KR101946121B1 - Anti-allergic composition and composition for improving atopic dermatitis using 5-hydroxy-6,7-dimethoxyphthalide - Google Patents

Abstract

The present invention discloses a composition for anti-allergy using 5-hydroxy-6,7-dimethoxyphthalide and a composition for ameliorating atopic dermatitis, which is isolated from P. falciparum extract.
The 5-hydroxy-6,7-dimethoxyphthalide has inhibitory activity on degranulation when treated with an adipose cell line RBL-2H3 (rat basophilic leukemia) activated with gE and antigen (DNP-BSA) The SCORAD index is significantly lowered in clinical trials in patients.

Description

TECHNICAL FIELD The present invention relates to an anti-allergic composition and a composition for improving atopic dermatitis using 5-hydroxy-6,7-dimethoxytryptamide isolated from P. falciparum extract, 7-dimethoxyphthalide}

The present invention relates to a composition for anti-allergy using 5-hydroxy-6,7-dimethoxyphthalide and a composition for improving atopic dermatitis.

Recently, allergic diseases are increasing rapidly. The number of patients with allergies in Korea is more than 6 million, and the number of patients continues to increase due to the increase in westernized living environment, pollution, and use of various chemicals. The global market for allergies in 2012 is estimated at W20trn and the global market for functional foods with allergies is estimated at W5trn in 2012 (Nutrition Business Journal 'U.S. Market Overview' Vol. XVII, No.5 / 6 (2012)).

Allergy refers to that the immune system in the body is excessively adversely affected by external foreign matter or external environmental stimulus, causing pollenosis, urticaria, rhinitis, asthma, atopy and the like. For others, problematic substances can cause allergies to certain people. It is also classified as an environmental disease because it is caused by external stimuli.

Allergic reactions are classified into five types: anaphylaxis type, type cell type, immune complex type, type cell mediated type and type stimulation type hypersensitivity type. Most of them belong to the formulations. (Roitt et al., Immunology 6th eds USA, Mosby 323-383 (2001)).

Mast cells and basophils are known to be the major cells causing allergic diseases through secretion of allergen-induced inflammatory mediators by antigen stimulation (Blank, U. et al., Allergy 68 (9): 1093 (101) (2013); Schroeder JT Adv Immunol 101: 123-61 (2009); Galli, SJ New Eng J. Med. 328: 257-265 (1993)). Formulation Allergies are induced by specific antigens such as food, house dust mite, ticks, and pollen. When the human body is exposed to the antigen, T-helper cells and B cells are sequentially activated to produce IgE. IgE binds to the mast cell receptor (FcRI) to form a complex, which when re-sensitized to the same antigen and cross-linked with IgE activates mast cells and initiates an allergic reaction (Metcalfe DD et al., Crit. Rev. Immunol 3: 23-74 (1981); Church MK and Levi-Schaffer FJ Allergy Clin. Imm. 99: 155-60 (1997)). When mast cells are activated, the concentration of Ca ++ in the cytoplasm increases and acts on various kinds of intracellular regulators such as cholesterol and cytoskeleton of the cell membrane to move the granules to the cell membrane, Fusion and fusion of granules and cell membranes lead to degranulation. These allergic reactions occur in two stages: within 5-10 minutes after antigen challenge, secrete already produced mediators such as histamine, -hexosaminidase, and serotonin from mast cells and secrete newly synthesized lipid mediators to produce urticaria, The smooth muscle reaction with the ischemic vasculature causes a major hypersensitivity reaction, which is called an early phase reaction. After 2-4 hours, inflammatory cells such as T-helper type 2 (Th2), eosinophils, neutrophils, and mononuclear cells are activated to produce a late phase reaction to synthesize and secrete cytokines, thereby sustaining inflammatory infiltration Chang, TW and Shiung Y. J Allergy Clin Immunol 117 (6): 1203-12 (2006); Abul K. et al., Cellular Molecular Immunology, 6th Edition, Seoul: 2008); Schwartz LB et al., J Immunol. 126 (4): 1290-4 (1981)).

