Anti-allergic oral composition

Abstract

To provide a technology for anti-allergic purposes.SOLUTION: An anti-allergic oral composition comprising a sulfated polysaccharide derived from Aphanothece sacrum.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present disclosure relates to oral antiallergic compositions. [Background technology]

[0002] Allergies are typically broadly classified into type I allergies, type II allergies, type III allergies, and type IV allergies. Type I allergy is an allergy caused by the release of chemical mediators such as histamine from mast cells or basophilic cells (degranulation). Type II allergy is an allergy caused by the reaction of an antigen or hapten with an antibody, followed by the binding of complement thereto. Type III allergy is an allergy caused by a complex between an antigen and an antibody (so-called immune complex). Type IV allergy is an allergy caused by the release of cytokines from sensitized T cells as a result of the reaction between sensitized T cells and an antigen.

[0003] Type I allergy is associated with, for example, atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, urticaria, allergic conjunctivitis, allergic dermatitis, anaphylactic shock, and the like (Patent Document 1). Type II allergy is associated with, for example, Hashimoto's disease (chronic thyroiditis), Goodpasture's syndrome, organ transplant rejection, etc. (Patent Document 2). Type III allergy is associated with, for example, systemic vasculitis, cryoglobulinemia, glomerulonephritis, etc. (Patent Document 2). Type IV allergic diseases are associated with, for example, contact dermatitis, organ transplant rejection, etc. (Patent Document 2).

[0004] Of these, for type I allergies, known active ingredients of oral compositions for anti-type I allergies include, for example, at least one selected from the group consisting of Satsuma mandarin peel and Ponkan mandarin peel, and β-lactoglobulin (Patent Document 3), and a decomposed extract of bagasse (Patent Document 4).

[0005] Meanwhile, the freshwater cyanobacterium Aphanothece sacrum and polysaccharides (sugar derivatives) derived therefrom are known. These polysaccharides (sugar derivatives) are known to be used for the treatment and prevention of keratoconjunctival disorders (Patent Document 5). Furthermore, a skin composition containing water, the sugar derivatives, glycerin, and a phospholipid polymer has been developed (Patent Document 6). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent No. 7372112 [Patent Document 2] Patent No. 5885154 [Patent Document 3] Patent No. 6000206 [Patent Document 4] Patent No. 7208735 [Patent Document 5] Patent No. 6779599 [Patent Document 6] Patent No. 6336355 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present disclosure is to provide a technology for anti-allergy. [Means for solving the problem]

[0008] The present inventors have conducted extensive research to solve the above problems and have found that the above problems can be solved by orally ingesting a specific polysaccharide.

[0009] <1> An anti-allergic oral composition containing sulfated polysaccharides derived from Aphanothece sacrum. <2> The allergy is type I allergy. <1> The oral composition according to claim 1. <3> The type I allergy is atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, hives, allergic conjunctivitis, allergic dermatitis, or anaphylactic shock. <2> The oral composition according to claim 1. <4> An oral composition for suppressing IgE production, containing sulfated polysaccharides derived from Aphanothece sacrum. <5> An oral composition for suppressing rectal temperature drop, comprising sulfated polysaccharides derived from Aphanothece sacrum. <6> An oral composition for inhibiting the release of chemical mediators, comprising sulfated polysaccharides derived from Aphanothece sacrum. <7> The chemical mediator is histamine, β-hexosaminidase, or leukotriene. <6> The oral composition according to claim 1. <8> A pharmaceutical composition, <1> ~ <7> The oral composition according to any one of the preceding claims. <9> A food composition, <1> ~ <7> The oral composition according to any one of the preceding claims. <10> A feed composition, <1> ~ <7> The oral composition according to any one of the preceding claims. [Effects of the Invention]

[0010] The present disclosure can provide a technology for anti-allergy. [Brief explanation of the drawings]

[0011] [Figure 1] 1 is a graph showing the results of a Passive Cutaneous Anaphylaxis (PCA) reaction test (part 1) according to one embodiment of the present disclosure. [Figure 2] 1 is a graph showing the results of a test to evaluate the inhibition of IgE production according to one embodiment of the present disclosure. [Figure 3] 1 is a graph showing the results of a test to evaluate inhibition of rectal temperature reduction according to one embodiment of the present disclosure. [Figure 4] 1 is a graph showing the results of a Passive Cutaneous Anaphylaxis (PCA) reaction test (part 2) according to one embodiment of the present disclosure. [Figure 5] 1 is a graph showing the results of a Passive Cutaneous Anaphylaxis (PCA) reaction test (part 3) according to one embodiment of the present disclosure. [Figure 6] 1 is a graph showing the results of a histamine release inhibition evaluation test (part 1) according to one embodiment of the present disclosure. [Figure 7] 1 is a graph showing the results of a histamine release inhibition evaluation test (part 2) according to one embodiment of the present disclosure. [Figure 8] 1 is a graph showing the results of a β-hexosaminidase release inhibition evaluation test (part 1) according to one embodiment of the present disclosure. [Figure 9] 1 is a graph showing the results of a β-hexosaminidase release inhibition evaluation test (part 2) according to one embodiment of the present disclosure. [Figure 10] 1 is a graph showing the results of a β-hexosaminidase release inhibition evaluation test (part 3) according to one embodiment of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0012] In the present disclosure, the terms "ingestion," "ingest," "ingested," etc. may be used interchangeably with the terms "administration," "administered," "administered," etc. The terms "administration," "administered," "administered," etc. may be terms used for, for example, feed compositions and pharmaceutical compositions.

[0013] One aspect of the present disclosure is an anti-allergic oral composition containing a sulfated polysaccharide derived from Aphanothece sacrum.

[0014] The "sulfated polysaccharides derived from Aphanothece sacrum" of the present disclosure are a type of sulfated polysaccharide derived from the freshwater cyanobacterium Aphanothece sacrum and having an average molecular weight of 2,000,000 or more. Specifically, they are sugar derivatives having a repeating structure of sugar chain units in which sugar structures having hexose structures and sugar structures having pentose structures are linked in a linear or branched chain via α-glycosidic or β-glycosidic bonds, the sugar chain units containing sulfated sugars, and in which 2.7 or more hydroxyl groups per 100 hydroxyl groups are sulfated, or in which sulfur accounts for 1.5% by weight or more of the total elements. Hereinafter, they may be referred to as "polysaccharides of the present disclosure" or "sugar derivatives," etc.

[0015] The provision "derived from Aphanothece saccharin" is part of the provision for identifying the sugar derivative as a substance, and does not limit the source of the sugar derivative or the method of production.

