KR20240112687A - Composition for Anti-Allergy Using an Extract of Caragana sinica - Google Patents

Abstract

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

Description

Composition for anti-allergy using an extract of Caragana sinica}

The present invention relates to an anti-allergy composition using Caragana sinica extract.

Allergy is a compound word created by combining the Greek words 'allos', meaning 'change', and 'ergo', meaning 'action', and has the meaning of abnormal biological reaction. As the etymology suggests, an allergy is a substance that causes no reaction in normal people, an allergen, which causes hypersensitivity reactions such as hives, itchiness, runny nose, and cough in certain people due to abnormalities in the innate or acquired immune function. Typical allergens include pollen, drugs, vegetable fibers, bacteria, mold, house dust mites, food (ovalbumin, milk protein, peanuts), dyes, and chemicals. Recently, the increase in indoor living, new materials, and chemicals can cause allergies. These allergic diseases are increasing worldwide due to an increase in substances, environmental pollution, decreased immune function due to stress, and changes in dietary habits.

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 irritant hypersensitivity reaction type. Most of them belong to type I, and generally When referring to allergic reaction, it 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, and mast cells are known to be important effector cells in these allergic diseases (Wills-Karp M., et al., Science, 282:2258-61, 1998; Grunig G., et al., Science, 282:2261-2263, 1998). Mast cells are cells widely distributed in organs throughout the body, such as the skin, respiratory tract, lymphatic vessels, gastrointestinal mucosa, blood vessels, and brain (Ishizaka t., et al. J. Immunol. 119:1589, 1997).

Allergic reactions are induced through activation of intracellular signaling pathways when allergens bind to IgE bound to the IgE high-affinity receptor (FcεRI) on the surface of these mast cells. Specifically, when allergens invade the body, T-helper cells and B cells are sequentially activated to produce IgE. Allergens that invade the body are recognized by antigen-presenting cells, and these antigen-presenting cells differentiate Th0 (T helper cell type 0) into Th2 cells through allergen presentation. These differentiated Th2 cells secrete cytokines such as IL-4, IL-5, and IL-13, which not only promote the development of acid leukocytes in the bone marrow and induce acid leukocytes into inflamed tissues, but also induce B cells. It acts on and induces the production of IgE and IgG1.

The IgE produced in this way binds to the high-affinity IgE receptor (FcRIα) on the mast cell membrane, and when allergens such as mites and pollen bind to it and form a cross-link, the receptor (FcRIα) aggregates. Mast cells are activated by cross-linking and aggregation of the receptor (FcRIα), and the concentration of calcium ions (Ca ⁺⁺ ) in the cytoplasm increases. The increase in calcium ion concentration acts on various types of intracellular regulatory factors such as cholesterol in the cell membrane and the cytoskeleton system, causing granules to move to the cell membrane, and at the same time, degranulation through vacuolization of granules, fusion of granules with each other, and fusion of granules and cell membranes. causes When degranulation occurs, preformed mediators previously stored in mast cells, such as histamine, are released, and newly synthesized prostaglandins and leukotrienes are also secreted. Chemical transmitters released through degranulation include histamine, serotonin, and leukotrienes, which have effects such as vasodilation, increased vascular permeability, contraction of bronchial smooth muscle, and hypersecretion, as well as NCF, which acts as a chemotactic factor for inflammatory cells such as eosinophils and neutrophils. , ECF, PAF, etc., causing allergic reactions (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; J Biol Chem 268:9049- 9054, 1993).

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

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

Currently, anti-allergy drugs include antihistamines (alkylamines, desloratadine, etc.), mast cell stabilizers (cromoglycate, etc.), leukotriene blockers (montelukast, etc.), and anti-IgE antibodies (Omalizumab). However, the effects of these drugs are often temporary and often have severe side effects. Therefore, it can be said that there is still a need to develop new substances that have the effect of preventing and improving allergic diseases while having fewer side effects and lasting effects. In particular, natural products have the advantage of being relatively less toxic.

The present invention discloses the anti-allergic activity of the extract of Bone Damascus.

The purpose of the present invention is to provide an anti-allergy composition using an extract of Bone Damascus.

Other or specific purposes of the present invention will be presented below.

The present invention confirms that, as confirmed in the examples and experimental examples below, the extract of Bone Tongue 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 this.

Considering the above, the present invention can be viewed as an anti-allergy composition or an antihistamine composition containing a bone marrow extract as an active ingredient.

