TW201902499A - 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 activation of terpenoid receptor 2

The present invention relates to a composition for the activation of a terpenoid receptor 2 .

The immune system of an organism can be divided into two types: innate natural immunity and acquired immunity. Natural immunity is widely preserved in the immune system of insects to humans, compared to the acquired immunity that exists only in higher organisms such as humans. Naturally immunized cells (macrophages, dendritic cells, etc.) quickly detect the invasion of foreign objects when pathogenic bacteria invade from the outside, and send out warning signals such as cytokines in addition to direct attacks such as phagocytosis. Thereafter, the activation-acquired immune system functions as a first stage of defense against infection. In the series of immune response systems, the receptor protein that is found to be the most active stage of the invasion of this task is a Toll-like receptor (referred to as "TLR" in this specification).

To date, TLR has been identified in 10 human species, and it is generally believed that the molecular structures that identify individual TLRs are different from each other. Among them, TLR2 is known to react with cell wall components such as peptidoglycan (PGN), palmitate, microbial lipoprotein, and zymosan, and then uses cell signal transmission as a medium (Non-Patent Document 1). For example, it has been confirmed that the intestinal barrier function can be improved by activating the TLR2 and protecting the cells by strengthening the tight junctions between the intestinal tracts (Non-Patent Document 2). [Previous Technical Paper] [Non-Patent Document]

[Non-Patent Document 1] Wetzler, L.M. Vaccine 21, S55-S60 (2003) [Non-Patent Document 2] Elke Cario et al., GASTROENTEROLOGY 127(1), pp224-238 (2004)

These cell signaling signals through TLR2 regulate various in vivo reactions led by the biological immune system. The team of the present invention focused on the applicability of the cell signal transduction system via TLR2, and focused on the food material that can activate TLR2. As a result, it was found that a strain belonging to Bifidobacterium (especially Bifid's OLB6378 strain) has potent TLR2 activation ability.

That is, the present invention has been made in an effort to provide a novel composition for the activation of the terpenoid receptor 2 and an activator for the terpenoid receptor 2.

The following invention can be provided in accordance with the present invention. [1] A TLR2 activating composition and a TLR2 activating agent (hereinafter sometimes referred to as "the composition and the agent of the present invention"), which are strains belonging to Bifidobacterium (hereinafter sometimes referred to as It is called "the Bifidobacterium of the present invention" or a processed product thereof as an active ingredient. [2] The composition according to the above [1], wherein the strain belonging to the species F. faecalis has a TLR2 activation ability of 2 times or more with respect to the strain of Lactobacillus brevis strain JCM1192T. [3] The composition according to the above [1] or [2], which belongs to the strain Bifidobacterium bifidum OLB6378 (Bifidobacterium bifidum OLB6378, accession number: NITE BP-31). [4] The composition according to any one of the above [1] to [3], which is used for improving the intestinal shielding function. [5] The composition according to the above [4], which is for use in a disease which can be treated, prevented or ameliorated by improving intestinal barrier function, or a treatment, prevention and/or improvement of symptoms. [6] The composition according to [5] above, wherein the disease is necrotic enteritis, ulcerative colitis, persistent diarrhea, allergic disease or food allergy. [7] The composition according to any one of the above [1] to [6], which is intended for a newborn, for a baby, for a child, and/or for an elderly person. [8] The composition according to any one of the above [1] to [7], wherein the bacteria belonging to the strain of Bifidobacterium are heat-treated. [9] A method for activating TLR2, a method for improving intestinal shielding function, and a method for treating, preventing and/or improving a disease or symptom which can be treated, prevented or ameliorated by improving intestinal shielding function, It is intended to contain an effective amount of a species belonging to Bifidobacterium or a treatment thereof, and to ingest a target or to be administered. [10] Use of a species belonging to Bifidobacterium or a treatment thereof for producing a TLR2 activator, for producing a intestinal barrier function improving agent, for TLR2 activation or for improving intestinal shielding function . [11] Use of a species belonging to Bifidobacterium or a treatment thereof for the manufacture of a therapeutic, prophylactic or ameliorating agent for treating, preventing or ameliorating a disease or symptom by improving intestinal barrier function Or a treatment, prevention, and/or amelioration of a disease or condition that can be treated, prevented, or ameliorated by improving intestinal barrier function. [12] A species belonging to Bifidobacterium or a treatment thereof for use in TLR2 activation, or for improving intestinal barrier function. [13] A species belonging to Bifidobacterium or a treatment thereof for use in the treatment, prevention and/or improvement of a disease or symptom which can be treated, prevented or ameliorated by improving intestinal barrier function.

