Novel bifidobacterium sp. strain, and composition for preventing, alleviating, or treating fatty liver diseases, comprising same as active ingredient

Abstract

The present application relates to; a pharmaceutical composition, a food composition and a feed composition for preventing or treating fatty liver diseases; and a method for treating fatty liver diseases by using same, the pharmaceutical composition comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof. The Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, according to the present invention, alleviates inflammatory cell infiltration and fibrosis in the liver, and exhibits the effects of inhibiting cholesterol synthesis and inhibiting triglyceride accumulation in the liver, and thus can be used for treating fatty liver diseases, particularly nonalcoholic fatty liver disease.

Description

A novel strain of the genus Bifidobacterium, and a composition for preventing, improving, or treating fatty liver disease comprising the same as an effective ingredient

[0001] The present application relates to a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP and a composition for preventing, improving or treating fatty liver disease, including the same.

[0002]

[0003] As modern society has shifted to a meat-based diet, calorie intake has increased while exercise has decreased, leading to a rapid increase in the incidence of metabolic diseases, including obesity, diabetes, hyperlipidemia, non-alcoholic fatty liver disease, and dyslipidemia.

[0004] Fatty liver disease is defined as a condition in which the weight of the liver due to fat deposition is more than 5% of the weight of the liver. Among them, non-alcoholic fatty liver disease (NAFLD) refers to cases in which the cause of fatty liver is not due to viruses, drugs, genetics, or alcohol. It is a chronic liver disease known to be closely related to metabolic syndrome such as insulin resistance, obesity, hypertension, and dyslipidemia, and includes a series of processes ranging from simple steatosis to non-alcoholic steatohepatitis (NASH) and cirrhosis. The prevalence of non-alcoholic fatty liver disease is reported to vary worldwide depending on the population. In the West, it is known that approximately 20-30% of normal adults without specific causes of liver disease have non-alcoholic fatty liver disease, and in Korea, the prevalence of non-alcoholic fatty liver disease in adults is reported to be approximately 16-50%. Non-alcoholic fatty liver disease is generally known to have a higher prevalence in obese individuals, as insulin resistance caused by obesity is known to be a major cause of fat deposition in the liver. Mild fatty liver disease, such as simple fat deposition, which only has fat accumulated and no damage to liver cells, has a relatively good prognosis, but if left untreated, it can progress to cirrhosis, a serious liver disease. 4-27% of patients with cirrhosis develop liver cancer, and 30-40% of patients with cirrhosis die from liver disease complications and cardiovascular disease after 10 years. Therefore, it is a disease that requires active management from the early stages.

[0005] Accordingly, various treatments targeting non-alcoholic fatty liver disease (NAFLD) have been developed. For example, Korean Patent Publication No. KR 10-2016-0107420 A discloses a NAFLD treatment comprising ginsenoside F2. However, the cause of NAFLD remains unclear, making it challenging to develop effective treatments. Furthermore, because the liver, the target organ, is an organ where various detoxification processes are performed, using drugs with enhanced pharmacological activity carries the risk of damaging liver cells, making the development of effective drugs challenging.

[0006] Therefore, the development of an excellent treatment for the overall treatment of fatty liver disease is currently insufficient, and there is an urgent need for the development of a treatment that is both effective and safe.

[0007]

[0008] The problem to be solved by the present invention is to provide a pharmaceutical composition for preventing or treating fatty liver disease, a food composition, or a feed composition comprising a Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP; the strain, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0009]

[0010] One purpose of the present application is to provide a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP.

[0011] Another object of the present application is to provide a pharmaceutical composition for preventing or treating fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0012] Another purpose of the present application is to provide a food composition for preventing or improving fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0013] Another object of the present application is to provide a feed composition for preventing or improving fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0014]

[0015] The composition comprising the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP according to the present invention, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, improves inflammatory cell infiltration and fibrosis in the liver, and exhibits a remarkable effect on inhibiting cholesterol synthesis and neutral fat accumulation in the liver, and thus can be usefully applied to the treatment of non-alcoholic fatty liver disease.

[0016] In addition, there is a synergistic effect in reducing liver weight, suppressing fat accumulation in the liver, and improving inflammatory cell infiltration and fibrosis in the liver when combined with the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP according to the present invention and tazemetostat, so that their combined administration can be utilized for more effective treatment of fatty liver disease.

[0017]

[0018] Figure 1 is a diagram showing the results of analyzing the similarity between the 16S rRNA gene sequences of existing Bifidobacterium pseudolongum strains and the whole genome sequence of the Bifidobacterium pseudolongum strain deposited with accession number KCTC 15133BP isolated in Example 1.

[0019] Figure 2 is a diagram showing the results of verification of the NASH development inhibition effect of Bifidobacterium pseudolongum (KCTC 15133BP) strain in the NASH mouse model in Example 2. Specifically, Figure 2A is a schematic diagram showing the method for establishing the NASH diet mouse model used in Example 2. Figure 2B is a diagram showing the results of Sirius red, oil red o (ORO), hematoxylin and eosin (H&E) staining and F4 / 80 value analysis of the normal control group (NC), the NASH diet mouse group (NASH), and the group administered the KCTC 15133BP strain to the NASH diet mouse model (NASH+KGMB04).

[0020] Figure 3 shows the results of the metagenome analysis of the NASH diet mouse model, where NC represents the normal control group and NASH represents the NASH diet mouse model.

