JP2021155375A - Agent for improving intestinal bacterial flora - Google Patents

Abstract

To provide an agent for improving intestinal bacterial flora.SOLUTION: An agent for improving intestinal bacterial flora contains black soybean seed coat extract as an active ingredient.SELECTED DRAWING: None

Description

The present invention relates to oral compositions preferably used to improve the gut microbiota.

The intestinal flora, also called the intestinal flora, is an ecosystem in the body composed of bacteria in the intestine. The gut microbiota interacts with the host via the intestinal epithelium and may be involved in a variety of diseases such as constipation, diarrhea, infections, allergic diseases, inflammatory bowel disease, obesity and diabetes. It has become clear. It is known that the intestinal bacterial flora varies from individual to individual and can change depending on the health condition of the individual and the food and drink ingested. For example, it is known that when humans are obese, the composition ratio of bacteria belonging to the genus Bacteroidetes is low in the intestinal flora, and the composition ratio of bacteria belonging to Firmicutes is high. As the body weight is reduced, the composition ratio of bacteria belonging to the genus Bacteroidetes increases, and the composition ratio of bacteria belonging to Firmicutes decreases. Therefore, it is considered that the health condition of the individual can be improved to a good condition by improving the intestinal bacterial flora, and various studies for that purpose have been conducted (for example, Patent Document 1). ~ 4 mag).

By the way, short-chain fatty acids (SCFA) are organic acids produced by intestinal bacteria and are known to play an indispensable role in maintaining the health of mammals including humans. In the case of humans, three types of short-chain fatty acids, acetic acid, propionic acid, and butyric acid, are typical short-chain fatty acids, which are produced by intestinal bacteria in the human large intestine and absorbed from the mucosal epithelium of the large intestine. Butyric acid is used as an energy source for colonic epithelial cells, and butyric acid and propionic acid are mainly metabolized and used in the liver and muscles. In addition, short-chain fatty acids are known to have the effect of adjusting the balance of the intestinal bacterial flora by lowering the pH in the intestine in the large intestine, making it difficult for pathogenic microorganisms to grow. In addition, it strengthens the barrier function of the intestinal epithelium and helps prevent infection (above, Non-Patent Document 1), stimulates the intestinal tract to enhance the peristaltic movement of the intestine, and promotes bowel movement (Non-Patent Document 2). The effect is known. Focusing on the effects of such short-chain fatty acids, various studies aimed at increasing the production of short-chain fatty acids in the intestine have been conducted (for example, Patent Documents 5 to 6 and the like).

In addition, equol is a compound having various characteristics such as high estrogen-like action as compared with O-desmethylangolencin (O-DMA), which is a metabolite of daidzein, which is a typical isoflavone. It has been reported that it has a therapeutic and preventive effect on osteoporosis (for example, Patent Document 7), alleviation of menopausal symptoms, prevention of metabolic syndrome, improvement of wrinkles and sagging of skin, and preventive effect on breast cancer (for example, non-patent). Document 3 etc.). In addition, equol has also been reported to have antioxidant effects (improvement of stains, whitening) and antiandrogen effects (prevention of prostate cancer, improvement of hair loss).

Such equol is a metabolite produced by the metabolism of daidzein, which is an aglycone of isoflavone, by intestinal bacteria in the intestine. However, it is known that the amount of the intestinal flora and the amount of equol-producing bacteria in the intestine varies from person to person, and the ability of the body to produce equol varies from person to person. Therefore, attempts have been made to increase the production efficiency of equol by affecting the intestinal bacterial flora.

JP-A-2017-163980 Japanese Unexamined Patent Publication No. 2018-50526 Japanese Unexamined Patent Publication No. 2018-184363 Japanese Unexamined Patent Publication No. 2018-52896 Japanese Unexamined Patent Publication No. 2004-346043 JP-A-2019-170335 Japanese Unexamined Patent Publication No. 2005-232074

Fukuda S, Toh H, Hase K, et al., Nature. 2011: 469: 543-547 Journal of the Japanese Home Economics Vol. 44, 4245-4254 (1993) "Chemistry and Biology", published by Japan Society for Bioscience and Biotechnology, Vol. 44, No. 3, p. 151-153 (2006)

An object of the present invention is to provide an oral composition suitably used for improving the intestinal bacterial flora. More specifically, it improves the intestinal flora and at the same time increases the abundance of short-chain fatty acid-producing bacteria in the intestinal bacteria; increases the abundance of equol-producing bacteria in the intestinal bacteria; and / Alternatively, it is an object of the present invention to provide an oral composition capable of reducing the abundance ratio of pathogenic bacteria in the intestinal bacteria.

As a result of diligent studies to solve the above problems, the present inventors changed the bacterial composition ratio in the intestinal flora when ingesting the black soybean seed coat extract, and the presence of equol-producing bacteria in the intestinal bacteria. It has been found that the proportion increases, the abundance of short-chain fatty acid-producing bacteria in the intestinal bacteria increases, and / or the abundance of pathogenic bacteria in the intestinal bacteria decreases. From this, it is considered that ingestion of black soybean seed coat extract improves the intestinal bacterial flora and promotes health.

