WO2018190407A1 - COMPOSITION FOR ACTIVATING Toll-LIKE RECEPTOR 2 - Google Patents

Abstract

The purpose of the present invention is to provide a novel composition for activating a Toll-like receptor 2. The present invention provides a composition for activating a Toll-like receptor 2 (TLR2) that includes bacteria belonging to Bifidobacterium bifidum or a treated product thereof as an active ingredient.

Description

Composition for activating Toll-like receptor 2 Reference to related applications

This application enjoys the benefit of priority of Japanese Patent Application No. 2017-80361 (filing date: April 14, 2017), which is a prior Japanese application, the entire disclosure of which is incorporated herein by reference. To be part of

The present invention relates to a composition for activating Toll-like receptor 2.

There are two types of living immune systems: innate immunity and acquired immunity. In contrast to acquired immunity that exists only in higher organisms such as humans, innate immunity is an immune mechanism that is widely conserved from insects to humans. When cells in charge of innate immunity (macrophages, dendritic cells, etc.) enter the pathogenic bacteria from the outside world, they quickly detect them and emit warning signals such as cytokines in addition to direct attacks such as phagocytosis. It functions as the first step of infection protection by activating the acquired immune system. A receptor protein that plays a role in discovering bacteria as the first trigger in such a series of immune reaction systems is a Toll-like receptor (referred to herein as “TLR”).

10 types of TLRs have been identified in humans so far, and each TLR is considered to have a different molecular structure. Among them, TLR2 is known to mediate cell signaling in response to microbial cell wall components such as peptidoglycan (PGN), lipoteichoic acid, microbial lipoproteins, and zymosan (Non-Patent Document 1). For example, it has been confirmed that the activation of TLR2 can improve the intestinal barrier function through enhanced protection of tight junctions in the intestinal tract (Non-patent Document 2).

Thus, cell signaling via TLR2 governs various in vivo reactions including the biological immune system. The present inventors paid attention to the usefulness of the cell signal transmission system via TLR2, and conducted intensive studies on food materials capable of activating TLR2. As a result, it was found that bacteria belonging to Bifidobacterium bifidum (particularly Bifidobacterium bifidum OLB6378) have a strong TLR2 activation ability.

That is, an object of the present invention is to provide a novel Toll-like receptor 2 activation composition and Toll-like receptor 2 activator.

According to the present invention, the following inventions are provided.
[1] A composition for activating TLR2 and a TLR2 activator comprising a bacterium belonging to Bifidobacterium bifidum (hereinafter sometimes referred to as “bifidobacteria of the present invention”) or a processed product thereof as an active ingredient (Hereafter, it may be called "the composition and agent of this invention.").
[2] The composition according to [1] above, wherein the bacterium belonging to Bifidobacterium bifidum has a TLR2 activation ability twice or more that of Bifidobacterium breve JCM1192T strain.
[3] The composition according to [1] or [2] above, wherein the bacterium belonging to Bifidobacterium bifidum is Bifidobacterium bifidum OLB6378 strain ( Bifidobacterium bifidum OLB6378, accession number: NITE BP-31). object.
[4] The composition according to any one of the above [1] to [3] for use in improving the intestinal barrier function.
[5] The composition according to [4] above, which is used for treatment, prevention and / or amelioration of a disease or symptom which can be treated, prevented or ameliorated by improving intestinal barrier function.
[6] The composition according to [5] above, wherein the disease is necrotizing enterocolitis, ulcerative colitis, persistent diarrhea, allergic disease or food hypersensitivity.
[7] The composition according to any one of [1] to [6] above, which is for newborns, infants, children and / or elderly people.
[8] The composition according to any one of [1] to [7] above, wherein the bacteria belonging to Bifidobacterium bifidum are heat-treated cells.
[9] A method for activating TLR2 and improving intestinal barrier function, comprising ingesting or administering an effective amount of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof to a subject in need thereof Methods and methods for treating, preventing and / or ameliorating diseases or symptoms that can be treated, prevented or ameliorated by improving intestinal barrier function.
[10] Bacteria belonging to Bifidobacterium bifidum for producing TLR2 activator, for producing intestinal barrier function improving agent, for TLR2 activation, or for improving intestinal barrier function Or use of the processed product.
[11] For the manufacture of a therapeutic agent, preventive agent or ameliorating agent for a disease or condition that can be treated, prevented or ameliorated by improving the intestinal barrier function, or treating, preventing or ameliorating by improving the intestinal barrier function Use of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof for the treatment, prevention and / or amelioration of a possible disease or symptom.
[12] A bacterium belonging to Bifidobacterium bifidum or a processed product thereof for use in activating TLR2 or for improving intestinal barrier function.
[13] A bacterium belonging to Bifidobacterium bifidum or a processed product thereof for use in the treatment, prevention and / or amelioration of a disease or symptom which can be treated, prevented or ameliorated by improving the intestinal barrier function.