[0021] Figure 4 shows the results of short-chain fatty acids (SCFAs) analysis of cultures containing each strain of the genus B. pseudolongum.

[0022] Figure 5 is a diagram showing the results of measuring lipid (triglyceride, TG) levels in a strain of the genus B. pseudolongum. Figure 5A is a schematic diagram of the T3-L1 preadipocyte differentiation process, Figure 5B is a diagram showing the production of lipid droplets in 3T3-L1 adipocyte cells, and Figure 5C is a diagram showing the results of measuring TG levels in 3T3-L1 adipocyte cells.

[0023] Figure 6 is a diagram showing the relative expression levels of methionine cycle-related genes (Gnmt1, Ahcy) and citrulline precursor synthesis gene (Cps1), an important metabolite of the urea cycle, in the standard strain (KCTC 3224T, TT) and the strain of the present invention (KGMB04131, KK) treatment groups in the MCD diet steatohepatitis mouse model.

[0024] Figure 7 is a diagram showing the expression level of the histone methylase EZH2 gene in liver tissues derived from NASH patients of the Korean and European groups.

[0025] Figure 8 is a diagram confirming the effect of a group administered tazemetostat alone in a NASH mouse model, and specifically, Figure 8A is a diagram showing a method for establishing a NASH dietary mouse model, Figure 8B is a diagram showing the results of confirming the morphology of the liver, tissue staining (H&E), fat-specific staining (Oil red O staining), and the level of inflammatory cell infiltration (F5 / 80+), and Figure 8C is a diagram showing the weight and body weight of the liver.

[0026] Figure 9 is a diagram showing the results of a metagenomic analysis of a NASH diet mouse model. NC represents a normal control group, NASH represents a NASH diet mouse model, and TAZ represents a NASH diet mouse model administered with tazemetostat.

[0027] Figure 10A is a diagram showing a method for establishing a NASH dietary mouse model, and Figure 10B is a diagram showing the results of liver morphology and tissue staining (H&E) and fat-specific staining (Oil red O staining) and the level of inflammatory cell infiltration (F5 / 80+) when tazemetostat and KCTC 15133BP strain were administered together in a NASH mouse model.

[0028] Figure 11 is a diagram showing the results of analysis at the transcriptome level through RNA-seq of mouse liver tissues of the single administration group of tazemetostat and KCTC 15133BP strain and the combined administration group.

[0029] Figure 12 is a diagram showing the results of gene network analysis in a NASH diet mouse model.

[0030] Figure 13 is a diagram showing the relative expression levels of methionine cycle-related genes (Gnmt1, Ahcy) and citrulline precursor synthesis gene (Cps1), an important metabolite of the urea cycle, in the single administration group of tazemetostat and KCTC 15133BP strain and the combined administration group in a NASH diet mouse model.

[0031]

[0032] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in this application can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not limited by the specific descriptions described below. Furthermore, numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated into this specification in their entirety by reference to more clearly explain the level of the technical field to which this application belongs and the contents of this application.

[0033]

[0034] One aspect of the present invention provides a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP.

[0035] In the present invention, the term "Bifidobacterium" strain refers to a non-motile, anaerobic gram-positive bacterium belonging to the actinomycetes. Bifidobacterium strains are microorganisms included in the Ministry of Food and Drug Safety Notice 19 and have excellent stability. The Bifidobacterium strains include the Bifidobacterium pseudolongum strain of the present invention, and in addition, many strains such as Bifidobacterium longum, Bifidobacterium lactis, and Bifidobacterium bifidum belong to this group.

[0036] In the present invention, the term "Bifidobacterium pseudolongum" refers to one of the microorganisms belonging to the genus Bifidobacterium, which is generally distributed in the intestines of mammals. The Bifidobacterium pseudolongum strain may be used interchangeably as an abbreviated name for B. pseudolongum.

[0037] The Bifidobacterium pseudolongum strain of the present invention may be derived from a human body, and specifically, may be derived from the human intestinal microbiome, but is not limited thereto. In addition, the Bifidobacterium pseudolongum strain may be isolated from feces or the like, but it should be understood that the method of derivation and isolation is not limited as long as it has the characteristics of the strain.

[0038] The above Bifidobacterium pseudolongum strain may be a live strain or an attenuated strain (killed strain).

[0039] For the purposes of the present invention, the Bifidobacterium pseudolongum strain may be, but is not limited to, the strain deposited under the accession number KCTC 15133BP. In addition, the inventors of the present invention also named the KCTC 15133BP strain KGMB04131, and therefore, the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP may be used interchangeably under the names KCTC 15133BP and KGMB04131.

[0040]

[0041] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0042] The present invention discloses for the first time a novel use of the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP for the treatment of fatty liver disease.

[0043]

[0044] In the present invention, the term "culture" means a culture solution, a concentrated culture solution, a dried product of a culture solution, a culture filtrate, a concentrated culture filtrate, a dried product of a culture filtrate, a dilution, a concentrate, a fermentation product, etc., obtained by culturing a specific microorganism in a culture medium, and the culture solution means that it contains a specific bacteria, and the culture filtrate means that it does not substantially contain a specific bacteria (here, "substantially" means that a specific bacteria separated by filtration or the like is excluded, but does not mean that the bacteria are completely excluded from the filtrate).

