WO2021029659A1 - Composition for preventing or treating allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or salt thereof as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing or treating allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a salt thereof as an effective component. The composition comprising ethyl vanillin as an effective component, according to the present invention, has an effect of improving atopic dermatitis due to an allergic reaction, and may be used as a composition for preventing or treating various allergic diseases by reducing an inflammatory reaction.

Description

Composition for preventing or treating allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a salt thereof as an active ingredient

This application is for Korean Patent Application No. 10-2019-0101055 filed on August 19, 2019, Korean Patent Application No. 10-2019-0098384 filed on August 12, 2019, and August 12, 2019. Korean Patent Application No. 10-2019-0098385 is claimed as priority, and the entire specification is a reference of this application.

The present invention relates to a composition for preventing or treating allergic diseases or atopic dermatitis comprising ethyl vanilin, suberic acid, thiazole or a salt thereof as an active ingredient.

Atopy is derived from the ancient Greek word'atopia', which means'strange, abnormal'. In 1923, Cooke and Coca first began to refer to a tendency that could genetically trigger a specific immune antibody response to an allergen, called atopy. With a steadily increasing trend worldwide, the incidence rate is increasing in developed countries with industrial development.

The cause of the onset of atopy has not been accurately identified yet, but is not limited to one factor such as family history, changes in diet, penetration of allergens, and abnormal skin barriers. It occurs mainly in infancy and infancy and may persist or begin in adulthood. Typical symptoms of atopy appear on the hands, scalp, face, neck, and elbows, but there are differences in the patterns that appear by period. Symptoms of infancy are rough skin and dryness, dermatitis on the outside of limbs, cheeks and foreheads, and sores or scabs sit after scratching with hands. In childhood, it mainly appears on the folds of arms, legs and neck rather than the face, and the skin becomes dry. In puberty and adulthood, symptoms of thickening of the skin on areas such as the face and hands appear.

Local steroids, local immunomodulators, systemic steroids, systemic immunosuppressants, and antihistamines are used as atopic drugs. Topical steroids are used when atopic symptoms are severe and are the most basic method to manage bacterial or viral infections. However, steroids have been introduced in 1950 and have been used for many years, but long-term use is restricted due to problems of skin safety and tolerance depending on the frequency, concentration, and duration of use. In addition, potential skin side effects such as skin, atrophy, telangiectasia, and steroid acne, as well as inhibition of hypohalamic-pituitary-adrenal (HPA), and Cushing's syndrome. Sufficient care should be taken when using it as it may cause side effects.

The local immunomodulators tacrolimus and pimecrolimus are known to be used over a long period of time for the purpose of preventing recurrence of lesions as they have a relatively low possibility of side effects even when used for a long time, unlike conventional topical steroids. It is suitable for use as a treatment and maintenance therapy. However, tacrolimus may have side effects such as decreased kidney function, hand tremor, and hair loss, and pimecrolimus may have serious side effects such as acne and burning as well as skin cancer and lymphoma. When steroids worsen in the acute phase, local immunomodulators are suitable for use as treatment and maintenance therapy for mild atopy, but safety against side effects has not yet been secured, so there is an urgent need for alternative supplements.

Accordingly, as a result of the applicant's efforts to develop a material that has few side effects and is effective in relieving atopic dermatitis symptoms, Ethyl vanilin, Suberic acid, and Thiazole relieve atopic symptoms. The present invention was completed by confirming that it alleviates and improves the inflammatory response.

Prior art literature

Patent Document Korean Patent Application Publication No. 10-2018-0128602

Patent Document Korean Patent Application Publication No. 10-2018-0128602

Patent Document Korean Patent Application Publication No. 10-2018-0128602

An object of the present invention is for the prevention or treatment of allergic diseases or atopic dermatitis comprising ethyl vanilin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof as an active ingredient It is to provide a pharmaceutical composition.

Another object of the present invention is to provide a composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a salt thereof as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention is to solve the above-described problems, allergic diseases including ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole) or a pharmaceutically acceptable salt thereof as an active ingredient or It provides a pharmaceutical composition for the prevention or treatment of atopic dermatitis.

According to one aspect of the present invention, the allergic disease is edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, 　 contact It may be selected from the group consisting of sexual dermatitis, urticaria, pruritus, insect allergy, food allergy and drug allergy, but is not limited thereto.

According to an aspect of the present invention, the active ingredient expresses IL-4 (Interleukin-4), IL-13, TNF-α (tumor necrosis factor-alpha), IL-1β, IL-6, or IL-8. By reducing or reducing the secretion amount of histamine, it may exhibit anti-allergic effects, prevent or treat atopic dermatitis.

In addition, the present invention provides a quasi-drug composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a cosmetic composition for the prevention or improvement of allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a cosmetically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a perfume composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a cosmetically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food composition for the prevention or improvement of allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole, or a food acceptable salt thereof as an active ingredient.

In addition, the present invention provides a quasi-drug composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a cosmetic composition for the prevention or improvement of allergic diseases or atopic dermatitis comprising ethyl vanillin, suberic acid, thiazole or a cosmetically acceptable salt thereof as an active ingredient.

The composition comprising ethyl vanillin, suberic acid, and thiazole of the present invention as an active ingredient has an effect of improving atopic dermatitis caused by an allergic reaction, and for preventing or treating various allergic diseases by reducing the inflammatory response (for improvement) It can be used as a composition.

1 is a graph showing the change in the secretion amount of histamine in mast cells (RBL-2H3 cells) treated with ethylvanillin (each value is the mean ± SEM of three times obtained from three independent wells; the letter on the bar is P) <0.05 shows a statistically significant difference).

2 is a graph showing changes in the expression of inflammatory cytokine-related molecules (IL-4, IL-13, and TNF-α) in mast cells treated with ethylvanillin (each value is three times obtained from three independent wells. Mean ± SEM; letters on the bars represent statistically significant differences at P<0.05).

3 is a graph showing changes in the expression of inflammatory cytokines-related molecules (IL-1β, IL-6 and IL-8) in keratinocytes treated with ethylvanillin (each value is 3 obtained from three independent wells). Meeting mean ± SEM; letters on bars show statistically significant difference at P<0.05).

4 is a graph showing the change in the secretion amount of histamine in mast cells treated with suberic acid (RBL-2H3 cells) (each value is the mean ± SEM of three times obtained from three independent wells; the letters on the bar are It shows a statistically significant difference at P<0.05).

5 is a graph showing changes in expression of inflammatory cytokine-related molecules (IL-4, IL-13 and TNF-α) in mast cells treated with suberic acid (each value is 3 obtained from three independent wells). Meeting mean ± SEM; letters on bars show statistically significant difference at P<0.05).

6 is a graph showing changes in the expression of inflammatory cytokine-related molecules (IL-1β, IL-6 and IL-8) in suberic acid-treated keratinocytes (each value is obtained from three independent wells. Mean ± SEM of 3 times; letters on the bars show statistically significant differences at P<0.05).

