JP2010053125A - Anti-allergic agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a readily producible and easily ingestible agent having anti-allergic actions. <P>SOLUTION: A solvent extract of green soybeans is used as an anti-allergic agent. The solvent extract of the green soybeans is useful as an anti-allergic agent, because the solvent extract thereof has inhibitory actions on production of IgE, and actions of deflecting Th1/Th2 balance to the side of Th1 thereof. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an IgE production inhibitor, a Th1 / Th2 increasing agent and an antiallergic agent, and an antiallergic food and drink containing them, characterized by containing a solvent extract of green soybeans.

  Allergic diseases are one of the most common diseases in developed countries. Typical examples include hay fever, allergic conjunctivitis, bronchial asthma, atopic dermatitis and the like, as well as rheumatism, inflammatory bowel disease, food allergy and the like. Among these, the number of patients with hay fever and atopic dermatitis is increasing rapidly, which is a big social problem.

  Allergic reactions are roughly classified into four types. Among them, many allergic diseases are considered to be caused by type I allergy. Type I allergy, also called immediate allergy, is an allergen-specific IgE antibody overproduced from B cells, chemical mediators such as histamine and leukotriene released from mast cells and basophils, or helper T cells. It is said that a plurality of factors such as deviation of the functional balance of the subtypes Th1 cells and Th2 cells toward the Th2 side are complicatedly intertwined. That is, the allergen that has entered the body is taken up by the antigen-presenting cells and decomposed, and a part of the information is presented to the T cells. The T cells that have received the antigen are differentiated and activated into Th2 cells, and produce cytokines such as IL-4 to activate the B cells. Activated B cells produce IgE antibodies specific for allergens, and these IgE antibodies bind to Fc receptors present on the surface of mast cells and basophils. When the allergen enters the body again and crosslinks IgE antibodies on the surface of mast cells or basophils, a large amount of chemical mediators such as histamine are produced and released from the cells, and leukotrienes are produced on the cell surface. These produced substances increase vascular permeability and cause edema and excessive secretion of nasal discharge, and contract smooth muscles and cause airway contraction. On the other hand, it is becoming clear that the Th1-Th2 balance is chronically biased toward the Th2 side due to changes in lifestyle and dietary habits in recent years, contributing to an increasing tendency of allergic diseases.

  Currently, allergic symptoms can be improved by reducing the chances of contact with allergens such as pollen and house dust with a mask, goggles, air purifier, etc. Physical methods such as the method to prevent, histamine that inhibits the binding of free histamine to the receptor, steroid agents that suppress the inflammatory reaction, degranulation inhibitor or leukotriene synthesis inhibitor that has a cell membrane stabilizing action, etc. There are pharmacological methods such as a method of suppressing an allergic reaction with other antiallergic agents. However, it is extremely difficult to completely prevent contact with allergens, and anti-allergic agents often have side effects and have limitations in daily long-term application.

  On the other hand, instead of these drugs, attempts have been made to improve allergic symptoms by ingesting foods exhibiting anti-allergic effects. For example, foods supplemented with tea, perilla, β-glucan, etc. are known. It has been. However, although these foods have fewer side effects than pharmaceuticals, they are limited in allergy improvement and cannot be said to have sufficient effects, and none of them has been satisfactory.

  Therefore, an antiallergic food that can be taken on a daily basis and that can effectively relieve symptoms such as hay fever is desired.

By the way, soybean is well known as a raw material for tofu, soy sauce, natto and the like, and attempts have been made to obtain functional components from soybean. For example, Patent Document 1 discloses a composition for preventing / treating allergies comprising isoflavones and specific saponins as active ingredients, and describes that these ingredients can be extracted from soybeans. However, the amount of isoflavones and saponins contained in soybean is small, and it is necessary to extract from the hypocotyl site, which is considered to have a particularly high content in soybean, and to concentrate it by performing a more complicated purification process. In addition, isoflavones and saponins have strong bitterness and poor taste, cannot be continuously ingested, and may be toxic, so it is necessary to ingest them while closely controlling the intake interval and intake. There was a problem that it was difficult to ingest so as to obtain an effect.
Patent Document 2 discloses an antiallergic composition containing stachyose as an active ingredient, and describes that stachyose can be extracted from soybeans. However, the amount of stachyose contained in soybean is small, and an effective component cannot be obtained by a simple extraction operation, and it is necessary to perform a complicated purification process such as chromatography to concentrate it to a high concentration of 98%. Moreover, stachyose does not act on IgE and is effective only on type IV allergies among allergies, so although it can be expected to be effective on delayed type allergies involving immune cells, hay fever, allergenicity associated with IgE No effect can be expected for immediate allergies (type I, type II, type III allergy) such as pruritus (itch) symptoms of conjunctivitis, bronchial asthma, and atopic dermatitis.