[0045] In the present invention, the term "crusted material" refers to a crusted material of the deposited Bifidobacterium pseudolongum strain of the present invention, and may include all useful substances present in the strain that can be released by crusting the strain.

[0046] In the present invention, the term "extract" refers to an extract obtained by extracting the deposited Bifidobacterium pseudolongum strain of the present invention using a solvent. Any known solvent may be included without limitation, and the extraction method may also be included without limitation. In addition, the extract may include an extract obtained by extracting the deposited Bifidobacterium pseudolongum strain itself, or an extract obtained by extracting a culture of the strain or a lysate of the strain.

[0047]

[0048] The present inventors have found that the Bifidobacterium pseudolongum strain, particularly the novel strain KCTC 15133BP, has a remarkable effect on the treatment of fatty liver disease.

[0049] The term "fatty liver disease" as used herein refers to a condition in which fat accumulates in excess of 5% of the liver, compared to a normal liver. Fatty liver disease can be broadly divided into alcoholic fatty liver disease caused by excessive drinking and non-alcoholic fatty liver disease. Specifically, the fatty liver disease in the present invention may be non-alcoholic fatty liver disease.

[0050] The term "non-alcoholic fatty liver disease (NAFLD)" in the present invention refers to a fatty liver disease not caused by alcohol, and includes a series of processes ranging from simple steatosis in the liver to steatohepatitis and cirrhosis. The causes of this include fat deposition in the liver due to insulin resistance, excessive nutritional intake, and genetic causes. In this case, the term "non-alcoholic fatty liver disease" may be used interchangeably with the terms NAFLD and MASLD (Metabolic dysfunction-associated steatotic liver disease).

[0051] Specifically, the non-alcoholic fatty liver disease may be at least one selected from the group consisting of non-alcoholic fatty liver, non-alcoholic steatohepatitis (NASH), and non-alcoholic liver cirrhosis, but is not limited thereto. In this case, the non-alcoholic steatohepatitis may be used interchangeably with the name NASH or MASH (metabolic dysfunction-associated steatohepatitis).

[0052]

[0053] In the present invention, the term "prevention" means any act of inhibiting or delaying the onset or progression of fatty liver disease by administering a pharmaceutical composition containing the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP of the present invention, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0054] As used herein, the term "treatment" refers to clinical intervention to alter the natural process of a subject or cell to be treated, and this can be performed during the progression of a clinical pathological condition or to prevent it. The desired therapeutic effect includes preventing the occurrence or recurrence of a disease, alleviating symptoms, reducing all direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, alleviating or temporarily alleviating the disease state, reversing it, or improving the prognosis. The present invention includes all acts of improving the course of fatty liver disease by administering a pharmaceutical composition comprising a Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0055]

[0056] The Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP of the present invention and the composition containing the same may improve inflammatory cell infiltration and fibrosis in the liver.

[0057] In one embodiment of the present invention, it was confirmed that the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP has excellent efficacy in improving steatosis, inflammatory cell infiltration, and fibrosis in the liver (Example 2).

[0058]

[0059] The Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP of the present invention and the composition containing the same may have an effect of inhibiting cholesterol synthesis in the liver and an effect of inhibiting neutral fat accumulation.

[0060] In one embodiment of the present invention, it was confirmed that the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP has excellent propionic acid production ability, excellent cholesterol synthesis inhibition ability in the liver, and excellent neutral fat accumulation inhibition effect, and thus has a significantly superior non-alcoholic fatty liver disease treatment effect compared to other Bifidobacterium pseudolongum strains (Example 4).

[0061]

[0062] The Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP of the present invention and the composition containing the same may increase the expression of the GNMT (glycine N-methyltransferase) gene, which are genes related to the methionine cycle in the liver, the AHCY (adenosylhomocysteinase) gene, and / or the CPS1 (Carbamoyl-Phosphate Synthase 1) gene, which is a precursor synthesis gene of citrulline, which is an important metabolite of the urea cycle, or restore the decreased expression.

[0063] In one embodiment of the present invention, it was confirmed that the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP had a superior effect on restoring GNMT, AHCY, and CPS1 gene expression compared to the standard strain (Example 5).

[0064]

[0065] As described above, the present invention has the major significance in that it has been confirmed for the first time that the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP improves steatosis, inflammatory cell infiltration, and fibrosis in the liver, has excellent propionic acid production ability, and has an inhibitory effect on neutral fat accumulation, and thus can be applied to the prevention and treatment of non-alcoholic fatty liver disease.

[0066]

[0067] In one specific example, the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof in the pharmaceutical composition of the present invention may be included in an amount of 0.00001 wt% to 99.99 wt% based on the total weight of the pharmaceutical composition, specifically 0.1 wt% to 90 wt%, more specifically 0.1 wt% to 70 wt%, and even more specifically 0.1 wt% to 50 wt%, but is not limited thereto, and may be variously changed depending on the condition of the administration subject, the type and degree of specific symptom, etc. If necessary, it may also be included in the total content of the pharmaceutical composition.

[0068] In one specific example, in the pharmaceutical composition of the present invention, the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP may be included in the composition in a therapeutically effective amount or nutritionally effective concentration, for example, about 1x10 1 cfu to about 1x 10 20 The composition may be included as a content of cfu or as a culture of an equivalent number of live or dead cells.

[0069]

[0070] In one specific embodiment, the pharmaceutical composition of the present invention may be co-administered with Tazemetostat.

