one of an excipient, a diluent, and a carrier; Preferably, the pharmaceutical composition further includes auxiliary materials; Preferably, the auxiliary materials include at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a fragrance; Preferably, the pharmaceutical composition is a dosage form suitable for infants, children or adults; Preferably, the pharmaceutical composition is a dosage form for gastrointestinal administration or non-gastrointestinal administration; Preferably, the pharmaceutical composition is an oral preparation or an injection; Preferably, 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 pill, an injection, a suppository, an aerosol, an oral liquid, an ointment, an emulsion, and an irrigation solution; Preferably, the strain can at least partially proliferate in the intestine of the subject; Preferably, the pharmaceutical composition further includes a product instruction manual.

6. Pharmaceutical combination product, Characterized in that, Comprising independently: A first product, wherein the first product comprises the Parabacteroides strain according to claim 1, or comprises the metabolite of the Parabacteroides strain according to claim 1, or comprises the culture according to claim 2, or comprises the microbial agent according to claim 3; A second product, wherein the second product comprises a combined medicine; Preferably, the combined medicine comprises at least one of a tumor drug, a metabolic disease drug, an inflammatory or autoimmune disease drug, an infectious disease drug, and a central nervous system disease drug; Preferably, the tumor drug comprises at least one of a chemotherapy drug, a photosensitizer, a photothermal agent, and an immunotherapy drug; Preferably, the chemotherapy drug comprises at least one of paclitaxel, camptothecin, 5-fluorouracil, cisplatin, doxorubicin, mitomycin, and epirubicin; Preferably, the photosensitizer comprises at least one of boron dipyrrole, chlorin, and eosin B; Preferably, the photothermal agent comprises at least one of noble metal nanoparticles, organic polymers, carbon-based nanomaterials, magnetic nanomaterials, and semiconductor nanomaterials; Preferably, the immunotherapy drug comprises at least one of an immune cell therapy drug and an immune checkpoint inhibitor; Preferably, the immune cell therapy drug comprises at least one of a T cell therapy drug, a tumor infiltrating lymphocyte therapy drug, and an NK cell therapy drug; Preferably, 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, CD27, CD28, CD70, CD80, CD86, CD137, CD276, KIRs, TNFRSF4, GITR, GITRL, 4-1BBL, A2aR, VTCN1, IDO, and KLRA; Preferably, the first product comprises 1×10 3 CFU / g to 10 13 CFU / g or 1×10 3 CFU / mL to 10 13 CFU / mL of the strain of Parabacteroides; Preferably, each gram or milliliter of the first product comprises at least 1×10 3 、2×10 3 、5×10 3 、1×10 4 、2×10 4 、5×10 4 、1×10 5 、2×10 5 、5×10 5 、1×10 6 、2×10 6 、5×10 6 、1×10 7 、2×10 7 、5×10 7 、1×10 8 、2×10 8 、5×10 8 、1×10 9 、2×10 9 、5×10 9 、1×10 10 、2×10 10 、5×10 10 、1×10 11 、2×10 11 、5×10 11 、1×10 12 、2×10 12 、5×10 12 、1×10 13 colony-forming units (CFU) of the Parabacteroides strain; Preferably, the Parabacteroides strain in the first product is a live bacterium, an attenuated bacterium, a killed bacterium, a freeze-dried bacterium, or an irradiated bacterium; Preferably, the first product and / or the second product further comprises at least one of an excipient, a diluent, and a carrier; Preferably, the first product and the second product each independently further comprises at least one auxiliary material; Preferably, the auxiliary material comprises at least one of a lubricant, a wetting agent, a binder, an emulsifier, a suspension stabilizer, a solubilizer, a preservative, a sweetener, and a fragrance; Preferably, the pharmaceutical combination product is in a dosage form suitable for infants, children, or adults; Preferably, the first product and the second product are each independently a dosage form for gastrointestinal administration or a dosage form for non-gastrointestinal administration; Preferably, the first product and the second product are each independently an oral preparation or an injection; Preferably, the first product and the second product are each independently any one of a powder, a granule, a tablet, a capsule, a gel, a suspension, a drop pill, an injection, a suppository, an aerosol, an oral liquid, an ointment, an emulsion, and an irrigation solution; Preferably, the Parabacteroides strain can at least partially proliferate in the intestine of a subject; Preferably, the pharmaceutical combination product further comprises a product instruction manual.