JP 2007-197398 A JP 2003-321372 A

  Therefore, it can be ingested safely, inexpensively and easily on a daily basis, and has a remarkable antiallergic action, and various immediate allergic diseases, particularly type I allergic diseases such as hay fever, allergies There is still a high demand for natural materials effective for the prevention and / or improvement of conjunctivitis, bronchial asthma, atopic dermatitis and the like.

  As a result of diligent research to solve the above problems, the present inventors have found that an extract of green soybean, preferably a composition simply extracted from green soybean using water, alcohol or hydrous alcohol, is used in addition to yellow soybeans and the like. Compared with soybeans of this type, it exhibits an extremely strong IgE-lowering effect, improves the Th1-Th2 balance predominantly (increases Th1 / Th2), and is useful for the prevention and / or improvement of various allergic diseases I found out. And when green soybean was used, even if it did not perform complicated concentration, it discovered that a highly effective antiallergic agent was obtained simply, and completed this invention.

That is, the present invention relates to the following matters.
(1) An IgE production inhibitor comprising a solvent extract of green soybeans.
(2) A Th1 / Th2 increasing agent characterized by containing a solvent extract of green soybeans.
(3) An antiallergic agent comprising a solvent extract of green soybeans.
(4) The agent according to any one of (1) to (3), wherein the green soybean solvent extract is a water extract.
(5) The agent according to any one of (1) to (3), wherein the solvent extract of green soybean is a hydrous ethanol extract.
(6) An antiallergic food, beverage, feed and cosmetic comprising the agent according to any one of (1) to (5).

  According to the present invention, it can be safely and inexpensively and easily ingested on a daily basis, and has a remarkable antiallergic action, such as hay fever, allergic conjunctivitis, bronchial asthma, and atopic dermatitis. An antiallergic agent effective for prevention and / or improvement of allergic diseases is provided. The antiallergic agent of the present invention suppresses the production of IgE at the cellular level and deflects the Th1-Th2 balance toward the Th1 side, so that the antiallergic agent exerts the effect even if it is spread throughout the body by oral ingestion or by local application. Therefore, an undesirable action can be minimized, and the effect can be continuously felt, which is extremely useful.

Hereinafter, preferred embodiments of the present invention will be specifically described.
In the present invention, antiallergic action refers to prevention, treatment, and prevention of various adverse effects caused by Th2 side bias (Th2 predominance) by deflecting the Th1-Th2 balance of helper T cells to Th1 side (Th1 predominance). An action that reduces or mitigates. In the present invention, the antiallergic action refers to an action for preventing, treating, reducing or alleviating various harmful effects caused by IgE antibodies. Furthermore, it is induced by a Th2 dominant state or a chemical mediator released from mast cells by IgE antibody, etc., caused by smooth muscle contraction, enhanced vascular permeability, neutrophil migration, platelet aggregation, etc. It also includes the action of preventing, treating, reducing or alleviating various symptoms.

  In the present invention, the Th1-Th2 balance of helper T cells is biased to the Th1 side, so that the immunosuppressive state caused by the tumor is improved, and the immunocompromised state caused by the anticancer drug treatment or the radiotherapy is improved. , Prevention or improvement of acquired immune deficiency syndrome (AIDS), prevention or treatment of infectious diseases caused by various bacteria, prevention or improvement of various symptoms derived from type I allergy, improvement of immunity decline associated with aging and pathological conditions, etc. It is useful for improving various symptoms, problems, improving health, and promoting nutrition and tonic.

  The Th1-Th2 balance can be measured by a method of measuring and comparing the expression level and secretion level of IFN-γ secreted by Th1 cells with the expression level and secretion level of IL-4 secreted by Th2 cells. it can. The action caused by IgE antibody is measured by a method of directly measuring the amount of IgE in blood, a method of measuring the amount of chemical mediator such as histamine released from mast cells, basophils, etc. by IgE. be able to.

  As the antiallergic agent of the present invention, those containing a solvent extract of green soybean as an active ingredient are preferable. Soybean is also called field meat and contains abundant protein, and it is one of the few foods that are eaten worldwide as an important source of nutrients. Can be taken continuously. Moreover, the active ingredient of the antiallergic agent of the present invention is economically superior because it can be easily extracted from green soybeans only by solvent extraction and has a high effect.