[0071] In the present invention, the term "tazemetostat" refers to a type of anticancer agent that acts as an EZH2 inhibitor and has a structure represented by the following chemical formula 1. In the present invention, the tazemetostat can be understood as a concept that includes all compounds represented by the following chemical formula 1 or pharmaceutically acceptable salts thereof. Tazemetostat is also known as a therapeutic agent for epithelioid sarcoma. Meanwhile, tazemetostat has the IUPAC name of N-[(4,6-Dimethyl-2-oxo-1H-pyridin-3-yl)methyl]-3-[ethyl(oxan-4-yl)amino]-2-methyl-5-[4-(morpholin-4-ylmethyl)phenyl]benzamide. Such a compound may be commercially available through a known synthetic method.

[0072]

[0073]

[0074]

[0075] Specifically, the tazemetostat may be in the form of itself, a salt thereof (e.g., a pharmaceutically acceptable salt), or a solvate thereof. Furthermore, tazemetostat may be in any pharmaceutically acceptable form.

[0076] The type of the above salt is not particularly limited. However, it is preferable that it be in a form that is safe and effective for an individual, such as a mammal, but is not particularly limited thereto.

[0077] As used herein, the term "pharmaceutically acceptable salt" includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, and ammonium.

[0078] Additionally, the term "solvate" used in the present invention refers to a peptide, compound or salt thereof according to the present invention formed in a complex with a solvent molecule.

[0079]

[0080] In the present invention, the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP can have a synergistic effect on the prevention or treatment of fatty liver disease when administered in combination with tazemetostat.

[0081] In one embodiment of the present invention, when a combination of the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP and tazemetostat was co-administered to an individual, it was confirmed that the size and weight of the liver were significantly reduced, and fat accumulation and inflammatory cell infiltration in the liver were significantly improved compared to when each component was administered alone. This confirmed a remarkable synergistic effect in the treatment of fatty liver disease when the KCTC 15133BP strain and tazemetostat were co-administered.

[0082]

[0083] In the present invention, the terms "combined administration," "combined use," or "using in combination" should be understood not only to mean simultaneous administration, but also to mean an administration form in which the pharmaceutical composition containing the strain of the present invention and tazemetostat act together in a subject so that each substance can perform a level equivalent to or higher than its original function. Therefore, when the term "combined use" is used herein, it should be understood that it refers to simultaneous, separate, sequential, or reverse administration, and the order is unlimited. When the administration is sequential, reverse, or separate, the order of administration is not particularly limited, but the interval between administrations of two or more components should be such that the beneficial effects of the combination are not lost.

[0084]

[0085] In one specific example, when the tazemetostat is included in the same composition as the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP and administered simultaneously, it may be included in an amount of 0.00001 wt% to 80 wt% based on the total weight of the pharmaceutical composition, but is not limited thereto. In this case, as an example, the pharmaceutical composition of the present invention may further include tazemetostat.

[0086]

[0087] A composition comprising the pharmaceutical composition of the present invention may additionally contain one or more active ingredients exhibiting the same or similar function.

[0088] The pharmaceutical composition of the present invention may additionally include a pharmaceutically acceptable carrier in addition to the pharmaceutical composition of the present invention.

[0089]

[0090] The pharmaceutical composition according to the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods, and may include appropriate carriers, excipients, or diluents conventionally used in the manufacture of pharmaceutical compositions for formulation.

[0091] The carrier or excipient or diluent may include various compounds or mixtures including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0092] When formulating, it can be manufactured using diluents or excipients such as fillers, weighting agents, binders, wetting agents, disintegrants, and surfactants that are commonly used.

[0093] Solid dosage forms for oral administration can be prepared by mixing the strain with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

[0094] Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives.

[0095] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, and glycerol gelatin.

[0096] The preferred dosage of the pharmaceutical composition and concomitant ingredients according to the present invention varies depending on the patient's condition, body weight, severity of the disease, drug form, route of administration, and duration of administration, but can be appropriately selected by those skilled in the art. However, for desirable effects, the dosage may be 0.0001 to 2,000 mg / kg per day, specifically 0.001 to 2,000 mg / kg. The dosage may be administered once a day or divided into several doses. However, the scope of the present invention is not limited by the above dosage.

[0097] The pharmaceutical composition according to the present invention can be administered by intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, or rectal administration.

[0098]

[0099] Another aspect of the present invention provides a kit for preventing or treating fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, and tazemetostat as active ingredients.

[0100] The above tazemetostat, Bifidobacterium pseudolongum strain, culture, lysate, extract pharmaceutical composition, prevention and treatment, etc. are defined in the same manner as described above.

[0101]

[0102] In the present invention, the term "kit" refers to a collection of parts that can be assembled to create something, and in the present invention, refers to a collection of experimental supplies provided to prevent or treat fatty liver disease.

[0103]

[0104] Another aspect of the present invention provides a method for treating non-alcoholic fatty liver disease, comprising administering to a subject a pharmaceutically effective amount of a pharmaceutical composition comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0105] Specifically, the above treatment method may further include a step of co-administering tazemetostat to the subject.

[0106] The above tazemetostat, Bifidobacterium pseudolongum strain, culture, lysate, extract pharmaceutical composition, combined administration, treatment, etc. are defined in the same manner as described above.

[0107] The term "subject" in the present invention means all animals, including rats, mice, livestock, etc., including humans, in which fatty liver disease is induced or can be induced, and may be animals other than humans, but is not limited thereto.