According to the composition and the preparation of the present invention, activation of TLR2 and its related effects can be expected in ingestion subjects and administration subjects. Bacteria belonging to Bifidobacterium bifidum, which is an active ingredient of the present invention, are bacteria contained in food materials that have been ingested by mankind for many years. Therefore, the compositions and preparations of the present invention have no side effects. This is advantageous in that activation of TLR2 can be achieved.

It is the figure which showed the measurement result (Average value +/- standard deviation, detection limit 15.6ng / mL) of TLR2 activation ability (SEAP density | concentration) of Bifidobacterium bifidum OLB6378 strain and another test strain.

Detailed description of the invention

The active ingredient of the present invention is a bacterium belonging to Bifidobacterium bifidum . In the present invention, the bacterium belonging to Bifidobacterium bifidum has a strong TLR2 activation ability. That is, in the present invention, the Bifidobacterium having Toll-like receptor 2 activation ability higher than the Toll-like receptor 2 activation ability of the general-purpose strain Bifidobacterium breve JCM1192T strain. Bacteria belonging to bifidum can be used as an active ingredient, preferably 2 times or more (preferably 2.2 times or more, more preferably 2.4 times or more) with respect to Bifidobacterium breve JCM1192T strain Bacteria belonging to Bifidobacterium bifidum having Toll-like receptor 2 activation ability can be used as active ingredients. Bifidobacterium breve JCM1192T strain can be obtained from RIKEN, BioResource Center, Microbial Materials Development Office. The degree of TLR2 activation ability can be evaluated using the concentration of alkaline phosphatase (SEAP) secreted by TLR2 activation as an index.

In a preferred embodiment of the present invention, as bacteria belonging to the Bifidobacterium bifidum, Bifidobacterium bifidum OLB6378 strain (Bifidobacterium bifidum OLB6378, Accession Number: NITE BP-31) can be used. This strain is deposited with the Patent Microorganism Depositary Center for Product Evaluation Technology. The contents specifying the deposit are described below.

(1) Depositary Institution: National Institute of Technology and Evaluation, Patent Microorganism Depositary Center (2) Contact: 2-5-8 Kazusa Kamashi, Kisarazu City, Chiba Prefecture 292-0818 (phone number 0438-20-5580)
(3) Accession number: NITE BP-31
(4) denotation for identification: Bifidobacterium bifidum OLB6378
(5) Date of original deposit: October 26, 2004 (6) Date of transfer to deposit under the Budapest Treaty: January 18, 2006

Bifidobacterium bifidum OLB6378 is a Gram-positive obligate anaerobe derived from human infant feces. When this bacterium is applied on a plate of BL agar medium (Eiken Chemical Co., Ltd.) and cultured at 37 ° C for 48 hours in anaerobic condition using AnaeroPack Kenki (Mitsubishi Gas Chemical Co., Ltd.), an opaque circular hemispherical gloss is obtained. A colony is formed. In addition, amplification products are observed by PCR using specific primers of Bifidobacterium bifidum (Intestinal Flora Symposium 8, Molecular Biology Detection and Identification of Intestinal Flora, Toshihiro Mitsuoka, Takahiro Matsumoto). Moreover, it has fermentability with respect to galactose, glucose, fructose, lactose, and gentiobiose.