  The green soybean used in the present invention is not a yellow soybean generally used for food, but has a blue to blue-green to green appearance after ripeness. It is called double blue, single blue, kaori beans, dacha beans, green beans, aobata, green beans, etc. There are varieties of blue soybeans that are blue only in the seed coat and varieties that are all blue in the contents, the latter being more preferred. In addition, what is eaten as edamame is often boiled with a blue-colored fruit before completion of ripeness of yellow soybean, which is common, but this does not correspond to the green soybean referred to in the present invention.

Hereinafter, the manufacturing method of the solvent extract of green soybeans used for this invention is demonstrated in detail. The extraction method is not particularly limited, but the above-described green soybean as it is or after being cut or pulverized, dried, pulverized after drying, squeezed and extracted juice is immersed in an extraction solvent, stirred or A known method such as a reflux method and a supercritical fluid extraction method can be used.

  The solvent that can be used for the extraction is preferably water or an organic solvent. Specific examples of water include pure water, distilled water, tap water, acidic water, alkaline water, and neutral water. Examples of organic solvents include methanol, ethanol, n-propanol, isopropanol, and n-butanol. Alcohols which are liquid at room temperature such as polyhydric alcohols such as 1,3-butylene glycol, propylene glycol and glycerine; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; acetone And ketones such as ethyl methyl ketone; hexane; and chloroform. These may be used alone or in combination of two or more. Among the above organic solvents, it is preferable to use alcohols that are liquid at room temperature, for example, lower alcohols having 1 to 4 carbon atoms, from the viewpoint of operability and environmental properties. More preferably, ethanol is used.

  In the present invention, the organic solvent includes a water-containing organic solvent in which an aqueous component is further contained in the organic solvent. From the viewpoint of maintaining high extraction efficiency, the content of the aqueous component in the water-containing organic solvent is usually 80% by volume or less, preferably 65% by volume or less, and more preferably 50% by volume or less. As the water-containing organic solvent, a water-containing alcohol in which an aqueous component is further contained in the alcohol as described above, more preferably water-containing ethanol is used.

  As specific extraction methods, green soybean is added to a room temperature or warmed solvent under reduced pressure, normal pressure or increased pressure, and extracted while being immersed or stirred, and extracted while refluxing in a solvent. Is mentioned. At that time, it is appropriate that the extraction temperature is 5 ° C. to the boiling point of the solvent or less, and the extraction time varies depending on the type of solvent used, the extraction conditions, and the content of aqueous components in the case of a water-containing organic solvent. However, about 30 minutes to 72 hours is appropriate. When performing extraction by refluxing, it is preferable to use a low boiling point solvent so that the extract of soybeans does not denature or undergo thermal decomposition. Further, the extraction operation can also be performed by a supercritical fluid extraction method using carbon dioxide or the like.

  Next, the mixture containing the extract and the residue is subjected to filtration, centrifugation, or the like, if necessary, to remove the solid component that is the residue to obtain an extract. The removed solid component can be used again for the extraction operation, and this operation may be repeated several times.

  The extract thus obtained may be used as it is as a solvent extract of green soybeans, and further, if necessary, used as a dried or powdered product by a method such as concentration or freeze-drying or spray-drying. May be.

  In the case of drying the extract, any specific drying method may be used as long as the solvent extract of green soybean can be performed under conditions that do not cause denaturation or thermal decomposition. An excipient is added, and methods such as filtration, centrifugation, centrifugal filtration, spray drying, spray cool, drum drying, vacuum drying, freeze drying and the like can be mentioned, and these methods can be used alone or in combination.

The agent of the present invention is characterized in that it contains the solvent extract of green soybean obtained by the above-mentioned process, but it is continuously used alone or in various forms such as pharmaceuticals, foods or feeds. When it is ingested or continuously applied in the form of a cosmetic, Th1 / Th2
Is increased, and the amount of IgE is decreased. Therefore, it is useful for prevention and / or improvement of various disorders and diseases such as hay fever, atopy, and autoimmune diseases. Further, since the agent of the present invention is made from safe green soybeans, it is highly safe and has a good flavor. Therefore, it can be taken orally as it is or applied as a cosmetic. Continuous administration of the period is possible. Furthermore, long-term continuous intake is easy as various pharmaceutical, food and feed forms.

  The agent of the present invention is usually applied in the range of 0.01 to 100 g per day for adults based on the dry soybean solvent extract obtained by the above process. In the case of oral administration, the general daily intake is 0.1 to 50 g, but since the extract is highly safe, the intake can be further increased. The daily intake may be taken once, but may be taken in several divided doses.