Therefore, a substance that inhibits the activation of mast cells, inhibits histamine release, or inhibits the synthesis of prostaglandins and leukotrienes, can be an effective therapeutic drug for allergies.

Currently, anti-allergic drugs are used such as alkylamines, desloratadine, mast cell stabilizer (cromoglycate etc.), leukotriene blockers (montelukast etc.) and anti-IgE antibodies (Omalizumab) And most of the side effects are severe. Therefore, it is necessary to develop a new substance which has the effect of prevention and improvement of allergic diseases, and the side effect is small and the effect is constant, and the technology using natural substances with low toxicity is being developed.

Atopic dermatitis is a chronic or recurrent dermatitis that can occur in infants and children, and it is characterized by symptoms such as pruritus, dry skin, swelling around the hair follicle, hypophytosis, ecchymosis, and eczema lesions (Korean J Invest Dermatol. 67-72, 2007). A person suffering from atopic dermatitis usually exhibits an allergic reaction to foreign substances such as house dust, ticks, animal hair, food, pollen, mold and the like, and atopic dermatitis is a cause of allergic rhinitis, asthma, conjunctivitis (J Clin Invest. 113 (5): 651-7, 2004; J Allergy Clin Immunol. 2003; 112 (2) ): 252-62; Kor J Pharmacogn, 43: 59-65, 2012).

The cause of atopic dermatitis has not yet been clarified yet, but it is known that atopic dermatitis is caused by complex interactions such as genetic and environmental factors and mental stress in modern medicine (Korean Society of Dermatology 2008; 46 (2 Immunologic factors include an abnormal increase in IgE, a central role in cellular immunity, and an increase in serum IgE levels. Th1 / Th2 imbalance due to the increase in number of Th2 cells compared to Th1 cells, and the increase in number of mature cells and eosinophils, increase in number of CD4 + T lymphocytes, etc. (J Invest Dermatol., 96: 523-526,1991; J Invest Dermatol., 97: 389-394,1991; Immunol., 11: 81-88,1999; Curr Drug Targets Inflamm Allergy., 2 : 199-120, 2003; J Allergy Clin Immunol., 107: 871-877, 2001; Adv Immun International Journal of Immunology, 14 (7): 767-773, 2002; Pediatr Allergy Immunol., 19: 605-613, 2008; Kor J Pharmacogn, 43: 59-65, 2012).

The present invention discloses the anti-allergic activity of 5-hydroxy-6,7-dimethoxyphthalide isolated from P. falciparum extract and the activity of improving atopic dermatitis, one of the allergic diseases.

The present invention provides a composition for anti-allergy using 5-hydroxy-6,7-dimethoxyphthalide isolated from P. falciparum extract.

Another object of the present invention is to provide a composition for improving atopic dermatitis using 5-hydroxy-6,7-dimethoxyphthalide isolated from P. falciparum extract.

Other and further objects of the present invention will be described below.

The present invention is based on the finding that 5-hydroxy-6,7-dimethoxyphthalide of the following formula (1) isolated from P. falciparum extract is converted into IgE and antigen (DNP-BSA) (RBL-2H3, rat basophilic leukemia) inhibited degranulation, and significantly lowered the SCORAD index in clinical trials in 100 men and women with atopic dermatitis symptoms It is completed by confirmation. In the granules of mast cells, β-hexosaminidase is also stored in addition to histamine. When mast cells produce degranulation, β-hexosaminidase is released together with histamine. Therefore, β-hexosaminidase inhibits mast cell 48 (4): 315-321 (2005)), the SCORAD index is used as an index substance for evaluating the antitumor activity and antihistamine activity (Immunol., 123: 1445-1450 (1970); J Korean Soc Appl Biol Chem. Is an objective assessment index of atopic dermatitis developed by The European Task Force on Dermatitis (Dermatology. 1997; 195: 10-9).