As a medium for culturing the bifidobacteria of the present invention, a medium usually used for a bifidobacteria medium can be used. That is, the medium that can be used in the present invention is not particularly limited, and any medium can be used as long as it contains a nitrogen source, an inorganic substance, and other nutrients in a predetermined range in addition to the main carbon source. As the carbon source, lactose, glucose, sucrose, fructose, starch hydrolysate, molasses, etc. can be used according to the assimilation ability of the bacteria used. As the nitrogen source, organic nitrogen-containing substances such as casein hydrolyzate, whey protein hydrolyzate, and soy protein hydrolyzate can be used. In addition, meat extract, fish extract, yeast extract, etc. can be used as a growth promoter.

The bifidobacteria of the present invention is preferably cultured under anaerobic conditions, and a known method such as a method of culturing while aeration of carbon gas can be applied. Culture can also be performed using other techniques such as air conditions or batch culture conditions. The culture temperature is preferably 25 to 50 ° C., and particularly preferably 35 to 42 ° C. However, the present invention is not limited to this, and other temperature conditions may be used as long as the bacteria can grow. The pH of the medium during the culture is preferably maintained at 6.0 to 7.0, but may be other pH conditions as long as the bacteria can grow. The culture time is preferably 3 to 48 hours, more preferably 8 to 24 hours, and particularly preferably 10 to 20 hours. However, other culture time may be used as long as the bacteria can grow.

The obtained microbial cells can be contained in the composition and the preparation of the present invention as a processed product subjected to the following treatment. The processed product of bifidobacteria of the present invention includes a culture that is still after completion of the culture, a culture that has been subjected to centrifugation or filtration after the completion of the culture, a concentrate thereof, and further processed into a paste, various methods Dried products (spray-dried products, freeze-dried products, vacuum-dried products, drum-dried products, etc.), liquids dispersed in media, diluents with diluents, heat-treated foods (heat-treated cells), ultraviolet rays And / or radiation-treated products (light-treated cells) treated with radiation, drug-treated products (drug-treated cells) treated with chemicals (bactericides, antibacterial agents, bacteriostatic agents), dried products with a mill, etc. The crushed crushed material is included. Centrifugation, filtration, and concentration are performed by commonly used techniques. Drying can be performed by, for example, vacuum drying, spray drying, freeze drying, drum drying, or the like. In the present specification, these may be simply abbreviated as “processed products”.

Bacteria belonging to Bifidobacterium bifidum obtained by the above method and processed products thereof are live bacteria, or heat-treated bacteria, and then processed as a crushed or unground product, or a mixture of plural kinds Can be contained in the composition and the preparation of the present invention. If it is a living bacterium, effects such as growth and colonization in the intestine after ingestion or administration can be expected, and further, the effects of the present invention can be expected to be sustained. In addition, 0.1 mL of a heat-treated cell (for example, a suspension (dispersion) of the heat-treated cell) is sampled and smeared on a petri dish containing a medium in which bifidobacteria can grow. If the colony is not formed even if cultured, it is not necessary to consider the characteristics of bifidobacteria that it is difficult to survive in the presence of oxygen, and the composition and agent of the present invention This is preferable because the application range is expanded.

When the heat-treated bifidobacteria of the present invention is used in the present invention, for example, the heating temperature is usually 60 to 300 ° C., preferably 60 to 200 ° C., more preferably 60 to 150. ° C, more preferably 60-140 ° C, more preferably 60-130 ° C, more preferably 60-120 ° C, more preferably 60-110 ° C, more preferably 60-100 ° C, more preferably 70-100 ° C, More preferably, it is 80 to 100 ° C, particularly preferably 85 to 95 ° C. It is preferable to set the heat treatment condition at 60 ° C. or higher because vegetative cells of the fungus are sterilized. Further, it is preferable that the temperature of the heat treatment is 300 ° C. or lower because the bacteria remain without being carbonized.

The heat treatment time is, for example, 0.01 to 120 minutes, preferably 0.015 to 60 minutes, more preferably 0.02 to 40 minutes, still more preferably 0.025 to 30 minutes, and still more preferably 0. 0.03 to 25 minutes, particularly preferably 0.03 to 20 minutes. It is preferable to set the heat treatment time to 0.1 minutes or longer, because the vegetative cells of Bifidobacterium are sterilized. In addition, it is preferable that the heat treatment time is 120 minutes or less because heat denaturation can be suppressed and vegetative cells can be sterilized efficiently.