[0016] The sulfated saccharide may be a lactated sulfated saccharide or a non-lactated sulfated saccharide, and the sulfated saccharide may be sulfated muramic acid or sulfated N-acetylmuramic acid, or may be one or more of these.

[0017] The sugar derivatives include at least glucose, galactose, mannose, galactosamine, xylose, arabinose, glucuronic acid, galacturonic acid, fucose, and rhamnose, and may have a functional group selected from a group of functional groups including at least a sulfate group, a lactic acid group, and a methyl group bound to any binding position in these sugar structures.

[0018] A part of the sugar structure constituting the sugar derivative may further be bound to a peptide or a lipid.

[0019] The average molecular weight of the sugar derivative may be 20,000,000 or more. The molar ratio of the main sugar structures constituting the sugar derivative may be arabinose 1.1:fucose 3.7:rhamnose 15.4:xylose 17.0:mannose 10.5:galactose 12.3:galacturonic acid 4.6:glucuronic acid 4.7:glucose 28.8:galactosamine 2.03.

[0020] The average molecular weight of the sugar derivative is, for example, 2,000,000 or more, 5,000,000 or more, 10,000,000 or more, 20,000,000 or more, and 30,000,000 or less, 28,000,000 or less, 25,000,000 or less, 22,000,000 or less, etc. It may also be a compatible combination thereof, for example, 2,000,000 to 30,000,000, 5,000,000 to 28,000,000, 10,000,000 to 25,000,000, 20,000,000 to 22,000,000, etc.

[0021] The sugar derivative may be extracted, isolated, or purified from Aphanothece sacrum. The method for extracting the sugar derivative from Aphanothece saccharin is not particularly limited, and may be, for example, Examples of such methods include conventional methods such as those described in Patent Document 5. Specifically, the following method can be mentioned: A suitable amount of Aphanothece sacrum is frozen and thawed, then washed with water to remove water-soluble pigments. The fat-soluble pigments are then removed by stirring overnight with an organic solvent such as ethanol. After the organic solvent is separated from the Aphanothece sacrum from which the pigments have been removed, the Aphanothece sacrum is dissolved in, for example, a 0.1-0.6 N aqueous sodium hydroxide solution, heated to, for example, 40-80°C, with stirring for, for example, 5-12 hours. The aqueous solution of the sugar derivative extracted by this procedure is neutralized, for example, with concentrated hydrochloric acid. Desalting may be performed by immersing the solution in, for example, 60-80% isopropyl alcohol, or this may not be necessary. The sugar derivative is then precipitated in a gel form by pouring it into, for example, 100% isopropanol with stirring, followed by drying with hot air. The hot air drying may be performed at, for example, 85-100°C or higher for, for example, 6-24 hours. In another example, sugar derivatives are extracted from A. sacchariflorus by heating an aqueous dispersion of A. sacchariflorus in an autoclave at 135°C for 30 minutes. The extracted sugar derivatives may be purified by centrifugation, filtration, alcohol washing, etc. Alternatively, before extracting the sugar derivatives from A. sacchariflorus, the A. sacchariflorus may be frozen and thawed, followed by a step of removing pigments.

[0022] The sugar derivatives used may be commercially available products.

[0023] Furthermore, the sugar derivative may be contained in crushed dried Amorphophallus saccharinus (sometimes referred to as "crushed dried Amorphophallus saccharinus" in the present disclosure) as long as it exhibits an anti-allergic effect. Therefore, the present disclosure can also provide an anti-allergic oral composition containing the crushed dried matter of Aphanothece sacrum containing the sugar derivative. The crushed dried Amorphophallus saccharinus containing the sugar derivative may be, for example, a crushed freeze-dried Amorphophallus saccharinus containing the sugar derivative, a crushed sun-dried Amorphophallus saccharinus containing the sugar derivative, or a crushed hot-air-dried Amorphophallus saccharinus containing the sugar derivative, or may be one or more of these. The crushed dried A. saccharinum containing the sugar derivatives can be prepared by any conventional method. For example, the crushed freeze-dried A. saccharinum containing the sugar derivatives can be prepared by freezing the A. saccharinum while it still contains water in a low-temperature freezer, removing the water contained within the algae by vacuum freeze-drying to obtain freeze-dried A. saccharinum, and crushing it (e.g., by grinding it with a mortar and pestle, or by powdering it with a grinding device). The crushed A. saccharinum can then be suspended in an appropriate solvent or solution.

[0024] Furthermore, the sugar derivative may be in a form contained in an extract of Aphanothece sacrum (sometimes referred to as "Aphanothece sacrum extract" in the present disclosure), as long as it exhibits an anti-allergic effect. Therefore, the present disclosure can also provide an anti-allergic oral composition containing an extract of Aphanothece sacrum containing the sugar derivative. The Aphanothece saccharin extract containing the sugar derivative can be prepared, for example, by the method described above.

[0025] The antiallergic oral composition of this embodiment may contain the polysaccharide of the present disclosure alone, or may contain, as another component, a component that is suitable for oral administration and that exhibits an antiallergic effect when in the form of an oral composition. When the composition of this embodiment contains such another component, the composition of this embodiment may be a mixture of the polysaccharide of the present disclosure and such another component, and these components may be homogeneous or heterogeneous.

[0026] The subject to ingest the composition of this embodiment is a mammal. Mammals include humans and non-human mammals. Non-human mammals include, for example, dogs. Examples include pet animals and livestock animals, and more specifically, examples include cows, horses, goats, sheep, pigs, monkeys, dogs, cats, rats, mice, hamsters, and guinea pigs.

[0027] The composition of this embodiment has an antiallergic effect. That is, for example, when a subject ingests the composition of this embodiment, an antiallergic effect is exhibited that is not exhibited when a placebo is ingested (or before ingesting the composition of this embodiment, or when not ingesting the composition of this embodiment). Note that when a subject ingests a placebo (or before ingesting the composition of this embodiment, or when not ingesting the composition of this embodiment), the antiallergic effect is usually not exhibited, but it is acceptable for the effect to be exhibited as long as it is exhibited more effectively when the composition of this embodiment is ingested. For these reasons, the polysaccharides of the present disclosure can be used as an active ingredient in an antiallergic oral composition. In other words, the composition of this embodiment can contain the polysaccharides of the present disclosure as an active ingredient.

[0028] The allergy in this embodiment may be type I allergy, type II allergy, type III allergy, or type IV allergy, and may be one or more of these.