The present invention has been completed based on these findings, and includes the following embodiments.
(I) An agent for improving the intestinal bacterial flora (I-1) An agent for improving the intestinal bacterial flora containing a black soybean seed coat extract as an active ingredient.
(I-2) The agent for improving the intestinal bacterial flora according to (I-1), which exerts at least one action and effect selected from the group consisting of the following (A) to (C):
(A) Increase in the abundance ratio of equol-producing bacteria in the intestinal bacteria,
(B) Increase in the abundance ratio of short-chain fatty acid-producing bacteria in the intestinal bacteria,
(C) Reduction of the abundance ratio of pathogenic bacteria in the intestinal bacteria.
(I-3) The equol-producing bacterium is a bacterium of the genus Adlercreutzia.
The short-chain fatty acid-producing bacterium is at least one bacterium selected from the group consisting of a bacterium belonging to the genus Parabacteroides, a bacterium belonging to the genus Blautia, and a bacterium belonging to the family Ruminococcaceae, and / or a bacterium whose pathogenic bacterium belongs to the family Desulfovibrionaceae. Is,
The agent for improving the intestinal bacterial flora according to (I-2).
(I-4) The agent for improving the intestinal flora according to any one of (I-1) to (I-3), which is used as an agent for increasing short-chain fatty acid-producing bacteria in the intestine.
The invention described in (I-4) can be rephrased as "an agent for increasing short-chain fatty acid-producing bacteria in the intestine, which contains a black soybean seed coat extract as an active ingredient".
(I-5) The agent for improving the intestinal bacterial flora according to any one of (I-1) to (I-3), which is used as a reducing agent for pathogenic bacteria in the intestine.
The invention described in (I-5) can be rephrased as "a reducing agent for pathogenic bacteria in the intestine containing a black soybean seed coat extract as an active ingredient".

(II) Method of using black soybean seed coat extract (II-1) For blending black soybean seed coat extract into an oral composition and imparting an effect of improving the intestinal bacterial flora to the oral composition. How to use black soybean seed coat extract.
(II-2) The improvement of the intestinal flora is to adjust the bacterial composition ratio of the intestinal flora so as to be at least one of the following (A) to (C) (II-1). how to use:
(A) The abundance ratio of equol-producing bacteria in the intestinal bacteria increased,
(B) The abundance ratio of short-chain fatty acid-producing bacteria in the intestinal bacteria increased,
(C) The abundance ratio of pathogenic bacteria in the intestinal bacteria is reduced.
(II-3) The equol-producing bacterium is a bacterium of the genus Adlercreutzia.
The short-chain fatty acid-producing bacterium is at least one bacterium selected from the group consisting of a bacterium belonging to the genus Parabacteroides, a bacterium belonging to the genus Blautia, and a bacterium belonging to the family Ruminococcaceae, and / or a bacterium whose pathogenic bacterium belongs to the family Desulfovibrionaceae. Is,
The usage method described in (II-2).

By orally ingesting the composition containing the black soybean seed coat extract of the present invention, the intestinal flora can be improved and the proportion of equol-producing bacteria present in the intestinal flora can be increased. As a result, in the presence of isoflavone, equol production in the intestine can be promoted, the amount of equol in the body can be increased, and the pharmacological action and health-promoting action of equol can be enjoyed.
In addition, by orally ingesting the composition containing the black soybean hull extract of the present invention, the intestinal flora is improved, and at the same time, short-chain fatty acid-producing bacteria in the intestine, particularly succinic acid-producing bacteria, acetic acid / lactic acid production. The abundance ratio of bacteria and butyric acid-producing bacteria can be increased. As a result, the amount of short-chain fatty acids produced in the intestine can be increased, the intestinal environment can be improved under weak acidity, and the pharmacological and health-promoting effects of the short-chain fatty acids can be enjoyed. Short-chain fatty acids can be metabolized by the gut microbiota to increase the gut microbiota.
Furthermore, by orally ingesting the composition containing the black soybean seed coat extract of the present invention, the intestinal flora can be improved and the abundance ratio of pathogenic bacteria in the intestine can be reduced. As a result, it is possible to suppress the adverse effects on the body caused by pathogenic bacteria in the intestine, and it is possible to contribute to the maintenance of health.

The abundance ratio (%) of equol-producing bacteria (genus Adlercreutzia) in the cecum contents of mice fed with black soybean seed coat extract is shown. Mean ± standard error (n = 5 or 6), Tukey HSD test shows that different letters are significantly different between groups (p <0.05) (same for Figures 2 and 3). The abundance ratio (%) of short-chain fatty acid-producing bacteria in the cecum contents of mice fed with black soybean seed coat extract is shown. (A) Intestinal abundance ratio of succinic acid-producing bacteria (bacteria of the genus Parabacteroides) (%), (B) Intraceal abundance ratio of acetic acid / lactic acid-producing bacteria (bacteria of the genus Blautia) (%), (C) Butyric acid production Intestinal abundance ratio (%) of bacteria (bacteria belonging to the family Ruminococcaceae). The cecal abundance ratio (%) of pathogenic bacteria in the cecal contents of mice fed with black soybean seed coat extract is shown.

