Pharmaceutical composition and food composition for treating allergy

Abstract

To provide a new antiallergic agent.SOLUTION: The problem can be solved by 3-methyl-6-hydroxyisocoumarin 8-O - β - D-glucoside, 7, 8-dihydro-8 - β - D-glucopyranosyloxy-4-methoxy-7-methyl - 5H - furo [2, 3-g] [2] benzopyran-5-one, 2-O-trans-caffeoyl hydroxycitric acid or 2-O-trans-caffeoyl hydroxycitric acid 6' - O-methyl ester.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition and a food composition for treating allergies. [Background technology]

[0002] For example, type I allergies are caused by IgE produced by B cells, a type of lymphocyte. In the case of hay fever, IgE is produced in the body, recognizing and specifically binding to pollen as an allergen (antigen). The IgE then binds to the surface of basophils and mast cells via the IgE receptor (FcεRI). When pollen enters the body, it binds to IgE, stimulating the transmission of signals within basophils and mast cells, resulting in the release of granules containing substances that cause allergic symptoms, such as histamine. This phenomenon is called degranulation. The histamine released extracellularly by degranulation irritates the mucous membranes, causing allergic symptoms. Therefore, inhibiting the release of granules by basophils and mast cells is important for preventing or treating allergic symptoms. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2020-26395 [Patent Document 2] Japanese Patent Application Publication No. 2019-127455 Summary of the Invention [Problem to be solved by the invention]

[0004] For example, Patent Document 1 describes an anti-type I allergy agent containing phlestoic acid, and Patent Document 2 describes an anti-allergy agent containing p-coumaric acid. However, the development of new anti-allergy agents is desired. An object of the present invention is to provide a novel antiallergic agent. [Means for solving the problem]

[0005] The present inventors have conducted extensive research into novel antiallergic agents and have surprisingly found that certain novel compounds exhibit antiallergic activity. The present invention is based on this finding. Therefore, the present invention provides [1] Formula (1): [ka] 3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside represented by [2] Formula (2): [ka] 7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one represented by [3] Formula (3): [ka] (In the formula, R 1 is a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R 2 are 1 to 5, each independently representing a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms, and R 3 , R 4 , R 5 , R 6 , and R 7 are each independently a hydrogen atom, a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms), [4] Formula (5): [ka] 2-O-trans-caffeoylhydroxycitric acid, represented by [5] Formula (6): [ka] 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester represented by [6] A pharmaceutical composition comprising, as an active ingredient, a compound selected from the group consisting of the compounds according to [1] to [5] or a salt thereof. [7] The pharmaceutical composition according to [6], which is for preventing or treating allergies. [8] The pharmaceutical composition according to [7], wherein the allergy is type I allergy. [9] The pharmaceutical composition according to any one of [6] to [8], which suppresses degranulation.

[10] A food composition for suppressing allergic symptoms, comprising, as an active ingredient, a compound selected from the group consisting of the compounds according to [1] to [5] or a salt thereof.

[11] The food composition according to

[10] , wherein the allergic symptoms are type I allergic symptoms; and

[12] The food composition according to

[10] or

[11] , wherein the suppression of allergic symptoms is suppression of degranulation. Regarding. [Effects of the Invention]

[0006] According to the present invention, it is possible to provide a novel pharmaceutical composition, particularly a pharmaceutical composition for preventing or treating allergies. The present invention also provides a food composition for suppressing allergic symptoms. [Brief explanation of the drawings]

