WO2006132293A1

WO2006132293A1 - Food composition having immunosuppressive effect by coating antigen structure of allergenic protein by polysaccharide modification

Info

Publication number: WO2006132293A1
Application number: PCT/JP2006/311468
Authority: WO
Inventors: Akio Kato
Original assignee: Yamaguchi University
Priority date: 2005-06-09
Filing date: 2006-06-01
Publication date: 2006-12-14
Also published as: JP2006340658A

Abstract

An object of the invention is to provide a method capable of realizing the reduction of allergen safely and a food composition having an allergy suppressive effect which can be orally taken safely. As a means for achieving the object, the invention provides a food composition utilizing oral immune tolerance, more particularly a food composition characterized by containing, as an active ingredient, an allergenic protein in which an antigen structure is coated by polysaccharide modification and inducing the oral immune tolerance in the gut immune system.

Description

According to the coating of the antigenic structure of the allergen protein by polysaccharide modification

Food composition having immunosuppressive effect

".Technical field

This is K>> Food Group Using Oral Immune Tolerance ^{: For} more details, please refer to the following. Immunity :: Inducing tolerance に関する Characteristic of food composition. '' Background technology

Allergic diseases such as hay fever are collectively called allergens .. Protein protein is contained in the body, and the whole body's immune system reacts excessively. It is a disease that causes P harm .. In recent years, an increasing trend has been seen. In particular, Japanese cedar pollinosis that occurs in early spring in Japan is also affected by the mass plantation of Japanese cedar and cypress that ignored the ecosystem in the Showa period.

It has become a disease that has a serious social impact · _r

': However, these diseases are due to the immune response inherent in humans: The fundamental cure has not been established so far. For this reason, drugs for the purpose of regulating the vaginal secretion system, such as '¾ histami / drug anti-allergic agent', have been used as treatment methods. ： In addition, Kinka also conducts research on the T allergen itself _: progress, allergen removal 'method symptomatic treatment with anti-inflammatory agents: ...., etc .: a method of reducing allergens by treating them with protein modifying agents, 'Traces': Allergens are injected several times and presented with a desensitization therapy that reduces allergic responses.:: Being: L'Er (Special 2 Q 0 5— 0 3 4 0 4 6, 'JP 2 Q 0 5 - 1 0 4 8 4 .7, JP 2 0 0, 2..' _^:: 2 4 9: 4: 4.2 ^ ₀ ':..

On the other hand, in recent years, the relationship between food and habits of allergies has started to be pointed out, such as drug therapy; 'allergic allergies ^' measures to deal with diseases are gradually being considered. If you are able to prevent allergic disease in your diet, you can suffer from these diseases. “It ’s very desirable, but so far, · There is no notification of food compositions that have been demonstrated to have a markedly allergic inhibitory effect. ...

• Invention disclosure.

'In view of ^the':..: Upper serial瑰拔', ^-: Ho present inventors',' allergen Tampa protein structure:. A: related to such their functions.

'Ru'. Research progressed, 'Polysaccharide A c JI protein known as a causative agent of cedar pollinosis .. and ί Goyori:;.: Allergen and.:; Can be reduced, 'and find out. ·' (^ Ι · sui ·

'Μ, ·-a "ϊ Ό'. '' A. ^;: - _: E" ^: t-. 'A ： 1.'('2: 0 0 3): Β i' osci '. B io' ech ii o 'l. B i oche m. 6 7 (1 1): 2 4 2 5-3 0)). In the present invention, the present inventors are able to bind one type of polysaccharide, galactomannan, and CJI by Maillard reaction, and in an experimental system using human serum, galactomannan-one CJI complex is more human than CJI. It was reported that the reactivity with IgE was significantly reduced (allergens were reduced) (Figs. 1 and 2).

Regarding the coating of allergen proteins with polysaccharides, a method is disclosed in which the polysaccharide pullulan and cedar pollen allergen are covalently bound with cyanuraseton chloride and used as a desensitizer (Patent No. 2 5 9 4 1 2 3). This method, however, treats allergens with chemicals and is difficult to apply to foods that enter the mouth. In view of these current situations, the present invention provides a method that can safely reduce allergens, and a food composition that has an allergy suppression effect that is safe and safe even when taken orally. Let it be an issue. The present inventors have further researched on the reduction of allergens by polysaccharides, and in particular introduced the viewpoint of “reduced allergen that can be taken orally” to complete the present invention. In other words, an ingestible polysaccharide and an allergen protein are combined under the dry conditions using the Maillard reaction and introduced into the intestinal tract immune system to establish a system that induces immune tolerance. The invention was completed.