[0029] The type I allergy may be atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, urticaria, allergic conjunctivitis, allergic dermatitis, or anaphylactic shock, or may be one or more of these. That is, the type I allergy in this embodiment may be a specific embodiment of type I allergy or a specific symptom caused by type I allergy. Allergens that cause type I allergies may be, for example, house dust, dirt, pollen, mites, mold, bacteria, etc., and may be one or more of these.

[0030] The type II allergy may be Hashimoto's disease (chronic thyroiditis), Goodpasture's syndrome, or organ transplant rejection, or may be one or more of these. That is, the type II allergy in this embodiment may be a specific embodiment of type II allergy, or a specific symptom caused by type II allergy.

[0031] The type III allergy may be systemic vasculitis, cryoglobulinemia, or glomerulonephritis, or may be one or more of these. That is, the type III allergy in this embodiment may be a specific embodiment of type III allergy, or a specific symptom caused by type III allergy.

[0032] The type IV allergy may be contact dermatitis or organ transplant rejection, or may be one or more of these. That is, the type IV allergy in this embodiment may be a specific embodiment of type IV allergy, or a specific symptom caused by type IV allergy, etc.

[0033] The composition of this embodiment is also effective in suppressing IgE production, suppressing a decrease in rectal temperature, or suppressing the release of chemical mediators. Therefore, the composition of this embodiment is preferably used for suppressing IgE production, suppressing a decrease in rectal temperature, suppressing the release of chemical mediators, etc. The composition may be used for one or more of these purposes.

[0034] The chemical mediator may be histamine, β-hexosaminidase, leukotriene, prostaglandin, protease, serotonin, peptide, cytokine, or the like, and may be one or more of these. For example, when the allergy in this embodiment is type I allergy, the chemical mediator may be a chemical mediator released from cells (e.g., mast cells, basophils, etc., and may be one or more) by degranulation, Specifically, the substance may be histamine, β-hexosaminidase, leukotriene, prostaglandin, protease, serotonin, peptide, cytokine, or the like, and one or more of these may be used. The chemical mediator may be a chemical mediator in a part of the subject's body (e.g., tissue; organ such as ear, skin, or organ; organ system, etc.), blood (preferably serum), etc., and may be one or more.

[0035] The composition of this embodiment has an antiallergic effect and can therefore be used to prevent or treat (including alleviate) allergies.

[0036] The composition of this embodiment may be used for non-therapeutic purposes or for therapeutic purposes. "Non-therapeutic purposes" are purposes that do not include medical procedures. In other words, purposes that do not include treatment of a subject through therapy. Examples include uses for maintaining or promoting health.

[0037] When the composition of this embodiment is used for non-therapeutic purposes, it can be administered to healthy individuals. A "healthy individual" may be an individual who does not have an allergy, an individual who is concerned about an allergy, or an individual who is prone to an allergy at the time of taking the composition of this embodiment. When the composition of this embodiment is used for non-therapeutic purposes, it becomes possible to prevent allergies in healthy individuals.

[0038] When the composition of this embodiment is used for therapeutic purposes, it can be used in unhealthy individuals. The "unhealthy individual" may be a person who is allergic at the time of taking the composition of this embodiment. When the composition of this embodiment is used for therapeutic purposes, it becomes possible to treat allergies in unhealthy individuals.

[0039] The content of the polysaccharides of the present disclosure in the composition of this embodiment is appropriately determined depending on the embodiment of the composition, but is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.01% by mass or more, in total, and is preferably 100% by mass or less, more preferably 75% by mass or less, even more preferably 50% by mass or less. Consistent combinations thereof are also acceptable. For example, the total amount is 0.0001% by mass to 100% by mass, 0.001% by mass to 75% by mass, 0.01% by mass to 50% by mass, etc.

[0040] The intake amount (effective amount) of the composition of this embodiment is appropriately determined depending on the form of the composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as the anti-allergic effect is exhibited in the subject who ingests it. The total amount of the polysaccharides of the present disclosure per kg body weight per day is preferably 10 mg or more, more preferably 20 mg or more, even more preferably 30 mg or more, and preferably 70 mg or less, more preferably 60 mg or less, and even more preferably 50 mg or less. A compatible combination thereof is also acceptable. For example, the total amount per kg body weight per day is 10 mg to 70 mg, 20 mg to 60 mg, 30 mg to 50 mg, etc.

[0041] The composition of this embodiment can be taken once a day or in divided doses. It may also be taken once every few days or weeks, but is preferably taken daily. For example, daily intake for 4 days or more, daily intake for 5 days or more, daily intake for 7 days or more, daily intake for 14 days or more, daily intake for 21 days or more, daily intake for 28 days or more, etc.

[0042] The composition of this embodiment is intended to be taken orally.

[0043] The composition of this embodiment contains the polysaccharide of the present disclosure and has an anti-allergic effect. Although not particularly limited, it may be, for example, a crushed dried product of Aphanothece sacrum or an extract of Aphanothece sacrum. That is, the present disclosure can provide an oral anti-allergic crushed dried Aphanothece sacrum extract containing the polysaccharide of the present disclosure, and an oral anti-allergic Aphanothece sacrum extract containing the polysaccharide of the present disclosure. The description of the crushed dried Aphanothece sacrum and the Aphanothece sacrum extract has been given above.

[0044] The composition of this embodiment can be used as, for example, a food or drink composition, a feed composition, a pharmaceutical composition, etc. For example, it can be provided as an "antiallergic food and drink composition (antiallergic oral food and drink composition) containing the polysaccharide of the present disclosure." Hereinafter, this may be referred to as the "food and drink composition of this embodiment." The "food and drink composition of this embodiment" includes supplements. It can also be provided as an "antiallergic feed composition (antiallergic oral feed composition) containing the polysaccharide of the present disclosure." Hereinafter, this may be referred to as the "feed composition of this embodiment." It can also be provided as an "antiallergic oral pharmaceutical composition containing the polysaccharide of the present disclosure." Hereinafter, this may be referred to as the "pharmaceutical composition of this embodiment."

[0045] <Food and drink composition of this embodiment> When the polysaccharides of the present disclosure are used as ingredients for the food and beverage composition of this embodiment, they can be used as general food and beverage compositions, as well as foods for specified health uses, nutritional supplements, functional foods, foods for the sick, food additives, etc. (These also include beverages.) The food and beverage composition may be, for example, formed into an edible form, such as granules, particles, tablets, capsules, or pastes, using conventional means after adding appropriate auxiliary agents, and then provided for consumption. It may also be added to various foods, such as processed meat foods such as ham and sausage, processed seafood foods such as kamaboko and chikuwa, bread, confectionery, butter, powdered milk, and fermented dairy products, or to beverages such as water, fruit juice, milk, and soft drinks.

