JP2022184424A - Visceral fat reducing agent - Google Patents

Abstract

To provide a visceral fat reducing agent useful for reducing visceral fat.SOLUTION: A visceral fat reducing agent contains Blautia bacteria as an active ingredient.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to a visceral fat reducing agent.

Visceral fat is an adipose tissue distributed between the stomach and the liver and around the liver, and its excessive accumulation is known to be a risk of arteriosclerotic lesions. Visceral adiposity is now the primary criterion for the diagnosis of metabolic syndrome. Managing the accumulation of visceral fat is extremely important for extending healthy life expectancy.

More than 1000 kinds of bacteria, more than 100 trillion, live in the human gastrointestinal tract, forming an intestinal microflora. Recent studies have revealed that the intestinal microbiota is involved in the pathogenesis of various diseases such as metabolic diseases, immune diseases, and psychiatric diseases, in addition to gastrointestinal diseases. In particular, it has been suggested that the composition of the intestinal flora is closely related to obesity and obesity-induced metabolic diseases.
The intestinal flora inhabiting the gastrointestinal tract is dominated by two phyla, the phylum Firmicutes and the phylum Bacteroidetes. Comparing the intestinal microflora of obese and normal mice, it has been reported that obese mice have more bacteria from the phylum Firmicutes and less bacteria from the phylum Bacteroidetes, while normal mice have more bacteria from the phylum Bacteroidetes. there is Also in humans, it has been reported that the ratio of Firmicutes/Bacteroidetes is high in obese people. Furthermore, although no consistent correlation was found between the presence of bacteria in the phyla Firmicutes and Bacteroidetes and visceral fat, when the bacterial species were analyzed by "genus" smaller than the phylum, visceral fat area was found to increase regardless of gender. Small people have a high abundance ratio of B. brautia in their intestines, and it has been reported that among intestinal bacteria, B. brautia is related to the amount of visceral fat in both men and women (Non-Patent Document 1).

Brautia bacteria are major bacteria accounting for about 3 to 11% of the total intestinal bacteria of humans and animals. Brautia bacteria are known to produce short-chain fatty acids such as acetic acid, propionic acid, and butyric acid, which work to eliminate obesity in the body. It has been reported that Blautia bacterial strains, such as Blautia hydrogenotrophica, have also been shown to be effective in increasing or maintaining microbiota diversity and in treating brain injury. have been reported (for example, Patent Documents 1 and 2).
However, there is no report on the effect of ingestion of Brautia bacteria on visceral fat accumulation.

Japanese Patent Publication No. 2019-508363 Japanese Patent Publication No. 2020-520916

npj Biofilms and Microbiomes 2019 5:28

TECHNICAL FIELD The present invention relates to providing a visceral fat reducing agent useful for reducing visceral fat.

Means for Solving the Problems In view of the above problems, the present inventors conducted intensive studies and found that visceral fat is significantly reduced when Brautia bacteria are orally ingested.

That is, the present invention provides a visceral fat-reducing agent containing Brautia bacteria as an active ingredient.

The visceral fat-reducing agent of the present invention exhibits an excellent visceral fat-reducing effect. Therefore, it can be expected to prevent, ameliorate or treat the onset or progression of health disorders or diseases closely related to visceral fat accumulation.

Graphs showing total visceral fat weight, epididymal circumference weight, mesenteric weight, and perirenal/retroperitoneal weight in each experimental group.

In the present invention, Blautia bacteria are bacteria belonging to the genus Blautia classified into the phylum Firmicutes.
Examples of Blautia bacteria include Blautia hydrogenotrophica, Blautia stercoris, Blautia faecis, Blautia coccoides, and Blautia gluceracea. , Blautia hansenii, Blautia luti, Blautia producta, Blautia schinkii, Blautia wexlerae and the like. These can use 1 type(s) or 2 or more types. Among these, Brautia hensennii is preferable from the viewpoint of improving the effect of reducing visceral fat.
Brautia hensennii can be exemplified by strain JCM 14655 or bacteria identical thereto. The same bacterium is a homologous bacterium whose 16S rRNA gene nucleotide sequence has 98% or more, preferably 99% or more homology with the 16S rRNA gene nucleotide sequence of the cell line and has the same mycological properties. refers to bacteria.
The Brautia Hensenni JCM 14655 strain is generally available from the RIKEN BioResource Research Center Microbial Materials Development Office (RIKEN BRC) based on the JCM number of the strain described in the online catalog (for example, for the JCM 14655 strain , https://www.jcm.riken.jp/cgi-bin/jcm/jcm_number?JCM=14655).