Figure 7 is a graph showing the change in the secretion amount of histamine in mast cells treated with thiazole (RBL-2H3 cells) (each value is the mean ± SEM of three times obtained from three independent wells; the letter on the bar is P) <0.05 shows a statistically significant difference).

8 is a graph showing changes in the expression of inflammatory cytokine-related molecules (IL-4, IL-13, and TNF-α) in mast cells treated with thiazole (each value is three times obtained from three independent wells. Mean ± SEM; letters on the bars represent statistically significant differences at P<0.05).

9 is a graph showing changes in expression of inflammatory cytokine-related molecules (IL-1β, IL-6, and IL-8) in keratinocytes treated with thiazole (each value is 3 obtained from three independent wells). Meeting mean ± SEM; letters on bars show statistically significant difference at P<0.05).

The present inventors believe that ethylvanillin, suberic acid, or thiazole alleviates atopic symptoms, and IL-4, IL-13, TNF-α, IL-1β, IL-6, which are inflammatory cytokines secreted by the immune response of cells Alternatively, the present invention was completed by confirming that the expression of IL-8 is significantly reduced and the amount of histamine secreted is reduced.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for preventing or treating allergic diseases or atopic dermatitis, comprising ethyl vanilin, suberic acid, thiazole, or a salt thereof as an active ingredient.

Specifically, the ethyl vanilin (Ethyl vanilin) is a phenolic aldehyde (phenolic aldehyde)-based material of the molecular formula C ₉ H ₁₀ O ₃ , the molecular weight is 166.18, the structure is as shown in [Chemical Formula 1]. Ethyl vanillin is referred to as 3-ethoxy-4-hydroxybenzaldehyde and bourbonal.

Ethylvanillin is a white or pale white powder, insoluble in water and soluble in chloroform and ethanol. Ethyl vanillin is known as a scent component, and the anti-inflammatory substance is vanilla, specifically sweet, creamy, vanilla, and caramel odor. Ethyl vanillin is contained in Ageratum conyzoides, Billygoat weed, which is mainly grown in Brazil, and is contained in the fruit of Vanilla planifolia ( Flat-leaved vanilla, vanilla). It is known to exist 80%.

Ethyl vanillin is registered as a flavoring agent in the European COE (Council of Europe), Korea Food and Drug Administration (KFDA), and the Food and Drug Association (FDA) food additive database, and is used as a supplement, and is used as a sweetener for cosmetics by the Korean Cosmetics Association. It is approved and used for the purpose of blending flavors.

The physiological activity of ethylvanillin reported so far has antioxidant activity, and it was reported that the maximum antioxidant activity was observed 5 minutes after oral administration of ethylvanillin 32.7 mg/kg to ICR male mice. In addition, it was confirmed that ethyl vanillin may be useful when preserving or storing food through the results showing the bacteria-reducing efficacy when treated with 2 mg/mL of ethyl vanillin and pH 5.0 in the C. sakazakii cocktail strain. On the other hand, in the toxicity test of ethyl vanillin, the LD ₅₀ value was 3,500 mg/kg when ethyl vanillin was administered orally to rats, and the LD ₅₀ value was 7,940 mg/kg when ethyl vanillin was transdermally administered to rabbits. Reported.

[Formula 1]

Ethyl vanillin of the present invention may include ethyl vanillin hydrate, ethyl vanillin derivative, and the like within a range having the same efficacy as the ethyl vanillin, and may also include a solvate or stereoisomer thereof.

The method of obtaining the ethyl vanillin is not particularly limited, and may be isolated from a plant containing the ethyl vanillin, chemically synthesized using a known manufacturing method, or a commercially available one may be used.

Specifically, the suberic acid is a kind of dibasic acid, acicular crystal, Cas No. 505-48-6, the structural formula is C ₈ H ₁₄ O ₄ , and the molecular weight is 174.2 g/mol. The molecular structure of suberic acid is shown in Formula 2 below. Suberic acid is also referred to as Octanedioic acid, Cork acid, and 1,6-Hexanedicarboxylic acid. Suberic acid has a melting point of 141-144℃ and a boiling point of 230℃. Suberic acid is mainly contained in plants such as Ricinus communis L. (Castor oil plant, castor), Quercus suber (Cork tree, cork), and Vernonia galamensis (Ironweed, thistle flower).

Suberic acid is mainly used for industrial manufacturing purposes. For example, it is used in plastic lubricants, hydraulic fluids for hydraulic machinery, or in the manufacture of candles. There is no physiologically active function of suberic acid that has been reported so far, and mainly metabolites have been studied a lot. In the case of ketosis patients, it is reported that suberic acid is detected as metabolites in the urine and is used as an indicator of ketosis patients, and it is reported that high concentrations are also detected in the urine of diabetic patients. The LD ₅₀ value of suberic acid is reported to be more than 2000mg/kg when administered orally to rats.

[Formula 2]

Suberic acid of the present invention may include suberic acid hydrates, suberic acid derivatives, and the like within a range having the same efficacy as the suberic acid, and may also include a solvate or stereoisomer thereof.

The method of obtaining the suberic acid is not particularly limited, and may be isolated from a plant containing the suberic acid, chemically synthesized using a known manufacturing method, or a commercially available one may be used.

Specifically, the thiazole is a heterocyclic compound, and the IUPAC name is 1,3-thiazole, and thiazole, 5H-thiazole-1 -Has tinnitus such as 5H-Thiazol-1-ium and 5H-1,3-thiazole. The structural formula of thiazole is C ₃ H ₃ NS and the molecular weight is 85.12 g/mol, and the structure is as shown in [Chemical Formula 3] below.

Thiazole is a colorless or pale yellow transparent liquid with a fishy odor. Thiazole is known as a safe substance that can be used as a fragrance, and is a flavor agent by FEMA (Flavor and Extract Manufacturers' Association), FDA (Food and Drug Administration), and JECFA (Joint FAO/WHO Expert Committee on Food Additives). It is approved as an ingredient and has been used industrially for the purpose of giving taste and aroma.

[Formula 3]

The thiazole of the present invention may include a thiazole hydrate, a thiazole derivative, and the like within a range having the same effect as the thiazole, and may also include a solvate or stereoisomer thereof.

The method for obtaining the thiazole is not particularly limited, and may be isolated from a plant containing the thiazole, chemically synthesized using a known manufacturing method, or a commercially available one may be used.

In the present invention, the term “cosmetically acceptable salt”, “food pharmaceutically acceptable salt”, “pharmaceutically acceptable salt” or “salt thereof” may be an acid addition salt formed by a free acid. have. The acid addition salt can be prepared by a conventional method, for example, dissolving the compound in an excess aqueous acid solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. In addition, the same molar amount of the compound and an acid or alcohol (eg, glycol monomethyl ether) in water may be heated, and then the mixture may be evaporated to dryness, or the precipitated salt may be suction filtered.

As the free acid, an inorganic acid or an organic acid may be used. Non-limiting examples of the inorganic acid may be hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like, which may be used alone or in combination of two or more. Non-limiting examples of the organic acid include methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid (propionic acid). acid), citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid (glucuronic acid), aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like can be used. These may be used alone or in combination of two or more.