[0046] The food and beverage composition of this embodiment can be administered to any subject, including healthy and unhealthy individuals, but in the case of a food and beverage composition labeled for a specific use (particularly a health use) or function, it may be used for the above-mentioned non-therapeutic purposes.

[0047] The food and drink composition of this embodiment can contain water, proteins, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, and the like as main components. Examples of proteins include animal and plant proteins such as whole milk powder, skim milk powder, partially skim milk powder, casein, soy protein, egg protein, and meat protein, as well as hydrolysates of these proteins, and butter. Examples of carbohydrates include sugars, modified starch (dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), and dietary fiber. Examples of lipids include vegetable oils and fats such as lard, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, hydrogenated oils, and interesterified oils. Examples of vitamins include vitamin A, carotenes, B vitamins, vitamin C, D vitamins, vitamin E, K vitamins, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey minerals. Examples of organic acids include malic acid, citric acid, lactic acid, and tartaric acid.

[0048] The food and drink composition of this embodiment can be produced according to a conventional method. The food and beverage composition of this embodiment can be produced by adding the polysaccharide of the present disclosure to ordinary raw materials. Except for the addition of the polysaccharide of the present disclosure, the food and beverage composition can be produced in the same manner as ordinary food and beverage compositions. The amount, method, and timing of the polysaccharide of the present disclosure can be selected as appropriate. The food and beverage composition of this embodiment can be packaged in an appropriate container such as a bottle, bag, can, box, or pack, as needed.

[0049] The content of the polysaccharides of the present disclosure in the food and beverage composition of this embodiment is appropriately determined depending on the embodiment of the food and beverage composition, but is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.01% by mass or more in total, and is preferably 100% by mass or less, more preferably 75% by mass or less, even more preferably 50% by mass or less in total. A consistent combination thereof is also acceptable. For example, the total amount is 0.0001% by mass to 100% by mass, 0.001% by mass to 75% by mass, 0.01% by mass to 50% by mass, etc.

[0050] The intake amount (effective amount) of the food and beverage composition of this embodiment is appropriately determined depending on the form of the food and beverage composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as the anti-allergic effect is exhibited in the subject who ingests it. The total amount of the polysaccharides of the present disclosure per kg body weight per day is preferably 10 mg or more, more preferably 20 mg or more, even more preferably 30 mg or more, and preferably 70 mg or less, more preferably 60 mg or less, and even more preferably 50 mg or less. A compatible combination thereof is also acceptable. For example, the total amount per kg body weight per day is 10 mg to 70 mg, 20 mg to 60 mg, 30 mg to 50 mg, etc.

[0051] The food and drink composition of this embodiment can be taken once a day or in divided doses. It may also be taken once every few days or weeks, but is preferably taken daily. For example, daily intake for 4 days or more, daily intake for 5 days or more, daily intake for 7 days or more, daily intake for 14 days or more, daily intake for 21 days or more, daily intake for 28 days or more, etc.

[0052] When the subject is a mammal other than a human, the polysaccharides of the present disclosure can be used as a material for the feed composition of this embodiment. In this case, the feed ingredients and the polysaccharides of the present disclosure can be appropriately mixed depending on the type, developmental stage, region, and other rearing environments of the mammal. Examples of feed ingredients include grains or processed grains (corn, milo, barley, etc.), bran (bran, rice bran, corn gluten feed, etc.), vegetable oil cakes (soybean oil cake, sesame oil cake, cottonseed oil cake, etc.), animal ingredients (skim milk powder, fish meal, meat and bone meal, etc.), minerals (calcium carbonate, calcium phosphate, salt, silicic acid anhydride, etc.), vitamins, amino acids, yeasts such as brewer's yeast, and finely powdered inorganic substances (crystalline cellulose, talc, silica, etc.).

[0053] The feed may contain, in addition to the feed raw materials, feed additives commonly used in compound feed, such as excipients, bulking agents, binders, thickeners, emulsifiers, coloring agents, flavorings, food additives, and seasonings, as well as other ingredients (antibiotics, disinfectants, anthelmintics, preservatives, etc.) as desired.

[0054] The form of the feed is not particularly limited, and examples thereof include powder, granules, paste, pellets, capsules (hard capsules, soft capsules), tablets, etc., and the feed may be used as pet food for companion animals or feed for laboratory animals.

[0055] The content of the polysaccharides of the present disclosure in the feed composition of this embodiment is appropriately determined depending on the embodiment of the feed composition, but is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.01% by mass or more in total, and is preferably 100% by mass or less, more preferably 75% by mass or less, even more preferably 50% by mass or less in total. Consistent combinations thereof are also acceptable. For example, the total amount may be 0.0001% by mass to 100% by mass, 0.001% by mass to 75% by mass, For example, it is 0.01% by mass to 50% by mass.

[0056] The dosage (effective amount) of the feed composition of this embodiment is determined appropriately depending on the form of the feed composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as the anti-allergic effect is exerted in the subject to which it is administered. The total amount of the polysaccharides of the present disclosure per kg body weight per day is preferably 10 mg or more, more preferably 20 mg or more, even more preferably 30 mg or more, and preferably 70 mg or less, more preferably 60 mg or less, and even more preferably 50 mg or less. Consistent combinations thereof are also acceptable. For example, the total amount per kg body weight per day is 10 mg to 70 mg, 20 mg to 60 mg, 30 mg to 50 mg, etc.

[0057] The feed composition of this embodiment can be administered once a day or in divided doses. It may also be administered once every few days or weeks, but daily administration is preferred. Examples include daily administration for 4 or more days, daily administration for 5 or more days, daily administration for 7 or more days, daily administration for 14 or more days, daily administration for 21 or more days, and daily administration for 28 or more days.

[0058] When the polysaccharide of the present disclosure is used as a material for the pharmaceutical composition of this embodiment, the polysaccharide of the present disclosure, which is the active ingredient, can be mixed with a solid or liquid non-toxic pharmaceutical carrier suitable for oral administration and administered in the form of a conventional pharmaceutical preparation. Such formulations include, for example, solid formulations such as tablets, granules, powders, and capsules; liquid formulations such as solutions, suspensions, and emulsions; and freeze-dried formulations, and these formulations can be prepared by conventional pharmaceutical methods. Examples of the non-toxic pharmaceutical carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glycerides, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid esters, amino acids, gelatin, albumin, water, physiological saline, etc. Furthermore, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, and isotonic agents can also be added as needed.