[0007] [Figure 1] 1 is a graph showing the inhibitory effect of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside on degranulation of RBL-2H3 cells due to antigen stimulation. [Figure 2] 1 is a graph showing the inhibitory effect of 7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one on antigen-stimulated degranulation of RBL-2H3 cells. [Figure 3]Graph (A) shows the inhibitory effect of 2-O-trans-caffeoylhydroxycitric acid or 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester on the degranulation of RBL-2H3 cells, and graph (B) shows their cytotoxicity. [Figure 4] 1 is a graph showing the inhibitory effect of adenosine on degranulation caused by antigen stimulation. [Figure 5] 1 is a graph showing the inhibitory effect of maltotriose on degranulation caused by antigen stimulation. [Figure 6] 1 is a graph showing the inhibitory effect of DL-malic acid on degranulation caused by antigen stimulation. [Figure 7] 1 is a graph showing the inhibitory effect of L-malic acid on degranulation caused by antigen stimulation. [Figure 8] 1 is a graph showing the inhibitory effect of D-malic acid on degranulation caused by antigen stimulation. DETAILED DESCRIPTION OF THE INVENTION

[0008] [1] Pharmaceutical composition The 3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside of the present invention is represented by the following formula (1): [ka] The compound is a novel compound represented by the formula (hereinafter, sometimes referred to as Compound A), and is preferably (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside.

[0009] The 7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-fro[2,3-g][2]benzopyran-5-one of the present invention is a compound represented by the following formula (2): [ka] The compound is a novel compound represented by the formula (hereinafter, sometimes referred to as Compound B), and is preferably (7S,8R)-7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-fro[2,3-g][2]benzopyran-5-one.

[0010] The compound of the present invention is represented by the following formula (3): [ka] (In the formula, R 1 is a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and R 2 are 1 to 5, each independently representing a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms, and R 3 , R 4 , R 5 , R 6 , and R 7 and each independently represent a hydrogen atom, a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms (hereinafter, sometimes referred to as compound C). Compound C is water-soluble and can be easily dissolved in water. The compound C is preferably represented by the following formula (4): [ka] (In the formula, R 1is a hydrogen atom or an alkyl group having 1 to 6 carbon atoms). The compound C is more preferably 2-O-trans-caffeoyl hydroxycitric acid, which is represented by the following formula (5): [ka] The compound is a novel compound represented by the formula (hereinafter, sometimes referred to as compound C1).

[0011] The compound C is more preferably 2-O-trans-caffeoyl hydroxycitric acid 6'-O-methylester, which is represented by the following formula (6): [ka] The compound is a novel compound represented by the formula (hereinafter, sometimes referred to as compound C2).

[0012] The salt of the compound is not limited to a salt with an inorganic base or an organic base, or a salt with an acid, as long as it is acceptable as a pharmaceutical or food product. Specific examples of salts with inorganic bases or organic bases include salts with inorganic bases, organic bases, or metal alkoxides. The salts can be produced by mixing the compound with an inorganic base, organic base, or metal alkoxide. Inorganic bases capable of forming salts include hydroxides, carbonates, bicarbonates, acetates, or hydrides of alkali metals (e.g., lithium, sodium, or potassium); and hydroxides or hydrides of alkaline earth metals (e.g., magnesium, calcium, or barium). Organic bases capable of forming salts include dimethylamine, triethylamine, piperazine, pyrrolidine, piperidine, 2-phenylethylamine, benzylamine, ethanolamine, diethanolamine, pyridine, or collidine. Metal alkoxides include sodium methoxide, potassium tert-butoxide, or magnesium methoxide. Preferred salts of β-alanine are sodium salts, potassium salts, calcium salts, or combinations thereof. Specific examples of salts with acids include salts with inorganic acids and organic acids. Examples of inorganic acids that can form salts include hydrochloric acid.