[Chemical Formula 1]

In view of the foregoing, the present invention provides an antiallergic composition comprising 5-hydroxy-6,7-dimethoxyphthalide, an isomer thereof, a prodrug thereof, a hydrate thereof or a solvate thereof as an active ingredient (or an antihistamine Or a composition for improving atopic dermatitis.

In addition, 5-hydroxy-6,7-dimethoxyphthalide, an active ingredient of the present invention, was tested for inhibitory activity on the degranulation of the extract of Zygomyceta rhizoma, and then the above compound was isolated from the extract to confirm the degranulation inhibitory activity of the compound In the following Examples and Experimental Examples, the degranulation inhibitory activity of the extract of Zygomycorr japonicus was explicitly disclosed by experimental results in that the Zygomycetes mushroom extract contains the above-identified compounds, such as degranulation inhibitory activity It will be obvious to those skilled in the art that the extract of Zanthoxylum mushroom may have degranulation inhibitory activity and the like.

Therefore, the composition for anti-allergy (or the composition for antihistamine) or the composition for improving atopic dermatitis of the present invention may contain an extract of Mushroom mushroom as an active ingredient.

Also, as shown in the following Experimental Example, the active ingredient of the present invention, 5-hydroxy-6,7-dimethoxiphthalide, reduces the amount of transdermal water loss ("TEWL") when applied to patients with atopic dermatitis, .

Accordingly, the present invention can be also understood as a skin moisturizing composition comprising 5-hydroxy-6,7-dimethoxyphthalide or the like or an extract of Mushroom mushroom as an active ingredient in another aspect.

As used herein, isomers include not only essentially pure diastereomers but also optical isomers, as well as conformation isomers (i.e. isomers differing only in the angle of one or more chemical bonds), position isomers, (Especially tautomers) or geometric isomers (e. G., Cis-trans isomers).

In the present specification, the term " prodrug " refers to a drug that chemically changes a drug to control its physical and chemical properties. Although the drug itself does not exhibit physiological activity, it does not exhibit chemical or enzymatic action Which means that the drug can be converted into the original drug and can be used to produce the drug.

In the present specification, the term "hydrate" means a compound having water bonded thereto, and it is meant to include an inclusion compound having no chemical bonding force between water and the compound.

&Quot; Solvate " as used herein means a compound formed between a molecule or ion of a solute and a molecule or ion of a solvent.

In the present specification, " atopic dermatitis " is defined as including any disease classified as atopic dermatitis in the art, regardless of whether it occurs directly or indirectly. In general, atopic dermatitis is classified into infantile atopic dermatitis, infantile atopic dermatitis, adult atopic dermatitis and maternal atopic dermatitis according to the onset period or the object of the invention. In this specification, atopic dermatitis includes all these types of atopic dermatitis .

In the present specification, the term " improvement " is meant to include atopic dermatitis treatment, prevention and improvement (alleviation of symptoms).

In the present specification, the term "extract of Zygomycorrhizae mushroom" refers to an extract of Zygomycetes mushroom itself, its mycelia, or its fruiting body in water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol), methylene chloride, ethylene, acetone, The extract obtained by leaching using ether, chloroform, ethyl acetate, butyl acetate, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof , Carbon dioxide, pentane, or fractions obtained by fractionating the extract. The extraction method refers to a method of extracting a supernatant from a supernatant, such as ice, reflux, warming, ultrasonic irradiation , Supercritical extraction, and the like can be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and leaving with a solvent having a different polarity, the crude extract is adsorbed on a column packed with silica gel, and then a hydrophobic solvent, a hydrophilic solvent, Quot; means fractions obtained as a mobile phase. Also, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying and the like. Preferably an extract obtained by using water, ethanol or a mixed solvent thereof as an extraction solvent, and more preferably an extract obtained by using a mixed solvent of water and ethanol as an extraction solvent.

In the present specification, the term " active ingredient " alone means an ingredient which exhibits the desired activity or which can exhibit activity together with a carrier which is not itself active.

In the present specification, the term " anti-allergy " is meant to include improvement (symptom relief), treatment, prevention (prevention or delay of onset) of an allergic disease as defined below.

