KR20220076984A - Composition for Anti-Allergy Using an Extract of Persicaria longiseta - Google Patents

Abstract

The present invention discloses a composition for anti-allergy using an extract of the genus spp. which has an inhibitory activity of degranulation when treated with the mast cell line RBL-2H3 (rat basophilic leukemia) activated with IgE and antigen (DNP-BSA).

Description

Composition for Anti-Allergy Using an Extract of Persicaria longiseta

The present invention relates to a composition for anti-allergy using an extract of Gaeyeogwi ( Persicaria longiseta ).

Allergy is a compound word created by combining the Greek words 'allos' meaning 'change' and 'ergo' meaning 'action'. As the etymology is like this, an allergy refers to a hypersensitivity reaction such as urticaria, itchiness, runny nose, and cough in a specific person due to an abnormality in innate or acquired immune function to an allergen, a substance that does not show any reaction to ordinary people. Typical allergens include pollen, drugs, vegetable fibers, bacteria, mold, house dust mites, food (eg, egg albumin, milk protein, peanuts), hair dye, and chemicals. Due to an increase in substances, environmental pollution, a decrease in immune function due to stress, and changes in diet, these allergic diseases are increasing worldwide.

Allergic reactions are classified into five types: type I anaphylaxis type, type II cytolytic type, type III immune complex type, type IV cell mediated type, and type V stimulatory hypersensitivity type, most of which belong to type I, and generally Allergic reaction is used in the same sense as type I (Roitt I, Brostoff J, Male D. Immunology 6th eds. U.S.A. Mosby. 2001:323-383).

Representative allergic diseases include atopic dermatitis, bronchial asthma, allergic rhinitis, allergic keratitis, and skin urticaria. In these allergic diseases, mast cells are known as important effector cells (Wills-Karp M., et al., Science, 282:2258-61, 1998; Grunig G., et al., Science, 282:2261-2263, 1998). Mast cells are cells that are widely distributed in organs throughout the body, such as the skin, respiratory system, lymphatic vessels, mucous membranes of the gastrointestinal tract, blood vessels, and brain (Ishizaka t., et al. J. Immunol. 119:1589, 1997).

Allergic reactions are induced through activation of intracellular signaling pathways when an allergen binds to IgE, which is bound to the IgE high affinity receptor (FcεRI) on the surface of mast cells. Specifically, when an allergen invades a living body, T-helper cells and B cells are sequentially activated to generate IgE. Allergens that invade the living body are recognized by antigen-presenting cells, which differentiate Th0 (T helper cell type 0) into Th2 cells by presenting the allergen. The differentiated Th2 cells secrete cytokines such as IL-4, IL-5, and IL-13, which promote the development of acid leukocytes in the bone marrow and induce acid leukocytes to the inflamed tissue, as well as B cells. It induces the production of IgE and IgG1.

The generated IgE binds to the high-affinity IgE receptor (FcRIα) of the mast cell membrane, and when allergens such as mites and pollen bind to it and cross-link it, the receptor (FcRIα) aggregates. By crosslinking and aggregation of the receptor (FcRIα), mast cells are activated and the concentration of calcium ions (Ca ⁺⁺ ) in the cytoplasm, etc. rises. The increase in calcium ion concentration acts on several types of intracellular regulators such as cholesterol in the cell membrane and the cytoskeleton system, and moves the granules to the cell membrane, and at the same time degranulation through vacuolization of granules, fusion of granules with each other, and fusion of granules with cell membranes. causes When degranulation occurs, preformed mediators stored in mast cells, such as histamine, are released and, at the same time, newly synthesized prostaglandin and leukotriene are secreted together. Chemical transmitters liberated by degranulation include histamine, serotonin, and leukotriene, which have actions such as vasodilation, enhancement of vascular permeability, contraction or hypersecretion of bronchial smooth muscle, and NCF acting as a chemotactic factor of inflammatory cells such as eosinophils and neutrophils. , ECF, PAF, etc. cause an allergic reaction (Ennis M, et al., Br J Pharmacol 70:329-334, 1980; Metcalfe DD, et al., Crit Rev Immunol 3:23-74, 1981; Miyajima I, et al., J Clin Invest 99:901-914, 1997; Church MK, et al., J Allergy Clin Immunol 99:155-160, 1997; Jones DA, et al., J Biol Chem 268:9049- 9054, 1993).

Cyclooxygenase (COX) and lipoxygenase (LO) are known to catalyze a part of the process of producing prostaglandins and leukotrienes, which are allergy symptom-causing substances, from arachidonic acid (Sekiya K , et al., Biochem Biophys Acta 713:68-72, 1982)

Therefore, substances that inhibit the activation of mast cells, inhibit the release of histamine, or inhibit the synthesis of prostaglandins and leukotrienes can be effective therapeutic drugs for allergies.