[0013] The pharmaceutical composition of the present invention comprises, as an active ingredient, a compound selected from the group consisting of compounds represented by formulas (1) to (5) or a salt thereof. The pharmaceutical composition of the present invention is preferably a pharmaceutical composition for type I allergy and may be an agent having the effect of suppressing type I allergic symptoms. That is, it can alleviate, treat, or prevent symptoms caused by type I allergy, such as hay fever, urticaria, allergic rhinitis, or bronchial asthma. Furthermore, the effect of suppressing type I allergic symptoms may be the effect of suppressing degranulation of mast cells or basophils in the mechanism of type I allergic reactions. Therefore, the pharmaceutical composition of the present invention may be an agent for suppressing degranulation of mast cells or basophils. That is, the pharmaceutical composition of the present invention can inhibit degranulation of at least mast cells or basophils, and therefore can effectively suppress symptoms of type I allergy. The pharmaceutical composition of the present invention may contain the above-mentioned maltotriose, adenosine, malic acid, or a salt thereof as an active ingredient. In one embodiment, the pharmaceutical composition of the present invention contains one of the above compounds. In another embodiment, the pharmaceutical composition of the present invention contains two or more of the above compounds in combination. Preferably, the pharmaceutical composition of the present invention contains two or more of the above compounds in combination. Includes.

[0014] Maltotriose has the following formula (7): [ka] It is a trisaccharide formed by the α1-4 glycosidic bond of three glucose molecules, represented by the formula: It is produced, for example, by the degradation of amylose or starch by amylase.

[0015] Adenosine has the following formula (8): [ka] It is a nucleoside formed by the β-N9-glycosidic bond between adenine (a type of purine base) and ribose, and is used in the body as a component of DNA and RNA.

[0016] Malic acid has the following formula (9): [ka] L-Malic acid is a type of hydroxy acid represented by the following formula (10): [ka] D-malic acid is a compound represented by the following formula (11): [ka] Both L-malic acid and D-malic acid are effective as active ingredients in the antiallergic agent of the present invention. DL-malic acid, which is a mixture of L-malic acid and D-malic acid, may also be used as the active ingredient.

[0017] "allergy" The allergies targeted by the pharmaceutical composition of the present invention include, but are not limited to, type I allergies. Examples of type I allergies include atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, hives, food allergies, animal allergies, allergic conjunctivitis, anaphylactic shock, and allergic gastroenteritis. The pharmaceutical composition of the present invention has the effect of preventing the onset of allergic diseases or alleviating or treating allergic diseases that have developed in subjects who have developed or are at risk of developing allergic diseases.

[0018] Type I allergic reactions involve antigen-specific IgE antibodies and mast cells and basophils that possess the IgE-specific high-affinity IgE receptor FcεRI. When IgE bound to the surface of mast cells or basophils via FcεRI is crosslinked by an antigen, FcεRI is activated, transmitting a signal downstream, resulting in the release of intracellular granule contents such as histamine, leukotriene C4, PAF, or eosinophil chemotactic factor. This phenomenon is called degranulation. In tissues where degranulation occurs, smooth muscle contraction, increased vascular permeability, or increased glandular secretion occurs, resulting in the appearance of allergic symptoms. An increase in intracellular calcium ion concentration is also involved in degranulation. The pharmaceutical composition of the present invention can inhibit antigen-stimulated degranulation of mast cells and / or basophils. In particular, the pharmaceutical composition of the present invention has the effect of inhibiting the extracellular release of granules containing chemical mediators such as histamine and leukotrienes from mast cells or basophils (degranulation inhibitory effect). Therefore, the pharmaceutical composition of this embodiment can effectively suppress, treat, or prevent symptoms caused by type I allergic reactions.

[0019] The dosage form of the pharmaceutical composition of the present invention is not particularly limited, and includes oral and parenteral preparations, with oral preparations being preferred. Examples of the oral preparations include solid or powder preparations such as fine granules, granules, tablets, capsules, and pills, as well as liquid preparations such as suspensions, emulsions, syrups, and extracts. Examples of parenteral preparations include injections.

[0020] The pharmaceutical composition of the present invention may consist solely of the compound, or may contain the compound. When the pharmaceutical composition of the present invention contains the compound, it may contain other additives.