[0059] The pharmaceutical composition of this embodiment is administered to unhealthy subjects and used for the above-mentioned therapeutic purposes.

[0060] The content of the polysaccharide of the present disclosure in the pharmaceutical composition of this embodiment is appropriately determined depending on the embodiment of the pharmaceutical composition, but is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, even more preferably 0.01% by mass or more, in total, and is preferably 100% by mass or less, more preferably 75% by mass or less, even more preferably 50% by mass or less. Consistent combinations thereof are also acceptable. For example, the total amount is 0.0001% by mass to 100% by mass, 0.001% by mass to 75% by mass, 0.01% by mass to 50% by mass, etc.

[0061] The dosage (effective amount) of the pharmaceutical composition of this embodiment is appropriately determined depending on the form of the pharmaceutical composition, the method of use, the subject, the subject's age, sex, and other conditions, but is not particularly limited as long as the anti-allergic effect is exhibited in the subject to which it is administered. The total amount of the polysaccharide of the present disclosure per kg body weight per day is preferably 10 mg or more, more preferably 20 mg or more, even more preferably 30 mg or more, and preferably 70 mg or less, more preferably 60 mg or less, and even more preferably 50 mg or less. A compatible combination thereof is also acceptable. For example, the total amount per kg body weight per day is 10 mg to 70 mg, 20 mg to 60 mg, 30 mg to 50 mg, etc.

[0062] The pharmaceutical composition of this embodiment can be administered once a day or in divided doses. It may also be administered once every few days or weeks, but is preferably administered daily. For example, it may be administered once every four days. Daily administration for 5 days or more, daily administration for 7 days or more, daily administration for 14 days or more, daily administration for 21 days or more, daily administration for 28 days or more, etc.

[0063] In addition to the above-mentioned aspects, the present disclosure can also provide the following aspects, for example. Use of the polysaccharides of the present disclosure for the manufacture of an oral antiallergic composition. The polysaccharide of the present disclosure for oral use in the prevention or treatment of allergies. A composition containing the polysaccharide of the present disclosure for oral use in the prevention or treatment of allergies. 10. The oral non-therapeutic use of the polysaccharides of the present disclosure, wherein the polysaccharides of the present disclosure are used for the prevention of allergies. 10. The oral non-therapeutic use of a composition containing the polysaccharide of the present disclosure, wherein the composition is used for the prevention of allergies. A method for preventing or treating allergies, comprising orally administering to a subject in need thereof a prophylactically or therapeutically effective amount of a polysaccharide of the present disclosure. A method for preventing or treating allergies, comprising orally administering a prophylactically or therapeutically effective amount of a composition containing the polysaccharide of the present disclosure to a subject in need of prevention or treatment.

[0064] As mentioned above, the composition of this embodiment is also effective in suppressing IgE production, suppressing a decrease in rectal temperature, and suppressing the release of chemical mediators. For this reason, the present disclosure can also provide, as another aspect, an "oral composition for suppressing IgE production, containing the polysaccharide of the present disclosure." The present disclosure can also provide, as another aspect, an "oral composition for suppressing rectal temperature reduction, containing the polysaccharide of the present disclosure." The present disclosure can also provide, as another aspect, an "oral composition for suppressing chemical mediator release, containing the polysaccharide of the present disclosure." In either embodiment, the explanation of the "antiallergic oral composition containing the polysaccharide of the present disclosure" according to the previous embodiment is incorporated herein by reference.

[0065] Furthermore, one of the uses of anti-allergy, suppression of IgE production, suppression of rectal temperature decrease, and suppression of chemical mediator release can be used as a preferred embodiment for the other uses. For example, an "oral composition for suppressing IgE production containing the polysaccharide of the present disclosure" is preferably used for anti-allergy, suppression of rectal temperature decrease, or suppression of chemical mediator release, and these uses may be one or more. [Example]

[0066] The present invention will be specifically described below using examples, but the present invention is not limited to these examples. Each of the following examples and comparative examples is based on the results of n=3 to 5.

[0067] <Preparation of polysaccharides of the present disclosure> A solution containing the polysaccharide of the present disclosure was prepared according to a conventional method. Specifically, the procedure is as follows: an appropriate amount of Aphanothece sacrum was frozen, thawed, and washed with water to remove water-soluble pigments. The fat-soluble pigments were then removed by stirring overnight in ethanol. After separating the ethanol from the Aphanothece sacrum from which the pigments had been removed, the Aphanothece sacrum was heated to 40°C in a 0.4-0.6N aqueous sodium hydroxide solution and dissolved with stirring for approximately 5 hours. The aqueous solution of the sugar derivative extracted by this procedure was neutralized with concentrated hydrochloric acid, and this concentrated solution was poured into 100% isopropanol with stirring to precipitate a gel-like sugar derivative. The sugar derivative was then dried with hot air to obtain a fibrous sugar derivative. 995 parts by mass of purified water was added to 5.0 parts by mass of the dried Aphanothece sacrum-derived polysaccharide obtained in this manner, and the mixture was stirred at 75-80°C for at least 8 hours using a stirrer and a stirrer to prepare a solution (0.5% by mass) containing the polysaccharide of the present disclosure. The concentration of this solution was adjusted for use in the following tests. In Example 5-3, a suspension containing crushed freeze-dried A. sacrum was used. This suspension was prepared according to a conventional method. Specifically, A. sacrum was frozen in a low-temperature freezer at −45°C while still containing water. A vacuum freeze-dryer (FD-20BM, manufactured by Nippon Techno Service Co., Ltd.) was used to remove the water contained within the algae to obtain freeze-dried A. sacrum. This was then ground with a mortar and pestle, and 600 μg of the ground A. sacrum was suspended in 100 μL of 10% carboxymethylcellulose (CMC) solution to prepare a suspension containing crushed freeze-dried A. sacrum.

[0068] <Passive Cutaneous Anaphylaxis (PCA) Reaction Test (Part 1)> (Example 1-1) BALB / c mice (5 weeks old, female) that had been pre-bred for 6 days were orally administered 100 μl of the polysaccharide of the present disclosure (300 μg) adjusted to a concentration of 0.3% by mass with sterile water per day using a stomach tube for 4 consecutive days. Then, on the day after the final administration, 100 μl of anti-trinitrophenyl (TNP) IgE (anti-TNP-IgE antibody, BD Biosciences) (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered into the tail vein of the mice to sensitize them. Thirty minutes later, 10 μl of a 1.6% picryl chloride solution (picryl chloride dissolved in a 1:1 mixture of acetone and olive oil) containing the antigen (picryl chloride) was applied to the ear. Before and 2 hours after application, ear thickness was measured using a micrometer (dial thickness gauge, Model G-1A, Ozaki Seisakusho (PEACOCK)). The difference in ear thickness (μm) was calculated by subtracting the ear thickness (μm) before application from the ear thickness (μm) 2 hours after application. A larger difference in ear thickness indicates a more severe allergic reaction.