In the present specification, the term " allergic disease " means a pathological symptom caused by an allergic reaction mediated by mast cell activation such as degranulation of mast cells. Such allergic diseases include atopic dermatitis, asthma, Allergic rhinitis, allergic conjunctivitis, allergic dermatitis, allergic contact dermatitis, and the like. The term " allergic contact dermatitis "

In the present specification, " antihistamine " means physiological change due to excessive secretion of histamine or improvement of the disease caused by dysfunction. The physiological changes caused by the secretion of histamine or diseases caused by dysfunction include allergic diseases such as nasal obstruction, dizziness, vomiting, hyperacidity, gastroesophageal reflux disease, inflammation, hypotension associated with anapalacsis .

In the present specification, the term " anti-allergy " is meant to include improvement (symptom relief), treatment, prevention (prevention or delay of onset) of an allergic disease as defined below.

In the present specification, the term " allergic disease " means a pathological symptom caused by an allergic reaction mediated by mast cell activation such as degranulation of mast cells. Such allergic diseases include atopic dermatitis, asthma, Allergic rhinitis, allergic conjunctivitis, allergic dermatitis, allergic contact dermatitis, and the like. The term " allergic contact dermatitis "

In the present specification, " antihistamine " means physiological change due to excessive secretion of histamine or improvement of the disease caused by dysfunction. The physiological changes caused by the secretion of histamine or diseases caused by dysfunction include allergic diseases such as nasal obstruction, dizziness, vomiting, hyperacidity, gastroesophageal reflux disease, inflammation, hypotension associated with anapalacsis .

The active ingredient of the composition for anti-allergy, the composition for improving atopic dermatitis and the composition for skin moisturizing (hereinafter referred to as " the composition of the present invention ") of the present invention can be used as an active ingredient for anti- (Effective amount) depending on the intended use, formulation, compounding purpose and the like. A typical effective amount will be determined within the range of 0.001% by weight to 20.0% by weight based on the total weight of the composition. The term " effective amount " as used herein refers to an amount of an effective amount of the composition of the present invention when administered to a mammal, preferably a human, Refers to the amount of the active ingredient contained in the composition of the present invention, which can exhibit antiphlogistic effects. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

The composition of the present invention can be used not only for the effective ingredient but also for the enhancement of the anti-allergic activity, the activity for improving atopic dermatitis, the enhancement of the skin moisturizing activity, or the addition of the similar activity such as skin protecting activity (skin damaging by ultraviolet rays, May further comprise any compound or natural extract known to be safe in the art and known to have a corresponding activity in order to improve the convenience of taking or ingesting and applying the skin.

These compounds or extracts include the national pharmacopoeia (Korean pharmacopoeia), the national health functional food circulation in Korea (in Korea, the health functional food standard and specification), the functional cosmetics circulation in each country (in Korea, Compounds or extracts listed in the official documents such as "Functional Cosmetics Standards and Specifications"), compounds or extracts approved by the respective countries in accordance with the laws of each country that govern the manufacture and sale of pharmaceuticals (in Korea, "Pharmaceutical Affairs Law"), And compounds or extracts that are individually recognized as functional according to the laws of the respective countries that regulate the manufacture and sale of functional foods (in Korea, the "Act on Health Functional Foods"). For example, N-acetylglucosamine, glucosamine, and hyaluronic acid, which have skin moisturizing activity, and an active ingredient that has hypersensitive immune response mitigating activity, such as the Korean Functional Cosmetics Cycle, the Korean Health Functional Food Cycle, PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol) and complexes of Enterococcus faecalis heat-treated dry powder and guava leaf extract, condition L. sakei Probio 65, gamma-linolenic acid-containing maintained with improved activity, gwachae derived lactic acid bacteria (L. plantarum CJLP133), probiotics such as ATP, composites, such as Enterococcus faecalis heat-treated dry powder, a guava leaf extract having anti-allergic activity, Actinidia (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol) and the like, Collagen enzyme degrading peptide having a skin moisturizing activity, collactive collagen peptide, N-acetylglucoside, alpha-bisabolol, and oil-soluble licorice (Glycyrrhiza extract) Extracts of bamboo leaves, melon extracts, brambles, extracts of bamboo, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, antioxidants, acetylglucosamine, konjac potato extract, dandelion extract, rice bran extract, corn germ extract, low molecular collagen peptide, Such compounds or extracts may be such compounds as extract (powder), beeswax alcohol, ubiquinol, coenzyme Q10, tomato extract, grape seed extract, French seaweed bark extract, red ginseng (red ginseng concentrate) and the like.