The optimum heat treatment time is, for example, 0.2 to 120 minutes, preferably 0.2 to 60 minutes, more preferably 0.2 to 40 minutes in the heat treatment at a low temperature range (60 to 100 ° C.). More preferably, it can be 0.2 to 30 minutes, more preferably 0.2 to 25 minutes, and particularly preferably 0.2 to 20 minutes. In the heat treatment at a high temperature range (100 to 300 ° C.), for example, 0.01 to 0.5 minutes, preferably 0.015 to 0.5 minutes, more preferably It is 0.02 to 0.5 minutes, more preferably 0.025 to 0.5 minutes, further preferably 0.03 to 0.5 minutes, and particularly preferably 0.03 to 0.5 minutes. For example, the heat treatment is performed at 90 ° C. for 15 minutes.

The heat treatment method is not particularly limited. For example, the obtained microbial cells can be heated under predetermined conditions using a heat sterilizer such as a plate sterilizer, a tubular sterilizer, a direct heating sterilizer, a jacketed tank, or the like.

The composition and agent of the present invention can be provided by the bifidobacteria of the present invention or a processed product thereof alone, or can be provided by mixing the bifidobacteria of the present invention or the processed product thereof with other components. You can also. The compounding amount of the bifidobacteria of the present invention in the composition and the agent of the present invention can be arbitrarily determined according to the purpose, application, form, dosage form, symptom, body weight, etc., and the present invention is not limited thereto. However, the content can be blended at a content of 0.1 to 90% (w / w) relative to the total amount, more preferably at a content of 0.1 to 50% (w / w). Can be blended. In the present invention, the agent of the present invention comprises the bifidobacteria of the present invention or a processed product thereof, and the composition of the present invention comprises the bifidobacteria of the present invention or a processed product thereof and other components. It can be.

The bifidobacteria and its processed product of the present invention can activate TLR2. However, activation of TLR2 can improve intestinal barrier function through protection of intestinal tight junction (see Non-Patent Document 2). Therefore, the composition and agent of the present invention can improve the intestinal barrier function through TLR2 activation. That is, according to the present invention, an intestinal barrier function improving composition and an intestinal barrier function improving agent comprising the bifidobacteria of the present invention or a processed product thereof as an active ingredient are provided.

Here, “intestinal barrier function” refers to a function of protecting the intestine from food antigens, bacteria and viruses that enter the intestine together with food, intestinal bacteria, and harmful substances produced by these. In addition, “intestinal barrier function improvement” is used in the meaning including “recovery of intestinal barrier function” and “inhibition of enhancement of intestinal barrier permeability”. “Improving intestinal barrier function” is further used in the meaning including enhancing the lowered intestinal barrier function, suppressing the decrease in barrier function, and further enhancing the barrier function of the normal intestinal tract.

In the present invention, "recovery of the intestinal barrier function" means recovery from a state in which the permeability in the intestinal tract has increased beyond a normal range to a pathological or undesirable level for some reason, It is used not only to return to, but also to bring it close to a qualitatively normal state. In addition, for recovery, promotion of recovery that shortens the recovery period until recovery from a state where the permeability in the intestinal tract exceeds a normal range for some reason and is pathologically or undesirably increased, in other words, usually Compared to the case of taking the food, it is used in a sense including restoring to a normal state earlier than natural healing.

In the present invention, “inhibition of increased permeability of the intestinal barrier” means that the barrier function in the intestinal tract is destroyed due to an exogenous or intrinsic cause, and the permeability of the intestinal barrier is changed from a normal state to a pathological state. It is used in the meaning of preventing or suppressing the occurrence of the phenomenon and maintaining its permeability in a normal state. The enhancement of intestinal barrier permeability as used in the present invention includes not only epithelial cells, but also destruction, breakdown, weakness, and reduction of the intestinal barrier related to biological mechanisms that constitute other intestinal barriers such as intestinal mucosa. .