(I) Gut microbiota improving agent The gut microbiota improving agent of the present invention (hereinafter, also simply referred to as “the present improving agent”) is effective as an extract of black soybean seed coat, preferably an edible extract. It is characterized by being an ingredient.

The black soybean used in the present invention is a black seed (grain) (black soybean) of a short-day annual plant belonging to the genus Glycine max (L.) Merrill of the leguminous family. Black soybeans include, for example, varieties such as middle-aged light black, tokachikuro, Iwaikuro, Tamadai black, Tanba black, Shinano black, and ganku, but any kind of black soybean seeds may be used.

Black soybeans can be separated into seed coats and embryos (cotyledons and hypocotyls) by, for example, using a sorting machine or the like. In the present invention, the seed coat of black soybean obtained by the separation can be used as a processing raw material. At the time of processing, the seed coat of black soybean may be in a separated state (raw or dried), or in a crushed or crushed state (crushed, crushed, and powdered). Includes).

As a method for extracting from black soybean seed coat, a generally used method can be used. Without limitation, for example, a method of immersing raw or dried black soybean seed coat (as is, or crude, shredded, crushed, crushed) in a water-soluble solvent; a method of extracting with stirring if necessary. ; Or the percolation method and the like can be mentioned. The temperature conditions used for extraction are not particularly limited, and may be any of low temperature, room temperature, and heating conditions (including high temperature), but heating conditions (including high temperature) are preferable. More specifically, when extracting with a hydrous lower alcohol described later, the temperature is in the range of 30 ° C. or higher, preferably 40 ° C. to 60 ° C., and although not limited, extraction under such temperature conditions takes 60 minutes or longer, preferably 90 ° C. Perform for about 1 to 120 minutes. When extracting with an acidic aqueous solution, the temperature is in the range of 50 ° C. or higher, preferably 50 to 80 ° C., and although not limited, the extraction under such temperature conditions is carried out for 10 minutes or longer, preferably about 20 minutes to 120 minutes.

The water-soluble solvent used for extraction is not particularly limited, and examples thereof include water, lower alcohols, and mixtures thereof. Examples of the lower alcohol include lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol and isopropyl alcohol and butanol. Ethanol can be preferably used as the lower alcohol. The water-soluble solvent is preferably water or a hydrous lower alcohol (particularly hydrous ethanol), and more preferably water. When a hydrous lower alcohol is used as a solvent, the amount of lower alcohol contained therein is preferably 80% by volume or less.

The water-soluble solvent used for extraction is preferably adjusted to be acidic. Although not particularly limited, the pH range of the water-soluble solvent is preferably in the range of about pH 1 to 4, and particularly preferably in the range of pH 1 to 2. In order to adjust the pH of the water-soluble solvent within such a range, an appropriate acidic substance such as an organic acid or an inorganic acid can usually be used.

Specific examples of acidic substances include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, and boric acid; as well as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, 10-camphasphonic acid, and fluorosulfonic acid. Organic acids such as (above, sulfonic acid), formic acid, acetic acid, citric acid, and oxalic acid (above, carboxylic acid) can be mentioned. An acid having a sulfo group is preferable, and specific examples thereof include sulfuric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, 10-camphasphonic acid, and fluorosulfonic acid. Of these, sulfuric acid is preferable. The normality of the acid in the water-soluble solvent is not particularly limited as long as the water-soluble solvent is in the above pH range, but is preferably in the range of 0.01 to 0.5 N, more preferably 0.03. It is in the range of ~ 0.5N.

To the extent that the effects of the present invention are exhibited, the obtained extract is further subjected to purification treatment such as filtration, coprecipitation or centrifugation to remove solids, extraction treatment, adsorption treatment and the like, if necessary. May be good.

The black soybean seed coat extract thus prepared may be sterilized by a known method such as UHT sterilization or retort sterilization, if necessary, as long as the effect of the present invention is exhibited.

The present improving agent may be in any form as long as it is orally administered. In addition, as long as it has a form of oral administration (oral ingestion), it is classified according to its use (pharmaceutical products, quasi-drugs, foods and drinks [foods for specified health use, foods with functional claims, foods with nutritional function, etc. And supplements]) are not particularly limited. Foods and drinks are preferable, and foods for specified health use or foods with functional claims that can claim their actions and effects are more preferable. In addition, the present improving agent may be applied to feeds and pet foods for animals other than humans (non-human animals: including livestock, poultry, and pets). Further, the present improving agent may be an additive added to pharmaceutical products, quasi-drugs, foods and drinks, feeds or pet foods.

As an oral administration form, specifically, an extract prepared by the above extraction method is prepared in the form of a liquid agent (including an extract form or a syrup) or a jelly agent; the extract is in the form of powder or granules by a conventional method. Powders, fine granules, or granules formulated in; capsules filled with liquids, powders, or granules (hard capsules, soft capsules); or powders or granules are further tableted to form tablets. Can be mentioned (solid preparation).