7. Use of the Parabacteroides strain according to claim 1, the culture according to claim 2, the bacterial agent according to claim 3, the composition according to claim 4, the pharmaceutical composition according to claim 5 or the pharmaceutical combination product according to claim 6 in the preparation of a drug for preventing and / or treating a disease; Preferably, the disease includes at least one of tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, glandular tumors and metastatic tumors; Preferably, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic cancer, osteoma, soft tissue sarcoma, 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 a tumor in at least one of blood, lymph and bone marrow; Preferably, the tumor or cancer includes at least one of oral 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, appendiceal cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic cholangiocarcinoma, gallbladder cancer, hilar cholangiocarcinoma, distal cholangiocarcinoma, ampullary carcinoma of the hepatopancreatic ampulla, pancreatic cancer, gastric neuroendocrine tumor, neuroendocrine tumor of the duodenum and ampulla of Vater, jejuno-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, carcinosarcoma of the uterus, uterine corpus sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvic 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 cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma and leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or viruses include at least one of hepatitis B virus, hepatitis C virus, human papillomavirus, Epstein-Barr virus, Helicobacter pylori and Fusobacterium nucleatum; Preferably, the metabolic disease includes at least one disease with abnormal metabolism 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 psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, allergic dermatitis, acne, Crohn's disease, ulcerative colitis, bronchitis, allergic rhinitis, Graves' disease, Hashimoto's thyroiditis, rheumatoid arthritis, type 1 diabetes, systemic lupus erythematosus, scleroderma, glomerulonephritis, systemic vasculitis, Addison's disease, autoimmune myositis, and Sjogren's syndrome; Preferably, the infectious disease includes a disease caused by at least one infection 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, preventing and / or treating tumors is achieved by at least one of the following (a) to (l): (a) Inhibiting tumor volume growth; (b) Inhibiting tumor weight increase; (c) Inhibiting tumor cell growth; (d) Increasing the tumor treatment response rate; (e) Improving the therapeutic efficacy of immunosuppressive drugs, such as improving the therapeutic efficacy of anti-PD-1 antibody treatment; (f) Preventing and / or inhibiting the spread or metastasis of tumor cells; (g) Reducing tumor drug resistance; (h) Inhibiting HDAC activity by at least one of acetic acid or acetate, propionic acid or propionate, butyric acid or butyrate, and valeric acid or valerate in SCFA; (i) Inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) Regulating or activating the subject's immune system; (k) Promoting tumor cell senescence or apoptosis; (l) Inhibiting angiogenesis in tumor tissues.

8. Use of the Parabacteroides strain according to claim 1, the culture according to claim 2, the bacterial agent according to claim 3, the composition according to claim 4, the pharmaceutical composition according to claim 5, or the pharmaceutical combination product according to claim 6 in preventing and / or treating diseases; Preferably, the disease includes at least one of tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases, and central nervous system diseases; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, glandular tumors, and metastatic tumors; Preferably, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic cancer, osteoma, soft tissue sarcoma, 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 a tumor in at least one of blood, lymph, and bone marrow; Preferably, the tumor or cancer includes at least one of oral 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, appendiceal cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic cholangiocarcinoma, gallbladder cancer, hilar cholangiocarcinoma, distal cholangiocarcinoma, ampullary cancer of the hepatopancreatic ampulla, pancreatic cancer, gastric neuroendocrine tumor, neuroendocrine tumor of the duodenum and ampulla of Vater, jejuno-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, carcinosarcoma of the uterus, uterine sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney cancer, renal pelvic cancer, ureteral cancer, bladder cancer, urethral cancer, eyelid cancer, conjunctival cancer, conjunctival melanoma, uveal melanoma, retinoblastoma, lacrimal gland cancer, orbital sarcoma, lymphoma of the accessory organs of the eye, brain cancer, spinal cord tumor, differentiated thyroid cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, and leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or viruses include 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 with 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 psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, atopic 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 Sjogren's syndrome; Preferably, the infectious disease includes a disease 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, preventing and / or treating a tumor is achieved by at least one of the following (a) - (l): (a) Inhibiting tumor volume growth; (b) Inhibiting tumor weight increase; (c) Inhibiting the growth of tumor cells; (d) Increasing the tumor treatment response rate; (e) Enhancing the therapeutic efficacy of immunosuppressive drugs, such as enhancing the therapeutic efficacy of anti-PD-1 antibodies; (f) Preventing and / or inhibiting the spread or metastasis of tumor cells; (g) Reducing the drug resistance of anti-tumor drugs; (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 SCFAs; (i) Inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) Regulating or activating the immune system of a subject; (k) Promoting tumor cell senescence or apoptosis; (l) Inhibiting angiogenesis in tumor tissues.