[0108] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

[0109] The term "pharmaceutically effective amount" in the present invention means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined based on factors including the type and severity of the individual, age, sex, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0110]

[0111] Another aspect of the present invention provides a food composition for preventing or improving liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0112] The food composition of the present invention may further comprise tazemetostat.

[0113] At this time, the above-mentioned tazemetostat, Bifidobacterium pseudolongum strain, culture, lysate, extract pharmaceutical composition, liver disease, prevention, etc. are defined in the same manner as described above.

[0114] In the present invention, the term "improvement" means any act of improving or beneficially changing fatty liver disease by administering a composition containing the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP of the present invention, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0115] The food composition of the present invention includes forms such as pills, powders, granules, infusions, tablets, capsules, or liquids, and foods to which the composition of the present invention can be added include, for example, various foods, such as beverages, gum, tea, vitamin complexes, and health supplements.

[0116] In the food composition of the present invention, there are no particular limitations on other ingredients other than the Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof and / or tazemetostat, which are essential ingredients that can be included, and various herbal extracts, food additives, or natural carbohydrates, etc. can be included as additional ingredients, as in conventional foods.

[0117] In addition, as mentioned above, food auxiliary additives may be additionally added, and the food auxiliary additives include food auxiliary additives conventional in the art, such as flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, etc.

[0118] Examples of the above natural carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., common sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. In addition to those described above, natural flavoring agents (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used as flavoring agents.

[0119] In addition to the above, the food composition of the present invention may include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, it may include fruit pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination.

[0120] The above health supplements include health functional foods and health foods.

[0121] The above functional food is the same term as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition. Here, "functionality" means regulating nutrients for the structure and function of the human body or obtaining a useful effect for health purposes such as physiological action. The food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long period of time because it uses food as a raw material, and is highly portable, so the food composition of the present invention can be taken as a supplement to enhance the effect of preventing or improving fatty liver disease.

[0122]

[0123] Another aspect of the present invention provides a feed composition for preventing or improving liver disease, comprising a Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

[0124] The feed composition of the present invention may further comprise tazemetostat.

[0125] At this time, the above-mentioned tazemetostat, Bifidobacterium pseudolongum strain, culture, lysate, extract pharmaceutical composition, liver disease, prevention, improvement, etc. are defined in the same manner as described above.

[0126] The above feed composition may include, in addition to the Bifidobacterium pseudolongum strain deposited under the accession number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, tazemetostat, a known carrier, stabilizer, or additive acceptable for pharmaceutical, food, or feed use. For example, there are binders, emulsifiers, and preservatives added to prevent quality deterioration, and amino acids, vitamins, enzymes, flavoring agents, non-protein nitrogen compounds, silicates, buffers, extractants, oligosaccharides, and the like added to the feed to increase utility. In addition, feed admixtures may be additionally included, but are not limited thereto. The feed composition may also include various nutrients such as vitamins, amino acids, and minerals, antioxidants, and other additives, as needed, and may be in an appropriate form such as powder, granules, pellets, or suspension. The feed composition of the present invention can be supplied alone or mixed into feed for single animals. The feed of the present invention is not particularly limited, and is not limited to any feed for animals such as dogs, cats, horses, and cows. Any feed may be used, including powdered feed, solid feed, dry feed, wet feed, moist pellet feed, dry pellet feed, EP (Extruder Pellet) feed, and raw feed.

[0127]

[0128] Another aspect of the present invention provides a use of a Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, for preventing or treating liver disease.

[0129] Another aspect of the present invention provides a composition comprising a Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, for use in the prevention or treatment of liver disease. Specifically, the composition may further comprise tazemetostat.

[0130] Another aspect of the present invention provides a combination comprising a) a Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, and b) tazemetostat, and a use of the combination for preventing or treating liver disease.

[0131]

[0132] In the present invention, the term "combination" refers to a combined administration use of the Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat, and can be understood to have the same meaning as "combined use." This also includes, but is not limited to, pharmaceutical compositions and pharmaceutical kit forms characterized by combined use of the Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat.

[0133] The above combination is

[0134] a) administered as a mixture of Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat; or

[0135] b) The Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat may be administered in a separate form, but are not limited thereto.

[0136] When the Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat are in separate forms, the Bifidobacterium pseudolongum strain deposited under KCTC 15133BP and tazemetostat are each formulated as separate preparations, and the separate preparations can be administered simultaneously, separately, sequentially, or in reverse order.

[0137]

[0138] Another aspect of the present invention provides a composition for use in the prevention or treatment of liver disease, comprising a combination comprising a) a Bifidobacterium pseudolongum strain deposited under the registration number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, and b) tazemetostat.

[0139] The term "composition comprising a combination" in the present invention may be, but is not limited to, the combination itself or a composition comprising the combination and having a therapeutic use. The term "composition comprising a combination" may be used interchangeably with "composition" herein.

[0140] A composition comprising a combination according to the present invention is for co-administration of a Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain or a mixture thereof, and tazemetostat. The Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain or a mixture thereof, and tazemetostat may be formulated as a single formulation, or may be formulated individually.

[0141]

[0142] Another aspect of the present invention provides a method for preventing or treating liver disease, comprising administering and / or using in combination a composition comprising a pharmaceutically effective amount of a Bifidobacterium pseudolongum strain deposited under KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof, and a composition comprising a pharmaceutically effective amount of tazemetostar to a subject in need thereof.