Currently, anti-allergic drugs include antihistamines (alkylamines, desloratadine, etc.), mast cell stabilizers (cromoglycate, etc.), leukotriene blockers (montelukast, etc.), and anti-IgE antibody (Omalizumab). However, in most cases, the effect of these drugs is temporary and the side effects are severe in many cases. Therefore, it can be said that there is still a need to develop a new substance that has a preventive and ameliorating effect on allergic diseases, few side effects and a continuous effect, and in particular, natural products have the advantage of relatively low toxicity.

The present invention discloses anti-allergic activity of the extract

It is an object of the present invention to provide a composition for anti-allergy using the extract

Other objects or specific objects of the present invention will be set forth below.

As confirmed in the Examples and Experimental Examples below, it was confirmed that the present invention inhibits degranulation when treated with the mast cell line RBL-2H3 (rat basophilic leukemia) activated with IgE and antigen (DNP-BSA). It is completed by doing

In consideration of the above, the present invention can be identified as an anti-allergy composition or antihistamine composition comprising the extract of Gaejaekyi as an active ingredient.

In the present specification, the term "gaeyeogwi extract" refers to the stem, leaf, fruit, flower, root, outpost, above-ground part, underground part, or a mixture thereof, which is an extraction target, water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) ), methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or It refers to an extract obtained by leaching using a mixed solvent, an extract obtained by using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract, and the extraction method is the polarity of the active material, extraction degree, and preservation degree In consideration of this, any method such as chilling, reflux, warming, ultrasonic radiation, and supercritical extraction can be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by methods such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, it refers to 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 addition, as used herein, the term "active ingredient" refers to a component that alone exhibits the desired activity or can exhibit activity together with a carrier that has no activity by itself.

In addition, in the present specification, "anti-allergy" is meant to include improvement (relief of symptoms), treatment, and prevention (suppression or delay of onset) of allergic diseases as defined below.

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

The anti-allergic composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit the amelioration activity of the intended treatment according to the use, formulation, purpose of formulation, etc. It will be determined within the range of 0.001% to 15% by weight based on the total weight of the composition. Herein, the term "effective amount" refers to the intended medical and pharmacological effects, such as the improvement of allergic diseases, when the composition of the present invention is administered to a mammal, preferably a human subject to the application, during the administration period as suggested by a medical professional. It refers to the amount of the active ingredient included in the composition of the present invention. Such an effective amount can be determined empirically within the ordinary ability of one of ordinary skill in the art.

In addition to the active ingredient, the anti-allergic composition of the present invention may further include any compound or natural extract known to have anti-allergic activity and safety has already been verified to increase/reinforce anti-allergic activity.

Specifically, as such compounds or extracts, Enterococcus faecalis heat-treated dry powder, which has been individually recognized for its 'relieving hypersensitivity immune response' function according to the Korean 「Health Functional Food Act」, complexes such as guava leaf extract, Actinidia extract, leaf extract, Composite such as picaopreto powder, PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol), L. sakei Probio 65, which has been individually recognized for its 'improving sensitive skin condition' function, oil containing gamma-linolenic acid , fruit and vegetable-derived lactic acid bacteria ( L. plantarum CJLP133 ), probiotics ATP, and the like.

One or more of these compounds or natural extracts may be included in the anti-allergic composition of the present invention together with its active ingredient.

In a specific embodiment, the anti-allergic composition of the present invention can be grasped as a food composition. When the composition of the present invention is identified as a food composition, its use may be understood to inhibit allergic skin hypersensitivity reaction.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated drinks, and ion drinks, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, confectionery, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be manufactured.

In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to Korea's "Health Functional Food Act", or confectionery, beans, tea, and beverages according to each food type in the Food Ordinance of Korea "Food Sanitation Act" (Ministry of Food and Drug Safety Notification "Food Standards and Specifications") , special purpose food, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives can be generally understood as substances that are added and mixed or infiltrated into food in manufacturing, processing, or preserving food. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Ordinance in accordance with the laws of each country that regulates the manufacture and distribution of food (“Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Specifications” announced by the Ministry of Food and Drug Safety), food additives are classified into chemically synthetic products, natural additives, and mixed preparations in terms of ingredients. It is divided into agents, preservatives, emulsifiers, acidulants, thickeners, etc.

The sweetener is used to impart appropriate sweetness to food, and both natural and synthetic ones may be used in the composition of the present invention. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents may be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural product, the purpose of nutritional enhancement in addition to flavor may be concurrently used. The natural flavoring agent may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or obtained from green tea leaves, dandelion leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. Optionally, a synthetic flavoring agent may be used, and the synthetic flavoring agent may be an ester, an alcohol, an aldehyde, a terpene, or the like.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. are mentioned, and acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used as an acidulant. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.

As a thickening agent, a suspending agent, a settling agent, a gel former, a bulking agent, etc. can be used.

The food composition of the present invention may contain, in addition to the food additives described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as 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, dibenzoylthiamine, and the like. Minerals include calcium preparations such as calcium citrate, magnesium stearate Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.

In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the product type.

In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Ordinance of each country or the Food Additives Code.