According to the composition and the agent of the present invention, activation of TLR2 and effects related thereto can be expected for the subject to be ingested and the subject to be administered. Since the active ingredient of the present invention belongs to a species contained in a food material which has been ingested for many years by the human species of the F. faecalis strain, the composition and the agent of the present invention can be effectively utilized in the case of no side effects, and the TLR2 is planned. Activation.

[Specific Description of the Invention]

The active ingredient of the present invention belongs to the species Bifidobacterium bifidum. In the present invention, the strain belonging to the species Bifidobacterium has a strong TLR2 activation energy. That is, in the present invention, the ubiquinone receptor 2 of the more general strain Bifidobacterium breve JCM1192T strain can be activated, and the higher activation activity of the quinone receptor 2 belongs to Bieder The species of the bacteria is used as an active ingredient, and preferably, the activity of the steroid-like receptor 2 is 2 times or more (preferably 2.2 times or more, more preferably 2.4 times or more) relative to the strain of Lactobacillus brevis strain JCM1192T. The chemical energy belongs to the species of F. faecalis and is used as an active ingredient. And the short Bifidobacterium lactis JCM1192T strain can be purchased from the Bioresource Center of the Institute of Physical Chemistry. Further, the degree of TLR2 activation energy can be evaluated by using the alkaline phosphatase (SEAP) concentration secreted by TLR2 activation as an index.

In a preferred embodiment of the present invention, a strain belonging to Bifidobacterium can be used, and Bifidobacterium bibd 378 strain (Bifidobacterium bifidum OLB6378, accession number: NITE BP-31) can be used. The strain is currently deposited in the patent microbiology deposit center of the independent administrative legal person product evaluation technology base institution. The following are specific contents of the deposit.

(1) Name of the depository institution: Patent administrative microbial deposit center of the independent administrative legal person product evaluation technology base mechanism (2) Contact information: Postal code 292-0818, 2-5-8, Kurobetsu, Kisarazu, Chiba Prefecture (telephone number 0438-20 -5580) (3) Registration number: NITE BP-31 (4) Identification mark: Bifidobacterium bifidum OLB6378 (5) Original registration date: October 26, 2004 (6) Deposit-transfer date based on the Budapest Treaty: January 2006 18th

Beaver's OLB6378 strain is a Gram-positive facultative anaerobic bacterium from the feces of human infants. The bacterium was applied to a BL vegetable medium (manufactured by Eiken Chemical Co., Ltd.) on a flat plate, and an anaerobic gas bag (manufactured by Mitsubishi Gas Chemical Co., Ltd.) was used for culturing for 48 hours in an anaerobic state at 37 ° C to form an opaque circular shape. Hemispherical, glossy colonies. In addition, PCR amplification products using primers having the specificity of Bifidobacterium bifidum (Intestinal Flora Symposium 8, Intestinal Flora Molecular Biology Detection, Identification, Guanggang Zhizuo, Matsumoto Takahiro) were confirmed. In addition, it has fermentability to galactose, glucose, fructose, lactose, and gentiobiose.

As the medium for cultivating the Bifidobacterium of the present invention, a medium which is generally used for culturing Bifidobacterium can be used as the medium. That is, the medium which can be utilized in the present invention is not limited, and any medium may be used if the source of the main carbon is a medium having a nitrogen source, an inorganic substance, and other nutrients in a predetermined range. Lactose, fructose, sucrose, fructose, starch hydrolyzate, molasses, etc., which are carbon sources, can be used for the availability of the bacterium. As the nitrogen source, an organic nitrogen-containing substance such as a casein hydrolyzate, a whey protein hydrolyzate, or a soy protein hydrolyzate can be used. Other growth promoters may use meat extracts, fish extracts, yeast extracts, and the like.