In this context, immune tolerance is a type of immune response that is induced mainly by M cells (including dendritic cells and epithelial cells) in the intestinal tract and taking up substances that act as antigens (Fig. 3). In this system, it is becoming clear that continuous oral administration of antigen produces inhibitory T cells against the antigenic substance, and alleviates the immune response to these antigenic substances. is there. The biological significance of immune tolerance is considered to be an aspect of the biological response to external stimuli that “do not overreact to what we often say”. As a means of suppressing an allergic reaction without suppressing the immune reaction itself or treating the allergen with chemicals or the like as in the past, the present inventors dried allergen proteins and ingestible polysaccharides. The allergen protein can be used as a “food composition” by binding it under the conditions using the Maillard reaction and combining it with immune tolerance, particularly oral tolerance.

That is, the first aspect of the present invention provides a food composition characterized by having an allergen-inhibiting effect due to oral tolerance by using an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient.

According to a second aspect of the present invention, there is provided a food composition comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and having an allergy suppressing effect due to oral tolerance, wherein the modification Is a modification utilizing the Maillard reaction that occurs under natural conditions.

According to a third aspect of the present invention, an allergen protein whose antigenic structure is coated with a polysaccharide modification is used as an active ingredient, and has an allergy suppressing effect due to oral tolerance. A food composition, wherein the modification is a modification utilizing a Maillard reaction that occurs under dry conditions.

According to a fourth aspect of the present invention, there is provided a food composition comprising an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient, and having an allergy suppressing effect by oral tolerance. The modification is a modification using a Maillard reaction that occurs under dry conditions, and the reaction is a covalent bond formed between an amino group on the surface of a protein molecule and a carbonyl group at the reducing end of a polysaccharide. A reaction that occurs after mixing allergen proteins and polysaccharides, with temperature conditions in the range of 50 to 70 degrees Celsius, and relative humidity conditions in the range of 55 to 75 degrees Celsius. Provided is a food composition characterized in that it occurs without being catalyzed by.

According to a fifth aspect of the present invention, there is provided a food composition characterized by comprising an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient and having an allergy suppressing effect by oral tolerance. A food composition is provided, wherein the polysaccharide is a natural polysaccharide derived from animals and plants.

A sixth aspect of the present invention is a food composition comprising an allergen protein coated with an antigen structure by a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. The food composition is characterized in that the polysaccharide is a natural polysaccharide derived from animals and plants and is at least one of xyloglican, chitosan, alginic acid, and galactomannan.

According to a seventh aspect of the present invention, there is provided a food composition characterized by comprising an allergen protein 'whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. Provided is a food composition characterized in that the polysaccharide is galactomannan present in soybean seed coat.

An eighth aspect of the present invention is a food composition characterized in that it comprises an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and has an allergy suppressing effect due to oral tolerance. The polysaccharide is the polysaccharide according to any one of the fifth to seventh aspects, and the modification is the modification according to any one of the second to fourth aspects. A food composition is provided.

According to a ninth aspect of the present invention, there is provided a food composition comprising an allergen protein coated with an antigen structure by a polysaccharide modification as an active ingredient and having an allergy suppressing effect due to oral immune tolerance. The food composition is characterized in that the allergy suppressing effect is an inhibitory effect derived from immune tolerance in the intestinal tract immune system.

A tenth aspect of the present invention is a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. The allergy-suppressing effect stems from immune tolerance in the intestinal tract immune system, Provided is a food composition characterized by having an inhibitory effect that enables oral administration by suppressing anaphylaxis.

The first aspect of the present invention is a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. The allergic inhibitory effect is derived from immune tolerance in the intestinal tract immune system, and is an inhibitory effect that enables oral administration by suppressing anaphylaxis. The allergen protein is coated with a polysaccharide. It is an allergen protein that is difficult to be decomposed by proteases in the gastrointestinal tract and can reach the intestinal tract while retaining the antigen structure, and the allergen structure does not touch the mucous membrane before reaching the intestinal tract by the coating. It is an allergen protein that enables it to reach the intestinal tract It provides a food composition.

The 12th aspect of the present invention is a food composition characterized in that it comprises an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and has an allergy suppressing effect due to oral tolerance. The allergic inhibitory effect is derived from immune tolerance in the intestinal tract immune system and is an inhibitory effect that enables oral administration by suppressing anaphylaxis. Specifically, suppressive T cells are produced in the intestinal tract immune system. Allergen protein that is difficult to be decomposed by proteases in the gastrointestinal tract by being coated with a polysaccharide, and that can reach the intestinal tract while retaining the antigen structure. It is a protein, and the allergen structure does not touch the mucous membrane before reaching the intestinal tract due to the coating. 'Characterized in that an allergen protein made it possible to reach the provides a food composition.