[0021] When the pharmaceutical composition of the present invention is an oral preparation, other additives may include excipients, binders, disintegrants, emulsifiers, lubricants, flow enhancers, diluents, preservatives, colorants, flavorings, corrigents, stabilizers, moisturizers, antiseptics, antioxidants, or suspending agents, and specific examples thereof include gelatin, sodium alginate, starch, corn starch, sucrose, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid esters, talc, and magnesium stearate. , polyethylene glycol, magnesium silicate, anhydrous silicic acid, or synthetic aluminum silicate.

[0022] When the pharmaceutical composition of the present invention is a parenteral preparation, other additives may include water-soluble solvents such as physiological saline or Ringer's solution, water-insoluble solvents such as vegetable oils or fatty acid esters, isotonic agents such as glucose or sodium chloride, solubilizing agents, stabilizers, preservatives, suspending agents, or emulsifiers.

[0023] The pharmaceutical composition of the present invention can contain 90% by weight or more, 50% by weight or more, 10% by weight or more, or 1% by weight or more of the compound.

[0024] The dosage or intake of the pharmaceutical composition of the present invention can be adjusted appropriately depending on the formulation, as well as the age, sex, body weight, and severity of allergic symptoms of the subject. However, administration or ingestion of the pharmaceutical composition is preferably an amount that can prevent the onset of allergic diseases or alleviate or treat existing allergic diseases. Specifically, the amount of the compound added can be 0.01 to 1,000 mg / kg body weight / day, preferably 0.1 to 750 mg / kg body weight / day, more preferably 1 to 500 mg / kg body weight / day, even more preferably 5 to 400 mg / kg body weight / day, even more preferably 10 to 300 mg / kg body weight / day, even more preferably 15 to 200 mg / kg body weight / day, and most preferably 20 to 150 mg / kg body weight / day. Of course, the above-mentioned administration method is merely an example, and other administration methods may also be used. The administration method, dosage, administration period, and administration interval of the pharmaceutical composition to humans are preferably determined through controlled clinical trials.

[0025] The pharmaceutical composition of the present invention can be administered to humans, but may also be administered to animals other than humans, including pets such as dogs, cats, rabbits, hamsters, guinea pigs, and squirrels; livestock such as cows and pigs; laboratory animals such as mice and rats; and animals kept in zoos, etc.

[0026] The pharmaceutical composition of the present invention may be a pharmaceutical composition for preventing or treating allergies. The pharmaceutical composition includes pharmaceuticals and quasi-drugs. Examples of pharmaceuticals include herbal medicine preparations and herbal medicine preparations. Examples of quasi-drugs include nutritional drinks and health medicines containing herbal medicines.

[0027] Whether or not the pharmaceutical composition of the present invention has a degranulation inhibitory effect can be determined using cells, such as rat basophilic leukemia cells (RBL-2H3 cells), which release granulocytes containing histamine and the like extracellularly upon crosslinking of IgE bound to the cell surface by an antigen. Such cells can be stimulated with an antigen and confirmed by calculating the extent to which degranulation is inhibited in a sample to which the pharmaceutical composition has been added compared to a sample to which the pharmaceutical composition has not been added.

[0028] The pharmaceutical composition of the present invention may contain an anti-allergy ingredient other than maltotriose or adenosine, such as maltotriose, adenosine, malic acid, (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside, 7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one, 2-O-trans-caffeoylhydroxycitric acid, or 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester. The pharmaceutical composition of the present invention can contain two or more of these active ingredients, and by containing multiple active ingredients, allergic symptoms can be suppressed synergistically.

[0029] [2] Food composition for suppressing allergic symptoms In this specification, the term "food composition" refers to a food composition containing the compound or a salt thereof as an active ingredient. The food composition of the present invention can be used as a food composition for suppressing allergic symptoms. Examples of the food composition include foods and beverages.