The present improving agent is prepared in various dosage forms (oral administration form) by combining the above black soybean seed coat extract with conventionally known edible carriers, excipients, etc. that are pharmaceutically acceptable or acceptable as foods and feeds. You can also do it.

When the present improving agent is in the form of a liquid preparation, it may be cryopreserved, or it may be preserved after removing water by freeze-drying or the like. Freeze-dried preparations, dry syrups, etc. are used by adding sterilized water or the like at the time of use and dissolving them again.

When the present improving agent is in the form of a solid agent, for example, in the case of tablets, conventionally known carriers in the art can be widely used. Examples of such carriers include excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, and stearic acid; water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin. Binders such as solutions, carboxymethyl cellulose, cellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone, crystalline cellulose, hydroxypropyl cellulose, hypromerose, sodium alginate; dried starch, canten powder, laminaran powder, sodium hydrogen carbonate, polyoxyethylene sorbitan fatty acid Disintegrants such as esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, crospovidone, povidone, low-substituted hydroxypropyl cellulose; disintegration inhibitors such as stear, cocoa butter, hydrogenated oil; quaternary ammonium salts, lauryl Absorption enhancer such as sodium sulfate; Moisturizer such as glycerin; Adsorbent such as starch, lactose, kaolin, bentonite, colloidal silicic acid; Lubricating agent such as purified talc, stearic acid, boric acid powder, polyethylene glycol, etc. Can be used. Further, the tablet may be a tablet coated with a normal skin, for example, a sugar-coated tablet, a gelatin-encapsulated tablet, an enteric-coated tablet, a film-coated tablet or a double tablet, or a multi-layer tablet, if necessary. Further, the composition containing the active ingredient can be filled into a conventionally known capsule made of gelatin, pullulan, starch, gum arabic, hydroxypropylmethyl cellulose (HPMC) or the like as a raw material to obtain a capsule.

Further, in the form of a pill, a carrier conventionally known in the art can be widely used. Examples thereof include excipients such as glucose, lactose, starch, cacao butter, hardened vegetable oil, kaolin and talc, binders such as gum arabic powder, tragant powder, gelatin and ethanol, and disintegrants such as laminarin and canten. Can be used.

In addition to the above, as additives, for example, surfactants, absorption promoters, adsorbents, fillers, preservatives, stabilizers, emulsifiers, solubilizers, etc. may be appropriately selected and used according to the form of the preparation. can.

All of these forms can be prepared by using ordinary methods in the art. For example, tablets are mixed and dispersed well after adding the above active ingredients and other excipients necessary for obtaining tablets as appropriate. It can be obtained by locking. Further, the powder can be obtained by appropriately adding the above-mentioned active ingredient and other excipients necessary for obtaining the powder, and mixing and powdering by a suitable method.

The present improving agent may have a form of ordinary food or drink in addition to the above-mentioned pharmaceutical form. The food or drink can be produced by adding an additive containing the black soybean seed coat extract described above or the black soybean seed coat extract described later to various foods and drinks. The food or drink may be in a form that can be orally ingested, such as a solution, suspension, emulsion, jelly (gel), sol, powder, or solid molded product, and is not particularly limited. Specifically, for example, instant noodles, retort foods, canned foods, microwave foods, instant soups / miso juices, freeze-dried foods and other instant foods; soft drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea. Beverages, powdered beverages, concentrated beverages, nutritional beverages, alcoholic beverages and other beverages; bread, pasta, noodles, cake mixes, fried flour, bread flour and other wheat flour products; candy, caramel, chewing gum, chocolate, cookies, biscuits, cakes , Pies, snacks, crackers, Japanese sweets, desserts and other sweets; sauces, tomato processed seasonings, flavored seasonings, cooking mixes, sauces, dressings, soups, curry and stew ingredients Fats and oils such as processed fats and oils, butter, margarine, and mayonnaise; dairy beverages, yogurts, cheese, fermented milk, lactic acid bacteria beverages, ice creams, creams and other dairy products; processed egg products such as pudding and mayonnaise; fish ham Processed marine products such as sausages and pastes of marine products; Processed livestock products such as meat hams and sausages; Processed agricultural products such as canned agricultural products, jams and marmalades, pickles, boiled beans, cereals; frozen foods, nutritional foods, etc. .. Although not limited, it is preferably an acidic food from the viewpoint of suppressing precipitation and deterioration of color of polyphenols derived from black soybean seed coat extract.

Further, the present improving agent may have the form of ordinary feed or pet food. These feeds and pet foods can also be produced by adding the above-mentioned black soybean seed coat extract to various feeds and pet foods.