A thirteenth aspect of the present invention is a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and having an allergic suppression effect by oral tolerance. The allergic inhibitory effect is derived from immune tolerance in the intestinal tract immune system, and is an inhibitory effect that enables oral administration by suppressing anaphylaxis. Specifically, suppressive T cells are produced in the intestinal tract immune system. Allergen protein that is difficult to be decomposed by proteases in the gastrointestinal tract by being coated with a polysaccharide, and that can reach the intestinal tract while retaining the antigen structure. It is a protein, and the allergen structure does not touch the mucous membrane etc. before reaching the intestinal tract due to the coating. Wherein the polysaccharide is the polysaccharide according to any one of the fifth to seventh aspects, and the modification is the above-mentioned modification. There is provided a food composition characterized by being a polysaccharide according to any one of the second to fourth aspects.

A fourteenth aspect of the present invention is characterized in that an allergen protein whose antigenic structure is coated with a polysaccharide modification is an active ingredient, and has an allergy suppressing effect due to oral tolerance. A food composition, characterized in that the allergen is at least one of an animal allergen, a plant allergen, and a dietary allergen. A fifth aspect of the present invention is a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergic suppression effect by oral tolerance. Provided is a food composition characterized in that the allergen is at least one of pollen allergen or dietary allergen.

The 16th aspect of the present invention is a food composition characterized by comprising an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient, and having an allergic suppression effect by oral tolerance. There is provided a food composition characterized in that the allergen is a pollen allergen, and the pollen is selected from cedar pollen, cypress pollen, pine pollen, ragweed pollen, and mugwort pollen.

The i 7th aspect of the present invention is a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and having an allergy suppression effect by oral tolerance, A food composition is provided wherein the allergen is a cedar pollen allergen '. -The eighteenth aspect of the present invention is a food composition characterized by comprising an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient and having an allergy suppression effect by oral tolerance. The allergen is the allergen according to any one of the 14th to 17th aspects, and the polysaccharide is any one of the 5th to 7th aspects. The modification according to any one of the second to fourth aspects, wherein the allergy suppression by oral tolerance is the ninth to the 12th aspect. The food composition is characterized by having an effect described in any one of the above.

According to a ninth aspect of the present invention, there is provided a food composition comprising an allergen protein coated with an antigen structure by a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. Provided is a food composition, wherein the allergen is a dietary allergen and is selected from chicken eggs, soybeans, buckwheat, wheat, and seafood.

According to a 20th aspect of the present invention, there is provided a food composition characterized by comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy suppression effect by oral tolerance. The food composition according to any one of the above second to first aspects, wherein the allergen is a dietary allergen and is selected from chicken eggs, soybeans, buckwheat, wheat, and seafood. I will provide a.

By using the food composition provided by the present invention, allergen proteins capable of causing an allergic reaction are introduced into the intestinal tract immune system without causing anaphylaxis to induce immune tolerance and prevent allergic diseases. Is possible. In addition, the present invention effectively induces “oral immune tolerance” and feels stress such as drug injection in the normal diet. Without allergies, it becomes possible to prevent allergic diseases.

This specification includes the contents described in the claims, specification and drawings of Japanese Patent Application No. 20 0 5-1 6 9 1 96 which is the basis of the priority of the present application. Brief Description of Drawings

Fig. 1 shows the electrophoretic pattern (SDS-PAGE) of the cedar pollen allergen C J I I I galactomannan complex. In the figure, the arrow indicates the direction of migration, and lane 1 is the molecular weight marker 1 (from the top 96, 67, 42, 30, 20, 14 kDa). Lane 2 is isolated and purified itC J I, and lanes 3 and 4 show C J I-galactomannan complex (3 is the mixing weight ratio 1: 2, 4 is 1: 4).

FIG. 2 shows the results of comparison of the binding ability between human IgE antibody and CJI and CJI-galatatomannan complex. The vertical axis of the graph represents IgE bindingrate (relative value when binding to CJI is 10-0), lane 1 is CJI, lane 2 is CJI-galatamannan complex, lane 3 Represents a control. IgE antibodies from each patient bound to C.J, but the ability to bind to CJI-galactomannan complex was reduced to control levels.

Figure 3 shows a schematic diagram of the functions of the intestinal tract immune system. When antigen is taken up from M cells present in the intestinal epithelium, immune tolerance is induced by the action of inhibitory T cells.

Figure 4 shows the Maillard reaction between allergen protein and polysaccharide. The naturally occurring Maillard reaction under drying bonds the ε-amino group of the protein to the reducing end carbonyl group of the polysaccharide (Α), and several polysaccharides bind to the surface of the protein (Β). Α shows its chemical formula and B shows a schematic diagram. C shows a schematic diagram of CJI-galactomannan complex formation. Powder C J I and galactomannan (weight ratio 1: 2 or 1: 4) were mixed and reacted in a desiccator (including a saturated K I solution) at a relative humidity of 65% and 60 for 2 weeks.