The bifidobacteria of the present invention and the processed product thereof have actions such as improvement of diarrhea, improvement of constipation, suppression of growth of harmful bacteria in the intestine, production of vitamin B group, promotion of digestion and absorption by decomposition of lactose, etc. It has been known. Therefore, in addition to the activation of TLR2 and the effects related thereto, the effects and effects of the composition and agent of the present invention can be expected.

As described above, the bifidobacteria of the present invention and the processed product thereof can activate TLR2, and thus improve the intestinal barrier function. Here, it is known that necrotic enteritis, ulcerative colitis, persistent diarrhea, allergic disease and food hypersensitivity, etc. develop due to the breakdown or decrease of intestinal barrier function (Guamer F et al., Eur J Clin Nutr, 56 (Suppl 4), S34-38 (2002) etc.). Therefore, the composition and the agent of the present invention can be used for treatment, prevention and / or improvement of a disease or symptom which can be treated, prevented or ameliorated by improving intestinal barrier function. That is, according to the present invention, treatment, prevention and / or improvement of a disease or symptom which can be treated, prevented or ameliorated by improving the intestinal barrier function, comprising the bifidobacteria of the present invention or a processed product thereof as an active ingredient. And a therapeutic, preventive and / or ameliorating agent for the disease or condition. Diseases or symptoms that can be treated, prevented or ameliorated by improving intestinal barrier function include necrotizing enterocolitis, ulcerative colitis, persistent diarrhea, allergic diseases and food sensitivity.

The composition and agent of the present invention and the therapeutic agent, preventive agent and improver of the present invention can be provided in the form of a pharmaceutical (eg, pharmaceutical composition), quasi-drug, food, feed, etc. Can be performed as described.

The bifidobacteria and processed products thereof of the present invention can be orally administered to humans and non-human animals. Oral preparations include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. These preparations can be formulated using a pharmaceutically acceptable carrier by a technique usually performed in this field. Pharmaceutically acceptable carriers include excipients, binders, diluents, additives, fragrances, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, preservatives, etc. Is mentioned. In the present invention, the oral preparation may be provided as an enteric preparation so that it does not dissolve in the stomach but dissolves in the intestine. For example, it can be provided as a capsule, or the surface of a tablet or the like can be coated with an enteric film. .

The bifidobacteria of the present invention and the processed product thereof can be administered to humans and non-human animals in the body other than oral administration such as tube administration, nasal tube administration, infusion, suppository, etc. Is possible depending on. For example, a viscous liquid composition containing the bifidobacteria of the present invention or a processed product thereof, or a semi-solid composition containing the bifidobacteria of the present invention or a processed product thereof can be used for chewing and swallowing functions. Can also be administered to human and non-human animals that cannot be taken orally or. Even if the function of chewing or swallowing is reduced by aging or the like by ingesting or administering the composition and preparation of the present invention other than by oral ingestion, activation of TLR2 in these human and non-human animals Can be expected.

The bifidobacteria and processed products thereof of the present invention can be taken orally by humans and non-human animals. When the bifidobacteria of the present invention and a processed product thereof are orally ingested, the food or raw material of the food containing the bifidobacteria of the present invention or the processed product thereof, even in an isolated, purified or crudely purified form It may be a form.

When the bifidobacteria of the present invention and the processed product thereof are provided as food, the bifidobacteria of the present invention or the processed product thereof can be directly contained in the food, and the food contains the bifidobacteria of the present invention or the processed product thereof. It is a food containing an effective amount. Here, “containing an effective amount” of the bifidobacteria of the present invention and the processed product thereof means that the bifidobacteria of the present invention and the treatment thereof are within the range described later when an amount normally consumed in each food is ingested. Content that can be consumed. “Food” means health food, functional food, health functional food (for example, food for specified health use, functional nutrition food, functional indication food), food for special use (for example, food for infants, food for pregnant women, disease, etc.) Food).