[0030] Specific examples of foods include fresh prepared foods such as salads; cooked foods such as steak, pizza, and hamburger steak; stir-fried foods such as stir-fried vegetables; vegetables such as tomatoes, bell peppers, celery, bitter melon, carrots, potatoes, and asparagus, and processed foods made from these vegetables; meats such as beef, pork, and chicken, and processed foods made from these meats; seafood such as salmon, tuna, sea bream, flounder, trout, octopus, and squid, and processed foods made from these seafood; cookies, bread, biscuits, hardtack, cakes, rice crackers, yokan, pudding, jelly, Examples include confectioneries such as ice cream, chewing gum, crackers, chips, chocolate, and candy; noodles such as udon, pasta, and soba; fish paste products such as kamaboko, ham, and fish sausage; dairy products such as cheese, cream, and butter; condiments such as miso, soy sauce, dressing, ketchup, mayonnaise, soup base, noodle soup, curry powder, mirin, and roux; soy foods such as tofu; processed agricultural and marine products such as furikake, tsukudani, and cereals; fruits such as apples or grapes; and konjac.

[0031] Examples of beverages include coffee drinks; cocoa drinks; vegetable juices obtained from vegetables; fruit juice drinks such as grapefruit juice, orange juice, grape juice, apple juice, and lemon juice; tea drinks such as green tea, black tea, green tea, and oolong tea; alcoholic beverages such as beer, wine (red wine, white wine, sparkling wine, etc.), sake, plum wine, happoshu, whiskey, brandy, shochu, rum, gin, and liqueurs; dairy drinks; soy milk drinks; liquid diets; and sports drinks.

[0032] The food or beverage includes feed and beverage for animals, such as primates including humans, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, and mice.

[0033] These foods or beverages may contain, as desired, food additives and food ingredients, such as antioxidants, flavorings, acidulants, colorings, emulsifiers, preservatives, seasonings, sweeteners, spices, pH adjusters, stabilizers, vegetable oils, animal oils, sugars and sugar alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meat, seafood, etc. The amounts of these food ingredients and food additives to be added can be determined appropriately within the range that does not impair the object of the present invention.

[0034] These foods or beverages can be subjected to common sterilization processes, such as heat-pressure sterilization using retort and autoclave, batch sterilization, plate sterilization, electric heating sterilization, microwave heating sterilization, and steam sterilization such as injection and infusion.

[0035] Foods and beverages include functional foods (drinks) and health foods (drinks). As used herein, "health foods (drinks)" refers to foods or beverages that have or are expected to have some effect on health, and "functional foods (drinks)" refers to foods or beverages among the aforementioned "health foods (drinks)" that have been designed and processed to fully exhibit bioregulatory functions (i.e., the function of preventing the onset of allergic symptoms or alleviating or treating allergic symptoms). Functional foods and health foods can be in granular, solid, liquid, capsule, gel, or tablet form. The food composition of the present invention can be prepared by adding one or more of the above compounds to the above food or beverage.

[0036] 《Effect》 The mechanism by which the pharmaceutical composition and food composition for suppressing allergic symptoms of the present invention can suppress the symptoms of type I allergy has not been analyzed in detail, but can be assumed as follows. A variety of antigens, including hay fever and food, can cause type I allergies. However, the mechanism by which type I allergies develop is as follows: when IgE bound to the surface of mast cells or basophils via FcεRI is crosslinked by an antigen, FcεRI is activated, transmitting a signal downstream, resulting in the release of intracellular granule contents such as histamine, leukotriene C4, PAF, or eosinophil chemotactic factor. That is, although the causative antigens are different, the symptoms are caused by degranulation of mast cells or basophils. The allergy agent of the present invention inhibits degranulation from mast cells and basophils, which is involved in the development of symptoms common to type I allergies. Therefore, even if the antigens causing type I allergies are different, it is presumed that inhibiting degranulation can suppress symptoms of type I allergies such as atopic dermatitis, bronchial asthma, allergic rhinitis, hay fever, urticaria, food allergies, animal allergies, allergic conjunctivitis, anaphylactic shock, and allergic gastroenteritis. [Example]