  In that case, the solvent extract of green soybean obtained by the above-mentioned process can be used alone as it is, but as long as the effect of the present invention is not inhibited, additives, other known immunostimulatory substances, immunoregulators described later You may mix | blend a substance etc. individually or in combination.

  Examples of the dosage form when the agent of the present invention is used as a pharmaceutical composition include oral preparations such as tablets, capsules, granules, powders, syrups, dry syrups, solutions and suspensions, inhalants and suppositories. Enteral preparations such as ointments, creams, gels, patches, external preparations for skin, drops, injections and the like. Of these, oral agents are preferred.

  Such dosage forms include additives commonly used in the solvent extract of green soybean, which is an active ingredient, such as excipients, disintegrants, binders, lubricants, surfactants, alcohol, water, water-soluble Can be produced according to a conventional method by blending a functional polymer, sweetener, flavoring agent, acidulant and the like according to the dosage form. Liquid preparations such as liquids and suspensions may be dissolved or suspended in water or other appropriate medium immediately before taking. In the case of tablets and granules, the preparations may be prepared by well-known methods. The surface may be coated.

  The content of the solvent extract of green soybean in the pharmaceutical composition according to the present invention varies depending on the dosage form, but is usually 0.001 to 99% by mass, preferably 0.01 to 80% based on the dry mass. It is desirable that the dosage is in a mass% range so that the daily dose can be controlled so that the above-mentioned daily intake for adults can be taken.

  When the agent of the present invention is prepared as a food composition, its form is not particularly limited, and includes all foods that can be blended with the agent of the present invention in addition to health foods, functional foods, foods for specified health use, and the like. In the present invention, the food includes beverages. Specifically, various preparation forms such as liquid foods such as tablets, chewable tablets, powders, capsules, granules, drinks, and enteral nutrients can be used. A food composition in the form of a preparation can be produced in the same manner as a pharmaceutical preparation. Furthermore, the food composition includes tea drinks such as green tea, oolong tea and black tea, soft drinks, jelly drinks, sports drinks, milk drinks, carbonated drinks, fruit juice drinks, lactic acid bacteria drinks, fermented milk drinks, powdered drinks, cocoa drinks, purified water, etc. Beverages, butter, jam, sprinkles, margarine spreads, mayonnaise, shortening, custard cream, dressings, breads, cooked rice, noodles, pasta, miso soup, tofu, milk, yogurt, soup or sauces, confectionery ( For example, it may be prepared as biscuits, cookies, chocolate, candy, cake, ice cream, chewing gum, tablet) or the like.

  The food composition according to the present invention includes livestock, racehorses, feeds for appreciation animals, pet foods and the like. Since feed can be used in almost the same composition and form as food, the description regarding food in this specification can be applied to feed as well.

  The food composition further includes other food materials used in the production of food and feed, various nutrients, various vitamins, minerals, amino acids, various fats and oils, various additives (for example, flavoring ingredients, sweeteners, organic acids, etc. (Sour taste, surfactant, pH adjuster, stabilizer, antioxidant, pigment, flavor) and the like can be blended and manufactured according to conventional methods. Moreover, the food composition which concerns on this invention can also be manufactured by mix | blending the agent of this invention with the food normally eaten.

  In the food composition according to the present invention, the content of the solvent extract of green soybeans varies depending on the form of the food, but is usually 0.001 to 80% by mass, preferably 0.01 to, based on the dry mass. It is 50 mass%, More preferably, it is the range of 1-50 mass%. The daily intake may be taken once, but may be taken in several divided doses. It is preferable that the intake amount per day can be managed so that the intake amount per day for an adult can be consumed.

  Since the agent of the present invention has high safety, it can also be prepared as a cosmetic composition. Since the cosmetic composition according to the present invention can be continuously applied, it is particularly effective for preventing and / or improving atopic dermatitis.

  When preparing the cosmetics according to the present invention, the solvent extract of green soybean may be used as a cosmetic as it is, or the solvent extract of green soybean is blended by a general-purpose method, and the emulsion, cosmetic liquid, cream, lotion, essence , Packs and sheets, foundation, funny, blusher, lipstick, eye shadow, eyeliner, mascara, facial cleanser, skin cleanser, gel, gel, skin cleanser, body shampoo, shampoo, rinse hair Cosmetic compositions in the form of cosmetics, hair preparations, hair treatments, hair nourishing agents, bath preparations, ointments, quasi drugs, oil blotting paper, and the like may be prepared.