The form of “food” is not particularly limited. For example, it may be a liquid form such as a beverage or a liquid food, a paste form, a semi-liquid form, a gel form, a solid form or a powder form. Form may be sufficient. Regardless of the form of liquid, solid, powder, etc., the bifidobacteria of the present invention and processed products thereof are various foods (milk, soft drinks, fermented milk, yogurt, cheese, bread, biscuits, crackers, pizza crust, preparation Powdered milk, liquid food, special-purpose food, food for the sick, nutritional food, frozen food, processed food, and other commercially available foods). Moreover, when the usage form of a nutrient composition is a powder, it can manufacture by using means, such as spray drying and freeze-drying.

The bifidobacteria of the present invention or a processed product thereof may also be provided as a composition in which water, protein, carbohydrate, lipid, vitamins, minerals, organic acid, organic base, fruit juice, flavors and the like are mixed. . Examples of the protein include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate (WPC), whey protein isolate (WPI), α-casein, β-casein, κ -Casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and vegetable proteins, hydrolysates thereof; butter, milk minerals, cream, whey, non-protein nitrogen, sial Examples include various milk-derived components such as acids, phospholipids, and lactose. Examples of the saccharide include saccharides, processed starch (in addition to dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.) and dietary fiber. Examples of the lipid include animal oils such as lard, fish oil, etc., fractionated oils, hydrogenated oil, transesterified oil, etc .; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils. Examples of vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline. And folic acid. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. Any non-side effects that have been used as pharmaceuticals and foods are applicable. Moreover, these components can be used in combination of 2 or more types.

The daily intake or dose of the composition and the preparation of the present invention as a pharmaceutical or food is determined according to the pathological condition, age, symptom, body weight and use of the person to be ingested, and the composition and the preparation of the present invention are nutritious. Since it differs depending on whether it is the only thing, it is not particularly limited. In the case of ingestion and administration for the purpose of activation of TLR2 and its related effects, for example, the intake or dose of the bifidobacteria of the present invention and its processed product per day is 10 ⁸ per day, for example. ^above, 10 8 to ^{10 12,} 5 × ¹⁰ 8 to 5 × ^{10 11,} 10 9 to ^{10 11,} 5 × ¹⁰ 9 to 5 × ^{10 10} pieces, 6 × ¹⁰ 9 to 4 × ^{10 10,} 7 The number can be 10 × 10 ⁹ to 3 × 10 ¹⁰ , preferably 8 × 10 ⁹ to 2 × 10 ¹⁰ , and more preferably 9 × 10 ⁹ to 2 × 10 ¹⁰ . The intake amount and dose may be determined by the ingestion subject and the attending physician of the administration subject.

The composition and the preparation of the present invention are advantageous in that they can be provided as a composition and preparation for an age group that does not have sufficient physical strength because there is little risk of side effects. The composition and agent of the present invention can be provided, for example, as a composition and agent for newborns (especially, newborn infants after birth), infants, children and / or elderly people. Here, the term “newborn” refers to an infant that is shortly after birth, and is used to include healthy newborns, premature infants, premature infants, and low birth weight infants. The term “newborn” refers to a child in the neonatal period, and the term “newborn period” means a period just after birth. In the case of humans, the neonatal period usually falls within 4 weeks after birth.

The composition and agent of the present invention can be provided as a composition and agent for daily intake that is effective in activating TLR2 and exerting its related effects. In this case, the composition and the preparation of the present invention may be packaged so that the effective intake for one day of the bifidobacteria and the processed product of the present invention can be ingested. As long as it can be ingested, the packaging form may be one package or a plurality of packages. When provided in the form of a package, it is desirable that a description of the intake is provided on the package so that an effective intake for one day can be ingested, or a document with the description is provided together. Moreover, when providing the effective intake for one day with a plurality of packages, a plurality of packages with the effective intake for one day can be provided as a set for convenience of intake.

The packaging form for providing the composition and the agent of the present invention is not particularly limited as long as it is a form that defines a certain amount. For example, wrapping paper, bag, soft bag, paper container, can, bottle, capsule, etc. Can be used.

The composition and preparation of the present invention are preferably administered or ingested continuously for 1 week or more in order to exert their effects better, and the administration and ingestion period is more preferably 2 to 8 weeks, particularly preferably 2 ~ 4 weeks. Here, “continuously” means to continue administration or ingestion every day. When the composition and the agent of the present invention are provided in a package form, an effective intake for a certain period (for example, one week) may be provided as a set for continuous intake.