[0037] The present invention will be specifically described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0038] Example 1 In this example, a degranulation test was carried out using (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. (3S)-3-Methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside was dissolved in distilled water and adjusted to pH 7.0 before use. Rat basophilic cell line RBL-2H3 cells were cultured at a concentration of 4 × 10 4Cells were seeded into 96-well culture plates at a density of 100 cells / well and sensitized for 20 hours in 5% FBS-DMEM medium containing dinitrophenyl phosphate (DNP)-specific IgE. After washing twice with Tyrode's buffer, 120 μL / well of Tyrode's buffer containing various concentrations of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside was added and incubated at 37°C for 10 minutes. After 10 minutes, the antigen DNP-HSA was added and incubated at 37°C for 30 minutes to induce degranulation, and the supernatant was then collected.

[0039] The inhibitory effect of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside on degranulation was evaluated using the amount of β-hexosaminidase released from granules upon antigen stimulation as an index. Specifically, after collecting the supernatant, cells were lysed in 130 μL of Tyrode's buffer containing 0.1% Triton X-100 by sonication on ice for 5 seconds. Fifty μL of both the collected supernatant and the obtained cell lysate were transferred to each well of a new 96-well microplate and incubated at 37°C for 5 minutes. Then, 100 μL of 3.3 mM 4-nitrophenyl 2-acetamido-2-deoxy-β-D-glucopyranoside (substrate solution) in 0.1 M citrate buffer (pH 4.5) was added to each well and incubated at 37°C for an additional 25 minutes. The enzyme reaction was stopped by adding 100 μL of 2 M glycine buffer (pH 10.4). The absorbance of the reaction solution was measured at a wavelength of 415 nm using a microplate reader (iMark Microplate Reader, Bio-Rad). The β-hexosaminidase release rate (%), as an index of degranulation, was calculated according to the following formula: β-hexosaminidase release rate (%) = [(A 上清 -A 上清のブランク ) / {(A 上清 - A 上清のブランク )+(A 細胞溶解物 -A 細胞溶解物のブランク )}]×100 A: Absorbance at 415 nm for each well As shown in Figure 1, (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0040] Example 2 In this example, a degranulation assay was performed using (7S,8R)-7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one. The procedure of Example 1 was repeated, except that (7S,8R)-7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 2, (7S,8R)-7,8-dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0041] Example 3 In this example, degranulation tests and cytotoxicity tests were carried out using 2-O-trans-caffeoylhydroxycitric acid and 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester. Rat basophil cell line RBL-2H3 cells were cultured at 4 × 10 4 96-well plate with a cell density of 1000 cells / mL Cells were seeded onto culture plates. They were sensitized for 24 hours with 5% FBS-DMEM medium containing dinitrophenyl phosphate (DNP)-specific IgE. 2-O-trans-Caffeoylhydroxycitric acid and 2-O-trans-Caffeoylhydroxycitric acid 6'-O-methyl ester diluted with distilled water were added at 200 μL / well and incubated for 10 minutes. After 10 minutes, a solution of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside diluted with Tyrode's buffer was aspirated and discarded, and 200 μL / well of Tyrode's buffer was added. The antigen, DNP, was then added and incubated for 30 minutes to induce degranulation, after which the supernatant was collected.