[0143]

[0144] Hereinafter, the present application will be described in more detail through examples. These examples are intended to more specifically explain the present application, and the scope of the present application is not limited by these examples.

[0145]

[0146] Example 1. Genome analysis of Bifidobacterium pseudolongum KGMB04131 strain (KCTC 15133BP)

[0147]

[0148] A new strain of Bifidobacterium pseudolongum KGMB04131 was discovered and isolated from the feces of a healthy 21-year-old Korean male. Similarity with the 16S rRNA gene sequences of existing strains and complete genome analysis were performed. The results are shown in Fig. 1.

[0149]

[0150] As shown in Fig. 1, the similarity analysis between the 16S rRNA gene sequences of Bifidobacterium pseudolongum KGMB04131 strain and existing strains showed 99.86% similarity with the standard strain. In addition, the whole genome analysis of KGMB04131 showed that it consisted of a base sequence of 2,105,891 bp and showed a GC ratio of 63.25%. The isolated strain was named Bifidobacterium pseudolongum KGMB04131 and deposited with the Korea Research Institute of Bioscience and Biotechnology on October 12, 2022, and assigned the accession number KCTC 15133BP.

[0151]

[0152] Example 2. Verification of the NASH-inhibiting effect of Bifidobacterium pseudolongum (KCTC 15133BP) strain in a NASH mouse model.

[0153]

[0154] Next, we created a NASH model to verify the effectiveness of the Bifidobacterium pseudolongum (KCTC 15133BP) strain.

[0155]

[0156] The method for establishing a NASH diet mouse model is described in Fig. 2A, and the mouse model was created through this method. Specifically, in order to establish a mouse model similar to human NASH, mice were fed a NASH diet (protein, 9.8%, fat, 52.9%, carbohydrate, 37.3%, #TD.190142, Teklad Custom Diet) and sucrose solution (D-glucose = 18.9 g / L, fructose = 23.1 g / L) starting at 8 weeks of age, and 14 weeks later, 5X10 Bifidobacterium pseudolongum (KCTC 15133BP) strains were administered once a day by oral gavage. 8 CFU / day was administered. Sirius red, oil red o (ORO), hematoxylin and eosin (H&E) staining and F4 / 80 value analysis were performed on the experimental group, and the results are shown in Figure 2B.

[0157]

[0158] As a result, as can be seen in Fig. 2B, in the NASH diet mouse model, the group administered KCTC 15133BP strain together with the NASH diet (NASH_KGMB04) showed improvement in steatosis, inflammatory cell infiltration, and fibrosis compared to the group administered only the NASH diet (NASH), and this confirmed that the KCTC 15133BP strain has an excellent NASH development inhibitory effect.

[0159]

[0160] Example 3. Confirmation of the results of metagenome analysis in a NASH diet mouse model.

[0161]

[0162] Next, we aimed to analyze the intestinal microbiota of a NASH diet mouse model through metagenomic analysis. A NASH diet mouse model was created using the same process as in Example 2, and the results of the metagenomic analysis are shown in Figure 3 below.

[0163]

[0164] As can be seen in the above Figure 3, it can be confirmed that the Bifidobacterium pseudolungum strain is reduced in the intestinal flora of NASH diet mice compared to the normal control group.

[0165]

[0166] Example 4. Comparison of propionic acid production and neutral fat accumulation inhibition ability with Bifidobacterium pseudolongum strains of the same genus and species.

[0167]

[0168] Next, we attempted to measure and compare the propionic acid production ability, which is known to inhibit cholesterol synthesis in the liver, of the KCTC 15133BP strain of the present invention and other strains of the same genus.

[0169]

[0170] KGMB04129, KGMB04131 (KCTC 15133BP), and KGMB04804 are Bifidobacterium pseudolongum strains isolated from the feces of healthy Koreans, and KCTC 3224T was used as a standard strain and control. The results of short-chain fatty acid (SCFA) analysis of the culture broth of the B. pseudolongum strains are shown in Fig. 4.

[0171]

[0172] As can be seen in the above Figure 4, the results of short-chain fatty acids (SCFAs) analysis of the B. pseudolongum culture medium showed that the propionic acid production of the KGMB04131 (KCTC 15133BP) strain was significantly superior to other B. pseudolongum strains, and furthermore, it showed improved production compared to the standard strain (KCTC 3224T). From this, it was confirmed that the KCTC 15133BP strain had improved propionic acid production ability compared to other strains of the same genus and species, and thus had an excellent effect in inhibiting cholesterol synthesis in the liver.

[0173]

[0174] Next, we aimed to measure the neutral fat accumulation inhibition ability of the KGMB04131 (KCTC 15133BP) strain and other isogenic strains. A schematic diagram of the measurement process is illustrated in Figure 5A, and the measurement results are disclosed in Figures 5B and 5C.

[0175]

[0176] As can be seen in the above Figure 5, it can be confirmed that the extract of the KGMB04131 strain (KCTC 15133BP) has a significantly higher effect of inhibiting triglyceride accumulation in 3T3-L1 adipocytes than the standard strain (KCTC 3224T) and other B. pseudolongum strains.

[0177]

[0178] From the above results, it is suggested that the KCTC 15133BP strain has superior propionic acid production ability and neutral fat accumulation inhibition ability compared to other strains of the same genus and species, and thus has high utility as a NASH treatment.