In addition, the ethyl vanillin, suberic acid or thiazole can be made cosmetically or food acceptable metal salt using a base. The alkali metal or alkaline earth metal salt can be obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. As the metal salt, it is particularly preferable to prepare a sodium, potassium or calcium salt, but is not limited thereto. Further, the corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (eg, silver nitrate).

The salt of ethylvanillin, suberic acid or thiazole may include all salts of acidic or basic groups that may be present in the compound of ethylvanillin, suberic acid, or thiazole, unless otherwise indicated. For example, the salts of ethyl vanillin, suberic acid, or thiazole may include sodium, calcium, and potassium salts of the hydroxy group, and other cosmetically acceptable salts of the amino group include hardbromide, sulfuric acid, Hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salt, etc. And can be prepared through a method for preparing a salt known in the art.

In the present invention, "allergic disease" refers to a disease caused by an allergy reaction in which the body's immune response to an external antigen is excessive, and specifically, edema, hypersensitivity (anaphylaxis), allergic rhinitis (allergic) rhinitis), asthma, allergic conjunctivitis, allergic dermatitis, contact dermatitis, urticaria, pruritus, insect allergy, food allergy, and drug allergy. However, it is not limited thereto.

In the present invention, "atopic dermatitis" is one of allergic diseases and is a skin disease accompanied by symptoms such as itching, dry skin, increased skin thickness, and characteristic eczema.

The pharmaceutical composition for the prevention or treatment of allergic diseases or atopic dermatitis according to the present invention does not specifically limit the content as long as it contains ethyl vanillin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof, but preferably The dose of ethyl vanillin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof may be included in a concentration of 0.1 μM to 1000 μM, but is not limited thereto. At this time, when ethylvanillin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof is less than the above concentration range, there is a problem that it is difficult to exert a desirable preventive or therapeutic effect, and ethylvanillin, suberic acid, thiazole or its When the pharmaceutically acceptable salt exceeds the above concentration range, there may be concerns of toxicity, including cytotoxicity.

The pharmaceutical composition for the prevention or treatment of allergic diseases or atopic dermatitis according to the present invention, according to a conventional method, respectively, oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and It may be formulated and used in the form of a sterile injectable solution, and may include a suitable carrier, excipient, or diluent commonly used in the preparation of pharmaceutical compositions for formulation.

Examples of the carrier or excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecided And various compounds or mixtures including vaginal cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

In the case of formulation, it can be prepared using diluents or excipients such as fillers, weight agents, binders, wetting agents, disintegrants, and surfactants that are usually used.

Solid preparations for oral administration may be prepared by mixing at least one excipient such as starch, calcium bonate, sucrose or lactose, gelatin, etc. with the ethyl vanillin, suberic acid or thiazole. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous agents, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like can be used.

The preferred dosage of the pharmaceutical composition for the prevention or treatment of allergic diseases or atopic dermatitis according to the present invention varies depending on the patient's condition, weight, degree of disease, drug form, route and duration of administration, but may be appropriately selected by those skilled in the art. I can. However, for a desirable effect, it may be administered at 0.0001 to 2,000 mg/kg per day, preferably 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be divided several times. However, the scope of the present invention is not limited by the dosage.

The pharmaceutical composition for the prevention or treatment of allergic diseases or atopic dermatitis according to the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The composition comprising the active ingredient according to the present invention reduces the expression of IL-4, IL-13, TNF-α, IL-1β, IL-6, or IL-8 that is excessively secreted due to an immune response, or By reducing the amount of secretion, allergic diseases or atopic dermatitis symptoms can be improved and prevented.

Histamine, a representative itching mediator, was proposed by Lewis in 1927 to cause itching in inflammatory skin diseases. The main source of histamine in the skin is mast cells, and histamine, which is synthesized and stored, is secreted in response to stimulation. The secreted histamine causes erythema, pain, vasodilation and swelling due to direct action on blood vessels and release of neuropeptide from sensory nerves in addition to itching through histamine receptors present in keratinocytes.

In one embodiment of the present invention, ethyl vanillin, suberic acid or thiazole are allergic diseases or symptoms of atopic dermatitis by confirming that the secretion of histamine, a representative itching mediator, is significantly reduced as a result of treatment with ethyl vanillin, suberic acid, or thiazole. It was confirmed that it can improve and prevent.

In addition, in one embodiment of the present invention, as a result of treatment with ethyl vanillin suberic acid or thiazole, IL-4, IL-13, TNF-α, IL-1β, IL-, which are inflammatory cytokines secreted by immune responses of cells. By confirming that the expression of 6 and IL-8 was significantly reduced, it was found that ethylvanillin, suberic acid, or thiazole can suppress excessive immune responses and treat inflammation.

In addition, the present invention provides a composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanilin, suberic acid, thiazole, or a salt thereof as an active ingredient.

Details of the ethyl vanillin, suberic acid, and thiazole are as described above.

The composition for preventing or improving allergic diseases or atopic dermatitis of the present invention may be a health functional food composition, a cosmetic composition, a perfume composition, or a quasi-drug composition.

In the present invention, the term "health functional food" refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, "functionality" means obtaining useful effects for health purposes such as controlling nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food may be prepared without limitation as long as it is a formulation recognized as a health functional food. The health functional food composition of the present invention has the advantage of not having side effects that may occur when taking the drug for a long period of time, unlike general drugs, using food as a raw material, and is excellent in portability, enhancing the anti-allergic effect or the relief effect of atopic dermatitis symptoms. It can be ingested as a supplement to make.

In the health functional food for the prevention or improvement of allergic diseases or atopic dermatitis according to the present invention, when the ethyl vanillin, suberic acid or thiazole is used as an additive of a health functional food, it is added as it is or other food or food ingredient It can be used together, and can be suitably used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

Formulations of health functional foods may be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

The type of food is not particularly limited, and examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes, and may include all foods in the usual sense.

In general, when preparing food or beverage, the ethyl vanillin, suberic acid or thiazole may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on 100 parts by weight of the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and the present invention has no problem in terms of safety in terms of using fractions from natural products. It can also be used in the above amount.

Among the functional foods according to the present invention, the beverage may contain various flavoring agents or natural carbohydrates as an additional component, like a conventional beverage. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.

In addition to the above, the health functional food for the prevention or improvement of allergic diseases or atopic dermatitis according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloids. It may contain thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages. In addition, the health functional food composition for preventing or improving allergic diseases or atopic dermatitis of the present invention may contain flesh for the manufacture of natural fruit juice, fruit juice drink, and vegetable drink. These ingredients may be used independently or in combination. The ratio of these additives is not limited, but it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the functional food of the present invention.