FIG. 5 schematically shows a method for purifying cedar pollen allergen CJI. A and B are peaks at the purification stage, and C is an electrophoretogram (SDS-PAGE) of purified CJI.

FIG. 6 shows oral tolerance to mice. (Upper) Shows the schedule of immune tolerance induction experiments for mice. C J I -GM (20 μg / ά a y) was orally administered continuously for 3 weeks, after which allergen (5 / z g) was sensitized and resensitized one week later. (Lower) The amount of IgE production for allergen sensitization was compared between CJI-GM oral administration mice, CJI oral administration mice, and non-administration (Mock) mice. In C J I—GM-treated mice, IgE production was significantly reduced.

FIG. 7 shows oral tolerance to allergic mice. (Upper) The schedule of immune tolerance induction experiments for allergic mice is shown (symbols are the same as in Fig. 6). IgE production for allergen sensitization was determined using CJI-GM mice administered orally, CJI mice or Comparison with mice. Even in allergic mice, the amount of IgE produced decreased when CJI-GM was orally administered. .

Figure 8 shows oral tolerance in patients with cedar pollen allergy. (Upper) Shows the schedule for oral administration of CJI-GM (10 OmgZday) to 5 volunteers.

(Lower) The figure shows the number of cedar pollen scattered in Yamaguchi City in March 2005 (individual Z'cm ² , provided by the Yamaguchi Medical Association) plus a blood sampling schedule.

Figure 9 shows oral tolerance (2) for patients with cedar pollen allergy. CJI— The amount of IgE production before and after oral administration of GM was compared. The vertical axis of the graph shows the index (spot area) of IgE production, A, N, S, and K represent each patient. The graph shows the year when light gray is not given orally, and dark gray is given orally Each year is shown, and the arrow on the right side of the graph indicates the positive / negative threshold (4 cm ² ). Oral administration of CJI-GM significantly reduced IgE production.

FIG. 10 shows clinical findings related to the effects of the present invention. Each symptom of hay fever was divided into five stages, and the symptoms before and after oral administration of CJI GM were compared. All four patients felt a reduction in allergies, two of which were completely cured. Values in parentheses indicate clinical findings prior to tolerant administration. BEST MODE FOR CARRYING OUT THE INVENTION

The present invention is described in detail below. According to a first aspect of the present invention, there is provided a food composition comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and having an allergy suppressing effect by oral tolerance. The allergen protein here is a protein that can cause allergic diseases, and is not particularly limited as long as it is not toxic when taken orally. The polysaccharide in the present invention is also edible. If it is a thing, the kind in particular will not be limited.

The mechanism of inducing oral tolerance by ingesting the allergen-monosaccharide complex in the present invention is considered as follows. Orally administered complexes are partially free from degradation by digestive enzymes in the stomach, and the partially degraded products are recognized by the intestinal immune system. The allergen protein taken up here is recognized by the intestinal lymph apparatus (GAL T), and useful ones are accepted by the Peyer's patch at the center of the apparatus, and unnecessary ones are excluded. If allergens are taken orally continuously for a certain period of time, T cells induce inhibitory T cells in the intestinal tract immune system, and no longer directs antibody production to B cells even in the presence of foreign allergens. It is. As this shows, there is no problem as long as the allergen and polysaccharide usable in the present invention are safe even if they are taken orally.

In the second to fourth aspects of the present invention, an allergen protein whose antigenic structure is coated with a polysaccharide modification is used as an active ingredient, and has an allergic suppression effect by oral tolerance. A food composition is provided, wherein the modification is a modification utilizing a Maillard reaction that occurs under natural conditions. The modification here is more specifically a modification using the Maillard reaction that occurs under dry conditions. More specifically, the modification is a covalent bond formed between the amino group on the protein molecule surface and the carbonyl group at the reducing end of the polysaccharide. After mixing allergen protein and polysaccharide, the temperature condition is in the range of 50 degrees Celsius to 70 degrees Celsius (preferably from 55 degrees Celsius to 65 degrees Celsius, more preferably 60 degrees Celsius) Modification using the Maillard reaction that occurs when the relative moist conditions are in the range of 55% to 75% (preferably 60% to 70%, more preferably 65%). Yes (Figure 4). As described in the Examples, the modification of the present invention is performed by drying and heating a mixture of allergen protein powder and polysaccharide powder in a desiccator or the like at about 60 ° C. and a relative humidity of 65%. No catalyst is used. That is, the compound obtained by this modification is very characteristic in that it is safe to use as it is.