[0179]

[0180] Example 5. Results of gene expression analysis in the MCD diet mouse model

[0181]

[0182] A methionine-choline deficient diet (MCD diet) animal model was established to evaluate the effect on the expression of genes that play a key role in the methionine cycle of the KCTC 15133BP strain of the present invention.

[0183] To determine the effect of strains in a model mimicking NASH, C57BL / 6 mice were fed a methionine- and choline-deficient diet (MCD) for 6 weeks. The MCD diet, which is deficient in methionine and choline, induces lipid metabolism and inflammation through abnormalities in the hepatic methionine cycle and interferes with VLDL transport, leading to pathological changes similar to steatohepatitis. To apply the standard strain, microbial cultures were administered orally, and mice were allowed free access to food and water. Liver tissues were obtained from mice after 6 weeks of MCD diet, rapidly frozen, and stored at -70°C.

[0184] Afterwards, liver tissue was lysed for gene expression analysis. RNA was extracted using the traditional Trizol method to analyze changes in RNA. The extracted RNA was synthesized into cDNA using reverse transcriptase, and the amount of each gene was quantified using qRT-PCR. gapdh was used as a reference gene for calibration.

[0185]

[0186] As a result, as can be confirmed in Fig. 6, in the case of the treatment group of the standard strain (KCTC 3224T)(TT), the recovery of the expression of the GNMT and AHCY genes, which are key genes of the methionine cycle, was minimal, but in the case of the KCTC 15133BP strain of the present invention, it was confirmed that the recovery of the GNMT and AHCY genes in the methionine cycle was induced at a significant level.

[0187]

[0188] Example 6. Confirmation of the effect of combined administration of Bifidobacterium pseudolongum strain and tazemetostat.

[0189]

[0190] Next, the inventors of the present invention sought to identify a component that exhibits a significant synergistic effect on the treatment of liver disease when used in combination with the Bifidobacterium pseudolongum strain, in order to achieve a more effective treatment effect for fatty liver disease.

[0191]

[0192] Example 6-1. Confirmation of increased EZH2 gene expression levels in liver tissue derived from NASH patients.

[0193]

[0194] First, we confirmed that the expression level of the histone methylase EZH2 gene increased in liver tissues derived from NASH patients of Korean and European groups, and the results are shown in Fig. 7.

[0195]

[0196] As can be seen in Figure 7, it was confirmed that the expression of the EZH2 gene was significantly increased in liver tissues derived from all NASH patients compared to the normal group.

[0197]

[0198] Example 6-2. Confirmation of the therapeutic effect of tazemetostat monotherapy in a NASH mouse model.

[0199]

[0200] According to the results of Example 6-1 above, we sought to explore a component that could exhibit synergy through combination with the Bifidobacterium pseudolongum strain with respect to EZH2 gene expression.

[0201]

[0202] In a NASH mouse model, NASH was induced for 18 weeks using a high-fat diet, and the degree of disease amelioration was investigated by oral administration of tazemetostat, one of the EZH2 inhibitors, alone for the final 4 weeks. The morphological aspects of the liver were examined using tissue staining (H&E) and lipid-specific staining (Oil red O staining), and the level of inflammatory cell infiltration (F5 / 80+) was also determined. The results are shown in Figure 7.

[0203]

[0204] As shown in Figure 8, in the tazemetostat-administered (NASH+TAZ) experimental group, the size and weight of the hypertrophied liver due to the high-fat diet were significantly reduced in terms of liver morphology, and fat accumulation in the liver was improved in the drug-administered group. In addition, inflammatory cell infiltration (F5 / 80+) was also confirmed to be improved.

[0205]

[0206] Next, the results of the metagenomic analysis of the NASH diet mouse model are shown in Figure 9.

[0207]

[0208] As shown in Figure 9 above, tazemetostat was confirmed to partially correct the imbalance of intestinal microflora in NASH diet mice, but it did not restore the level of the beneficial Bifidobacterium pseudolungum strain to normal levels. Therefore, based on these results, we attempted to enhance the NASH treatment effect by co-administering B. pseudolungum with tazemetostat.

[0209]

[0210] Example 6-3. Confirmation of the therapeutic effect of combined administration of tazemetostat and Bifidobacterium pseudolongum strain in a NASH mouse model.

[0211]

[0212] NASH diet mice were co-administered with tazemetostat and Bifidobacterium pseudolongum (KCTC 15133BP) strains orally for 4 weeks, and the groups administered tazemetostat and each microbiota strain alone were administered in the same manner. Bifidobacterium pseudolongum strains were administered at a dose of 5X10 8 Tazemetostat was administered at a concentration of 33 mg / kg, with CFU / day.

[0213]

[0214] At this time, the morphological aspects of the liver were confirmed by tissue staining (H&E) and fat-specific staining (Oil red O staining), and the results of confirming the level of inflammatory cell infiltration (F5 / 80+) are shown in Figure 10 below.

[0215] As can be seen in Fig. 10, in the group co-administered with tazemetostat and Bifidobacterium pseudolongum (KCTC 15133BP) strain (TAZ+KGMB04), the size and weight of the liver were significantly reduced compared to the group administered alone, and fat accumulation and inflammatory cell infiltration in the liver were significantly improved, confirming the existence of a synergistic effect of co-administration.