The content of the black soybean seed coat extract in this improving agent may be appropriately set up to 100% by mass according to the type of form and the type of use (pharmaceuticals, foods, feed / pet food, etc.) described above. can. The dose (ingestion) of this improving agent may be appropriately changed depending on the type of human or animal, the sex and age of the subject, the condition of the subject and the degree of symptoms. Although not limited, for example, the daily dose (intake) for one human adult (body weight 50 kg) is usually 10 to 10 in terms of the amount of black soybean seed coat extract (dry amount) contained in this improving agent. About 500 mg can be mentioned. Usually, it is used in the form of oral administration (ingestion) once a day or divided into 2 to 3 times a day. The time of administration is not particularly limited, and for example, any one or more time zones of morning, noon, and evening meals can be exemplified. Also, without limitation, it may be with a meal or within 30 minutes before or after a meal.

(Action of intestinal bacterial flora improving agent)
The target audience for this improver is humans or non-human animals. It is preferably human. Non-human animals include pets (pets), laboratory animals, and animals kept in zoos and aquariums. By administering (ingesting) this improving agent to humans or non-human animals, the bacterial composition ratio of the intestinal flora of the human or non-human animals is adjusted, and the abundance ratio of bacteria beneficial to the living body is increased. It is possible to reduce the abundance of bacteria that have an adverse effect on the living body. In addition, by reducing the change in the bacterial composition ratio of the intestinal flora caused by ingesting a high-fat diet, especially the increase in the abundance of bacteria that have an adverse effect on the living body, before or near the intake of a high-fat diet. It is possible to return to a state or even better.

Ingestion of this improving agent induces, for example, the following changes in the intestinal flora.
Increase in the abundance of intestinal bacteria that are beneficial to the living body (A) Increase in the abundance of equol-producing bacteria in the intestinal bacteria Equol production in the intestine by ingestion of this improving agent by humans or non-human animals The abundance rate of bacteria can be increased. Equol-producing bacteria have the effect of converting isoflavones such as daidzein ingested into the body into equol with higher estrogen-like activity. That is, this improving agent promotes equol production by increasing the abundance ratio of equol-producing bacteria present in the intestine by being taken into the body, thereby promoting metabolism and activation of isoflavones. Examples of the equol-producing bacterium preferably include bacteria of the genus Adlercreutzia, and more preferably Adlercreutzia equoli faciens.

In addition, isoflavone is a kind of flavonoid and is a general term for compounds based on the aglycone skeleton of daidzein and its analogs glycitein and genistein. In addition to aglycone, isoflavones include glycosides, their acetylated products, and malonylated products. Specifically, daidzin, glycitein and genistein as isoflavone aglycon, daidzin, glycitin and genistine as isoflavone glycosides, acetyldaidzin, acetylglycitin and acetylgenistine as acetylated isoflavone glycosides, and malonyl of isoflavone glycosides. Examples include malonyldaidzin, malonylglycitin and malonylgenistine. These isoflavones may be used alone or in combination of two or more. Examples of edible materials containing isoflavones include isoflavone-containing sites (soybeans, soybean germ shafts, etc.) of plants containing isoflavones (for example, soybean plants such as soybeans), and processed products thereof (for example, dried products and extracts). , Those processed foods (for example, tofu, soymilk, fermented soymilk, natto, etc.) can be mentioned.

Thus, according to this improving agent, isoflavones are produced by improving the intestinal flora of human or non-human animals and adjusting the bacterial composition ratio of the intestinal flora so as to increase the abundance of equol-producing bacteria. It is possible to increase the production of equol and exert the action effect based on equol. The effects based on equol are not limited, but alleviate the symptoms of menopausal disorders (such as swelling / burning, headache, dizziness, autonomic imbalance-like symptoms, tachycardia, and blood glucose fluctuation); Suppression; Suppression of excess skin oil secretion (prevention or improvement of acne and pimples); Prevention of cancer (breast cancer, uterine body cancer, prostate cancer, stomach cancer, etc.); Type II diabetes (fasting blood glucose level, insulin resistance) Prevention or amelioration of lifestyle-related diseases (diabetes, hyperlipidemia, arteriosclerosis, high cholesterol, etc.); Prevention or improvement of skin firmness and elasticity, stains, wrinkles, sagging; improvement of blood sugar (prevention or improvement of coldness, stiff shoulders, tension headache, dullness and dryness of skin); prevention or improvement of male hair loss At least one action and effect selected from improvement and the like can be expected.

(B) Increasing the abundance ratio of short-chain fatty acid-producing bacteria in the intestinal bacteria Increase the abundance ratio of short-chain fatty acid-producing bacteria present in the intestine by ingesting this improving agent by humans or non-human animals. Can be done. Specifically, the short-chain fatty acid-producing bacteria are selected from the group consisting of bacteria of the genus Parabacteroides that produce succinic acid, bacteria of the genus Blautia that produce acetic acid and lactic acid, and bacteria of the family Ruminococcaceae that produce butyric acid. At least one can be mentioned. According to these short-chain fatty acid-producing bacteria, the abundance of short-chain fatty acids that exert useful functions in the intestine can be increased. In other words, according to this improver, the intestinal flora of human or non-human animals is improved, and the abundance of short-chain fatty acids that exert useful functions in the intestine is increased in the intestine. By adjusting the bacterial composition ratio, it becomes possible to exert an action effect based on short-chain fatty acids.
The effects of short-chain fatty acids in the intestine and their relationship to health are explained below.