In the fifth to eighth aspects of the present invention, there is provided a food composition comprising an allergen protein coated with an antigen structure by a polysaccharide modification as an active ingredient and having an allergy suppressing effect due to oral tolerance. A food composition is provided, wherein the polysaccharide is a natural polysaccharide derived from animals and plants. Specifically, the polysaccharide here is at least one of xyloglican, chitosan, alginic acid, and galatatomannan, and is preferably galactomannan present in soybean seed coat. As described above, the polysaccharide in the present invention is not a problem as long as it can be taken orally, but considering use as a food thread or a natural product, natural polysaccharides derived from animals and plants are preferable. In the examples of the present invention, galactomannan, which is present in a large amount in soybean seed coat and is guaranteed to be safe, is used as long as it is another natural polysaccharide and can cover the antigen structure. Is available. A food composition according to any of the fifth to eighth aspects of the present invention, wherein the modification is the modification according to any of the second to fourth aspects, is also to be included in the present invention.

In the ninth to thirteenth aspects of the present invention, a food composition comprising an allergen protein coated with an antigen structure by a polysaccharide modification as an active ingredient, and having an allergy suppressing effect due to oral tolerance. The food composition is characterized in that the allergy-suppressing effect is an inhibitory effect derived from immune tolerance in the intestinal tract immune system. The allergy-suppressing effect here is derived from immune tolerance in the intestinal tract immune system in detail, and is an inhibitory effect that enables oral administration by suppressing anaphylaxis. More specifically, the allergen protein described above is caused by polysaccharides. By being coated, it is difficult to be decomposed by proteases in the gastrointestinal tract, and it is possible for the protein component to retain its antigenic structure, and the allergen structure does not touch the mucous membrane etc. before reaching the intestinal tract by the coating. It is an allergen protein that is able to reach the level of the tumor, and more specifically, the antigen is presented by macrophages, dendritic cells, etc. in the intestinal tract immune system. This is an inhibitory effect due to the production of T cells. In order to introduce an allergen into the intestinal tract immune system, it must be taken orally and passed through the esophagus and stomach. However, when an uncoated allergen is taken orally as it is, it is abruptly called anaphylaxis in the esophageal mucosa. There is a risk that an allergic reaction will occur, and even if anaphylaxis does not occur, the allergen may be degraded in the stomach and not be introduced into the intestinal tract immune system. The allergen coating with a polysaccharide has two advantages of suppressing anaphylaxis and allowing the allergen to reach the intestinal tract, which is a unique viewpoint in the present invention. In the fourteenth to eighteenth aspects of the present invention, a food composition comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient and having an allergic suppression effect by oral tolerance. There is provided a food composition, wherein the allergen is at least one of an animal allergen, a plant allergen, and a dietary allergen. The allergen here is, more specifically, any one of pollen allergen or dietary allergen. More specifically, the pollen allergen is described in detail, for example, cedar pollen, cypress pollen, pine pollen, ragweed pollen, mugwort Pollen allergen derived from pollen, especially cedar pollen. That is, the food composition has an active ingredient in which the antigenic structure of the allergen protein contained in these pollen is coated with a polysaccharide modification. The allergen in the present invention may be any as long as it causes allergies and does not itself have toxicity. However, since the present invention provides a food composition, animal allergens, plant allergens, Dietary allergens are desirable. In the examples of the present invention, pollen allergens, more specifically cedar pollen allergens, were demonstrated, but any allergen that can be combined with polysaccharides by Maillard reaction is demonstrated. The present invention is applicable. In the 14th to 17th aspects of the present invention, a food composition that uses the polysaccharide described in any of the 5th to 7th aspects and performs the modification described in any of the 2nd to 4th aspects. A food composition having the effect of any one of the ninth to 12th aspects should also be included in the present invention.

In the nineteenth and twenty-fourth aspects of the present invention, a food composition comprising an allergen protein coated with an antigen structure by polysaccharide modification as an active ingredient and having an allergy-suppressing effect by oral tolerance. There is provided a food composition, wherein the allergen is a dietary allergen and is selected from eggs, soybeans, buckwheat, wheat, and seafood. That is, the food composition includes an active ingredient in which the antigenic structure of the allergen protein contained in these foods is coated with a polysaccharide modification. Food allergies, like pollen allergies, are becoming a serious problem in recent years, especially wheat and egg allergies that are included in many foods, which give great stress to the diet. It is considered that oral tolerance can be applied to these food allergies, and a food composition utilizing this is provided. A food composition obtained by adding the conditions in the second to the first to third aspects to the food composition is also included in the present invention. It is included.

In the second aspect of the present invention, there is provided a method for suppressing allergy by oral tolerance, comprising administering an effective amount of an allergen protein whose antigenic structure is coated with a polysaccharide modification. The allergy is not particularly limited, but can suppress pollen allergy and food allergy, for example. Allergen proteins are selected based on the allergy to be suppressed. That is, the allergen protein whose antigenic structure is coated with the polysaccharide modification in the present invention can be used as an active ingredient of an allergy inhibitor, for example, a pollen allergy inhibitor or a food allergy inhibitor.