The cells of the genus Brautia can be obtained by culturing according to a method commonly used for culturing the bacterium of the genus Brautia. For example, culture is performed at a culture temperature of 30 to 45° C. using GAM medium or the like. Cultivation is preferably performed under anaerobic conditions. After culturing, the cells can be separated by collection means such as centrifugation or membrane concentration.

The Brautia bacterium may be either viable or dead, but from the viewpoint of quality stability and safety to living organisms, it is preferable to use dead cells.
Examples of dead cells include heat-sterilized cells obtained by heat-sterilizing cultured cells. The dead cells may be in the form of a suspension in water or the like, or may be in the form of a dried product. Usual drying methods such as spray drying and freeze drying can be used.

As shown in the Examples below, ingestion of Brautia bacteria suppresses the accumulation of visceral fat promoted by ingestion of a high-fat diet. That is, Brautia bacteria have the effect of suppressing the accumulation of visceral fat and reducing visceral fat. It is thought that ingestion of Brautia bacteria affects the inhabitation status and number of Brautia bacteria existing in the intestine, and reduces visceral fat. Reduction of visceral fat makes it possible to prevent, improve or treat visceral fat-type obesity and metabolic syndrome (visceral fat syndrome) based on visceral fat accumulation.
Therefore, the Brautia bacterium can be a visceral fat-reducing agent that reduces visceral fat, and can be used to produce a visceral fat-reducing agent. Brautia bacteria can also be used to reduce visceral fat.
As used herein, the term “visceral fat” refers to fat accumulated in and around visceral tissues (eg, stomach, liver, kidney, etc.) in the abdominal cavity. "Reduction of visceral fat" means reducing the amount of visceral fat and suppressing accumulation of visceral fat.
"Use" can be administration or ingestion to animals, including humans, and can be therapeutic or non-therapeutic. "Non-therapeutic" means a concept that does not include medical practice, i.e., a concept that does not include methods of surgery, treatment or diagnosis of humans, and more specifically, surgical procedures performed on humans by a physician or a person under the direction of a physician. , is a concept that does not include methods of performing therapy or diagnosis.

The visceral fat-reducing agent of the present invention can be a human or animal drug, quasi-drug, food or feed that exerts the effect of reducing visceral fat when administered or taken to animals including humans, and the drug, drug It can be a material or formulation that is used by blending in quasi-products, food, or feed.

The drug (hereinafter, including quasi-drugs) contains Brautia bacteria as an active ingredient for reducing visceral fat. Furthermore, the drug may contain a pharmaceutically acceptable carrier or other active ingredient, pharmacological ingredient, etc., as necessary, as long as the function of the active ingredient is not impaired.
The medicament can be administered in any dosage form. Dosage forms include, for example, tablets (including chewable tablets), capsules, granules, powders, oral solid formulations such as lozenges, oral liquid formulations, oral liquid formulations such as syrups, injections, suppositories, and inhalation. Examples include parenteral preparations such as drugs, percutaneous absorption agents, and external preparations. A preferred mode of administration is oral administration. The size of the form can be arbitrarily adjusted according to the purpose of use.
Formulations of such various dosage forms may optionally contain pharmaceutically acceptable carriers such as excipients, binders, fillers, disintegrants, surfactants, lubricants, dispersants, Buffers, osmotic pressure adjusters, pH adjusters, emulsifiers, preservatives, stabilizers, preservatives, thickeners, fluidity improvers, flavoring agents, foaming agents, fragrances, coating agents, diluents, etc. It can be prepared according to a conventional method by appropriately combining with the medicinal ingredients of