[0216]

[0217] Next, the results of analysis at the transcriptome level through RNA-seq of mouse liver tissue are shown in Figure 11 below.

[0218] As shown in Figure 11, the results of transcriptome-level analysis using RNA-seq of mouse liver tissues confirmed that, compared to the single-administration group, co-administration of Tazemetostat and KGMB04131 strain resulted in a recovery of transcriptome levels closer to normal liver levels.

[0219]

[0220] Next, the results of gene network analysis comparing gene expression upon single and combined administration of tazemetostat and Bifidobacterium pseudolongum strains are shown in Figure 12. A gene network was constructed / analyzed by adding information on previously known protein-protein interactions and metabolic / regulatory pathways to the major genes and cell functions derived from the transcriptome analysis of Figure 11.

[0221] As a result, when Tazemetostat and the KGMB04131 strain were co-administered, the expression of genes related to amino acid metabolism and retinol metabolism, which had decreased in the NASH diet, was restored to normal, and the expression of genes related to inflammatory response and liver fibrosis was decreased. In particular, as shown in Fig. 11, most of the genes whose expression changed in the co-administration (1) were similar to the gene expression changes in the KGMB04131 strain single administration rather than in the Tazemetostat single administration (nodes marked as "B" in the legend of Fig. 11), or (2) appeared as a new synergistic effect in the co-administration (nodes marked as "S" in the legend of Fig. 11), thereby confirming the excellent NASH therapeutic effect of the B. pseudolongum KGMB04131 strain.

[0222]

[0223] Next, we aimed to determine the relative expression levels of genes that play key roles in the impaired methionine cycle in the NASH diet mouse model.

[0224] Specifically, to examine the combined effects of tazmetostat and Bifidobacterium pseudolongum in a model mimicking NASH, C57BL / 6 mice were fed a high-fat, high-fructose diet for 18 weeks. Cultures of tazmetostat and Bifidobacterium pseudolongum were administered orally for 4 weeks before sacrifice. During this period, the mice were allowed free access to food and water. Liver tissues were obtained from the mice, rapidly frozen, and stored at -70°C.

[0225] To analyze gene expression, liver tissue was lysed and RNA was extracted using the traditional Trizol method. The extracted RNA was synthesized into cDNA using reverse transcriptase, and the amount of each gene was quantified using qRT-PCR. gapdh was used as a reference gene for calibration.

[0226]

[0227] As a result, as can be confirmed in Fig. 13, it was confirmed that the Gnmt1 and Ahcy genes, which play a major role in the methionine cycle, and the Cps1 gene, which plays a major role in the urea cycle, were damaged in the MASLD environment. At this time, it was confirmed that the expression of the Gnmt1, Ahcy, and Cps1 genes was more increased in the KGMB04131 strain group administered alone than in the Tazemetostat single-administration group, and further, when Tazemetostat and the KGMB04131 strain were co-administered, the recovery of the expression of the Gnmt1 and Ahcy genes was particularly remarkable, and some additional increase in the expression of the Cps1 gene was also confirmed.

[0228]

[0229] From the above results, it was confirmed that the combination of tazemetostat and Bifidobacterium pseudolongum strains had a remarkable synergistic effect on the treatment of NASH.

[0230]

[0231] From the above description, those skilled in the art will understand that the present application can be implemented in other specific forms without altering its technical concept or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of this application should be interpreted to include all changes or modifications derived from the meaning and scope of the following claims and their equivalents, rather than the detailed description above.

[0232]

[0233]

Claims

1. Bifidobacterium pseudolongum strain deposited under accession number KCTC 15133BP.

2. A pharmaceutical composition for preventing or treating fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under the deposit number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

3. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the liver disease in paragraph 2 is non-alcoholic fatty liver disease (NAFLD).

4. A pharmaceutical composition for preventing or treating fatty liver disease, wherein in paragraph 3, the nonalcoholic fatty liver disease is at least one selected from the group consisting of nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, and nonalcoholic cirrhosis.

5. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition in paragraph 2 has at least one effect of inhibiting cholesterol synthesis in the liver and inhibiting neutral fat accumulation.

6. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition in the second paragraph increases the expression of the GNMT (glycine N-methyltransferase) gene or the AHCY (Adenosylhomocysteinase) gene.

7. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition in claim 2 further comprises a pharmaceutically acceptable carrier.

8. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition in paragraph 2 is administered in combination with Tazemetostat.

9. A pharmaceutical composition for preventing or treating fatty liver disease, wherein in paragraph 8, tazemetostat and Bifidobacterium pseudolongum strain are administered simultaneously or sequentially.

10. In paragraph 2, A pharmaceutical composition for preventing or treating fatty liver disease, wherein the composition further comprises tazemetostat.

11. In paragraph 2, A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition is administered by at least one administration method selected from the group consisting of intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration.

12. A pharmaceutical composition for preventing or treating fatty liver disease, wherein the pharmaceutical composition in claim 8 has at least one of the effects of reducing liver weight, inhibiting fat accumulation in the liver, and improving inflammatory cell infiltration and fibrosis in the liver.

13. A food composition for preventing or improving fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under the deposit number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

14. A feed composition for preventing or improving fatty liver disease, comprising a Bifidobacterium pseudolongum strain deposited under the deposit number KCTC 15133BP, a culture of the strain, a lysate of the strain, an extract of the strain, or a mixture thereof.

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