The term "cosmetic composition" used in the present invention is a composition including the compound, and the formulation may be in any form. For example, cosmetic products prepared using the above composition include creams such as nutritional cream, eye cream, massage cream, cleansing cream, packs, lotions such as nutritional lotion, essences, softening lotion, lotion such as nutritional lotion. Types, powders, foundations, makeup bases, and the like, and can be manufactured and commercialized in any of these formulations to achieve the object of the present invention, and are not limited to the above examples. In addition, the cosmetic composition according to the present invention can be formulated by a conventional cosmetic preparation method. Specifically, the cosmetics of the present invention are skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, pack, mask pack, mask sheet , Soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, and eye shadow.It may have any one formulation selected from the group consisting of.

The cosmetic composition of the present invention may contain other additives such as excipients, carriers, etc. in addition to ethyl vanillin, suberic acid, thiazole, or salts thereof, and it is possible to apply and mix the usual ingredients blended in general skin cosmetics as needed. .

Specifically, the cosmetic composition of the present invention may further include a transdermal penetration enhancer. The term used in the present invention, a transdermal penetration enhancer, is a composition that allows a desired component to penetrate into blood vessel cells of the skin at a high absorption rate. Preferably, other phospholipid components, liposome components, and the like used in lecithin cosmetics are included, but are not limited thereto.

In addition, as an oil that can be mainly used as an oily component, at least one selected from vegetable oils, mineral oils, silicone oils, and synthetic oils may be used. More specifically, mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, Vitis vinifera seed oil, macadamia nut oil, glyceryl octanoate, castor oil, ethylhexyl isononanoate, dime Chicon, cyclopentasiloxane, and sunflower seed oil can be used.

In addition, 0.1 to 5% by weight of a surfactant, a higher alcohol, and the like may be added to reinforce the emulsifying ability. As such surfactants, conventional surfactants such as nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, phospholipids, etc. may be used, and specifically, sorbitansquinoleate, polysorbate 60 , Glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, DIA-cetylphosphate, sorbitan stearate/sucrosecoate, glyceryl stearate/polyethylene glycol-100 Stearate, ceteareth-6 olivate, arachidyl alcohol/behenyl alcohol/arachidyl glucoside, polypropylene glycol-26-butes-26/polyethylene glycol-40 hydrogenated castor oil, etc. can be used. have. As the higher alcohol, an alcohol having 12 to 20 carbon atoms, such as cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol, and the like may be used alone or in combination of one or more.

The aqueous phase component may further add 0.001 to 5% by weight of one or more thickeners such as carbomer, xanthan gum, bentonite, magnesium aluminum silicate, cellulose gum, dextrin palmitate, etc. to adjust the viscosity or hardness of the aqueous phase.

In addition, the cosmetic composition of the present invention includes medicinal ingredients such as higher fatty acids and vitamins, sunscreen, and antioxidants (butylhydroxyanisole, propyl gallic acid, lysorbic acid, tocopheryl acetate, butylated hydroxy) as needed. Toluene), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, etc.) Sodium Malate, Pmalic Acid, Sodium Fmalate, Succinic Acid, Sodium Succinate, Sodium Hydroxide, Sodium Monohydrogen Phosphate, etc.), Moisturizing Agents (Glycerin, Sorbitol, Propylene Glycol, Butylene Glycol, Hexylene Glycol, Diglycerin , Betaine, glyceres-26, methylgluces-20, etc.), lubricants and the like can be added.

In addition, the cosmetic composition of the present invention further includes a substance capable of auxiliaryly providing essential nutrients to the skin, preferably, it may contain a natural scent, a cosmetic scent, or an auxiliary agent including, but not limited to, herbal medicines. have.

In the cosmetic composition of the present invention, the effective content of ethyl vanillin, suberic acid, thiazole, or a cosmetically acceptable salt thereof is not particularly limited, and may be included in an amount of 0.0001 to 20% by weight based on the total weight of the composition. Ethylvanillin, suberic acid, thiazole, or salts of less than 0.0001% by weight in the cosmetic may not have the expected improvement effect due to its small amount, and more than 20% by weight of ethylvanillin, suberic acid, thiazole, or a salt thereof Salts can exhibit known toxicity.

The term "fragrance composition" of the present invention may be formulated as a base for skin external use such as perfume, cosmetics, and bath, or food, pharmaceuticals, etc., and the blending amount is appropriately selected in order to achieve the desired effect according to conventional techniques in the art. Can be blended.

The formulation of the perfume 　 composition of the present invention is not particularly limited, but may be any one selected from powder, granule, liquid spray, solid and gel type formulations.

The 　 perfume 　 composition includes cosmetic products including perfume, soap cleaning products including bath soap, indoor cleaning products including glass cleaners, fragrance products including car air fresheners, bath products including herbal bath products, and stationery It can be used to manufacture a fragrance product, an environmental product including an office fragrance, or an industrial product including a synthetic resin.

The perfume composition of the present invention contains ethyl vanillin, suberic acid or thiazole, which are active ingredients, based on the total weight of the perfume composition, depending on the product type in which the perfume composition of the present invention is embodied, 0.00001% to 10% by weight, preferably Preferably, it may contain in the range of 0.00001% by weight to 1.0% by weight, more preferably 0.00001% by weight to 0.5% by weight.

Product types in which the fragrance composition is specified above include soap, cosmetics, bath agents, aroma oils, etc., specifically body lotion, shampoo, hair rinse, hair conditioner, hair treatment, antiperspirant, skin lotion, skin cream, deodorant , Perfume (spray or fumigant), lipstick, lip cream, bath agent, etc., but are not limited thereto.

These products may contain various additives such as blood circulation accelerators, anti-inflammatory agents, moisturizing agents, astringents, inorganic salts, organic salts, oily ingredients, surfactants, herbal medicines, colors, fragrances, sulfur, sinter deposits, disinfectants, etc. .

In particular, the perfume composition of the present invention will generally be used as an external application for the skin of cosmetic formulations such as makeup products, skin lotions, and skin creams. In this case, it may contain ingredients commonly used in these cosmetic formulations.

Particularly, the perfume composition of the present invention is preferably used by being incorporated into a bathing agent in that the active ingredients, ethyl vanillin, suberic acid, or thiazole have atopic dermatitis improvement activity. In this case, the active ingredient may preferably be included in the range of 0.00001 to 1% by weight, more preferably 0.0001 to 0.1% by weight, based on the total weight of the bath agent. When the fragrance composition of the present invention is incorporated into a bathing agent and used, the bathing agent may be added to the bath water in a concentration of 0.015 to 15 ppm and used.

The bath agent may contain inorganic salts, organic acids, oily ingredients, etc. in addition to the active ingredients of the perfume composition of the present invention.

As inorganic salts, sodium chloride, sodium hydrogen carbonate, sodium carbonate, borax, sodium sulfate, sodium sulfide, sodium sesquicarbonate, sodium nitrate, sodium thiosulfate, sodium polyphosphate, sodium phosphate, calcium oxide, magnesium oxide, calcium carbonate, magnesium carbonate, potassium chloride, sulfide Potassium and the like may be exemplified, and these may be used alone or as a mixture of two or more. These inorganic salts may be added to the bath agent in an amount of 5% by weight or more, preferably 10% by weight or more, based on the total weight of the bath agent.