[0069] (Comparative Example 1-1) After 6 days of preliminary rearing, BALB / c mice (5 weeks old, female) were orally administered 100 μl of sterilized water per day for 4 consecutive days using a gastric tube. Subsequently, on the day after the final administration, 100 μl of anti-TNP-IgE antibody (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered into the tail vein of the mice for sensitization. Subsequently, unlike Example 1-1, without applying a solution containing an antigen (picryl chloride) to the ear, the auricle thickness was measured in the same manner as in Example 1-1 at the time corresponding to before application and 2 hours after application in Example 1-1, and the "auricle thickness difference (μm)" was calculated.

[0070] (Comparative Example 1-2) The same procedure as in Example 1-1 was followed, except that sterilized water was used instead of the polysaccharide of the present disclosure.

[0071] (Results) The results are shown in Figure 1. * in the figure indicates a significant difference with a risk rate of 0.05% for Comparative Example 1-2.

[0072] <IgE Production Inhibition Evaluation Test> (Example 2-1) After 1 week of preliminary rearing, BALB / c mice (5 weeks old, female) were orally administered 100 μl of the polysaccharide of the present disclosure (300 μg) adjusted to a concentration of 0.3% by mass with sterilized water per day for 28 consecutive days using a gastric tube. 300 μl of ovalbumin (OVA) solution (PBS solution containing 10 μg of OVA and 1 mg of Al(OH)3 as an adjuvant) was administered intraperitoneally (Day 1), and then on Day 5, Day 10, and Day 1, 300 μl of OVA solution (PBS solution containing 10 μg of OVA and 0.5 mg of Al(OH)3 as an adjuvant) was administered intraperitoneally. Blood was collected from the tail vein on the day before (Day 0, Day 4, Day 9, Day 14) each day the OVA solution was administered (Day 1, Day 5, Day 10, Day 15). Serum was recovered from the collected blood, and the IgE level was measured using the serum by ELISA.

[0073] (Comparative Example 2-1) After one week of preliminary breeding, BALB / c mice (5 weeks old, female) were orally administered 100 μl of sterilized water per day using a stomach tube for 28 consecutive days. 300 μl of adjuvant solution (PBS solution containing 1 mg of Al(OH)3) was administered intraperitoneally (Day 1), followed by 300 μl of adjuvant solution (PBS solution containing 0.5 mg of Al(OH)3) administered intraperitoneally on Days 5, 10, and 15. Blood was collected from the tail vein on the day before (Day 0, Day 4, Day 9, Day 14) each day the adjuvant solution was administered (Day 1, Day 5, Day 10, Day 15). Serum was collected from the collected blood, and the IgE level was measured using the serum by ELISA.

[0074] (Comparative Example 2-2) The same procedure was followed as in Example 2-1, except that sterilized water was used instead of the polysaccharide of the present disclosure.

[0075] (result) The results are shown in Figure 2. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0076] <Rectal temperature drop suppression evaluation test> Example 3-1 After one week of preliminary breeding, BALB / c mice (5 weeks old, female) were orally administered 100 μl of the polysaccharide of the present disclosure (300 μg) adjusted to a concentration of 3 μg / μl with sterile water per day using a stomach tube for 28 consecutive days. 300 μl of ovalbumin (OVA) solution (PBS solution containing 10 μg of OVA and 1 mg of Al(OH)3 as an adjuvant) was administered intraperitoneally (Day 1), followed by 300 μl of OVA solution (PBS solution containing 10 μg of OVA and 0.5 mg of Al(OH)3 as an adjuvant) administered intraperitoneally on Days 5, 10, and 15. Then, on Day 20, the final day of administration, 100 μl of OVA solution (PBS solution containing 5 μg of OVA) was administered via the tail vein for antigen exposure, and rectal temperature was measured every 10 minutes for 90 minutes.

[0077] (Comparative Example 3-1) After one week of preliminary breeding, BALB / c mice (5 weeks old, female) were orally administered 100 μl of sterilized water per day using a stomach tube for 28 consecutive days. 300 μl of adjuvant solution (PBS solution containing 1 mg of Al(OH)3) was administered intraperitoneally (Day 1), followed by 300 μl of adjuvant solution (PBS solution containing 0.5 mg of Al(OH)3) administered intraperitoneally on Days 5, 10, and 15. Then, on Day 20, the final day of administration, 100 μl of adjuvant solution (PBS solution containing 1 mg of Al(OH) 3 ) was administered via the tail vein, and rectal temperature was measured every 10 minutes for 90 minutes.

[0078] (Comparative Example 3-2) The same procedure was followed as in Example 3-1, except that sterilized water was used instead of the polysaccharide of the present disclosure.

[0079] (result) The results are shown in Figure 3. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0080] Passive Cutaneous Anaphylaxis (PCA) Reaction Test (Part 2) )> (Example 4-1) After four days of preliminary breeding, BALB / c mice (4 weeks old, female) were orally administered 100 μl of the polysaccharide of the present disclosure (300 μg) adjusted to a concentration of 0.3% by mass with sterile water per day using a stomach tube for five consecutive days. As in Example 1-1, 100 μl of anti-TNP-IgE antibody (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered intravenously to the tail vein of each mouse on the day following the final administration to sensitize it. Then, 10 μl of a solution containing the antigen (picryl chloride) was applied to the ear. Before and 2 hours after application, ear thickness was measured using a micrometer (dial thickness gauge, Model G-1A, Ozaki Seisakusho (PEACOCK)). The difference in ear thickness (μm) was calculated by subtracting the ear thickness (μm) before application from the ear thickness (μm) 2 hours after application.

[0081] (Example 4-2) The same procedure as in Example 4-1 was carried out except that 100 μl of the polysaccharide of the present disclosure (100 μg) adjusted to a concentration of 0.1% by mass with sterilized water was used.