In this specification, "Goldamcho extract" refers to the stem, leaf, fruit, flower, root, underground part, above-ground part or a mixture thereof of the extract object, the stem, leaf, fruit, flower, root, or mixture thereof with water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), and methylene. Chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butylacetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or mixtures thereof. It refers to an extract obtained by leaching using a solvent, an extract obtained using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract. The extraction method is based on consideration of the polarity of the active substance, degree of extraction, and degree of preservation. Any method such as cold immersion, reflux, heating, ultrasonic radiation, or supercritical extraction can be applied. In the case of a fractionated extract, the extract is suspended in a specific solvent and then mixed with a solvent of different polarity to form a fraction obtained by adsorbing the crude extract onto a column filled with silica gel and then mixed with a hydrophobic solvent, a hydrophilic solvent, or a mixture of these. It is meant to include fractions obtained using the mobile phase. In addition, the meaning of the extract includes concentrated liquid extract or solid extract from which the extraction solvent has been removed by methods such as freeze-drying, vacuum drying, hot air drying, and spray drying. 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, in this specification, “active ingredient” refers to an ingredient that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active on its own.

Also, as used herein, “anti-allergy” refers to the inhibition of allergic reactions in allergic diseases involving allergic reactions mediated by mast cell activation, such as mast cell degranulation. These allergic diseases include atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, and allergic contact dermatitis. , allergic keratitis, etc.

The anti-allergy composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can show improvement activity for the allergic disease for which it is intended to be treated, depending on the use, formulation, purpose of formulation, etc., and a typical effective amount is It will be determined within the range of 0.001% by weight to 15% by weight based on the total weight of the composition. Here, “effective amount” means that when the composition of the present invention is administered to mammals, preferably humans, to which it is applied during the administration period recommended by medical experts, etc., it will exhibit the intended medical and pharmacological effects, such as the effect of improving allergic diseases. It refers to the amount of active ingredient contained in the composition of the present invention. Such effective amounts can be determined experimentally within the scope of the ordinary ability of those skilled in the art.

In addition to the active ingredients, the anti-allergy composition of the present invention may further include any compounds or natural extracts whose safety has already been verified and known to have anti-allergic activity in order to increase and enhance anti-allergic activity.

Specifically, such compounds or extracts include Enterococcus faecalis heat-treated dried powder, complexes such as guava leaf extract, Actinidia extract, lobule extract, Contains complexes such as picao preto powder, PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol), L. sakei Probio 65, which has been individually recognized for its function of ‘improving hypersensitive skin conditions’, and gamma-linolenic acid. , fruit and vegetable-derived lactic acid bacteria ( L. plantarum CJLP133), and probiotic ATP.

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

In a specific aspect, the anti-allergy composition of the present invention can be viewed as a food composition. When the composition of the present invention is considered a food composition, its use can be understood as suppressing allergic hypersensitivity reactions or suppressing allergic skin hypersensitivity reactions.

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

In addition, the food composition of the present invention can have any product classification in terms of legal and functional classification as long as it complies with the enforcement laws and regulations at the time of manufacture and distribution. For example, it is a health functional food according to the Korean 「Act on Health Functional Foods」, or confectionery, beans, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (Ministry of Food and Drug Safety Notification 「Food Standards and Specifications」) , special purpose food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated into food when manufacturing, processing, or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be guaranteed. In the food additive code of each country that regulates the production and distribution of food (in Korea, it is the Food Sanitation Act), food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korea Food Additive Code (Ministry of Food and Drug Safety Notification “Food Additive Standards and Specifications”), food additives are classified into chemical synthetics, natural additives, and mixed preparations in terms of composition. These food additives are classified into sweeteners and flavors in terms of function. It is classified into preservatives, emulsifiers, acidulants, thickeners, etc.

Sweeteners are used to impart an appropriate sweetness to food, and either natural or synthetic ones can be used in the composition of the present invention. Preferably, a natural sweetener is used, and natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones can be used. Preferably, natural products are used. When using natural products, it can serve the purpose of enhancing nutrition in addition to flavor. Natural flavoring agents may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, coriander leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. You can also use things obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo nuts. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, such as esters, alcohols, aldehydes, terpenes, etc.

Preservatives include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc., and emulsifiers include acacia gum, carboxymethyl cellulose, xanthan gum, Examples include pectin, and acidulants include acidic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. In addition to improving taste, acidulants may be added to ensure that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms.

As a thickening agent, a suspending agent, settling agent, gel forming agent, bulking agent, etc. may be used.

In addition to the food additives described above, the food composition of the present invention may contain bioactive substances or minerals known in the art and whose safety is guaranteed as food additives for the purpose of supplementing and reinforcing functionality and nutrition.

Such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, etc., and minerals include calcium preparations such as calcium citrate and magnesium stearate. Magnesium preparations such as iron citrate, iron preparations such as chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, etc. are included.

The food composition of the present invention may contain the above-described food additives in an appropriate amount to achieve the purpose of addition depending on the product type.

Regarding other food additives that may be included in the food composition of the present invention, each country's food code or food additive code can be referred to.

The composition of the present invention may be considered a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier in addition to the active ingredient and can be prepared into an oral formulation or a parenteral formulation depending on the route of administration by a conventional method known in the art. 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 a combination of two or more routes is when two or more types of drugs according to the administration route are combined, for example, when one drug is administered firstly through an intravenous route and the other drug is administered secondarily through a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and for specifics, reference can be made to the pharmacopoeia of each country, including the “Korean Pharmacopoeia.”