According to the present invention, it comprises ingesting or administering to a subject in need thereof an effective amount of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof or a composition containing the same. Provided are a TLR2 activation method, a method for improving intestinal barrier function, and a method for treating, preventing and / or improving a disease or symptom that can be treated, prevented or ameliorated by improving the intestinal barrier function. The subject of ingestion or administration is a mammal including a human, preferably a human. The method of the present invention can be carried out as described for the compositions and agents of the present invention.

According to the present invention, there is also provided use of a bacterium belonging to Bifidobacterium bifidum, a treated product thereof, or a composition containing the same for the production of a composition for activating TLR2. The present invention further provides use of a bacterium belonging to Bifidobacterium bifidum or a treated product thereof or a composition containing the same for the production of a TLR2 activator. Furthermore, according to the present invention, there is provided use of a bacterium belonging to Bifidobacterium bifidum or a treated product thereof or a composition containing the same for TLR2 activation. Furthermore, according to this invention, the microbe which belongs to Bifidobacterium bifidum, its processed material, or a composition containing it for using for TLR2 activation is provided. The use of the present invention and the bifidobacteria of the present invention and the treated product thereof and the composition containing them can be carried out according to the description relating to the composition and the preparation of the present invention.

According to the present invention, a bacterium belonging to Bifidobacterium bifidum for the production of an intestinal barrier function improving agent, for the production of an intestinal barrier function improving agent, or for the improvement of an intestinal barrier function. Alternatively, use of the treated product or a composition containing the same is provided. Furthermore, according to this invention, the microbe which belongs to Bifidobacterium bifidum, its processed material, or the composition containing it for using for intestinal barrier function improvement is provided. The use of the present invention and the bifidobacteria of the present invention and the treated product thereof and the composition containing them can be carried out according to the description relating to the composition and the preparation of the present invention.

Further according to the present invention, by improving the intestinal barrier function for the manufacture of a pharmaceutical composition for use in the treatment, prevention and / or improvement of a disease or condition that can be treated, prevented or ameliorated by improving the intestinal barrier function. Treatment or prevention of a disease or condition that can be treated, prevented or ameliorated for the manufacture of a therapeutic agent, preventive agent or ameliorating agent for a disease or condition that can be treated, prevented or ameliorated, or by improving intestinal barrier function The use of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof or a composition containing the same is provided for improvement. Furthermore, according to the present invention, a bacterium belonging to Bifidobacterium bifidum or a bacterium for use in the treatment, prevention and / or amelioration of a disease or symptom which can be treated, prevented or ameliorated by improving intestinal barrier function A treated product or a composition comprising the same is provided. The use of the present invention and the bifidobacteria of the present invention and the treated product thereof and the composition containing them can be carried out according to the description relating to the composition and the preparation of the present invention.

The use of the present invention and the method of the present invention are intended for both therapeutic and non-therapeutic uses among human and non-human animals. Here, “non-therapeutic” means not including an act of treating, treating or diagnosing a human (ie, a medical act on a human). Specifically, a doctor or a person who has received instructions from a doctor This means that it does not include methods for performing surgery, treatment or diagnosis on humans.

The present invention will be described more specifically based on the following examples, but the present invention is not limited to these examples.

Example 1: Evaluation of TLR2 activation ability of Bifidobacterium bifidum OLB6378 Strain Bifidobacterium bifidum OLB6378 (Accession number: NITE BP-31) (Reference strain), Bifidobacterium infantis strain, and Bifidobacterium longum strain.

Each test strain specimen for evaluating the TLR2 activation ability was prepared as follows. That is, each test strain was cultured for 18 hours at 37 ° C. under anaerobic conditions using GAM Broth “Nissui” (manufactured by Nissui Pharmaceutical, product code 05422). The bacterial culture (500 mL) was centrifuged (2,000 × G, 15 minutes, 4 ° C.), and the resulting precipitate was resuspended in physiological saline (washing step). This washing step was repeated three times, twice with physiological saline and once with distilled water, and distilled water was added to the resulting precipitate to give a 20 mL bacterial suspension. This bacterial suspension was heated at 80 ° C. for 30 minutes and then freeze-dried to obtain a dry bacterial powder. The dried bacterial powder was suspended in distilled water at a concentration of 2 μg / mL and used for evaluation of TLR2 activation ability.