[0082] (Comparative Example 4-1) After 4 days of preliminary breeding, BALB / c mice (4-week-old, female) were orally administered 100 μl of sterilized water per day using a stomach tube for 5 consecutive days. Then, on the day after the final administration, 100 μl of anti-TNP-IgE antibody (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered into the tail vein of the mice to sensitize them. Thereafter, unlike in Example 4-1, a solution containing the antigen (picryl chloride) was not applied to the ear, and the ear thickness was measured in the same manner as in Example 4-1 at times corresponding to before application and 2 hours after application in Example 4-1, and the "difference in ear thickness (μm)" was calculated.

[0083] (Comparative Example 4-2) The same procedure was followed as in Example 4-1, except that sterilized water was used instead of the polysaccharide of the present disclosure.

[0084] (result) The results are shown in Figure 4. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0085] <Passive Cutaneous Anaphylaxis (PCA) Reaction Test (Part 3)> (Example 5-1) After 5 days of preliminary breeding, BALB / c mice (4 weeks old, female) were orally administered 100 μl of the polysaccharide of the present disclosure (300 μg) adjusted to a concentration of 0.3% by mass with sterile water per day using a stomach tube for 4 consecutive days. As in Example 1-1, 100 μl of anti-TNP-IgE antibody (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered intravenously to the tail vein of each mouse on the day following the final administration to sensitize it. Then, 10 μl of a solution containing the antigen (picryl chloride) was applied to the ear. Before and 2 hours after application, ear thickness was measured using a micrometer (dial thickness gauge, Model G-1A, Ozaki Seisakusho (PEACOCK)). The difference in ear thickness (μm) was calculated by subtracting the ear thickness (μm) before application from the ear thickness (μm) 2 hours after application.

[0086] (Example 5-2) The same procedure as in Example 5-1 was carried out except that 100 μl of the polysaccharide of the present disclosure (30 μg) adjusted to a concentration of 0.03% by mass with sterilized water was used.

[0087] (Example 5-3) The same procedure was followed as in Example 5-1, except that a suspension (100 μL) containing crushed freeze-dried Aphanothece sacrum (600 μg) prepared above was used instead of the solution containing the polysaccharide of the present disclosure.

[0088] (Comparative Example 5-1) After 5 days of preliminary breeding, BALB / c mice (4-week-old, female) were orally administered 100 μl of sterilized water per day using a stomach tube for 4 consecutive days. Then, on the day after the final administration, 100 μl of anti-TNP-IgE antibody (2 μg) dissolved in phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) was administered into the tail vein of the mice to sensitize them. Thereafter, unlike in Example 5-1, a solution containing the antigen (picryl chloride) was not applied to the ear, and the ear thickness was measured in the same manner as in Example 5-1 at times corresponding to before application and 2 hours after application in Example 5-1, and the "difference in ear thickness (μm)" was calculated.

[0089] (Comparative Example 5-2) The same procedure was followed as in Example 5-1, except that sterilized water was used instead of the polysaccharide of the present disclosure.

[0090] (result) The results are shown in Figure 5. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0091] <Histamine release inhibition evaluation test (part 1)> Example 6-1 The amount of histamine in the ears of the mice after the test in Example 5-1 was measured using a Histamine ELISA Kit (Abnova, Catalog No. KA3844) according to the manufacturer's instructions. To measure histamine levels in the ear, ear tissues obtained from mice at autopsy and stored at -80°C were flash-frozen in liquid nitrogen, homogenized, and protein extracted with RIPA buffer containing protease inhibitors. Protein was quantified by the Lowry method, and the amount of histamine (ng) per μg of protein was calculated from the amount of histamine in the protein extract and the amount of protein.

[0092] (Comparative Example 6-1) The amount of histamine in the ears of the mice after the test in Comparative Example 5-1 was measured in the same manner as in Example 6-1.

[0093] (Comparative Example 6-2) The amount of histamine in the ears of the mice after the test in Comparative Example 5-2 was measured in the same manner as in Example 6-1.

[0094] (result) The results are shown in Figure 6. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0095] <Histamine release inhibition evaluation test (part 2)> Example 7-1 After the test in Example 5-1, the amount of histamine in the serum of the mice was measured using a Histamine ELISA Kit (Abnova, Catalog No. KA3844) according to the manufacturer's instructions. To measure the amount of histamine in the serum, blood collected from the mice was centrifuged to obtain serum, which was then diluted and used to measure the amount of histamine in the serum.

[0096] (Comparative Example 7-1) After the test in Comparative Example 5-1, the amount of histamine in the serum of the mice was measured in the same manner as in Example 7-1.

[0097] (Comparative Example 7-2) After the test in Comparative Example 5-2, the amount of histamine in the serum of the mice was measured in the same manner as in Example 7-1.

[0098] (result) The results are shown in Figure 7. There was a significant difference between the results marked with different signs in the figure (Tukey-Kramer test, p < 0.05).

[0099] <β-hexosaminidase release inhibition evaluation test (part 1)> (Reference example 1-1) HT-29 cells, a human colon adenocarcinoma cell line, were plated at 1.0 × 10 in the insert of a 12-well Transwell plate (Corning Costar, product number #3401). 5The cells were seeded at 4.0 × 10 cells / well and cultured until differentiation. Then, 4.0 × 10 cells of the rat basophilic leukemia cell line, RBL-2H3, were seeded on the basolateral side. 5 The cells were seeded at 100 cells / well and cultured overnight. After washing the cells with PBS, the polysaccharide of the present disclosure diluted with medium (final concentration: 0.75 mg / ml) was added to the apical side, and the cells were incubated for 4 hours. Thereafter, anti-TNP-IgE antibody was added to the basolateral side to a final concentration of 200 ng / mL, and the mixture was further incubated for 2 hours. The inserts were then removed, and the cells were washed with PBS. Degranulation of RBL-2H3 cells was then induced by adding TNP-BSA (20 ng / mL) dissolved in Shiraganian buffer (119 mM NaCl, 40 mM NaOH, 5 mM KCl, 1 mM CaCl, 0.4 mM MgCl, 25 mM PIPES, 5.6 mM glucose, 0.1% BSA, pH 7.2) and incubating for 30 minutes. Thereafter, the plate was placed on ice for 10 minutes to stop the degranulation reaction. 50 μl of the culture supernatant was transferred to a 96-well plate, and the same volume of substrate solution (2 mM p-nitrophenyl-N-acetyl-β-D-glucosaminide dissolved in 0.2 M citrate buffer, pH 4.5) was added and incubated for 1 hour. Thereafter, 100 μl of a stop solution (0.2 M glycine-NaOH, pH 13.0) was added, and the absorbance (wavelength 405 nm) was measured using a microplate reader.