  The cosmetic composition according to the present invention includes, in addition to a green soybean solvent extract, conventionally known excipients and fragrances according to the desired dosage form, fats and oils, surfactants, preservatives, metal ions. Add appropriate ingredients such as sequestering agents, water-soluble polymers, thickeners, pigments, UV protection agents, moisturizers, antioxidants, pH regulators, detergents, desiccants, emulsifiers, etc. Can be manufactured.

  The content of the green soybean solvent extract in the cosmetic composition according to the present invention is not particularly limited, but is usually 0.001 to 80% by mass, preferably 0.01 to 50% by mass based on the dry mass. Is within the range.

  Further, the agent, food, feed and cosmetics of the present invention include, for example, conjugated linoleic acid, taurine, glutathione, carnitine, creatine, coenzyme Q, glucuronic acid, glucuronolactone, pepper extract, ginger extract, cacao extract. , Guarana extract, garcinia extract, theanine, γ-aminobutyric acid, capsaicin, capsiate, various organic acids, flavonoids, polyphenols, catechins, xanthine derivatives, indigestible oligosaccharides such as fructooligosaccharides, polyvinylpyrrolidone, etc. May be.

  The compounding amount of these additives is appropriately determined according to the form of the composition, the type of additive and the amount of intake to be desired, but in the agent, food, feed and cosmetics of the present invention, 0.01 to 70 mass. %, Preferably in the range of 0.1 to 50% by mass.

  According to the present invention, various allergic diseases such as hay fever, atopy, and autoimmune diseases can be prevented and / or improved by increasing Th1 / Th2 and decreasing the amount of IgE. Furthermore, the solvent extract of green soybean used in the present invention has no side effects, has a good flavor and can be easily ingested, and can be easily ingested for a long period of time.

In addition, by administering the agent of the present invention as a food, feed, cosmetic or pharmaceutical composition in an effective amount as described above, prevention of various infectious diseases, hay fever, atopy, autoimmune diseases and / or An improvement method can be provided.
EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited only to a following example.

[Example 1] Manufacture of water extract 100 g of water was added to 10 g of crushed green soybean and heated at 90 ° C for 60 minutes for extraction. Insoluble matter was removed by filtration, and water was evaporated to obtain 2.2 g of powder.

[Example 2] Production of water-containing ethanol extract 100 g of water-containing ethanol (ethanol: water = 95: 5) was added to 10 g of crushed green soybeans, and the mixture was extracted by shaking (100 rpm) at room temperature for 24 hours. Insoluble matter was removed by filtration, and the solvent was evaporated to obtain 1.2 g of powder.

[Example 3] Production of water extract Extracted by adding 12 L of water to 1 kg of crushed green soybean (Echigomitori) and heating at 95 ° C for 60 minutes. The precipitate was removed by centrifugation, the supernatant was filtered to remove insoluble matters, and lyophilized to obtain 346 g of powder.

[Comparative Example 1]
In the same manner as in Example 2, extraction was performed from soybean (yellow soybean), black soybean, red pea, red kidney, carbaso using hydrous ethanol to obtain powder.

[Comparative Example 2] Production of water extract of yellow soybean 4L of water was added to 400 g of crushed yellow soybean (Fukuyutaka), and extraction was performed by heating at 95 ° C for 60 minutes. The precipitate was removed by centrifugation, the supernatant was filtered to remove insoluble matters, and lyophilized to obtain 145 g of powder.

[Test Example 1] IgE lowering action Intraperitoneal administration of 100 μL of antigen solution (1 μg of ovalbumin (hereinafter referred to as OVA) and 2 mg of adjuvant (aluminum hydroxide) as a suspension) to mice (Balb / c, rabbit) (First sensitization) One week later, 100 μL of the suspension was intraperitoneally administered (second sensitization).
After the second sensitization, blood was collected from the tail of the mouse, and the amount of OVA-specific IgE in the blood was measured using an enzyme antibody method measurement kit (DS mouse IgE ELISA (OVA); manufactured by Dainippon Sumitomo Pharma Co., Ltd.). It was divided into two groups, a control group and a green soybean administration group, so that the amount was not biased.
As test foods, AIN-76 feed (Table 1) was given to the control group, and AIN-76 feed containing 5% of the green soybean hydrous ethanol extract of Example 2 was fed to the green soybean administration group. . The administration start date of the test meal was defined as day 0, blood was collected from the tail after 0, 7, and 16 days, and the amount of OVA-specific IgE in the blood was measured. The results are shown in Table 2.