As the organic acid, succinic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid and the like can be exemplified, and these may be used alone or in a mixture of two or more. These organic acids may be added to the bath agent in the range of 0.1 to 50% by weight, based on the total weight of the bath agent.

Examples of the oily component include wax, hydrocarbon, higher fatty acid, higher alcohol, ester, and silicone oil.

The bath agent may also contain additional ingredients commonly used in the art. Examples of these components include inorganic acids such as boric acid, metasilicic acid, and silicic anhydride; Herbal powders such as fennel, ginkgo, ginger, citrus peel, valerian root, mint, ginseng, and oats; Natural pigments, such as coal tar dye, chlorophyll, riboflavin, safflower, and anthraquinone, which have been found to be harmless to the human body; Vitamins such as vitamin A, vitamin C, vitamin D, and vitamin E; Sulfur, mica powder, clay powder, loess powder, rice bran carbide, fungicides, and preservatives.

These bathing agents may be prepared in any form such as granules, tablets, liquids, powders, and the like.

The term "quasi-drug" as used in the present invention refers to items that are less effective than medicines among items used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals. According to this, quasi-drugs exclude items used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases of humans and animals, and products that have mild or no direct action on the human body.

The quasi-drug composition of the present invention is used for the purpose of preventing or improving allergic diseases or atopic dermatitis, and is not particularly limited in its formulation, for example, softening lotion, nutritional lotion, massage cream, nutrition cream, pack , A mask pack, a mask sheet, a gel or a cosmetic composition having a skin adhesive type cosmetic formulation, and may be a transdermal dosage form such as a lotion, ointment, gel, cream, patch, or spray.

In addition, in each formulation, the quasi-drug composition may be arbitrarily selected and blended according to the formulation or purpose of use of other quasi-drugs. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (inhibition or mitigation). For example, thickeners, stabilizers, solubilizers, conventional adjuvants such as vitamins, pigments and flavors, and carriers may be included.

The content of ethyl vanillin, suberic acid, thiazole or a salt thereof of the present invention is preferably 0.0001 to 20% by weight, respectively, based on the total weight of the quasi-drug composition. If it exceeds 20% by weight, color and stability may be deteriorated during the preparation of the composition, and if it is less than 0.0001%, the effect is negligible.

In addition, in the quasi-drug composition of each formulation, components other than the essential components described above may be appropriately selected and blended by a person skilled in the art without difficulty depending on the formulation or purpose of use.

Below, The present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Example 1: Evaluation of secretion and expression of inflammation-related cytokines after treatment with Ethyl vanilin on mast cells and keratinocytes

1-1. Experiment method

1) mast cell culture

Mast cells (rat basophilic leukemia cell line, RBL-2H3) were purchased and used from ATCC (Manassas, VA, USA). DMEM (Dulbecco modified eagle medium) (Gibco BRL, USA) containing 10% heat-inactivated feta bovine serum (FBS) (Gibco BRL, USA) and 1% penicillin and streptomycin (Gibco BRL, USA) Cells were cultured in the culture medium. Cells were cultured under conditions of 37° C. and 5% CO ₂ for experiments. The mast cells were suspended in DMEM containing 10% FBS and then dispensed into a 6 well plate (Corning, USA) to a cell number of 1×10 ⁶ cells/ml.

Then, it was sensitized with anti DNP (dinitrophenyl)-IgE (100 ng/ml) and incubated in an incubator at 37° C. and 5% CO ₂ for 24 hours. After washing twice with PBS (phosphate buffered saline), DNP-HSA (dinitrophenylated human serum albumin; 100 ng/ml) was treated for 1 hour to induce an immune response. At this time, in order to evaluate the atopic improvement effect by ethyl vanilin, anti-DNP (dinitrophenyl)-IgE was treated and ethyl vanilin (100 μM) was treated and incubated for 24 hours, and then the immune response was induced with DNP-HSA. I did. In the case of negative control cells, they were treated with DMSO instead of ethyl vanillin and cultured for 24 hours, and then an immune response was induced with DNP-HSA.

2) Cell culture of keratinocytes

Keratinocytes (human keratinocyte cell line, HaCaT) were purchased and used from ATCC (Manassas, VA, USA). Cells were cultured using a DMEM culture medium containing 10% FBS (fetal bovine serum) and antibiotics. The culture vessel was a 75T-flask and a 6-well plate, and cultured in a 37°C incubator supplied with 5% CO2. The culture medium was changed every 3 to 4 days, and when cells were excessively proliferated, they were subcultured. The dispensed HaCaT cells (5×10 ⁵ /well) were cultured for 24 hours and washed with PBS. To induce an immune response to keratinocytes, TNF-α (tumor necrosis factor-α) (10 ng/ml) and IFN-γ (interferon gamma) (10 ng/ml) were treated together in DMEM medium without FBS. And incubated for 24 hours. At this time, 100 μM of ethyl vanillin was treated with the immune response inducer at the same time to evaluate the effect of improving the immune response by ethyl vanillin, and in the case of negative control cells, DMSO instead of ethyl vanillin was simultaneously treated with the immune response inducing material and incubated for 24 hours .

3) Histamine release assay

In order to measure the concentration of histamine secreted from mast cells that induced the immune response, the immune response was terminated by standing in an ice bath for 10 minutes. After centrifuging the culture solution at 10,000 xg for 10 minutes, the supernatant was taken. Transferred to a 96-well plate. Histamine was quantified using ENZO's histamine release assay ELISA kit, and was measured at 450 nm wavelength of Infinite M200 pro microplate reader (TECAN).

4) RNA isolation and real-time PCR (quantitative reverse-transcription polymerase chain reacion) using the Trizol method

After removing the supernatant of the mast cells that induced the immune response, add 1 ml of trizol, leave for 2 minutes, add chloroform, vortex for 10 seconds, and centrifuge at 2,400 xg for 10 minutes, and then the supernatant The solution was taken, and the same amount of isopropanol was mixed and shaken. The supernatant was removed by centrifugation at 16,000 xg for 25 minutes, and the pellet was dissolved in 20 μl of DEPC (diethyl pyrocarbonate)-DW and stored at -20°C before use in the experiment. RT-PCR was reacted at 45°C for 30 minutes and 94°C for 5 minutes using a one-step RT-PCR PreMix kit (iNtRON Biotechnology, Korea), followed by denaturation at 94°C for 30 seconds, and 30 at 55°C. After annealing for a second, the cycle of stretching at 72°C for 1 minute was repeated 32 times, and the last extension was performed at 72°C for 5 minutes, followed by PiQ SYBR green supermix (Bio-Rad ) And CFX Connect™ Real-Time PCR Detection System (Bio-Rad) were used to perform quantitative PCR, and the primer sequence used at this time is as shown in [Table 1].