The food contains Brautia bacteria as an active ingredient for reducing visceral fat.
Foods include foods that claim to reduce visceral fat and for which labeling to that effect has been permitted or notified as necessary (foods for specified health uses, foods with function claims). An example of the display is "reduce visceral fat". Foods for which function labeling has been permitted or notified can be distinguished from ordinary foods.
The form of food can be solid, semi-solid or liquid (eg beverage). Examples include various food compositions (breads, cakes, noodles, confectionery, frozen foods, ice creams, candies, sprinkles, soups, dairy products, shakes, beverages, seasonings, etc.), and further , and nutritional supplement compositions in the same form as the above-described oral administration formulations (solid formulations such as granules, powders, tablets, capsules, microcapsules, and lozenges).
Various forms of food may be prepared by adding Brautia bacteria to any food ingredient, or other active ingredient, or additive acceptable to food (e.g., solvents, softeners, oils, emulsifiers, preservatives, acidulants, sweeteners). Ingredients, bittering agents, pH adjusters, stabilizers, coloring agents, UV absorbers, antioxidants, humectants, thickeners, sticking agents, dispersants, fluidity improvers, wetting agents, fragrances, seasonings, It can be prepared according to a conventional method by appropriately combining with a flavor modifier) and the like.

The feed contains Brautia bacteria as an active ingredient for reducing visceral fat.
The form of feed is preferably in the form of pellets, flakes, mash, or liquid. Examples include feed, pet food used for dogs, cats, small birds, and the like.
The feed contains Brautia bacteria, other feed ingredients such as meat, protein, grains, bran, meal, sugars, vegetables, vitamins, minerals, gelling agents, shape retention agents, pH adjusters, It can be prepared according to a conventional method by appropriately combining seasonings, preservatives, nutritional supplements, and the like.

The content of Brautia bacteria in the visceral fat-reducing agent of the present invention may vary depending on the dosage form and form thereof, but can be appropriately set in consideration of ease of administration or intake.

The dosage or intake of the visceral fat-reducing agent of the present invention may vary according to the species, body weight, sex, age, condition, or other factors of the subject of administration or intake. The dosage, route, interval of administration, and the amount and interval of intake can be appropriately determined by those skilled in the art. From the viewpoint of improving the effect, the number of Brautia bacteria is preferably 1×10 ⁴ or more, more preferably 1×10 ⁵ or more, and still more preferably 1×10 ⁷ or more. The number of bacteria is preferably 1×10 ¹⁵ or less, more preferably 1×10 ¹² or less, and still more preferably 1×10 ⁹ or less. The dosage or intake of the visceral fat-reducing agent of the present invention is preferably 1×10 ⁴ to 1×10 ¹⁵ bacteria, more preferably 1×10 ⁵ to 1 bacteria, as Brautia bacteria. ×10 ¹² , more preferably in the range of 1×10 ⁷ to 1×10 ⁹ bacteria.
In the present invention, such an amount is preferably divided into 1, 2 or 3 or more times per day for oral administration or oral intake.
In addition, in the examples described later, it is explained that the Brautia bacteria administered to mice has an effect of reducing visceral fat, but from the viewpoint of ethical and safety considerations, , daringly used mice, which have been established in official test methods or academic papers as hypothetical human models. Therefore, the visceral fat-reducing effect in the present invention is a concept indicating not only the visceral fat-reducing effect for mice but also activity for humans in general.

The visceral fat-reducing agent of the present invention can be administered or taken according to any schedule.
The administration or intake period is not particularly limited, and single administration or intake may be performed, or repeated/continuous administration or intake may be performed. In the case of repeated/continuous administration or ingestion, it is preferable to administer or ingest continuously for 2 weeks or longer, 4 weeks or longer, and 8 weeks or longer.