表２から明らかなように、本発明の抗アレルギー剤により、ＯＶＡにより感作された血液中のＩｇＥ増加量が低下することがわかる。

As can be seen from Table 2, the antiallergic agent of the present invention decreases the amount of IgE increase in blood sensitized by OVA.

[Test Example 2] IgE-lowering action Mice (Balb / c, rabbit, 6 weeks old) were administered intraperitoneally with 200 μL of antigen solution (1 μg of OVA and 2 mg of adjuvant (aluminum hydroxide) as a suspension) (sensitization 1). 2 weeks later, 200 μL of the suspension was intraperitoneally administered again (second sensitization).
One week after the second sensitization, blood was collected from the tail of the mouse, and the amount of OVA-specific IgE in the blood was measured using a measurement kit (DS mouse IgE ELISA (OVA); manufactured by Dainippon Sumitomo Pharma). The group was divided into three groups: a control group, a blue soybean water extract (Example 3) administration group, and a yellow soybean water extract (Comparative Example 2) so that there was no bias in the amount of serum IgE.
The grouping date was 0 days, and the test meal was started after 2 days. The control group was given the AIN-76 feed shown in Table 1, and the test group was extracted with 5% of the extract of Example 3 and Comparative Example 2, respectively. Was fed with a feed added so that the content of
Blood was collected midway after 14 days, dissected after 28 days, and the amount of OVA-specific IgE in the blood was measured. Also, in order to see the effect on other classes of antibodies, the total IgG amount after 28 days was measured. The results are shown in Table 3.

  As is apparent from Table 3, the antiallergic agent (green soybean extract) of the present invention remarkably suppresses an increase in IgE in blood sensitized by OVA, and its effect is higher than that of yellow soybean. It turns out that it is extremely excellent. Moreover, there is no change in the amount of IgG as compared with the control group, and it can be seen that the antiallergic agent of the present invention specifically acts on the IgE antibody and causes no side effects.

[Test Example 3] IgE-lowering action Mice (Balb / c, rabbit, 6 weeks old) were intraperitoneally administered with 200 μL of antigen solution (1 μg of OVA and 2 mg of adjuvant (aluminum hydroxide) as a suspension) (sensitization 1). 1 week later, 200 μL of the suspension was intraperitoneally administered again (second sensitization).
One week after the second sensitization, blood was collected from the tail of the mouse, and the amount of OVA-specific IgE in the blood was measured using a measurement kit (DS mouse IgE ELISA (OVA); manufactured by Dainippon Sumitomo Pharma). The group was divided into three groups: a control group, a green soybean water-containing ethanol extract (Example 2) administration group, and a green soybean water extract (Example 1) so that there was no bias in the serum IgE amount.
As the test food, the AIN-76 feed shown in Table 1 was given to the control group, and the test group was fed with the feed to which the extracts of Examples 1 and 2 were added to a content of 5% in the feed. did. The administration start date of the test meal was defined as day 0, and dissection was performed 24 days later to measure the amount of OVA-specific IgE in the blood. The results are shown in Table 4.

表４から明らかなように、本発明の抗アレルギー剤により、ＯＶＡにより感作された血液中のＩｇＥ増加量が低下すること、特に水抽出物が高い効果を示すことがわかる。

As is apparent from Table 4, it can be seen that the antiallergic agent of the present invention decreases the amount of IgE increase in blood sensitized by OVA, and in particular, the water extract shows a high effect.

[Test Example 4] IFN-γ gene expression increasing action Cell lines (Jurkat cells) were cultured in 10% FBS-added DMEM medium (GIBCO) under conditions of 37 ° C and 5% CO2. It diluted with the culture solution so that it might become 3 * 10 < ⁵ > cells / mL, and this was dispensed to a 24-well plate by 1 mL of each well. 100 μL of the water-containing ethanol extracts of various beans of Example 2 and Comparative Example were added to the culture solution to a final concentration of 500 μg / mL, and cultured for 48 hours.
After completion of the culture, the intracellular IFN-γ gene expression was measured by a quantitative PCR measurement apparatus (Thermal cycler
Dice TP-800 (manufactured by Takara Bio Inc.) was used for measurement. The results are shown in Table 5 as relative values with the control with no solvent extract added being taken as 1.