a. Strengthening the barrier function from harmful substances Acetic acid is said to have the function of enhancing the barrier function of the large intestine. Butyric acid is also said to have the effect of promoting the secretion of mucin, a mucosal substance, and protecting the large intestine by activating the MUC2 gene in intestinal cells.

b. Cancer prevention Short-chain fatty acids are said to help prevent colorectal cancer because they make the intestines weakly acidic, making it difficult to produce harmful secondary bile acids. In addition, butyric acid is said to suppress the onset of colorectal cancer by suppressing the abnormal proliferation of large intestine cells, promoting apoptosis, and suppressing the lesions of large intestine cells.

c. Prevention of obesity Short-chain fatty acids act on short-chain fatty acid receptors in adipocytes to suppress the uptake of energy into adipocytes and prevent adipocyte enlargement. It also acts on short-chain fatty acid receptors in nerve cells and promotes energy consumption via the sympathetic nervous system, and has the function of adjusting the energy balance.

d. Prevention of diabetes Butyric acid acts on L cells in the intestinal tract to promote the secretion of the intestinal hormone GLP-1. GLP-1 has the effect of preventing and ameliorating diabetes, suppressing the decrease in the number of pancreatic β-cells that secrete insulin, and promoting insulin secretion.

e. Suppression of appetite Butyric acid secretes GLP-1 as well as intestinal hormones such as PYY from L cells in the intestinal tract. GLP-1 and PYY are known to act on the brain to suppress appetite, maintain a feeling of fullness, and prevent overeating.

f. Regulation of immune function It is said that about 60% of the immune cells in the whole body are concentrated in the intestine, and the imbalance of the intestine (especially excessive immune response) affects the whole body. Butyric acid has the effect of increasing immune cells called Treg cells, which suppress the excessive immune response, and it is known that butyric acid promotes the acetylation of histones in colonic epithelial cells. Butyric acid is also said to be useful for inflammatory bowel disease, which is an immune disease of the intestine. Furthermore, it is said that lactic acid bacteria existing in the intestine are useful for prevention and improvement of autoimmune diseases.

g. Others By increasing the proportion of lactic acid bacteria present in the intestine, in addition to the prevention or amelioration of autoimmune diseases mentioned above, prevention or amelioration of cold illness, increase of good bacteria and suppression of bad bacteria are used to regulate the intestine. , Diarrhea, bowel movements, etc. can be expected to improve.

Reduction of the abundance ratio of intestinal bacteria that have an adverse effect on the living body (C) Reduction of the abundance ratio of pathogenic bacteria in the intestinal bacteria When humans or non-human animals ingest this improving agent, pathogens existing in the intestine The abundance ratio of sex bacteria can be reduced. Therefore, according to this improving agent, maintenance and promotion of health can be achieved by improving the intestinal flora of human or non-human animals and adjusting the bacterial composition ratio of the intestinal flora so as to reduce pathogenic bacteria. It becomes possible to plan.
Specific examples of pathogenic bacteria include bacteria of the family Desulfovibrionaceae. It is known that the intestinal bacterial flora is disrupted or disturbed by ingestion of a high-fat diet. As shown in the experimental example (see FIG. 3) described later, bacteria of the family Desulfovibrionaceae in the intestinal flora are destroyed by ingesting the primary bile acid seen in a high-fat diet. Will also increase. Even in such a state, by taking this improving agent, bacteria of the family Desulfovibrionaceae in the intestinal flora can be reduced, and the intestinal flora can be restored to its original state or better. It can be improved.

The subject to whom this improving agent is applied is not particularly limited as long as it is necessary to enjoy the above-mentioned effects. Although not limited, as an example, it can be broadly targeted to those who ingest a high-fat diet, which is a factor that alters the bacterial composition of the gut microbiota. Those who prefer to consume a high-fat diet are preferable, and those who are obese or prone to obesity are more preferable. A fat diet is a diet containing a large amount of fat and is not strictly defined, but a diet in which fat accounts for approximately 30 to 40% or more of the total energy intake (fat energy ratio) is mentioned. be able to. For example, meat such as beef rose, beef loin, ground beef, con beef (cow), pork rose, pork loin, bacon (pork), sausage (pork); Kind; Eggs and dairy products such as egg yolk, fresh cream (milk fat), and Camambale cheese; Grains such as croissant, Danish pastry, corn snack, potato chips; Macademian nuts, peanuts, almonds, cashew nuts, etc. Nuts: A diet rich in fat-containing ingredients such as fried, kin flour, mayonnaise, French dressing, olive oil, sesame oil, salted butter, and margarine tends to be a high-fat diet. Obesity is judged based on BMI (Body Mass Index: body weight [kg] / height [m] squared), and a BMI of 30 or more is evaluated as obesity, and 25 or more and less than 30 is evaluated as obese (slightly obese). Can be done.