[Table 1]

Primer sequence used for RT-PCR

1-2. Experiment result

1) Changes in histamine secretion

In order to confirm the effect of ethylvanillin on histamine secreted by degranulation from activated mast cells, the concentration of histamine in the cell culture was measured by an enzyme immunoreaction method. As a result, the histamine concentration was significantly increased in control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA compared to normal mast cells. On the other hand, as a result of treatment with ethyl vanillin while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, the concentration of histamine was significantly reduced (FIG. 1).

2) Changes in inflammatory cytokine production in mast cells

In control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA, the expression of inflammatory cytokines IL-4, IL-13, and TNF-α was significantly increased compared to normal mast cells. . On the other hand, as a result of treatment with ethylvanillin while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, increased inflammatory cytokines (IL-4, IL-13 and TNF-α) All expressions were significantly reduced (Fig. 2).

3) Changes in inflammatory cytokine production in keratinocytes

In control cells induced immune response with TNF-α and IFN-γ, the expression of inflammatory cytokines IL-1β, IL-6 and IL-8 was significantly increased compared to normal keratinocytes. On the other hand, as a result of treatment of keratinocytes with ethyl vanillin along with TNF-α and IFN-γ, the expression of inflammatory cytokines (IL-1β, IL-6 and IL-8) increased due to the immune response were all significantly. Decreased (Fig. 3).

Example 2: Evaluation of secretion and expression of inflammation-related cytokines after suberic acid treatment on mast cells and keratinocytes

2-1. Experiment method

1) mast cell culture

Mast cells (rat basophilic leukemia cell line, RBL-2H3) were purchased and used from ATCC (Manassas, VA, USA). DMEM (Dulbecco modified eagle medium) (Gibco BRL, USA) containing 10% heat-inactivated feta bovine serum (FBS) (Gibco BRL, USA) and 1% penicillin and streptomycin (Gibco BRL, USA) Cells were cultured in the culture medium. Cells were cultured under conditions of 37° C. and 5% CO ₂ for experiments. The mast cells were suspended in DMEM containing 10% FBS and then dispensed into a 6 well plate (Corning, USA) to a cell number of 1×10 ⁶ cells/ml.

Then, it was sensitized with anti DNP (dinitrophenyl)-IgE (100 ng/ml) and incubated in an incubator at 37° C. and 5% CO ₂ for 24 hours. After washing twice with PBS (phosphate buffered saline), DNP-HSA (dinitrophenylated human serum albumin; 100 ng/ml) was treated for 1 hour to induce an immune response. At this time, in order to evaluate the atopy improvement effect by Suberic acid, anti-DNP (dinitrophenyl)-IgE was treated and at the same time suberic acid (100 μM) was treated and incubated for 24 hours, and then the immune response with DNP-HSA Was induced. In the case of negative control cells, they were treated with DMSO instead of suberic acid and cultured for 24 hours, and then an immune response was induced with DNP-HSA.

2) Cell culture of keratinocytes

Keratinocytes (human keratinocyte cell line, HaCaT) were purchased and used from ATCC (Manassas, VA, USA). Cells were cultured using a DMEM culture medium containing 10% FBS (fetal bovine serum) and antibiotics. The culture vessel was a 75T-flask and a 6-well plate, and cultured in a 37°C incubator supplied with 5% CO2. The culture medium was changed every 3 to 4 days, and when cells were excessively proliferated, they were subcultured. The dispensed HaCaT cells (5×10 ⁵ /well) were cultured for 24 hours and washed with PBS. To induce an immune response to keratinocytes, TNF-α (tumor necrosis factor-α) (10 ng/ml) and IFN-γ (interferon gamma) (10 ng/ml) were treated together in DMEM medium without FBS. And incubated for 24 hours. At this time, to evaluate the effect of improving the immune response by suberic acid, 100 μM of suberic acid was treated simultaneously with the immune response inducing substance, and in the case of negative control cells, DMSO instead of suberic acid was treated with the immune response inducing substance at the same time for 24 hours. During incubation.

3) Histamine release assay

In order to measure the concentration of histamine secreted from mast cells that induced the immune response, the immune reaction was terminated by standing in an ice bath for 10 minutes, and the culture was centrifuged at 10,000 xg for 10 minutes, and the supernatant was taken. Transferred to a 96-well plate. Histamine was quantified using ENZO's histamine release assay ELISA kit, and was measured at 450 nm wavelength of Infinite M200 pro microplate reader (TECAN).

4) RNA isolation and real-time PCR (quantitative reverse-transcription polymerase chain reacion) using the Trizol method

After removing the supernatant of the mast cells that induced the immune response, add 1 ml of trizol, leave for 2 minutes, add chloroform, vortex for 10 seconds, and centrifuge at 2,400 xg for 10 minutes, and then the supernatant The solution was taken, and the same amount of isopropanol was mixed and shaken. The supernatant was removed by centrifugation at 16,000 xg for 25 minutes, and the pellet was dissolved in 20 μl of DEPC (diethyl pyrocarbonate)-DW and stored at -20°C before use in the experiment. RT-PCR was reacted at 45°C for 30 minutes and 94°C for 5 minutes using a one-step RT-PCR PreMix kit (iNtRON Biotechnology, Korea), followed by denaturation at 94°C for 30 seconds, and 30 at 55°C. After annealing for a second, the cycle of stretching at 72°C for 1 minute was repeated 32 times, and the last extension was performed at 72°C for 5 minutes, followed by PiQ SYBR green supermix (Bio-Rad ) And CFX Connect™ Real-Time PCR Detection System (Bio-Rad) were used to perform quantitative PCR, and the primer sequence used at this time is as shown in [Table 2].

[Table 2]

Primer sequence used for RT-PCR

2-2. Experiment result

1) Changes in histamine secretion

In order to confirm the effect of suberic acid on the histamine secreted by degranulation from activated mast cells, the concentration of histamine in the cell culture was measured by the enzyme immunoreaction method. As a result, the histamine concentration was significantly increased in control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA compared to normal mast cells. On the other hand, as a result of treatment with suberic acid while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, the concentration of histamine was significantly reduced (FIG. 4).

2) Changes in inflammatory cytokine production in mast cells

In negative control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA, the expression of inflammatory cytokines IL-4, IL-13 and TNF-α significantly increased compared to normal mast cells. I did. Meanwhile, as a result of treatment with suberic acid while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, increased inflammatory cytokines (IL-4, IL-13 and TNF-α) due to the immune response All of the expression of was significantly reduced (Fig. 5).

3) Changes in inflammatory cytokine production in keratinocytes

The expression of inflammatory cytokines IL-1β, IL-6, and IL-8 was significantly increased in negative control cells induced immune response with TNF-α and IFN-γ compared to normal keratinocytes. On the other hand, as a result of treatment of suberic acid with TNF-α and IFN-γ on keratinocytes, the expression of inflammatory cytokines (IL-1β, IL-6 and IL-8) increased due to the immune response was significant. Decreased to (Fig. 6).