9. A method for treating a disease, comprising administering to a subject an effective dose of the Bacteroides parathensis strain according to claim 1, the culture according to claim 2, the bacterial agent according to claim 3, the composition according to claim 4, the pharmaceutical composition according to claim 5 or the pharmaceutical combination product according to claim 6; Preferably, the disease includes at least one of tumors or cancers, metabolic diseases, inflammatory or autoimmune diseases, infectious diseases and central nervous system diseases; Preferably, the tumor or cancer includes at least one of solid tumors, soft tissue tumors, hematopoietic tumors, glandular tumors and metastatic tumors; Preferably, the solid tumor includes at least one of head and neck cancer, upper gastrointestinal cancer, lower gastrointestinal cancer, hepatobiliary system cancer, neuroendocrine tumor, thoracic cancer, osteoma, soft tissue sarcoma, 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 tumors in at least one of blood, lymph and bone marrow; Preferably, the tumor or cancer includes at least one of oral 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, appendiceal cancer, colon cancer, rectal cancer, anal canal cancer, liver cancer, intrahepatic cholangiocarcinoma, gallbladder cancer, hilar cholangiocarcinoma, distal cholangiocarcinoma, ampullary cancer of the hepatopancreatic ampulla, pancreatic cancer, gastric neuroendocrine tumor, neuroendocrine tumor of the duodenum and ampulla of Vater, jejuno-ileal neuroendocrine tumor, appendiceal neuroendocrine tumor, colorectal neuroendocrine tumor, pancreatic neuroendocrine tumor, thymic cancer, lung cancer, malignant pleural mesothelioma, angiosarcoma, desmoid tumor, Ewing sarcoma, fibrosarcoma, gastrointestinal stromal tumor, Kaposi sarcoma, leiomyosarcoma, liposarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, cutaneous malignant melanoma, vulvar cancer, vaginal cancer, cervical cancer, uterine corpus cancer, carcinosarcoma of the uterus, uterine sarcoma, ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gestational trophoblastic tumor, penile cancer, prostate cancer, testicular cancer, kidney 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 accessory organs of the eye, brain cancer, spinal cord tumor, differentiated thyroid cancer, undifferentiated thyroid cancer, medullary thyroid cancer, parathyroid cancer, adrenocortical cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, cutaneous lymphoma, plasma cell myeloma, and leukemia; Preferably, the tumor or cancer includes a tumor or cancer caused by bacteria or viruses; Preferably, the bacteria or viruses include 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 with 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 psoriasis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, chronic obstructive pulmonary disease, glomerulonephritis, myocarditis, dry eye, uveitis, Behcet's disease, asthma, atopic 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 Sjogren's syndrome; Preferably, the infectious disease includes a disease 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, preventing and / or treating a tumor is achieved by at least one of the following (a) to (l): (a) Inhibiting tumor volume growth; (b) Inhibiting tumor weight increase; (c) Inhibiting the growth of tumor cells; (d) Improving the response rate of tumor treatment; (e) Enhancing the therapeutic efficacy of immunosuppressive drugs, such as enhancing the therapeutic efficacy of anti-PD-1 antibodies; (f) Preventing and / or inhibiting the spread or metastasis of tumor cells; (g) Reducing the drug resistance of anti-tumor drugs; (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 SCFAs; (i) Inhibiting HDAC activity and / or promoting IFNβ transcriptional activity; (j) Regulating or activating the immune system of a subject; (k) Promoting the senescence or apoptosis of tumor cells; (l) Inhibiting angiogenesis in tumor tissues; Preferably, the number of administrations of the Parabacteroides strain, the culture, the bacterial agent, the composition, the pharmaceutical composition or the pharmaceutical combination product is at least once a day; Preferably, when the number of administrations is more than twice a day, the same dose is administered each time; Preferably, when the number of administrations is more than twice a day, different doses are administered each time; Preferably, the administration routes include at least one of oral administration, sublingual administration, nasal administration, rectal administration, inhalation administration, transdermal administration, intraperitoneal injection, subcutaneous injection, intradermal injection, intramuscular injection, intravenous injection.

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

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