The subject to which the food composition and the allergy inhibitor of the present invention are administered is not limited to, for example, mammals such as humans, domestic animals (such as sushi, horses, hudges, etc.), pet animals (Inu, Cats) and laboratory animals (mouse, rat, hamster, etc.). '

The food composition and allergy inhibitor of the present invention preferably contain a pharmaceutically acceptable carrier or additive together with an allele.gen protein whose antigenic structure is coated by polysaccharide modification. Examples of such carriers and additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, sodium alginate, water-soluble dextran, sodium carboxymethyl starch , Pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, 、 serine, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, pharmaceutical supplement In addition to surfactants that are acceptable as substances, artificial cell structures such as ribosomes can be mentioned.

When an allergy inhibitor is administered, it can be a solid preparation such as a tablet, granule, powder or pill, or a liquid preparation such as a liquid or syrup. These dosage forms may contain additives such as binders, excipients, lubricants, disintegrants, wetting agents, stabilizers, buffering agents, flavoring agents, preservatives, fragrances, and coloring agents.

The amount of allergen protein whose antigenic structure is coated by polysaccharide modification in the allergic inhibitor described above varies depending on the application, dosage form, administration route, etc., but is 1 to 5% by weight based on the total weight, preferably 2 to 3% by weight. In addition, the effective dose varies depending on the age of administration subject, administration route, and number of administration, and can vary widely. For example, the active ingredient is 1 to 100 mg / kg body weight per day, and can be administered from several times a day to once every several weeks.

Hereinafter, the present invention will be described specifically by way of examples. However, the technical scope of the present invention is not limited by the examples. Example Example 1

Examples of this invention will be described below. In this example, the effect of the present invention was demonstrated for cedar pollen allergen, but the present invention is not limited to the example. The demonstration experiment of the present invention was conducted with the permission of Yamaguchi University “Research Ethics Committee for Humans”.

(Materials, Reagents, etc.) Japanese cedar pollen was provided by Shinkyo Sangyo Co., Ltd. (Yamaguchi Prefecture). Antiserum for cedar pollinosis patients was provided in cooperation with Dr. M Muto, Department of Dermatology, Yamaguchi University Hospital. Biotinylated anti-human IgG antibody is ICN Pharmaeceutica 1s (CA, USA), anti-human IgE antibody is Chemicon International (CA, USA), HRP-streptavidin Those manufactured by Ve ctor Laboratory (CA, USA) were used.

(Purification of C J I) Purification of C J I, a squirrel allergen protein, was performed according to the method of Y a sue ed a H. e t a l. Figure 5 shows an overview of the preparation method. In order to remove fat from cedar pollen, 100 g of pollen was dissolved in 25 Om 1 dimethyl ether and stirred vigorously, after which dimethyl ether was removed. Treat this pollen with 2000 m 1 of 0.1 25M ammo ni um bicarbonate (pH 8.0, RT) for 48 hours, then centrifuge (1500 0 X g, 15 min) to separate the supernatant, This was called crudeextract (CE). Ammonium sulfate was added to CE until 80% saturation (56 1 g / 1, 4 ° C., ov er i n g ht), and then centrifuged (1 8500 X g, 40 m i n) to collect the precipitate. The precipitate was dissolved in 200 ml of 0.05 M Tris-HC 1 (pH 7.8) and dialyzed. The dialyzed CE was centrifuged (1 0000 X g, 15 m in), the supernatant was removed, and the mixture was diluted 2-fold with 0.05 M Tris_HC 1 u f fer (pH 7.8). The diluted solution was applied to a DAE A Toyo p aar 1 column, and the effluent was collected and dialyzed against 0.01 M acte te bu f f e r (pH 5.0, hereinafter abbreviated as AB). The 5-fold diluted solution was applied to a CM—T o yo pear 1 column flattened with 0.0 1 M AB (pH 5.0). The column was washed with A B, and the adsorbed protein was eluted with Na C 1 (in 0. 0 1M AB) with a 0-0.4M concentration gradient. The peak obtained by absorption at 280 nm was collected, dialyzed with distilled water, and lyophilized. This sample was dissolved in 4 ml 1 distilled water and equilibrated with 0.05 M ammo ni um bicarbonate (pH 8.0). A Sephadex G—1 00 column (2.6 X 1 00 cm, Pharmacia) I was desperate. Tanha. The protein was eluted with 0.05 M ammoni bc ar bona te solution at 20 m 1 / h, and the peak obtained at an absorbance of 280 nm was collected, dialyzed with distilled water, and lyophilized. Fig. 5 shows an electrophoretogram (SDS-PAGE) of the purified CJI.