The composition of the present invention can be identified as a food composition in a specific embodiment.

The food composition of the present invention can be prepared in any form and can be used in various forms such as beverages such as tea, juice, carbonated beverage, ionic drink, processed milk such as milk and request route, gum, rice cake, Such as confectionery, cotton, etc., tablets, capsules, rings, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars and the like. In addition, the food composition of the present invention may be classified into any product category as long as it meets the laws and regulations on the time of manufacture and distribution in the legal and functional category. For example, it is a health functional food according to the "Health Functional Food Act" in Korea or the food standard (standard and standard of food notices) of the Food Sanitation Act of Korea. Beverages, special-purpose foods, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives are generally understood to be substances that are added to foods and mixed or infiltrated into food in the manufacture, processing or preservation of food, and their safety must be ensured since they are ingested daily with food and for long periods of time. Food additives according to national laws regulating the manufacture and distribution of food (in Korea, the "Food Sanitation Act") are stipulated as safety-guaranteed food additives in terms of ingredients or function. In the Food Additives Code of Korea ("Food Additives Standards and Standards"), food additives are defined as chemical compounds, natural additives and mixed preparations in terms of ingredients. Preservatives, emulsifiers, acidulants, and thickeners.

The sweetener is used for imparting a sweet taste suitable for food, and both natural and synthetic sweeteners can be used in the food composition of the present invention. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavors are used to improve taste and flavor, and natural and synthetic flavors can be used. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. If desired, a synthetic flavor agent may be used. As the synthetic flavor agent, esters, alcohols, aldehydes, terpenes and the like may be used.

As the preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid) and the like can be used, and as the emulsifier, acacia gum, carboxymethyl cellulose, xanthan gum, pectin And acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like may be used as the acidulant. The acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste. Examples of the thickening agent include suspending agents, sedimentation agents, gel-forming agents, bulking agents and the like.

The food composition of the present invention may contain physiologically active substances or minerals which are known in the art and which are stable as a food additive, for the purpose of supplementing and reinforcing the functionality and nutrition, in addition to the above-mentioned food additives.

Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E and vitamin B12, tocopherol, dibenzoyl thiamine, etc. Examples of minerals include calcium preparations such as calcium citrate, magnesium stearate , Iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc and the like.

The food composition of the present invention may contain an appropriate amount of the above-mentioned food additives according to the product type so as to achieve the purpose of addition thereof.

With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Code or the Food Additive Code of the respective country.

In another specific embodiment, the composition of the present invention can be identified as a pharmaceutical composition.

The pharmaceutical composition of the present invention may be prepared into oral formulations or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredient. Where the route of administration may be any suitable route including local routes, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucosal tissues, and combinations of two or more routes may be used. An example of a combination of two or more routes is a combination of two or more formulations of the drug according to the route of administration, for example, one drug is administered intravenously and another drug is administered via a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration and formulation, and specific reference may be made to the pharmacopoeia of each country, including the " Korean Pharmacopoeia ".