[0100] (Reference example 1-2) The same procedure as in Reference Example 1-1 was carried out, except that the final concentration of the polysaccharide of the present disclosure was 1.5 mg / ml.

[0101] (Reference example 1-3) HT-29 cells (1.0 × 10 cells) were cultured in a 12-well Transwell plate insert (Corning Costar, product number #3401). 5The cells were seeded at 4.0 × 10 cells / well and cultured until differentiation. 5 The cells were seeded at 100 cells / well and cultured overnight. After washing the cells with PBS, the same volume of medium containing no polysaccharide of the present disclosure was added to the apical side instead of the polysaccharide of the present disclosure used in Reference Example 1-1, and the cells were incubated for 4 hours. Thereafter, instead of the anti-TNP-IgE antibody used in Reference Example 1-1, the same amount of medium was added to the basolateral side, and the mixture was further incubated for 2 hours. Thereafter, the same procedure as in Reference Example 1-1 was carried out.

[0102] (Reference example 1-4) Except that the polysaccharides of the present disclosure were replaced with the same amount of medium without the polysaccharides of the present disclosure. The same procedure was followed as in Reference Example 1-1.

[0103] (result) The results are shown in Figure 8. The results of Reference Examples 1-4 are used as the standard. There was a significant difference between the results marked with different signs in the figure (test by the Tukey-Kramer method, p < 0.05).

[0104] <β-hexosaminidase release inhibition evaluation test (part 2)> (Reference example 2-1) 4 × 10 RBL-2H3 cells were cultured in a 96-well plate. 4 The cells were seeded at 100 cells / well and incubated at 37°C overnight. After washing the cells with PBS, the polysaccharide of the present disclosure diluted with the medium (final concentration: 0.5 mg / ml) was added and the cells were incubated for 4 hours. Thereafter, anti-TNP-IgE antibody was added to a final concentration of 200 ng / ml, and the mixture was further incubated for 2 hours. The cells were then washed with PBS, and degranulation was induced in RBL-2H3 cells by adding TNP-BSA (20 ng / mL) dissolved in Shiraganian buffer (119 mM NaCl, 40 mM NaOH, 5 mM KCl, 1 mM CaCl, 0.4 mM MgCl, 25 mM PIPES, 5.6 mM glucose, 0.1% BSA, pH 7.2) and incubating for 30 minutes. Thereafter, the plate was left standing on ice for 10 minutes to stop the degranulation reaction. 50 μl of the culture supernatant was transferred to a 96-well plate, and the same volume of substrate solution (2 mM p-nitrophenyl-N-acetyl-β-D-glucosaminide dissolved in 0.2 M citrate buffer, pH 4.5) was added and incubated for 1 hour. Thereafter, 100 μl of a stop solution (0.2 M glycine-NaOH, pH 13.0) was added, and the absorbance (wavelength 405 nm) was measured using a microplate reader.

[0105] (Reference example 2-2) The same procedure as in Reference Example 2-1 was carried out, except that the final concentration of the polysaccharide of the present disclosure was 0.3 mg / ml.

[0106] (Reference example 2-3) The same procedure as in Reference Example 2-1 was carried out, except that the final concentration of the polysaccharide of the present disclosure was 0.15 mg / ml.

[0107] (Reference example 2-4) The same procedure as in Reference Example 2-1 was carried out, except that the final concentration of the polysaccharide of the present disclosure was set to 0.075 mg / ml.

[0108] (Reference example 2-5) 4 × 10 RBL-2H3 cells were cultured in a 96-well plate. 4 The cells were seeded at 100 cells / well and incubated at 37°C overnight. After washing the cells with PBS, the same volume of medium containing no polysaccharide of the present disclosure was added in place of the polysaccharide of the present disclosure used in Reference Example 2-1, and the cells were incubated for 4 hours. Thereafter, the same amount of medium was added in place of the anti-TNP-IgE antibody used in Reference Example 2-1, and the mixture was further incubated for 2 hours. Thereafter, the same procedure as in Reference Example 2-1 was carried out.

[0109] (Reference example 2-6) The same procedure was followed as in Reference Example 2-1, except that the same amount of medium not containing the polysaccharide of the present disclosure was used instead of the polysaccharide of the present disclosure.

[0110] (result) The results are shown in Figure 9. The results of Reference Example 2-6 are used as the standard. There was a significant difference between the results (Tukey-Kramer test, p < 0.05).

[0111] The results of Figures 8 and 9 show that when RBL-2H3 cells are present together with HT-29 cells, which are intestinal epithelial cells, the polysaccharide of the present disclosure inhibits degranulation from RBL-2H3 cells, but does not inhibit it in the absence of HT-29 cells.

[0112] <β-hexosaminidase release inhibition evaluation test (part 3)> (Reference example 3-1) The same procedure as in Reference Example 1-1 was carried out except that Caco2 cells, which are also human colon adenocarcinoma-derived cells, were used instead of HT-29 cells and the final concentration of the polysaccharide of the present disclosure was set to 1.5 mg / ml.

[0113] (Reference example 3-2) The same procedures as in Reference Example 1-3 were carried out, except that Caco2 cells similar to those described above were used instead of HT-29 cells.

[0114] (Reference example 3-3) The same procedures as in Reference Example 1-4 were carried out, except that Caco2 cells similar to those described above were used instead of HT-29 cells.

[0115] (result) The results are shown in Figure 10. The results of Reference Example 3-3 are used as the standard. There was a significant difference between the results marked with different signs in the figure (test by the Tukey-Kramer method, p < 0.05).

Claims

1. An anti-allergic oral composition containing sulfated polysaccharides derived from Aphanothece sacrum.

2. The oral composition of claim 1 , wherein the allergy is a type I allergy.

3. The oral composition according to claim 2, wherein the type I allergy is atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, urticaria, allergic conjunctivitis, allergic dermatitis, or anaphylactic shock.

4. An oral composition for suppressing IgE production, comprising sulfated polysaccharides derived from Aphanothece sacrum.

5. An oral composition for suppressing rectal temperature drop, comprising sulfated polysaccharides derived from Aphanothece sacrum.

6. An oral composition for inhibiting the release of chemical mediators, comprising sulfated polysaccharides derived from Aphanothece sacrum.

7. The oral composition of claim 6, wherein the chemical mediator is histamine, β-hexosaminidase, or leukotriene.

8. The oral composition according to any one of claims 1 to 7, which is a pharmaceutical composition.

9. The oral composition according to any one of claims 1 to 7, which is a food composition.

10. The oral composition according to any one of claims 1 to 7, which is a feed composition.

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