Whether or not the intestinal bacterial flora is improved by taking (administrating, ingesting) this improving agent can be evaluated and confirmed by analyzing the intestinal bacteria contained in feces. When evaluating using non-human animals, it can be evaluated and confirmed by performing intestinal bacterial flora analysis using the contents of the cecum, as shown in the experimental examples described later.

(II) Method of using black soybean seed coat extract The present invention also provides a method of using black soybean seed coat extract.
One of the methods of use is a method of using a black soybean seed coat extract for imparting an action of improving the intestinal bacterial flora to an oral composition.
The improvement of the intestinal flora targeted by the present invention is specifically that the bacterial composition ratio in the intestinal flora is (A) an increase in the abundance ratio of equol-producing bacteria in the intestinal bacteria, (B). It means that the intestinal flora is adjusted so that the abundance of short-chain fatty acid-producing bacteria in the intestinal bacteria increases and / or (C) the abundance of pathogenic bacteria in the intestinal bacteria decreases. do.
Examples of the equol-producing bacterium preferably include bacteria of the genus Adlercreutzia, and more preferably Adlercreutzia equoli faciens. Examples of short-chain fatty acid-producing bacteria include at least one selected from the group consisting of bacteria of the genus Parabacteroides that produce succinic acid, bacteria of the genus Blautia that produce acetic acid and lactic acid, and bacteria of the genus Ruminococcaceae that produce butyric acid. be able to. Specific examples of pathogenic bacteria include bacteria of the family Desulfovibrionaceae.

The method can be carried out by blending the black soybean seed coat extract with the target oral composition. In addition, instead of the black soybean seed coat extract, the above-mentioned improving agent containing the black soybean seed coat extract as an active ingredient can also be used.

Target oral compositions include compositions that are orally administered to humans or compositions that are ingested by humans, specifically oral medicines, quasi-drugs, and foods and drinks. Food and drink are preferable. When targeting non-human animals, feed or pet food can be used as the oral composition.

Kind of black soybean used as a raw material of black soybean seed coat extract used in the method of the present invention, method for obtaining black soybean seed coat, black soybean seed coat extract, particularly black soybean seed coat acidic extract which is a preferable aspect of the extract. The method for preparing the above is as described in (I) above, and can be incorporated in this column. The blending amount of the black soybean seed coat extract with respect to the oral composition is not particularly limited as long as it can impart an action of enhancing the equol-producing ability by the intestinal bacteria to the oral composition. The evaluation was made when the subject was continuously ingested an oral composition containing the black soybean seed coat extract (target composition) and when the subject was not mixed with the black soybean seed coat extract (comparative composition). It can be carried out by comparing the bacterial composition ratio of the intestinal flora with that of the case of continuous ingestion. The bacterial composition of the intestinal flora when the target composition is continuously ingested is compared with the bacterial composition of the intestinal flora when the comparative composition is continuously ingested, as described above ( When at least one of the tendencies A) to (C), preferably two or more, and more preferably three, is observed, the oral composition can be mixed with the black soybean seed coat extract to the oral composition. Therefore, it can be judged that the effect of improving the intestinal bacterial flora is imparted.

As described above, in the present specification, the terms "contains" and "contains" include the meanings of "consisting of" and "consisting of substantially".

Hereinafter, the present invention will be described with reference to experimental examples in order to help understanding the structure and effects of the present invention. However, the present invention is not limited by these experimental examples. The following experiments were performed under room temperature (25 ± 5 ° C.) and atmospheric pressure conditions, unless otherwise noted. Unless otherwise specified, "%" described below means "mass%" and "parts" means "parts by mass".

Experimental Example 1 Black soybean seed coat extract intake test (effect on intestinal bacterial flora)
A test animal (mouse) was fed with a black soybean seed coat extract to measure body weight, food intake, and cecal content, and genomic DNA was extracted from the cecal content to analyze the intestinal flora.

(1) Test animal Animal: Male C57BL / 6J mouse 8 weeks old (purchased from Japan SLC)
Breeding environment: Room temperature 25 ° C, humidity 55%, lighting was done by turning on the fluorescent lights in the room every 12 hours from 7:00 am to 7:00 pm.
Breeding period: After the animals were brought in, after a 2-week acclimation period with normal food solid samples, the animals were divided into the following test groups (a) to (d) so that the average body weight of each group would be equal (each). Group n = 6-7). Free intake of each feed and drinking water during the breeding period

Test area:
(A) Control group: Feed (normal diet: solid sample D12450J: Research Diet, the same applies hereinafter) + drinking water (distilled water, the same applies hereinafter) (b) Control group A: feed (normal diet) + 0. Ingest 3% cholic acid-added drinking water (c) Black soybean seed coat extract group: Feed (normal diet + 1.7% black soybean husk extract) + 0.3% cholic acid-added drinking water

Chronocare (manufactured by Fujicco Co., Ltd.) was used as the "black soybean seed coat extract". Chronocare is an acidic extract of black soybean seed coat containing 58% by mass or more of black soybean seed coat polyphenol, and contains excipients in a proportion of about 15% by mass. Black soybean seed coat polyphenols include, for example, Procyanidin B2, Procyanidin C1, Procyanidin A2, Procyanidin B5, epicatechin, and cyanidin-3-glucoside. The "1.7%" of the above "1.7% black soybean seed coat extract" is an amount converted into the content of the black soybean seed coat extract contained in chronocare.