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, it can be prepared as powder, granules, tablets, pills, sugar-coated tablets, capsules, solutions, gels, syrups, suspensions, and wafers along with a suitable carrier according to methods known in the art. It can be manufactured in a dosage form 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, and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol. etc. can be mentioned. When formulating, appropriate binders, lubricants, disintegrants, colorants, diluents, etc. can 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, and wax. As a lubricant, sodium oleate is used. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polydethylene glycol, etc. Disintegrants include starch, methyl cellulose, Examples include agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Additionally, diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories along with a suitable carrier according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension can be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose can be used. . When formulated for transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, paste preparations, linear preparations, and aerol preparations. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide. When formulated as a suppository, the carrier is Wethepsol ( witepsol), Tween 61, polyethylene glycols, cocoa fat, laurel paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, etc. can be used.

Regarding the specific formulation of pharmaceutical compositions, it is known in the art, and references can be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). The above documents are considered a part of this specification.

The 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, gender, age, patient's severity, and administration route. It may be in the /kg range. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the invention in any respect.

In another specific aspect, the composition of the present invention can be considered a cosmetic composition. When the composition of the present invention is considered a cosmetic composition, its use can be understood as suppressing allergic skin troubles, alleviating allergic skin irritation, and suppressing allergic skin hypersensitivity reactions.

Even if the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and by law, and is specifically classified into functional cosmetics with uses such as improving skin troubles and atopic dermatitis, and non-functional cosmetics. It may be general cosmetics, etc. The product form can take any product form, 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 or spray. Specific product forms may include softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder formulation.

In addition to the active ingredient, the cosmetic composition of the present invention may contain ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, conventional auxiliaries such as colorants and fragrances, and carriers.

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

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the formulation is a spray, chlorofluorohydrocarbon and propane may be used as carrier ingredients. /May contain propellants such as butane or dimethyl ether.

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

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

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

The cosmetic composition of the present invention can be prepared according to a cosmetic composition manufacturing method commonly used in the art, except that it contains an active ingredient that exhibits anti-allergic activity.

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

The anti-allergy composition of the present invention can be commercialized into foods, cosmetics, drugs, etc. for purposes such as improving allergic diseases.

Figure 1 shows the results showing the degranulation inhibitory activity of the IgE and antigen (DNP-BSA) extracts in the mast cell line RBL-2H3 (rat basophilic leukemia).

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 > Preparation of Goldamnia extract

1 L of 70% ethanol was added to 100 g of dried Caragana sinica root powder, immersed and extracted once for 24 hours at room temperature, and then filtered through filter paper. The obtained 70% ethanol filtrate was concentrated under reduced pressure and then freeze-dried to obtain an extract of Bone Damascus.

< Experiment example > Anti-allergy Activity test - β-hexosaminidase assay

The RBL-2H3 cell line, a rat basophilic leukemia cell line, was cultured in MEM medium containing 15% FBS, 100 unit/ml of penicillin and streptomycin, and 2×10 ⁵ /well in a 24-well plate under conditions of 37°C and 5% CO ₂ . After dispensing, the cells were cultured 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-cultivation, the samples of the examples were treated for 20 minutes at each concentration and then reacted with DNP-BSA for 20 minutes. After leaving on ice for 10 minutes to terminate the reaction, the supernatant was placed in a 96 well plate, 1mM P-nitrophenyl-acetyl-β-D-glucosaminid was added, and reacted at 37°C for 1 hour. Stop solution (0.1M NaHCO ₃ , O.1M Na ₂ CO ₃ ) was added to terminate the reaction, and the absorbance was measured with an ELISA reader in the 405 nm wavelength range. Ketifen (100 μg/ml) was used as a positive control.

The results are shown in Figure 1, and it can be seen that the sample of the example inhibited degranulation in a concentration-dependent manner, and the IC ₅₀ was 78.55 ㎍/mL. In the case of ketotifen, a positive control, IC ₅₀ was 37.49 ㎍/mL. Ketotifen, used as a positive control, is an antihistamine that is used for allergic rhinitis, allergic skin diseases such as atopy, and allergic asthma.

Claims (5)

An anti-allergy composition containing an extract of Bone Damascus as an active ingredient.
According to paragraph 1,
The anti-allergy composition, wherein the extract is obtained by extracting the roots of the root with water, ethanol, or a mixed solvent thereof.
According to claim 1 or 2,
An anti-allergy composition, characterized in that the composition is a pharmaceutical composition.
According to claim 1 or 2,
An anti-allergy composition, characterized in that the composition is a food composition.
According to claim 1 or 2,
The composition is an anti-allergy composition, characterized in that it is a cosmetic composition.