The culture of the present invention is preferably carried out under anaerobic conditions, and a well-known method such as culturing a gasified carbon dioxide gas at the same time may be used, and micro aerobic conditions such as liquid static culture, which are generally used, or batches may be used. Other methods are used for the cultivation under the secondary culture conditions. The culture temperature is preferably 25 to 50 ° C, more preferably 35 to 42 ° C, but the present invention is not limited thereto, and other temperature conditions may be used for the temperature at which bacteria can grow. The pH of the medium in the culture is preferably maintained at 6.0 to 7.0, and other pH conditions may be used for the time when the bacteria can grow. The culture time is preferably from 3 to 48 hours, more preferably from 8 to 24 hours, and particularly preferably from 10 to 20 hours, and other culture time can be used if the bacteria can grow.

The obtained bacterial material can be contained in the composition and the agent of the present invention by the following treatment. The treated product of the present invention includes a culture after termination of culture, a culture obtained by centrifugation or filtration after termination of culture, a concentrate thereof, and further processed into a paste, and various uses. Method of drying (spray dried product, freeze-dried product, vacuum dried product, drum dried product, etc.), liquid substance dispersed in a medium, diluted with a diluent, heat treated heat treatment (heating Treatment of the cells), treatment with light (UV-treated cells) treated with ultraviolet rays and/or radiation, drug treatments treated with drugs (bactericides, antibacterial agents, bacteriostatic agents) (drug-treated cells), The crushed product of the dried product or the like is crushed using a bead mill or the like. It can be carried out by a generally used method such as centrifugation, filtration, or concentration. Further drying can be carried out by vacuum drying, spray drying, freeze drying, drum drying or the like. In the present specification, these may be simply referred to as "processed materials".

The bacteria belonging to the Bifidobacterium and the processed material thereof obtained by the above method can be directly used as a bacteria or a heat-treated bacteria, as a crushed or unpulverized treated material, alone or in a plurality of mixtures. It is contained in the composition and the agent of the present invention. When it is a bacterium, it is expected that it can be proliferated and fixed in the intestine after ingestion or administration, and it is expected that the effect of the present invention can be continuously exhibited. In addition, if the heat-treated cells (for example, 0.1 mL of the suspension (dispersion) liquid from the heat treatment) are taken and applied to a plate medium containing a medium in which F. faecalis can grow, anaerobic After the culture under the condition, the colony type cannot be formed, and the composition of the composition and the agent of the present invention can be expanded because it is not necessary to consider the characteristics of the F. faecalis which is difficult to survive in the presence of oxygen. good.

In the present invention, when the heat treated P. bisporus of the present invention is used, the heat treatment conditions are, for example, a heating temperature of generally 60 to 300 ° C, preferably 60 to 200 ° C, more preferably 60 to 150 ° C, and further 60 to 60. 140 ° C, then better 60 ~ 130 ° C, 60 ~ 120 ° C is also better, 60 ~ 110 ° C, 60 ~ 100 ° C, 70 ~ 100 ° C, 80 ~ 100 ° C, the best system 85 ~ 95 ° C. By setting the heating condition to 60 ° C or higher, it is preferable to sterilize the vegetative cells of the bacteria. Further, by setting the heating condition to 300 ° C or lower, it is preferable that the bacteria do not remain 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, 0.025 to 30 minutes, 0.03 to 25 minutes, or preferably 0.03 to 20 minutes. By setting the heat treatment time to 0.1 minute or more, it is preferable to sterilize the vegetative cells of the bacteria. Further, by setting the heat treatment time to 120 minutes or less, it is preferable to efficiently sterilize the vegetative cells by suppressing heat denaturation.