Example 2

(Formation of CJI-galactomannan complex) The binding between galactomannan and CJI It was carried out using the Maillard reaction under dry conditions (Ka to A. eta 1. (199 0) Agric. Biol. Chem. 54: 107-112). A mixture of CJI and galactomannan powder (weight ratio 1: 2 and 1: 4) was dissolved in water to 10% (w / v) and freeze-dried. Place the CJI-galactomannan mixture on the top of the desiccator, and adjust the humidity with a saturated KI (potaci um iodide) solution at the bottom, and heat for 2 weeks under conditions of relative humidity (RH) 65% and 60 ° C. Dried. Under low water activity, the Maillard reaction between the ε-ami / group of the protein and the reducing terminal carbonyl group of the polysaccharide was promoted to form a CJI-galactomannan complex (CJI-GM) ( (Figure 4).

(Purification of CJI-galactomannan complex) In order to remove CJI not bound to galactomannan, the complex was applied to an ion exchange column (CM—Toyo pea 1650) and purified. Elution was performed with Na C 1 (in 0.01 M AB, pH 5.0) with a concentration gradient of 0 — 0.05 M. The C J I _GM peak was collected, dialyzed against distilled water, and then lyophilized. When the CJI_GM thus formed was compared with purified CJI by electrophoresis, it was found that its molecular weight was 50-100 kDa (Fig. 1). Example 3

(CJI-Galatatomannan complex and IgE binding, (Us ui M, Ka to A. eta 1. (2003) supra)) CJI and Galatatomannan binding to IgE antibody recognition In order to confirm the influence, we analyzed by the comp etitive EL IZA method. As a result of experiments using antibodies provided by three patients, as shown in Figure 2, CJI-GM showed a 95-98% decrease in the ability to bind to human IgE antibodies compared to CJI alone. It was. This indicates that galactomannan hardly binds to the human antibody as a result of coating the antigen structure of CJI.

Example 4

During BAB L / C mice diet (oral tolerance to mouse), 20 mu such that _§ 'CJI- GM was added per day, was given three consecutive weeks. Thereafter, the allergen was dissolved in 100 μl phosphate buffer along with 5 μg of alum and sensitized, and resensitized one week later (FIG. 6, upper panel). The lower part of Figure 6 shows the amount of IgE produced against allergens in mouse blood before and after sensitization. It was shown that IgE production was strongly suppressed in the CJI-GM administration group.

Example 5

(Tolerance of oral immunity to allergic mice) Allergic mice were prepared, and CJI-GM was orally administered according to the same schedule as in Example 4. Thereafter, allergens were similarly sensitized to examine IgE production (Fig. 7). ). As in Example 4, IgE production was suppressed in the CJI_GM administration group, and oral allergic tolerance was induced in allergic mice, and allergy was suppressed. Example 6

(Tolerance of oral immunity to cedar pollen allergic patients) For oral administration, 100 mg of CJI-GM (1: 4 by weight) packed in soluble capsules (or wrapped in oblate), volunteer cedar allergy patients Five patients were orally administered once daily during the period from late February to early February (Figure 8, upper panel). Thereafter, blood was collected in March, and blood Ϊ g E concentration was measured. In 2005, blood was collected on March 18th, when the second wave of the scattering cycle arrived, and blood I g E concentration was expected to increase. We compared blood IgE levels between the year when CJI and GM were administered orally and when not. The lower part of Fig. 8 shows the blood collection schedule added to the data on the amount of cedar pollen scattered in Yamaguchi City in March (provided by the Yamaguchi Medical Association), which has the highest amount of cedar pollen.

(Oral tolerance in patients with cedar pollen allergy 2) Figure 9 shows the effect of oral tolerance on oral administration of CJI-GM. 1 O Omg Z day of CJI-GM until just before pollen scatter January and half (45 days) Shows the amount of IgE produced in patients who were orally administered compared to the year when they were not administered orally . The amount of IgE production was calculated as the area of a spot formed on the skin by performing a scratch test on the skin using CJI. The threshold for allergic episodes is 4 cm ² , 3 out of 4 are below the threshold, and 1 person (severe hay fever) is higher than that before oral administration. The value is shown.

(Clinical findings) Clinical findings are an important indicator of allergic reactions. The results of the clinical findings of each item for the four patients who received CJI-GM trans-ro administration are shown in Fig. 10 in comparison with the year before administration. All four patients felt a reduction in allergies (light natsu), and two of them showed “complete cure” with no symptoms.