When the pharmaceutical composition of the present invention is prepared into an oral formulation, it may be formulated into powder, granules, tablets, pills, sugar tablets, capsules, solutions, gels, syrups, suspensions, wafers And the like. Examples of suitable carriers include starches such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, corn starch, potato starch and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Hydroxypropylmethylcellulose and the like; polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol Serol, and the like. In case of formulation, suitable binders, lubricants, disintegrants, coloring agents, diluents and the like may be included as needed. Examples of suitable binders include starch, magnesium aluminum silicate, starch pellets, gelatin, methyl cellulose, sodium carboxymethyl cellulose, polyvinyl pyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax and the like. Examples of the disintegrating agent include starch, methylcellulose, magnesium stearate, magnesium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine and the like.

When the pharmaceutical composition of the present invention is prepared into a parenteral dosage form, it may be formulated in the form of an injection, transdermal drug delivery, nasal aspirate and suppository together with a suitable carrier according to methods known in the art. As the carrier suitable for injection preparation, aqueous isotonic solutions or suspensions may be used. Specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, and isotonic solution such as 5% dextrose may be used . When formulated with a transdermal preparation, it can be formulated in the form of ointments, creams, lotions, gels, external liquids, pastes, liniments, and air-lozenges. Nasal inhalers may be formulated in the form of aerosol sprays using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, witepsol, tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, and sorbitan fatty acid esters.

The formulation of pharmaceutical compositions is well known in the art and can be found, for example, in Remington ' s Pharmaceutical Sciences (19th ed., 1995). This document is considered part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / day, depending on the patient's condition, body weight, sex, age, / kg < / RTI > The administration can be carried out once or several times a day. Such dosages should in no way be construed as limiting the scope of the invention.

The composition of the present invention may be regarded as a cosmetic composition in another specific embodiment.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified into any product category according to the use of the cosmetic composition. Specifically, the cosmetic composition may be a functional cosmetic having a use for improving skin wrinkles, have. Specific examples of the product form include a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax Foundation, spray or the like. In a specific product form, it may be a form of flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

The cosmetic composition of the present invention may contain, in addition to its active ingredient, conventional additives such as stabilizers, solubilizing agents, surfactants, vitamins, colorants and antioxidants, and carriers commonly used in cosmetic compositions.

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a carrier, such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention can be produced by a method for producing a cosmetic composition which is usually carried out in the art, except that it contains the active ingredient.

As described above, according to the present invention, a composition for antiallergy using 5-hydroxy-6,7-dimethoxytryptamide isolated from P. falciparum extract and a composition for improving atopic dermatitis can be provided. In addition, according to the present invention, it is possible to provide a composition for skin moisturizing using 5-hydroxy-6,7-dimethoxyphthalide isolated from P. falciparum extract.

These compositions of the present invention can be commercialized as foods, medicines (medicines, quasi-drugs), cosmetics and the like.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic illustration of the separation of 5-hydroxy-6,7-dimethoxyphthalide.
2 is ¹ H NMR data of 5-hydroxy-6,7-dimethoxyphthalide.
3 is ¹³ C NMR data of 5-hydroxy-6,7-dimethoxyphthalide.
4 is COZY-NMR data of 5-hydroxy-6,7-dimethoxyphthalide.
5 is HMBC-NMR data of 5-hydroxy-6,7-dimethoxyphthalide.
6 is HSQC-NMR data of 5-hydroxy-6,7-dimethoxyphthalide.
7 is ESI-MS data of 5-hydroxy-6,7-dimethoxyphthalide.
FIG. 8 shows the results of β-hexosaminidase assay of 5-hydroxy-6,7-dimethoxyphthalide.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited thereto.

< Example > From the production of the mushroom extract and from the 5- Hydroxy -6,7- Dimethoxyphthalide  Isolation and identification

< Example  1> From the production of the mushroom extract and from the 5- Hydroxy -6,7- Dimethoxyphthalide  detach

Dried mushrooms ( Sparassis crispa , G60) powder was added to 5 kg of 70% ethanol and extracted three times at room temperature. After the obtained extract was filtered, the filtrate was subjected to rotary evaporator to remove the extraction solvent, and the powdery extract was obtained.