When the most suitable heat treatment time is set to a heat treatment in a low temperature range (60 to 100 ° C), for example, it can be set to 0.2 to 120 minutes, preferably 0.2 to 60 minutes, more preferably 0.2 to 40 minutes, or 0.2 to 0.2. 30 minutes, 0.2 to 25 minutes, and the best system is 0.2 to 20 minutes. Further, when the most suitable heat treatment time is set in the heat treatment in the high temperature range (100 to 300 ° C), for example, it can be set to 0.01 to 0.5 minutes, preferably 0.015 to 0.5 minutes, more preferably 0.02 to 0.5 minute, 0.025~0.5 minutes, 0.03~0.5 minutes, and the best system is 0.03~0.5 minutes. For example, the heat treatment is carried out at 90 ° C for 15 minutes.

The method of heat treatment is not particularly limited. For example, a heat sterilization device such as a plate sterilizer, a cylindrical sterilizer, a direct heating sterilizer, or a sandwich boiler can be used for the obtained cells, and heating can be performed under predetermined conditions.

The composition and the agent of the present invention may be provided separately with the F. faecalis or the treated product thereof, or a mixture of the F. faecalis or the treated product thereof and other components may be provided. The composition of the present invention and the agent thereof may be optionally formulated according to the purpose, use, form, dosage form, symptoms, body weight, etc., and the present invention is not limited to the following conditions. However, the content thereof is blended with 0.1 to 90% (w/w) of the total amount, and more preferably 0.1 to 50% (w/w). The present invention consists of the present invention for use in the present invention, and the composition of the present invention may contain the presently described Bifidobacterium or the treated product thereof and other components.

The Bifidobacterium and the treated product thereof can activate TLR2, and the intestinal barrier function can be improved by the activation of TLR2 and by protecting the cells to strengthen the tight junction between the intestinal tract (refer to Non-Patent Document 2). Therefore, the composition and the agent of the present invention can be expected to improve intestinal barrier function by activation of TLR2. In other words, according to the present invention, it is possible to provide a composition for improving the intestinal tract function comprising the F. faecalis or the treated product thereof as an active ingredient of the present invention, and an intestinal tract function improving agent.

Here, the "intestinal shielding function" protects the intestines from the bacteria, viruses, intestinal bacteria and harmful substances produced by the food antigens and foods entering the intestines. It is also used to include "improving intestinal barrier function", "returning intestinal barrier function" and "obstructing intestinal barrier permeability". "Improving the intestinal barrier function" is used to improve the bowel function of the previously lowered intestinal tract, to suppress the reduction of the intestinal tract shielding function, and to improve the shielding function of the normal intestinal tract.

In the present invention, "returning the intestinal tract shielding function" means the permeability of the intestinal tract, the disease state exceeding the normal range for unknown reasons, or the state of recovery to an unexpected degree, not only returning to the normal state, but also returning to the normal state, Use is meant to be close to the normal state of nature. In addition, the reply means that the permeability of the intestinal tract is shortened from a disease state in which the unexplained cause exceeds the normal range, or the response during the reply period from the advance to the unexpected state, in other words, including use and ingestion. In general, the condition of the food is compared to the condition of returning to a normal state earlier than the natural cure.

In the present invention, "blocking intestinal permeability hyperactivity" means that the shielding function of the intestinal tract is destroyed due to external or internal factors, and the permeability of the intestinal barrier is prevented or inhibited from being changed from a normal state to a disease. State, including maintaining intestinal permeability in a normal state. In the present invention, the permeability of the intestinal barrier is increased not only in the epithelial cells but also in the destruction, damage, weakness, and function of the intestinal shield associated with the biological mechanism that constitutes the shield of the intestinal tract.

The Bifidobacterium and the treated product thereof are known to have effects of improving squatting, improving constipation, inhibiting proliferation of harmful bacteria in the intestine, producing a vitamin B group, and promoting digestion and absorption by decomposition of lactose. Therefore, in addition to the effects related to the activation of TLR2, the composition and the agent of the present invention can be expected to have such effects.