The agent for reducing visceral fat of the present invention is preferably administered or ingested at the time of eating/feeding, before eating/feeding, or after eating/feeding. When administered or taken before or after eating/feeding, it is preferably administered or taken within 10 minutes before or after eating/feeding. The content of eating (feeding) is not particularly limited, and includes meals or feeds containing proteins, carbohydrates (including carbohydrates), and lipids.
Subjects for administration or intake are not particularly limited as long as they are humans or non-human animals that require or desire it. Preferred examples of subjects include humans with a high accumulation of visceral fat (for example, humans with a visceral fat area of 100 cm ² or more), humans who tend to eat fatty foods, and the like.

Test example 1
1. Test feed and method Cells of Brautia hensennii strain JCM 14655 (Riken BRC) were sterilized by heating in an autoclave. After that, it was suspended in distilled water, centrifuged and washed repeatedly, and the residue was freeze-dried and powdered.
Three-week-old male normal mice (C57BL/6J mice) were preliminarily fed for one week, and then experimentally fed for 9 weeks with free intake of food and water. As for the feed, they were given a normal diet of AIN-93G, which is the basic diet specified by the US National Institute of Nutrition, and a high-fat diet with 20% by mass of lipid prepared with reference to the composition of the basic diet.
The experimental groups consisted of a normal diet control group (n = 8), a high-fat diet control group (n = 8), and a high-fat diet (Blautia bacteria added) group (n = 8) so that the initial body weight was almost constant. It was performed by dividing into 3 groups. The dose of Brautia bacteria was 1×10 ⁹ bacteria/one animal/day. During the experimental breeding period, body weight and water intake were measured, and feces were collected.
After the end of experimental breeding, animals were euthanized by cervical dislocation and then dissected. As visceral fat, perididymal fat, mesenteric fat, and perirenal/retroperitoneal fat were removed, and the weights of various organs were measured. The sum of these values was taken as the total weight of visceral fat.

2. Method for Identifying Brautia in Foods Added with Brautia A real-time PCR method can be used to identify Brautia. Specific examples are shown below.
Procedure 1. DNA extraction/extraction method Pretreatment/crude extraction: Suspend each sample in 4 M guanidine thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA, and use a cell disruptor (FastPrep FP100A (MP Biomedicals, USA). etc.) to pretreat the sample. Additionally, DNA can be extracted from bead-treated suspensions using the Magtration System 12 GC and GC series MagDEA DNA 200 (Precision System Science, Japan).

Procedure 2. PCR Conditions Amplify DNA fragments. As a kit, Blautia Hansenii Detection Kit (manufactured by Techno Suruga Lab Co., Ltd.) can be used.

Step 3. Real-Time PCR Method As a reagent used in the real-time PCR method, for example, TB Green Premix Ex Taq 2 (Tli RNase H Plus) (manufactured by Takara Bio Inc.) can be used. As an apparatus, for example, Rotor-Gene Q (manufactured by QIAGEN) can be used. As conditions for PCR, for example, the method described in Dig Dis Sci, 2015, 60 (9) 2654-61, etc. can be used.

A T-test was used for the test of significant difference. The obtained numerical values are shown as mean±standard error.

2. Results FIG. 1 shows the total weight of visceral fat, the weight of the epididymal perimeter, the mesentery, and the perirenal/retroperitoneal weight of each group. Different letters (a, b, c) in FIG. 1 indicate a significance level of P<0.05, and # indicates P=0.08 in comparison with normal diet. As a result, in the high-fat diet (addition of B. brautia) group, a decrease in visceral fat weight was observed compared to the high-fat diet control group. It was confirmed that it has an inhibitory effect.

Claims (3)

A visceral fat reducing agent containing Brautia bacteria as an active ingredient. 2. The visceral fat-reducing agent according to claim 1, wherein the Brautia bacterium is Brautia hensennii. 3. The visceral fat-reducing agent according to claim 1 or 2, which is administered or taken in an amount of 1×10 ⁴ to 1×10 ¹⁵ bacteria per day as Brautia bacteria.

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