The deagglutination inhibitory activity of these extracts was confirmed by the following β-hexosaminidase assay, and the extracts were subjected to Diaion HP-20 column chromatography using methanol / distilled water (0%, 30%, 70%, 100% MeOH) And divided into four small fractions (G60-1 to G60). 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90% of the second small fraction (G60-2) using methanol / distilled water as a mobile phase. (RP-C ₁₈ ) preparative high-performance chromatography was carried out with 10% smaller fractions (G60-2-1 to 10). 10%, 20%, 30%, 40%, 50%, 60%, 70%, and 80% of the fourth small fraction (G60-2-4) using water / acetonitrile as a mobile phase. (RP-C ₁₈ ) preparative high performance chromatography was carried out with 10% (G60-2-4-1 to 10) of the reverse phase column (RP-C ₁₈ ) (G60-2-4-9) was subjected to Sephadex LH-20 column chromatography (50, 60, 70% MeOH mobile phase system) to separate and purify compound 1. Figure 1 shows the schematic diagram of the separation of active compounds.

< Example  2> 5- Hydroxy -6,7- Dimethoxyphthalide  Sympathy

The separated Compound 1 was subjected to physicochemical and spectroscopic analyzes.

White amorphous powder;

ESI-MS m / z, 233.1642 [M + Na] ^&lt; ^{+ &gt};;

¹ H-NMR (700 MHz, acetone-d ₆ )? 6.78 (1H, s, H-4), 5.14 (1H, d, J = 1.4 Hz, H- ), 3.85 (3H, s, H-6).

^{13 C-NMR (175 MHz,} acetone-d 6) δ 68.2 (C-2), 145.0 (C-3), 103.8 (C-4), 157.1 (C-5), 140.1 (C-6), 152.3 (C-7), 109.0 ( C-8), 60.7 (OCH 3), 61.6 (OCH 3).

Compound 1 was found to be in the form of a white amorphous powder from ESI-MS (233.1642 [M + Na] ⁺ ) and the molecular formula was C ₁₀ H ₁₀ O _5. The ¹ H-NMR data and ¹³ C- From the COZY-NMR data, the HMBC-NMR data, and the HSQC-NMR data of Examples 1 to 6, the active compound was identified as 5-hydroxy-6,7-dimethoxyphthalide having the structure of Formula 1 above.

< Experimental Example > Anti-allergy  Activation Experiment and Atopic Dermatitis Improvement Experiment

< Experimental Example  1> Anti-allergy  Active experiment - β- hexosaminidase  assay

The RBL-2H3 cell line, a Rat basophilic leukemia cell line, was cultured in MEM medium containing 15% FBS, 100 units / ml penicillin and streptomycin at 37 ° C and 5% CO ₂ in a 24-well plate at 2 × 10 ⁵ / well Followed by incubation for 24 hours.

After removal of the supernatant, the MEM medium containing 25 ng / ml anti-DNP IgE was added, incubated for 4 hours, washed twice with PIPES buffer 500, and incubated with PIPES buffer containing 4 mM MgCl ₂ , 5.6 mM glucose, 180 and incubated for 10 minutes.

The supernatant was removed and MEM medium containing 25 ng / ml anti-DNP IgE was added and cultured for 4 hours. After incubation, the cells were washed twice with PIPES buffer (25 mM PIPES, 119 mM NaCl, 5 mM KCl, 1 mM CaCl ₂ , 0.4 mM MgCl ₂ , 40 mM NaOH, 5.6 mM glucose, 0.1% BSA) and preincubated for 10 minutes. After the pre-incubation, the samples of the examples were treated for 20 minutes by concentration and then reacted for DNP-BSA for 20 minutes. Ice was left for 10 minutes to terminate the reaction. The supernatant was added to a 96-well plate, and 1 mM P-nitrophenyl-acetyl-β-D-glucosaminyl was added thereto, followed by reaction at 37 ° C. for 1 hour. stop solution (0.1 M NaHCO ₃ , 0.1 M Na ₂ CO ₃ ), and the absorbance was measured with an ELISA reader at a wavelength of 405 nm. Ketotifen was used as a positive control. As a positive control, ketotifen is an antihistamine drug used for allergic asthma such as allergic skin diseases such as allergic rhinitis and atopy.