As described above, the Bifidobacterium and the treated material thereof can activate TLR2, thereby improving the intestinal shielding function. Here, it is known that due to damage or functional decline of the intestinal tract, it causes necrotic enteritis, ulcerative colitis, persistent diarrhea, allergic diseases and food allergies (refer to 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 treat, prevent or ameliorate the disease by improving the intestinal shielding function, or can be used for treating, preventing and/or improving symptoms. That is, according to the present invention, it is possible to provide a composition comprising the Bifidobacterium or the treated body thereof as an active ingredient, which can treat, prevent or ameliorate the disease, or the treatment, prevention and/or treatment of the symptoms by improving the intestinal shielding function. Or improving the composition, and the treatment, prevention, and/or amelioration agent of the condition or symptom. The diseases or symptoms which can be treated, prevented or ameliorated by improving the intestinal shielding function include necrotic enteritis, ulcerative colitis, persistent diarrhea, allergic diseases and food allergies.

The composition and the agent of the present invention, and the therapeutic, prophylactic and improving agents of the present invention can be provided as a pharmaceutical (for example, a pharmaceutical composition), a quasi-drug, a food, a feed, etc., and can be described below. Method implementation.

The Bifidobacterium and its treated lines of the present invention can be administered orally to humans and non-human animals. The oral preparations may, for example, be granules, powders, lozenges (including sugar-coated tablets), pills, capsules, syrups, emulsions, suspensions. These preparations can be formulated using a pharmaceutically acceptable carrier by a method generally used in the related art. The pharmaceutically acceptable carrier may, for example, be an excipient, a binder, a diluent, an additive, a fragrance, a buffer, a tackifier, a colorant, a stabilizer, an emulsifier, a dispersant, a suspending agent, a preservative, or the like. The present invention can also be provided as an enteric preparation in order to make the oral preparation insoluble in the stomach and soluble in the intestine, and for example, it can be provided as a capsule, or can be coated with an enteric film on the surface of a tablet or the like.

The Bifidobacterium and the treated material thereof of the present invention can be administered to humans and non-human animals by oral administration, nasal administration, drip, and medicinal administration, etc., and can be administered to the body. The shape of the composition and the agent of the present invention is affected. For example, by making the Bifidobacterium or the treated product thereof of the present invention into a viscous liquid composition, or by containing the F. faecalis or the treated material thereof of the present invention, it is semi-solid. The composition can be administered to humans and non-human animals that have reduced chewing and swallowing functions, are unable to be orally administered, or are not orally administered. The subject or the agent of the present invention can be ingested by a method other than oral ingestion, or by administration, even if the function of chewing and swallowing is lowered due to factors such as ageing, human and non-human animals can be expected. Activation of TLR2.

The Bifidobacterium and the treated body thereof of the present invention can be administered orally to humans and non-human animals. When the Bifidobacterium and the treated material thereof of the present invention are orally ingested, the form may be isolated, purified or crudely purified, or may be a food containing the F. faecalis or the treated product thereof of the present invention. Or the form of the raw material of the food.

When the F. faecium and the treated product thereof are provided as a food, the food may directly contain the F. faecalis or the treated product thereof, and the food product contains the F. faecalis of the present invention or It treats an effective amount of food. Here, the "contained effective amount" of the food of the F. faecalis and the treated product thereof refers to the ingestion of the F. faecalis of the present invention in the following range when the general dietary amount is ingested in each food. The content of the processed material. In addition, "food" is used to include healthy foods, functional foods, health functional foods (for example, specific health foods, nutritious foods, functional foods), and special purpose foods (such as children's foods, maternal foods, The patient uses food and so on.

The form of the "food" is not particularly limited. For example, it may be in the form of a liquid such as a drink or a liquid food, and may be in the form of a paste, a semi-liquid or a gel, or may be in the form of a solid or a powder. In addition, the F. faecalis and the processed product thereof can be added to various foods (milk, refreshing drink, fermented milk, yogurt, cheese, bread, biscuit, soda, etc.) in a liquid, solid, powder or the like. Biscuits, pizza crusts, prepared milk powder, liquid foods, special-purpose foods, patient foods, nutritious foods, frozen foods, processed foods, and other commercially available foods, etc., are then ingested by subjects. Further, when the use form of the nutritional composition is a powder, it can be produced by a method such as spray drying or freeze drying.