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

The pharmaceutical composition of the present invention may be prepared as an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of the combination of two or more routes is a case in which two or more formulations of drugs according to the route of administration are combined. For example, one drug is first administered by an intravenous route and the other drug is secondarily administered by a local route.

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

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art together with a suitable carrier It can be prepared in a formulation such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol and the like. In the case of formulation, an appropriate binder, lubricant, disintegrant, colorant, diluent, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and lubricants include sodium oleate. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium salt and calcium salt, polyethylene glycol, etc., and the disintegrant includes starch, methyl cellulose, 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 for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or isotonic solution such as 5% dextrose may be used. . When formulated for transdermal administration, it can be formulated in the form of an ointment, a cream, a lotion, a gel, an external solution, a pasta agent, a liniment agent, an air roll, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like can be used.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.

A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and the route of administration. It can be in the range /kg. Administration may be performed once or divided into several times a day. These dosages should not be construed as limiting the scope of the invention in any respect.

In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition. When the composition of the present invention is identified as a cosmetic composition, its use may be understood as a use for suppressing allergic skin troubles, alleviating allergic skin irritation, and the like.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in terms of its use or by law, and specifically functional cosmetics, non-functional products having uses such as improvement of skin troubles and improvement of atopic dermatitis It may be general cosmetics and the like. In terms of product form, it may take any product form, and specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can take the form of products such as foundation and spray. In the specific product form, it may be in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and fragrances, and carriers.

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

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 particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc. may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.

The cosmetic composition of the present invention can be prepared according to the method for preparing a cosmetic composition conventionally performed in the art, except that it contains the active ingredient exhibiting anti-allergic activity.

As described above, according to the present invention, it is possible to provide a composition for anti-allergy using an extract of the sagebrush.

The composition for anti-allergy of the present invention may be commercialized as food, cosmetics, drugs, etc. for the purpose of improving allergic diseases.

1 is a result showing the degranulation inhibitory activity in the mast cell line RBL-2H3 (rat basophilic leukemia) activated with IgE and antigen (DNP-BSA) of Gaeyeogwi extract.

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

< Example > Manufacture of extract

1 L of 50% ethanol was added to 100 g of dried persicaria longiseta above ground powder, followed by immersion extraction once for 24 hours at room temperature, followed by filtration with filter paper. The obtained 50% ethanol filtrate was concentrated under reduced pressure, and then freeze-dried to obtain an extract of the succulents.

< Experimental example > anti-allergy Activity test - β-hexosaminidase assay

Rat basophilic leukemia cell line, RBL-2H3 cell line, in MEM medium containing 15% FBS, 100 unit/ml penicillin and streptomycin and 2×10 ⁵ /well in a 24 well plate at 37°C and 5% CO ₂ condition. After dispensing, culture was performed for 24 hours.

After removing the supernatant, 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 (25mM PIPES, 119mM NaCl, 5mM KCl, 1mM CaCl ₂ , 0.4mM MgCl ₂ , 40mM NaOH, 5.6mM glucose, 0.1% BSA) and pre-incubated for 10 minutes. After pre-incubation, the samples of Example were treated for 20 minutes by concentration and then reacted with DNP-BSA for 20 minutes. After terminating the reaction by standing on ice for 10 minutes, the supernatant was placed in a 96 well plate, 1 mM P-nitrophenyl-acetyl-β-D-glucosaminide was added, and the reaction was carried out at 37° C. for 1 hour. Stop solution (0.1M NaHCO ₃ , 0.1M Na ₂ CO ₃ ) was added and the reaction was terminated, and absorbance was measured with an ELISA reader at a wavelength of 405 nm. As a positive control, ketotifen (100 μg/ml) was used.

The results are shown in FIG. 1, and it can be seen that the sample of Example inhibits degranulation in a concentration-dependent manner, and the IC ₅₀ is 40.75 μg/mL. The positive control, ketotifen, showed an IC ₅₀ of 38.13 μg/mL. Ketotifen, used as a positive control, is an antihistamine, which is used for allergic skin diseases such as allergic rhinitis, atopic dermatitis, and allergic asthma.

Claims (6)

A composition for anti-allergy comprising an extract of Gaeji-gyeon as an active ingredient.
The method of claim 1,
The anti-allergic composition, characterized in that the extract is an extract obtained by extraction with water, ethanol, or a mixed solvent thereof.
The method of claim 1,
The anti-allergic means the improvement, treatment of allergic diseases, inhibition or delay of the onset of such diseases,
The allergic disease is atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, allergic contact dermatitis (allergic contact dermatitis) ), and an anti-allergic composition, characterized in that it is one selected from the group consisting of allergic keratitis.
4. The method according to any one of claims 1 to 3,
The composition is an anti-allergic composition, characterized in that it is a pharmaceutical composition.
4. The method according to any one of claims 1 to 3,
The composition is an anti-allergic composition, characterized in that it is a food composition.
4. The method according to any one of claims 1 to 3,
The composition is an anti-allergy composition, characterized in that it is a cosmetic composition.

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