The results are shown in Fig. Referring to FIG. 8, it can be seen that the compounds of the above Examples inhibit degranulation in a concentration-dependent manner. These results were superior to the ketotifen used as a positive control (see FIG. 8).

< Experimental Example  2> Clinical trial for atopic dermatitis improving activity

Nutritional lotions were prepared with the ingredients and contents described below for the clinical trial for the improvement of atopic dermatitis.

Nutrition lotion ingredients and content

The compound of Example 2 2.0 wt%

2.0% by weight stearic acid

Cetyl alcohol 2.0 wt%

2.0% by weight of glyceryl monostearate

0.5% by weight of polyoxyethylene sorbitan monostearate

0.5% by weight of sorbitan sesquioleate

Glyceryl monostearate / glyceryl stearate /

1.0% by weight of polyoxyethylene stearate

Wax 1.0 wt%

Liquid paraffin 4.0 wt%

Squalane 4.0 wt%

Caprylic / capric triglyceride 4.0 wt%

Carboxyvinyl polymer 0.3 wt%

Butylene glycol 5.0 wt%

Glycerin 3.0 wt%

0.5% by weight triethanolamine

Purified water balance

The nutritional lotion was used to evaluate atopic dermatitis improving activity. As the nutrition lotion of the comparative example, the nutrition lotion containing the same amount of purified water instead of the active ingredient of the present invention as the active ingredient of the present invention was used in the components and contents of the nutrition lotion.

The atopic dermatitis improving activity was divided into two groups of 100 men and women having symptoms of atopic dermatitis over 5 years old and then the nutrition lotion of the present invention and the nutritional lotion of the comparative example were applied to each group for 2 to 3 times a day for 8 weeks .

(TEWAMET TM300, C + K electronic GmbH, Germany), and the SCORAD index (which is higher at higher levels of atopic symptoms) and transdermal water loss ("TEWL" (Corneometer CM825, C + K electronic GmbH, Germany) were measured.

The results are shown in [Table 1] to [Table 3] below as mean standard deviation.

SCORAD SCORAD 0 parking 8 parking Example 1 30.3 15.3 * Comparative Example 32.4 29.5 * p < 0.05

TEWL TEWL 0 parking 8 parking Example 1 23.22 13.50 * Comparative Example 22.95 22.55 * p < 0.05

Moisture content Moisture content 0 parking 8 parking Example 1 34.20 39.0 * Comparative Example 34.78 35.2 * p < 0.05

Statistical processing

For the analysis and analysis of the results of the clinical trial, Excel program and statistical software SPSS Window. Version 10.1 was used. For statistical significance, a significance level of 0.05 was set. Statistical methods used for analysis were analyzed by the independent sample T-test and the corresponding sample T-test.

Claims (9)

5-hydroxy-6,7-dimethoxyphthalide, a hydrate thereof or a solvate thereof as an active ingredient.
The method according to claim 1,
The anti-allergy means improvement, treatment, inhibition or retardation of the onset of an allergic disease,
Wherein the allergic disease is selected from the group consisting of atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis and allergic contact dermatitis.
The method according to claim 1,
The anti-allergy means improvement, treatment, inhibition or retardation of the onset of an allergic disease,
Wherein the allergic disease is atopic dermatitis.
delete 4. The method according to any one of claims 1 to 3,
Wherein the composition is a pharmaceutical composition.
5-hydroxy-6,7-dimethoxyphthalide, a hydrate thereof or a solvate thereof as an active ingredient.
5-hydroxy-6,7-dimethoxyphthalide, a hydrate thereof or a solvate thereof as an active ingredient.
5-hydroxy-6,7-dimethoxyphthalide, a hydrate thereof or a solvate thereof as an active ingredient.
5-hydroxy-6,7-dimethoxyphthalide, a hydrate thereof or a solvate thereof as an active ingredient.

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