The Bifidobacterium or the treated product thereof may further be provided as a composition mixed with water, protein, saccharide, lipid, vitamins, minerals, organic acids, organic bases, fruit juices, spices, and the like. The protein may, for example, be a 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, egg protein, meat and protein, and other animal and plant proteins, such hydrolysates; tallow, milk Minerals, butter, whey, non-protein nitrogen, sialic acid, phospholipids, lactose and other ingredients derived from milk. Further, examples of the saccharide include saccharides, processed starch (except dextrin, soluble starch, English starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like. Examples of the lipid include animal fats such as pig fat, fish oil, and the like, such as fractionated oil, hydrogenated oil, and transesterified oil; palm oil, eucalyptus oil, corn oil, rapeseed oil, coconut oil, and the like. Vegetable oils such as fractionated oil, hydrogenated oil, and transesterified oil. 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, nicotine, pantothenic acid, biotin, muscle. Alcohol, choline, folic acid, etc. Examples of the minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. The organic acid may, for example, be malic acid, citric acid, lactic acid or tartaric acid. It is applicable to any of pharmaceuticals and foods that have a proven track record and no side effects. These components can be used in combination of two or more kinds.

The intake or administration amount of the pharmaceutical or food of the composition and the agent of the present invention per day is determined by the condition, age, symptom, body weight, use of the subject, and the composition and the agent of the present invention. The only nutrients vary, and are not particularly limited. In the case of ingestion and administration for the purpose of TLR2 activation and its related effects, for example, the intake or administration amount of Pfidae and its treated product per adult in Japan, for example, 10 ⁸ per day. or more, it may be 10 ⁸ to 10 ^{12, 5 × 10 8 ~ 5 ×} 10 11 , 10 ⁹ to 10 ^{11, 5 × 10 9 ~ 5 ×} 10 10 ^{months, 6 × 10 9 ~ 4 ×} 10 10 One, 7 × 10 ⁹ ~ 3 × 10 ¹⁰ , can also use 8 × 10 ⁹ ~ 2 × 10 ¹⁰ , and more preferably 9 × 10 ⁹ ~ 2 × 10 ¹⁰ . The amount of intake and the amount of administration may also be determined by the ingesting subject and the attending physician of the subject.

Since the composition and the agent of the present invention are less likely to cause side effects, it is advantageous to provide the composition and the agent to an age layer which does not have sufficient physical strength. The composition and the agent of the present invention can be provided, for example, as a composition for newborns (especially newborn newborns), for infants, for children, and/or for elderly people. Agent. Here, "newborn" refers to a newborn baby, which is used to include healthy newborns, under-term infants, premature infants, and low birth weight infants. In addition, "newborn" refers to a child in the neonatal period, and the neonatal period refers to the period of birth, and the condition of the human is generally referred to as the neonatal period within 4 weeks after birth.

The composition and the agent of the present invention can provide a composition and an agent which are effective for one day of the TLR2 activation and the related effects. In this case, the composition and the agent of the present invention can be packaged so as to be able to take up the daily intake of the F. faecalis and the treated product thereof, and if it is an ingestible one-day effective intake, the package type The state can be a single package or a plurality of packages. When the package type is provided, it is desirable to record the relevant intake amount of the intake that can be ingested for one day on the package, or to provide the document with the record. When the effective intake is provided for one day in a plurality of packages, a plurality of packages of one-day effective intake in a kit mode may be provided for the convenience of ingestion.

The packaging form of the composition and the agent of the present invention is not particularly limited as long as it is a predetermined amount, and examples thereof include a packaging paper, a bag, a soft bag, a paper container, a can, a bottle, a capsule, and the like. Container for storage.

In order to more effectively exert the effects of the composition and the agent of the present invention, it is preferred to continue administration or ingestion for one week or longer, and the administration and ingestion period are preferably 2 to 8 weeks, and the special system is 2 to 4 weeks. . Here, "continuously" means continuous daily administration or ingestion. When the composition and the agent of the present invention are provided in a package form, in order to be stably ingested, an effective intake amount for a certain period (for example, one week) may be provided in a kit.