(2) Test method and its results 1. Body weight, food intake, cecal content The body weight, cecal content, and daily food intake (g / day / mice) of the test animals in each test group after free intake of each feed and drinking water for 2 weeks. ) Was measured. The results are shown in Table 1 as the average value of each group.

As shown in Table 1 above, the amount of cecal content tended to increase significantly in the (c) black soybean hull extract group as compared with both the (a) control group and the (b) control A group. Was done. On the other hand, there was no significant difference in body weight and food intake between the groups.

2. Analysis of intestinal bacterial flora in the contents of the cecum DNA was extracted from the contents of the cecum collected after free intake of each feed and drinking water for 2 weeks for the test animals in each test group, and DNA was extracted according to a conventional method. The V3-V4 region of the 16S rRNA gene was amplified and meta 16S flora analysis was performed by Illumina MiSeq. A total of 871,376 reads (51,258 reads on average) of the 17 samples obtained were used for bacterial flora analysis using QIIME (Quantitave Insights Into Microbial Ecology). From the obtained bacterial flora analysis, the bacterial flora composition at the family / genus level was determined.

The abundance ratio (%) of bacteria of the genus Adlercreutzia, which is an equol-producing bacterium, in the cecum is shown in FIG. Similarly, among the short-chain fatty acids, the abundance ratio (%) in the cecum of bacteria producing succinic acid (bacteria of the genus Parabacteroides) is shown in FIG. The internal abundance ratio (%) is shown in FIG. 2 (B), and the abundance ratio (%) of butyric acid-producing bacteria (bacteria belonging to the family Ruminococcaceae) in the cecum is shown in FIG. 2 (C). In addition, the abundance ratio (%) in the cecum of bacteria belonging to the family Desulfovibrionaceae, which is a pathogenic bacterium, is shown in FIG.

As shown in FIG. 1, equol-producing bacteria in the intestine were significantly increased in (c) black soybean seed coat extract group as compared with (a) control group and (b) control A group. Was confirmed. Further, as shown in FIGS. 2 (A) to 2 (C), (c) black soybean hull extract group is a short-chain fatty acid in the intestine as compared with (a) control group and (b) control A group. It was confirmed that the number of producing bacteria (succinic acid-producing bacteria, acetic acid / lactic acid-producing bacteria, butyric acid-producing bacteria) increased remarkably. On the other hand, as shown in FIG. 3, the pathogenic bacteria in the intestine were significantly reduced in (c) black soybean seed coat extract group as compared with (a) control group and (b) control A group. It was confirmed that there was.

It is known that the intestinal bacterial flora collapses when a high-fat diet is continuously ingested. In this experiment, instead of ingesting a high-fat diet, the test animals ((b) control group A, (c) black soybean hull extract group) were allowed to ingest drinking water containing primary bile acid (cholic acid). , The intestinal flora was disrupted or could be disrupted. As shown in FIGS. 1 to 3, even in such a state, by ingesting the black soybean seed coat extract, useful bacteria such as equol-producing bacteria and short-chain fatty acid-producing bacteria in the intestine increased remarkably (FIGS. 1 and 2). (See), on the other hand, the pathogenic bacteria increased by ingestion of primary bile acids were significantly reduced by ingestion of black soybean seed coat extract (see FIG. 3). These results suggest that oral ingestion of black soybean seed coat extract can improve the bacterial composition ratio of the intestinal flora and thereby promote health accordingly.

Claims (5)

An agent for improving the intestinal bacterial flora containing black soybean seed coat extract as an active ingredient. The agent for improving the intestinal bacterial flora according to claim 1, which exhibits at least one action and effect selected from the group consisting of the following (A) to (C):
(A) Increase in the abundance ratio of equol-producing bacteria in the intestinal bacteria,
(B) Increase in the abundance ratio of short-chain fatty acid-producing bacteria in the intestinal bacteria,
(C) Reduction of the abundance ratio of pathogenic bacteria in the intestinal bacteria. The equol-producing bacterium is a bacterium of the genus Adlercreutzia.
The short-chain fatty acid-producing bacterium is at least one bacterium selected from the group consisting of a bacterium belonging to the genus Parabacteroides, a bacterium belonging to the genus Blautia, and a bacterium belonging to the family Ruminococcaceae, and / or a bacterium whose pathogenic bacterium belongs to the family Desulfovibrionaceae. Is,
The agent for improving the intestinal bacterial flora according to claim 2. Used as an enhancer for short-chain fatty acid-producing bacteria in the intestine and / and
Used as a reducer for pathogenic bacteria in the intestines,
The agent for improving the intestinal bacterial flora according to any one of claims 1 to 3. A method for using a black soybean seed coat extract for blending a black soybean seed coat extract with an oral composition to impart an action of improving the intestinal bacterial flora to the oral composition.

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