[0042] The inhibitory effect of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside on degranulation was evaluated using the amount of β-hexosaminidase released from granules upon antigen stimulation as an index. Specifically, after collecting the supernatant, cells were lysed in 130 μL of modified Tyrode's buffer containing 0.1% Triton X-100 by sonication on ice for 5 seconds. Both the collected supernatant and the obtained cell lysate were transferred to each well of a new 96-well microplate (50 μL / well) and incubated at 37°C for 5 minutes. Then, 100 μL of 3.3 mM 4-nitrophenyl 2-acetamido-2-deoxy-β-D-glucopyranoside (substrate solution) dissolved in 0.1 M citrate buffer (pH 4.5) was added to each well and incubated at 37°C for an additional 25 minutes. The enzyme reaction was stopped by adding 100 μL of 2 M glycine buffer (pH 10.4). The absorbance of the reaction solution was measured at 405 nm using a microplate reader (SH-8000Lab, Corona Electric). The β-hexosaminidase release rate (%), as an index of degranulation, was calculated according to the following formula: β-hexosaminidase release rate (%) = [(A 上清 -A 上清のブランク ) / {(A 上清 -A 上清のブランク )+(A 細胞溶解物 -A 細胞溶解物のブランク )}]×100 A: Absorbance at 405 nm for each well As shown in Figure 3(A), 2-O-trans-caffeoylhydroxycitric acid and 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner. Furthermore, when the cytotoxicity of the compound was measured by the WST-8 method, the compound did not exhibit cytotoxicity at a concentration that inhibited degranulation, as shown in FIG. 3(B).

[0043] Example 4 In this example, a degranulation test was performed using adenosine. The procedure of Example 1 was repeated except that maltotriose was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 4, adenosine inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0044] Example 5 In this example, a degranulation test was performed using maltotriose. The procedure of Example 1 was repeated, except that maltotriose was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 5, maltotriose inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0045] Example 6 In this example, a degranulation test was performed using DL-malic acid. The procedure of Example 1 was repeated, except that DL-malic acid was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 6, DL-malic acid inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0046] Example 7 In this example, a degranulation test was performed using L-malic acid. The procedure of Example 1 was repeated, except that L-malic acid was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 7, L-malic acid inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner.

[0047] In this example, a degranulation test was performed using D-malic acid. The procedure of Example 1 was repeated, except that D-malic acid was used instead of (3S)-3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside. As shown in Figure 8, D-malic acid inhibited degranulation of RBL-2H3 cells in a concentration-dependent manner. [Industrial Applicability]

[0048] The pharmaceutical and food compositions of the present invention can alleviate, treat, or prevent symptoms caused by type I allergies such as hay fever, urticaria, allergic rhinitis, or bronchial asthma.

Claims

【Request Item 1】 【Chemistry 1】 The following formula (1): 3-methyl-6-hydroxyisocoumarin 8-O-β-D-glucoside represented by the formula:

2. The following formula (2): 【Chemistry 2】 7,8-Dihydro-8-β-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-furo[2,3-g][2]benzopyran-5-one represented by the formula:

3. The following formula (3): 【Transformation 3】 (In the formula, R １ is a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, R ２ are 1 to 5, each independently representing a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms; R ３ , R ４ , R ５ , R ６ , and R ７ are each independently a hydrogen atom, a hydroxy group, a carboxy group, or an alkyl group having 1 to 6 carbon atoms. A compound represented by the formula:

4. The following formula (5): 【Chemistry 4】 2-O-trans-caffeoylhydroxycitric acid represented by the formula:

5. The following formula (6): 【Transformation 5】 2-O-trans-caffeoylhydroxycitric acid 6'-O-methyl ester represented by the formula:

6. A pharmaceutical composition comprising, as an active ingredient, a compound selected from the group consisting of the compounds according to claims 1 to 5 or a salt thereof.

7. The pharmaceutical composition according to claim 6, which is for preventing or treating allergies.

8. The pharmaceutical composition according to claim 7, wherein the allergy is type I allergy.

9. The pharmaceutical composition according to any one of claims 6 to 8, which inhibits degranulation.

10. A food composition for suppressing allergic symptoms, comprising a compound selected from the group consisting of the compounds according to claims 1 to 5 or a salt thereof as an active ingredient.

11. The food composition according to claim 10, wherein the allergic symptoms are type I allergic symptoms.

12. The food composition according to claim 10 or 11, wherein the suppression of allergic symptoms is suppression of degranulation.

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