表５から、青大豆以外の大豆はＩＦＮ−γにほとんど作用しないことが分かる。一方、本発明の抗アレルギー剤により、Ｔｈ１の指標であるＩＦＮ−γの発現が顕著に増加し、Ｔｈ１／Ｔｈ２バランスをＴｈ１側へ偏向させる作用を有することが分かる。

From Table 5, it can be seen that soybeans other than green soybean have little effect on IFN-γ. On the other hand, it can be seen that the antiallergic agent of the present invention significantly increases the expression of IFN-γ, which is an index of Th1, and has an action of deflecting the Th1 / Th2 balance toward the Th1 side.

[Test Example 5] IL-4 gene expression inhibitory action In the same manner as in Test Example 4, a cell line (Jurkat cell) is cultured, dispensed into a 24-well plate, and the aqueous soybean soybean water extract of Example 2 is terminated. 100 μL was added to a concentration of 250 and 500 μg / mL, and cultured for 48 hours.
After completion of the culture, intracellular IL-4 gene expression was measured using a quantitative PCR measurement apparatus (Thermal cycler Dice TP-800; manufactured by Takara Bio Inc.). The results are shown in Table 6 as relative values with the control with no solvent extract added as 1.

表６から、本発明の抗アレルギー剤により、Ｔｈ２の指標であるＩＬ−４の発現が顕著に低下し、Ｔｈ１／Ｔｈ２バランスをＴｈ１側へ偏向させる作用を有することが分かる。

From Table 6, it can be seen that the antiallergic agent of the present invention significantly reduces the expression of IL-4, which is an index of Th2, and has an effect of deflecting the Th1 / Th2 balance toward the Th1 side.

[Test Example 6] Inhibition of IgE and IL-4 Amounts Intraperitoneal administration (sensitization) of 100 μL of antigen solution (1 μg of OVA and 2 mg of adjuvant (aluminum hydroxide) as a suspension) to mice (Balb / c, sputum) The first suspension), and again 14 days later, 100 μL of the suspension was intraperitoneally administered (second sensitization).
After the second sensitization, blood was collected from the tail of the mouse, and the amount of OVA-specific IgE in the blood was measured using an enzyme antibody method measurement kit (DS mouse IgE ELISA (OVA); manufactured by Dainippon Sumitomo Pharma Co., Ltd.). The groups were divided into three groups, a control group, a green soybean administration group, and a soybean administration group so that the amount was not biased.
As a test meal, AIN-76 feed was given to the control group, AIN-76 feed containing 5% of the green soybean hydrous ethanol extract of Example 2 was given to the green soybean administration group, and the soybean administration group was AIN-76 feed containing 5% of the soybean water-containing ethanol extract of Comparative Example 1 was fed and reared.
The administration start date of the test meal was defined as day 0, and the IL-4 gene expression level on Peyer's patch was measured using a quantitative PCR measuring apparatus (Thermal cycler Dice TP-800; manufactured by Takara Bio Inc.) after 3 weeks. The results are shown in Table 7 as relative values with the control with no solvent extract added as 1.
In addition, spleen cells were collected and cultured for 72 hours, and the amount of OVA-specific IgE in the culture supernatant was measured using a measurement kit (DS mouse IgE ELISA (OVA); manufactured by Dainippon Sumitomo Pharma Co., Ltd.) by the enzyme antibody method. . The results are shown in Table 8.

  From Table 7, the anti-allergic agent of the present invention has the effect of significantly reducing the expression of IL-4, which is an index of Th2, compared to soybeans other than green soybeans, and deflecting the Th1 / Th2 balance toward the Th1 side. I understand. Similarly, it can be seen from Table 8 that the antiallergic agent of the present invention reduces the amount of IgE increase in blood sensitized by OVA as compared to soybeans other than green soybeans.

[Example 4] Manufacture of tablets 84 g of water extract of green soybean obtained in the same manner as in Example 1, 10 g of crystalline cellulose (Asahi Kasei) and 5 g of polyvinylpyrrolidone (BASF) were mixed, and 30 mL of ethanol was added thereto. Then, granules were produced according to a conventional method by a wet method. After the granules were dried, 1.2 g of magnesium stearate was added to give granules for tableting, and tableting was performed using a tableting machine to produce 100 tablets each having 1 g.

[Example 5] Manufacture of granules A mixture of 100 g of hydrous ethanol extract of green soybean obtained in the same manner as in Example 2, 100 g of lactose (DMV) and 40 g of crystalline cellulose (Asahi Kasei) is mixed with 130 mL of ethanol. The mixture was added to the kneader and kneaded for 5 minutes by a usual method. After kneading, the mixture was sieved with 10 mesh and dried at 50 ° C. in a dryer. After drying, the particles were sized to obtain 240 g of granules.