Example 3: Evaluation of secretion and expression of inflammation-related cytokines after treatment with thiazole on mast cells and keratinocytes

3-1. Experiment method

1) mast cell culture

Mast cells (rat basophilic leukemia cell line, RBL-2H3) were purchased and used from ATCC (Manassas, VA, USA). DMEM (Dulbecco modified eagle medium) (Gibco BRL, USA) containing 10% heat-inactivated feta bovine serum (FBS) (Gibco BRL, USA) and 1% penicillin and streptomycin (Gibco BRL, USA) Cells were cultured in the culture medium. Cells were cultured under conditions of 37° C. and 5% CO ₂ for experiments. The mast cells were suspended in DMEM containing 10% FBS and then dispensed into a 6 well plate (Corning, USA) to a cell number of 1×10 ⁶ cells/ml.

Then, it was sensitized with anti DNP (dinitrophenyl)-IgE (100 ng/ml) and incubated in an incubator at 37° C. and 5% CO ₂ for 24 hours. After washing twice with PBS (phosphate buffered saline), DNP-HSA (dinitrophenylated human serum albumin; 100 ng/ml) was treated for 1 hour to induce an immune response. At this time, in order to evaluate the atopic improvement effect by Thiazole, anti-DNP (dinitrophenyl)-IgE was treated and thiazole (100 μM) was treated and incubated for 24 hours, and then an immune response was induced with DNP-HSA. . In the case of negative control cells, they were treated with DMSO instead of thiazole and incubated for 24 hours, and then an immune response was induced with DNP-HSA.

2) Cell culture of keratinocytes

Keratinocytes (human keratinocyte cell line, HaCaT) were purchased and used from ATCC (Manassas, VA, USA). Cells were cultured using a DMEM culture medium containing 10% FBS (fetal bovine serum) and antibiotics. The culture vessel was a 75T-flask and a 6-well plate, and cultured in a 37°C incubator supplied with 5% CO2. The culture medium was changed every 3 to 4 days, and when cells were excessively proliferated, they were subcultured. The dispensed HaCaT cells (5×10 ⁵ /well) were cultured for 24 hours and washed with PBS. To induce an immune response to keratinocytes, TNF-α (tumor necrosis factor-α) (10 ng/ml) and IFN-γ (interferon gamma) (10 ng/ml) were treated together in DMEM medium without FBS. And incubated for 24 hours. At this time, in order to evaluate the immune response improvement effect by thiazole, 100 μM of thiazole was treated with the immune response inducer at the same time, and in the case of negative control cells, DMSO instead of thiazole was treated with the immune response inducing material at the same time and cultured for 24 hours. .

3) Histamine release assay

In order to measure the concentration of histamine secreted from mast cells that induced the immune response, the immune reaction was terminated by standing in an ice bath for 10 minutes, and the culture was centrifuged at 10,000 xg for 10 minutes, and the supernatant was taken. Transferred to a 96-well plate. Histamine was quantified using ENZO's histamine release assay ELISA kit, and was measured at 450 nm wavelength of Infinite M200 pro microplate reader (TECAN).

4) RNA isolation and real-time PCR (quantitative reverse-transcription polymerase chain reacion) using the Trizol method

After removing the supernatant of the mast cells that induced the immune response, add 1 ml of trizol, leave for 2 minutes, add chloroform, vortex for 10 seconds, and centrifuge at 2,400 xg for 10 minutes, and then the supernatant The solution was taken, and the same amount of isopropanol was mixed and shaken. The supernatant was removed by centrifugation at 16,000 xg for 25 minutes, and the pellet was dissolved in 20 μl of DEPC (diethyl pyrocarbonate)-DW and stored at -20°C before use in the experiment. RT-PCR was reacted at 45°C for 30 minutes and 94°C for 5 minutes using a one-step RT-PCR PreMix kit (iNtRON Biotechnology, Korea), followed by denaturation at 94°C for 30 seconds, and 30 at 55°C. After annealing for a second, the cycle of stretching at 72°C for 1 minute was repeated 32 times, and the last extension was performed at 72°C for 5 minutes, followed by PiQ SYBR green supermix (Bio-Rad ) And CFX Connect™ Real-Time PCR Detection System (Bio-Rad) was used to perform quantitative PCR, and the primer sequence used at this time is as shown in [Table 3].

[Table 3]

Primer sequence used for RT-PCR

3-2. Experiment result

1) Changes in histamine secretion

To confirm the effect of thiazole on histamine secreted by degranulation from activated mast cells, the concentration of histamine in the cell culture was measured by an enzyme immunoreaction method. As a result, the histamine concentration was significantly increased in control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA compared to normal mast cells. On the other hand, as a result of treatment with thiazole while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, the concentration of histamine was significantly reduced (FIG. 7).

2) Changes in inflammatory cytokine production in mast cells

In control cells sensitized with anti-DNP (dinitrophenyl)-IgE and induced an immune response with DNP-HSA, the expression of inflammatory cytokines IL-4, IL-13, and TNF-α was significantly increased compared to normal mast cells. . On the other hand, as a result of treating thiazole while inducing an immune response by anti-DNP (dinitrophenyl)-IgE and DNP-HSA, increased inflammatory cytokines (IL-4, IL-13 and TNF-α) All expressions were significantly reduced (Fig. 8).

3) Changes in inflammatory cytokine production in keratinocytes

In control cells induced immune response with TNF-α and IFN-γ, the expression of inflammatory cytokines IL-1β, IL-6 and IL-8 was significantly increased compared to normal keratinocytes. On the other hand, as a result of treating keratinocytes with thiazole along with TNF-α and IFN-γ, the expression of inflammatory cytokines (IL-1β, IL-6 and IL-8) increased due to the immune response were all significantly. Decreased (Fig. 9).

Hereinafter, examples of preparation of pharmaceuticals, foods, or cosmetics containing ethyl vanillin, suberic acid or thiazole according to the present invention as an active ingredient will be described, but the present invention is not intended to limit them, but is intended to be described in detail. . Using ethyl vanillin, suberic acid, or thiazole having excellent preventive and therapeutic (or preventive and improving effect) effects of the allergic disease or atopic dermatitis, according to the following composition components and composition ratios, Preparation Examples 1 to 3 Pharmaceutical, food or cosmetic compositions were prepared according to conventional methods.

[Production Example 1] Preparation of pharmaceutical composition

<1-1> Preparation of powder

Ethylvanillin, suberic acid or thiazole 20 mg

100 mg of lactose hydrate

Talc 10 mg

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<1-2> Preparation of tablets

Ethylvanillin, suberic acid or thiazole 10 mg

Corn starch 100 mg

100 mg of lactose hydrate

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet preparation method.

<1-3> Preparation of capsules

Ethylvanillin, suberic acid or thiazole 10 mg

Microcrystalline cellulose 3 mg

Lactose hydrate 14.8 mg

Magnesium stearate 0.2 mg

After mixing the above ingredients, the capsules were prepared by filling them into gelatin capsules according to a conventional capsule preparation method.

<1-4> Preparation of injection

Ethylvanillin, suberic acid or thiazole 10 mg

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Sodium monohydrogen phosphate 26 mg

After mixing the above ingredients, it was prepared with the above ingredient content per ampoule (2 mL) according to a conventional method for preparing injections.