For evaluation of TLR2 activation ability, TLR2 Reporter Cell Line (manufactured by Novusbio, model number NBP2-26274), which is HEK293 cells into which each gene of TLR2 / NFκB / SEAP was introduced, was used. That is, TLR2 Reporter Cell Line was seeded in a 48-well plate at a concentration of 1.5 × 10 ⁵ cells / mL in a volume of 250 μL, and cultured in a high glucose DMEM (10% FCS added) medium containing 4 mM glutamine for 24 hours. Thereafter, 250 μL each of the suspension (2 μg / mL) of each test strain prepared as described above was added (final concentration 1 μg / mL). The culture supernatant 24 hours after the addition of the test strain was collected, and the concentration of alkaline phosphatase (SEAP) secreted by TLR2 Reporter Cell Line was quantified. Quantification was performed using SEAPorter Assay Kit (Novusbio, model number 10055K). The SEAP concentration at this time was used as an index of the TLR2 activation ability of each test strain.

The measurement result of TLR2 activation ability was as shown in FIG. The Bifidobacterium bifidum OLB6378 strain had a significantly higher TLR2 activation ability than other test strains, and was about 2.4 times higher than the Bifidobacterium breve JCM1192T strain (reference strain).

Claims (13)

1. A composition for activating Toll-like receptor 2 (TLR2), comprising a fungus belonging to Bifidobacterium bifidum or a processed product thereof as an active ingredient.
2. 2. The composition according to claim 1, wherein the bacterium belonging to Bifidobacterium bifidum has TLR2 activation ability more than twice that of Bifidobacterium breve JCM1192T strain.
3. The composition according to claim 1 or 2, wherein the bacterium belonging to Bifidobacterium bifidum is Bifidobacterium bifidum OLB6378 strain ( Bifidobacterium bifidum OLB6378, accession number: NITE BP-31).
4. The composition according to any one of claims 1 to 3, for use in improving the intestinal barrier function.
5. The composition according to claim 4, for use in the treatment, prevention and / or amelioration of diseases or symptoms that can be treated, prevented or ameliorated by improving the intestinal barrier function.
6. The composition according to claim 5, wherein the disease is necrotizing enterocolitis, ulcerative colitis, persistent diarrhea, allergic disease or food hypersensitivity.
7. The composition according to any one of claims 1 to 6, which is for newborns, infants, children and / or elderly people.
8. The composition according to any one of claims 1 to 7, wherein the bacteria belonging to Bifidobacterium bifidum are heat-treated cells.
9. TLR2 activation method, intestinal barrier function improving method, and intestinal tract comprising ingesting or administering an effective amount of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof to a subject in need thereof A method of treating, preventing and / or ameliorating a disease or condition that can be treated, prevented or ameliorated by improving barrier function.
10. Bacteria belonging to Bifidobacterium bifidum or treatment thereof for production of TLR2 activator, for production of intestinal barrier function improving agent, for TLR2 activation, or for improvement of intestinal barrier function Use of things.
11. Diseases that can be treated, prevented or ameliorated for the manufacture of therapeutic agents, prophylactic or ameliorating agents for diseases or symptoms that can be treated, prevented or ameliorated by improving intestinal barrier function, or which can be treated, prevented or ameliorated by improving intestinal barrier function Use of a bacterium belonging to Bifidobacterium bifidum or a processed product thereof for the treatment, prevention and / or amelioration of symptoms.
12. Bacteria belonging to Bifidobacterium bifidum or processed products thereof for use in activating TLR2 or for improving intestinal barrier function.
13. Bacteria belonging to Bifidobacterium bifidum or processed products thereof for use in the treatment, prevention and / or improvement of diseases or symptoms that can be treated, prevented or ameliorated by improving the intestinal barrier function.

Images