[Example 6] Manufacture of syrup agent While boiling 400 g of purified water, 750 g of white sugar and 100 g of the green soybean aqueous extract obtained in the same manner as in Example 1 were added and dissolved, and the cloth was heated when heated. The syrup was prepared by adding purified water to the total volume of 1000 mL.

[Example 7] Manufacture of liquid foods 700 g of pure water at about 65 ° C, 40 g of sodium caseinate (DMV), 160 g of maltodextrin (Sanwa starch) and hydrous ethanol extract of green soybean obtained in the same manner as in Example 2 50 g was added and dissolved, and then each component mixture of vitamin mix and trace minerals was added. The obtained mixed solution was put into a homomixer and roughly emulsified at about 8,000 rpm for 15 minutes. The obtained emulsified liquid is cooled to about 20 ° C., and after adding a fragrance, the final mess up is carried out. Sterilization was performed to obtain a liquid food.

[Example 8] Production of bread 160 g of wheat flour (strong flour) and 2 g of dry yeast were mixed. Separately, 5 g of the green soybean aqueous extract obtained in the same manner as in Example 1, 25 g of sugar, 3 g of sodium chloride and 6 g of skim milk powder were dissolved in 70 g of hot water, and 1 chicken egg was added and mixed well. Add this to the above mixture of flour and dry yeast, knead by hand, knead well by adding about 40 g of butter, make 20 roll bread dough, then ferment these bread dough, Eggs were spread on and then baked in an oven at 180 ° C. for about 15 minutes to prepare a roll.

[Example 9] Manufacture of confectionery Margarine and sugar were mixed and stirred well using a mixer to prepare whipped. Half of this was added to make a cream. The dough was prepared by adding the water extract of green soybean obtained in Example 3 to this and mixing lightly. The dough was formed and baked in an oven at 150 ° C. for 25 minutes to prepare a confectionery.

[Example 10] Manufacture of confectionery Whole eggs were loosened with a whisk, and sweeteners for 90 g of sugar were added and mixed well. To this, 10 g of the green soybean aqueous extract obtained in Example 3, 40 g of wheat flour and baking powder were added, mixed with stirring, and further mixed with butter and rum to prepare a dough. The dough was put into a mold and baked in an oven at 170 ° C. for 15 minutes to prepare a confectionery.

[Example 11] Manufacture of retort rice 2 grams of green soybean soy extract obtained in Example 3 was added to a general amount of water using 2 rice and cooked, and this was retort according to a conventional method. After filling the pack, it was sealed with nitrogen substitution and sterilized at 121 ° C. for 15 minutes to obtain retort rice.

[Example 12] Manufacture of sauce for pasta One serving (150 g) of meat sauce for pasta was put in a pan, and 1 g of a hydrous ethanol extract of green soybean obtained in the same manner as in Example 2 was added to the mixture and heated. did. After filling this pouch with a pouch, the pouch was sealed with nitrogen substitution and sterilized at 121 ° C. for 15 minutes to obtain a meat sauce for pasta.

[Example 13] Manufacture of vegetable juice To a commercially available vegetable juice, a hydrous ethanol extract of green soybean obtained in the same manner as in Example 2 was added and mixed so as to be 5% by mass to prepare a vegetable juice.

[Example 14] Manufacture of consomme soup Each slice of onion 100g, carrot 100g, long onion 100g, celery 50g, and tomato 100g was put in a pan, 500g of minced beef meat, 2 egg whites, 1kg beef bouillon And boil on fire, reduce the heat, boil on low heat for 1 hour while removing ak and fat floating on the surface, add 50 g of green soybean aqueous extract obtained in the same manner as in Example 1 And boiled for another 30 minutes and rubbed with a cloth to obtain a consomme soup.

[Example 15] Production of ointment (mass%)

  B is mixed while heating and dissolving at 70 ° C. to obtain an oil phase. A was mixed while being heated and dissolved at 70 ° C., and the oil phase of B was added thereto for mixing and emulsification. After that, the mixture was cooled and mixed well while adding C to obtain an ointment.

Claims (6)

  An antiallergic agent comprising a solvent extract of green soybeans.   The antiallergic agent according to claim 1, which suppresses IgE production.   The antiallergic agent according to claim 1, which increases Th1 / Th2.   The agent according to any one of claims 1 to 3, wherein the green soybean solvent extract is a water extract.   The agent according to any one of claims 1 to 3, wherein the solvent extract of green soybean is a hydrous ethanol extract.   An antiallergic food, beverage, feed and cosmetic comprising the agent according to any one of claims 1 to 5.