According to the present invention, it is possible to provide a TLR2 activation method comprising a species belonging to Bifidobacterium or a processed material thereof, or a composition containing the same, and allowing a subject in need thereof to be ingested or administered. A method for improving intestinal shielding function and a method for treating, preventing or ameliorating the treatment, prevention and/or improvement of a disease or symptom by improving intestinal shielding function. Ingested or administered to a subject comprising a human mammal, preferably a human. The method of the present invention can be carried out in accordance with the description relating to the composition and the agent of the present invention.

According to the present invention, it is possible to provide a composition for producing a composition for TLR2 activation, and containing a composition belonging to a species of Bifidobacterium or a treated product thereof. According to the present invention, there is further provided a use of a composition for producing a TLR2 activating agent and containing a species belonging to Bifidobacterium or a treated product thereof. Further, according to the present invention, the use of a composition belonging to a species belonging to Bifidobacterium or a processed material thereof for the activation of TLR2 can be further provided. According to the present invention, a composition for inactivating TLR2 and containing a species belonging to Bifidobacterium or a processed product thereof may be further provided. The use of the present invention and the inclusion of the F. faecalis or the treated product thereof and the compositions of the present invention can be carried out in accordance with the description relating to the composition and the agent of the present invention.

According to the present invention, it is possible to further provide a composition for improving intestinal barrier function, for producing an improved intestinal barrier function agent, or for improving intestinal barrier function, and containing a species belonging to Bifidobacterium or a treatment thereof Or use of its constituents. According to the present invention, it is further provided to contain a species belonging to Bifidobacterium or a treatment thereof or a composition containing the same for use in improving intestinal barrier function. The use of the present invention and the Bifidobacterium or the treated product thereof and the composition containing the same according to the present invention can be carried out in accordance with the description relating to the composition and the agent of the present invention.

According to the present invention, in order to manufacture a pharmaceutical composition for treating, preventing or ameliorating the treatment, prevention or amelioration of a disease or a symptom by improving the intestinal shielding function, in order to treat, prevent or ameliorate the disease by improving intestinal shielding function Or a therapeutic agent, a prophylactic agent and/or an ameliorating agent, or a Bifidobacterium bacterium which can provide treatment, prevention or improvement for treating, preventing or ameliorating a disease or symptom by improving intestinal shielding function Use of a strain or a treatment thereof or a composition containing the same. According to the present invention, it is possible to provide a species belonging to Bifidobacterium or a treatment thereof or a treatment thereof for treating, preventing or ameliorating the treatment, prevention and/or improvement of a disease or symptom by improving the intestinal barrier function. Composition. The use of the present invention and the Bifidobacterium or the treated product thereof and the composition containing the same according to the present invention can be carried out in accordance with the description relating to the composition and the agent of the present invention.

The use of the invention and the method of the invention can be used on human or non-human animals, and can also be used therapeutically or non-therapeuticly. "Non-therapeutic" as used herein means not including the operation, treatment or diagnosis of a human being (ie, medical treatment of a human being), specifically, it does not include a physician or a physician's instruction, Humans perform methods such as surgery, treatment or diagnosis. [Examples]

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

Example 1: Evaluation of TLR2 activation energy of Bifid's OLB6378 strain TLR2 activation energy of Beid's OLB6378 strain (Accession number: NITE BP-31), and Lactobacillus brevis strain JCM1192T strain (standard strain) , Bifidobacterium infantis strain, Bifidobacterium longum.

To evaluate the TLR2 activation energy, each test strain sample was prepared as follows. Namely, each test strain was cultured for 18 hours under anaerobic conditions and at 37 ° C using GAM Broth "Nissui" (manufactured by Nippon Pharmaceutical Co., Ltd., product No. 05422). The culture solution (500 mL) was centrifuged (2,000 × G, 15 minutes, 4 ° C), and the precipitate was resuspended in physiological saline (washing step). The washing step was repeated twice with physiological saline, distilled water once for 3 times, and the obtained precipitate was added to distilled water to prepare a 20 mL bacterial suspension. The bacterial suspension was heated at 80 ° C for 30 minutes, and then freeze-dried to obtain dried bacterial cells of the cells. The suspension was dried in distilled water to a concentration of 2 μg/mL, and used for evaluation of TLR2 activation energy.