<1-5> Preparation of liquid formulation

10 mg of ethyl vanillin, suberic acid or thiazole

Isomerized sugar 10 mg

Mannitol 5 mg

Purified water appropriate amount

Lemon flavor appropriate amount

The above ingredients are dissolved by adding each ingredient to purified water according to a conventional manufacturing method, added an appropriate amount of lemon flavor, adjusted to 100 mL by adding purified water, sterilized, and filled in a brown bottle to prepare a liquid formulation.

[Production Example 2] Preparation of health food

<2-1> Manufacture of health supplement food

10 mg of ethyl vanillin, suberic acid or thiazole

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

Vitamin B ₁₂ 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinic acid amide 1.7 mg

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 30 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health food, but it may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. , To prepare granules, and can be used to prepare a health food composition according to a conventional method.

<2-2> Manufacture of health drinks

10 mg of ethyl vanillin, suberic acid or thiazole

15 g of vitamin C

100 g of vitamin E (powder)

19.75 g of iron lactate

Zinc oxide 3.5 g

3.5 g of nicotinic acid amide

0.2 g of vitamin A

0.25 g of vitamin B1

0.3 g of vitamin B2

Purified water quantification

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 liter container, sealed and sterilized, and then stored in a refrigerator. It is used in the manufacture of health beverage composition.

The composition ratio is a mixture of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

[Production Example 3] Preparation of cosmetic composition

Hereinafter, a preparation example of a cosmetic composition containing the extract of the present invention will be described, but the present invention is not intended to limit it, but is intended to be described in detail.

<3-1> Nutritional lotion (milk lotion)

2.0% by weight of ethyl vanillin, suberic acid or thiazole

5.0% by weight of squalane

4.0 wt% beeswax

1.5% by weight of polysorbate 60

1.5% by weight of sorbitansquioleate

0.5% by weight of liquid paraffin

Caprylic/Capric Triglyceride 5.0% by weight

3.0% by weight of glycerin

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

0.1% by weight of carboxyvinyl polymer

0.2% by weight of triethanolamine

Preservative, color, and fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for a relatively nutrient lotion in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

<3-2> Flexible lotion (skin lotion)

2.0% by weight of ethylvanillin, suberic acid or thiazole

3.0% by weight of glycerin

2.0% by weight of butylene glycol

2.0 wt% propylene glycol

0.1% by weight of carboxyvinyl polymer

PEG 12 nonylphenyl ether 0.2% by weight

Polysorbate 80 0.4% by weight

Ethanol 10.0% by weight

0.1% by weight of triethanolamine

Preservative, color, and fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for a relatively soft lotion in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

<3-3> Nutrition cream

2.0% by weight of ethylvanillin, suberic acid or thiazole

Polysorbate 60 1.5% by weight

0.5% by weight of sorbitansquioleate

PEG60 hydrogenated castor oil 2.0% by weight

Liquid paraffin 10% by weight

5.0% by weight of squalane

Caprylic/Capric Triglyceride 5.0% by weight

5.0% by weight of glycerin

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

0.2% by weight of triethanolamine

Appropriate amount of preservative

Appropriate amount of pigment

Suitable amount of fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for a nutritional cream in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

<3-4> Massage cream

1.0% by weight of ethylvanillin, suberic acid or thiazole

10.0 wt% beeswax

Polysorbate 60 1.5% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitansquioleate 0.8% by weight

Liquid paraffin 40.0% by weight

5.0% by weight of squalane

Caprylic/Capric Triglyceride 4.0% by weight

5.0% by weight of glycerin

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

0.2% by weight of triethanolamine

Preservative, color, and fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for a massage cream in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

<3-5> pack

1.0% by weight of ethylvanillin, suberic acid or thiazole

13.0% by weight of polyvinyl alcohol

Sodium carboxymethylcellulose 0.2% by weight

5.0% by weight of glycerin

0.1% by weight allantoin

Ethanol 6.0% by weight

PEG 12 nonylphenyl ether 0.3% by weight

Polysorbate60 0.3% by weight

Preservative, color, and fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for the pack in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

<3-6> gel

0.5% by weight of ethyl vanillin, suberic acid or thiazole

Sodium ethylenediamine acetate 0.05% by weight

5.0% by weight of glycerin

0.3% by weight of carboxyvinyl polymer

Ethanol 5.0% by weight

PEG 60 hydrogenated castor oil 0.5% by weight

0.3% by weight of triethanolamine

Preservative, color, and fragrance

Purified water to 100% by weight

The above blending ratio is a mixture of ingredients suitable for a gel in a preferred embodiment, but the blending ratio may be arbitrarily modified, and can be prepared according to a conventional manufacturing method in the cosmetic field.

The above blending ratio is a mixture of ingredients suitable for a cosmetic composition in a preferred embodiment, but can be applied to cosmetics for various purposes including other colored cosmetics, and a drug that can be applied thinly to the human body according to its efficacy, that is, It can be used for manufacturing as an ointment, and the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the purpose of use.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not limiting.

Claims (9)

1. Ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole) or a pharmaceutical composition for the prevention or treatment of allergic diseases or atopic dermatitis comprising as an active ingredient a pharmaceutically acceptable salt thereof.
2. The method of claim 1,

   The allergic diseases include edema, anaphylaxis, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, 　 contact dermatitis, urticaria, pruritus, insects. Allergic disease or atopic dermatitis prevention or treatment pharmaceutical composition comprising at least one selected from the group consisting of allergy, food allergy and drug allergy.
3. The method of claim 1,

   The active ingredient is an allergic disease or atopic dermatitis, characterized in that it reduces the expression of IL-4, IL-13, TNF-α, IL-1β, IL-6 or IL-8, or reduces the amount of histamine secretion. Pharmaceutical composition for the prevention or treatment of.
4. A quasi-drug composition for preventing or improving allergic diseases or atopic dermatitis comprising ethyl vanilin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof as an active ingredient.
5. Ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole) or a cosmetic composition for the prevention or improvement of allergic diseases or atopic dermatitis comprising as an active ingredient a cosmetically acceptable salt thereof.
6. Ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole) or a perfume composition for the prevention or improvement of allergic diseases or atopic dermatitis, comprising as an active ingredient a cosmetically acceptable salt thereof.
7. Ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole), or a dietary functional food composition for the prevention or improvement of allergic diseases or atopic dermatitis containing as an active ingredient a food acceptable salt thereof.
8. Allergic disease comprising the step of administering or taking a composition comprising ethyl vanilin, suberic acid, thiazole, or a pharmaceutically acceptable salt thereof as an active ingredient to an individual; or How to prevent or treat atopic dermatitis.
9. Ethyl vanilin (Ethyl vanilin), suberic acid (Suberic acid), thiazole (Thiazole) or a composition containing a pharmaceutically acceptable salt thereof as an active ingredient for the prevention or treatment of allergic diseases or atopic dermatitis.

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