(Supplementary note) As reported in the news, etc., the year 2005 is a year in which the amount of cedar pollen scattered is unusually high as it is rarely seen in recent years. The value of (~ 80 000 cm 3) was shown. It is a noteworthy result that the food composition of the present invention showed a reduction in cedar pollen allele in the last two decades. Industrial applicability

By using the present invention, it is possible to provide a safe and highly effective food composition having an allergy control effect. It is said that there are over 10 million people in Japan (Reference: Ministry of Agriculture, Forestry and Fisheries website http: // ww w. Maff. Go. Jp /) “Functional food” for Japanese cedar pollinosis patients It can be provided in the form of `` health food '' and can be used in the food industry. (In addition to CJI, cedar pollen allergens can contain CJ 2 in trace amounts, but can also be used as a crude extract of plants. This is contained, and if this crude extract is modified with polysaccharides, it can coat the CJ2 antigen structure and produce oral tolerance). Similarly, food compositions containing other allergens to which the present invention is applied as active ingredients can also be used in the food industry. it is conceivable that.

All publications, patents and patent applications cited in this specification are incorporated herein by reference in their entirety.

Claims

A food composition comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy-suppressing effect due to oral immune tolerance. The food composition according to claim 1, wherein the modification is modification using a Maillard reaction that occurs under natural conditions. The food composition according to claim 1, wherein the modification is a modification using a Maillard reaction that occurs under dry conditions. The reaction is a covalent bond formed between the amino group on the surface of the protein molecule and the carbonyl group at the reducing end of the polysaccharide. After mixing the allergen protein envelope and the polysaccharide, the temperature condition is from 50 degrees Celsius to 70 degrees Celsius. The reaction occurs within a range of up to 5% and the relative humidity condition is within a range of 55% to 75%, and occurs without being catalyzed by chemicals. The food composition according to Item 3. The food composition according to any one of claims 1 to 4, wherein the polysaccharide is a natural polysaccharide derived from animals or plants. 6. The food composition according to claim 5, wherein the polysaccharide is at least one of xyloglican, chitosan, alginic acid, and galatatomannan. The food composition according to claim 5, wherein the polysaccharide is galactomannan present in soybean seed coat. An allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergic inhibitory effect due to oral immune tolerance, wherein the polysaccharide comprises the claims 5 to The polysaccharide according to any one of claims 7 to 7, wherein the modification is the modification according to any one of claims 2 to 4. object. The food composition according to any one of claims 1 to 8, wherein the allergy suppression effect is an suppression effect derived from immune tolerance in the intestinal tract immune system.

10. The food composition according to claim 9, wherein the allergy-inhibiting effect is an inhibitory effect that enables oral administration by inhibiting anaphylaxis.

11. The allergen protein, which is difficult to be decomposed by a protease in the gastrointestinal tract or the like by being coated with a polysaccharide, and that can reach the intestinal tract while retaining the antigen structure. The food composition according to claim 10, characterized in that the allergen protein can reach the intestine without touching the mucous membrane before reaching the intestine. . 12. The food composition according to claim 11, wherein the allergy inhibitory effect is an inhibitory effect caused by production of inhibitory T cells in the intestinal tract immune system.

13. The polysaccharide is characterized in that it is the polysaccharide according to any one of claims 5 to 7, wherein the modification is any of claims 2 to 4. The food composition according to claim 12, wherein the food composition is the decoration described in claim 1.

14. The food composition according to any one of claims 1 to 13, wherein the allergen is at least one of animal allergens, plant allergens, and dietary allergens. object.

15. The food composition according to claim 14, wherein the allergen is any one of a pollen allergen and a dietary allergen.

16. The food composition according to claim 15, wherein the allergen is a pollen allergen, and the pollen is selected from cedar pollen, cypress pollen, ragweed pollen, and mugwort pollen. object.

17. The food composition according to claim 16, wherein the allergen is a cedar pollen allergen.

18. A food composition comprising, as an active ingredient, an allergen protein coated with an antigen structure by polysaccharide modification, and having an allergic suppression effect by oral immune tolerance, wherein the allergen comprises the claim 1 The polysaccharide according to any one of claims 4 to 17, wherein the polysaccharide is the polysaccharide according to any one of claims 5 to 7. And the modification is any one of claims 2 to 4 Wherein the allergy suppression by oral tolerance is the effect according to any one of claims 9 to 12. Food composition.

19. The food composition according to claim 14, wherein the allergen is a dietary allergen and is selected from eggs, soybeans, buckwheat, wheat, and seafood.

20. A food composition comprising an allergen protein whose antigenic structure is coated with a polysaccharide modification as an active ingredient, and having an allergy suppressing effect by oral immune tolerance, wherein the allergen is a dietary allergen, The food composition according to any one of claims 2 to 12, wherein the food composition is selected from chicken eggs, soybeans, buckwheat, wheat, and seafood.

21. A method for suppressing allergy by oral tolerance, comprising administering an effective amount of an allergen protein whose antigenic structure is coated with a polysaccharide modification.