For the evaluation of TLR2 activation energy, TLR2 Reporter Cell Line (manufactured by Novus Bio, model NBP2-26274) of HEK293 cells into which each gene of TLR2/NFκB/SEAP was introduced was used. That is, TLR2 Reporter Cell Line cells were seeded at a concentration of 1.5 × 10 ⁵ cells/mL in a 48-well plate at 250 μL per well, and a medium containing 4 mM glutamic acid in high glucose DMEM (10% FCS) was added. Cultivate for 24 hours. Thereafter, a suspension (2 μg/mL) of each test strain prepared as described above was added to a flat plate (final concentration: 1 μg/mL) in an amount of 250 μL per well. The culture supernatant was collected 24 hours after the addition of the test strain, and the concentration of alkaline phosphatase (SEAP) secreted by the TLR2 Reporter Cell Line was quantified. The quantification was carried out using the SEAPorter Assay Kit (manufactured by Novusbio, model 10055K). The SEAP concentration at this time was used as an indicator of the TLR2 activation energy of each test strain.

The measurement results of TLR2 activation energy are shown in Fig. 1. The TLR2 activation of Beaver's OLB6378 strain was significantly higher than that of the other test strains, and was about 2.4 times higher than that of the short Bifidobacterium breve JCM1192T strain (standard strain).

Fig. 1 is a graph showing the results of measurement of TLR2 activation energy (SEAP concentration) of the Bidder's OLB6378 strain and other test strains (mean±standard deviation, detection limit: 15.6 ng/mL).

Claims (13)

A composition for activation of a terpenoid receptor 2 (TLR2), which comprises a species belonging to Bifidobacterium or a treated substance thereof as an active ingredient. The composition of the first aspect of the patent application, wherein the strain belonging to the species F. faecalis has a TLR2 activation ability of 2 times or more relative to the strain of the B. brevis L. JCM1192T. The composition of claim 1 or 2, which belongs to the strain Bifidobacterium bifidum OLB6378 (Bifidobacterium bifidum OLB6378, accession number: NITE BP-31). The composition of any one of claims 1 to 3 is for use in improving intestinal shielding function. The composition of claim 4 is for use in the treatment, prevention or amelioration of a disease which can be treated, prevented or ameliorated by improving the intestinal barrier function, or the treatment, prevention and/or improvement of symptoms. The composition of claim 5, wherein the disease is necrotic enteritis, ulcerative colitis, persistent diarrhea, allergic disease or food allergy. The composition of any one of the first to sixth aspects of the patent application, which is targeted for newborns, for infants, for children, and/or for elderly people. The composition according to any one of claims 1 to 7, wherein the bacteria belonging to the species of Bifidobacterium are heat-treated. A method for activating TLR2, a method for improving intestinal shielding function, and a method for treating, preventing and/or improving a disease or symptom which can be treated, prevented or improved by improving intestinal shielding function, An effective amount of a species belonging to Bifidobacterium or a treatment thereof, which allows a subject in need thereof to be ingested or administered. A species belonging to Bifidobacterium or a treatment thereof for producing a TLR2 activator, for producing a intestinal barrier function improving agent, for TLR2 activation, or for improving intestinal barrier function. Use of a species belonging to Bifidobacterium or a treatment thereof for the manufacture of a therapeutic, prophylactic or ameliorating agent for treating, preventing or ameliorating a disease or condition by improving intestinal barrier function, or For the treatment, prevention and/or improvement of diseases or symptoms that can be treated, prevented or ameliorated by improving the intestinal barrier function. A species belonging to Bifidobacterium or a treatment thereof for use in TLR2 activation, or for improving intestinal barrier function. A species belonging to Bifidobacterium or a treatment thereof for use in the treatment, prevention and/or improvement of a disease or symptom which can be treated, prevented or ameliorated by improving intestinal barrier function.