KR102017002B1 - Anti-atopic composition comprising extract of Geomgangsong thinning by-product as effective component - Google Patents

Abstract

The present invention relates to an anti-atopic composition containing the extract of Geumgangsong thinning by-products, the extract of Geumgangsong thinning by-products of beta-hexosaminidase (β-hexosaminidase) by T-cell activity compared to pine needles and pine needles Significantly reduce the expression and excellent effect of inhibiting the expression of histamine, the composition containing the extract of the Geumsong pine by-product of the present invention as an active ingredient is a functional food for the prevention or improvement of atopic dermatitis, cosmetics or atopic dermatitis for atopic dermatitis It can be usefully used as a therapeutic agent for dermatitis.

Description

Anti-atopic composition comprising extract of Geomgangsong thinning by-product as effective component

The present invention relates to an anti-atopic composition containing the extract of Geumgangsong thinning by-products as an active ingredient.

Atopic dermatitis is an allergic disease caused by genetic, environmental, and immunological causes and abnormalities in the stratum corneum that acts as a protective barrier on the outermost surface of the skin. Atopic dermatitis has a prevalence rate of about 10-20% worldwide, and it occurs mainly in infants between 1 month and 1 year of age, and is relieved to some extent when they are 3 years old, but may recur due to living conditions and individual differences.

The main symptoms of atopic dermatitis are severe itching, dry skin, rashes, rashes, crusts, and scaly skin (human scales), mainly accompanied by chronic dermatitis. Itching, which causes feelings to be scratched, is induced by an itching mediator such as histamine, which in normal people can easily relieve the symptoms, but in atopic patients, the more they scratch, the more they feel they want to scratch. It will scratch. This scratching behavior of atopic patients causes the skin barrier to collapse and cause secondary infections, further exacerbating inflammation. Atopic dermatitis may be temporarily alleviated, but may be recurred and exacerbated by irritants such as the environment and foods, and the deterioration and alleviation are repeated.

The cause of such atopic dermatitis is not well known, but it has been found to be mainly related to genetic factors and immune response.In addition, it is easy to feel skin itch compared to dry skin, normal people, infections caused by bacteria, viruses and fungi, Emotional and environmental factors appear to be the result of a complex interaction. Inflammation associated with atopic dermatitis is due to excessive immune cell action, which causes inflammatory cytokines and hydroxys such as tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1). Mass production of reactive oxygen species (ROS), such as radicals (OH), superoxide radicals (O ₂ −), and hydrogen peroxide (H ₂ O ₂ ), can cause damage to surrounding tissues.

In addition, mast cells are known as key cells that cause acute and chronic inflammatory diseases in most allergic diseases, including atopic dermatitis. When mast cells are activated, they produce inflammatory cytokines such as TNF-α, IL-1, and IL-6, as well as pruritogens such as histamine. In this way, mast cell-derived inflammatory cytokines further promote the inflammatory response and at the same time promote the secretion of itching agents such as histamine, thereby causing the skin barrier to collapse. Therefore, finding a substance that can suppress inflammatory cytokines and itching mediators without side effects will be effective in controlling various allergic diseases including atopic dermatitis.

Prescriptions for atopic dermatitis are mainly drug therapy such as steroids, antihistamines and antibiotics. Steroids (adrenal cortical hormones) are largely anti-inflammatory and immunosuppressive and have excellent effects, but when applied for a long time, side effects such as skin weakness, systemic hormonal symptoms, and addictive effects may occur. The treatment direction for atopic dermatitis, which is being studied recently, is to use immunosuppressants and new antihistamines. However, antihistamines alone do not completely block allergic reactions. This is because histamine is not the only chemical carrier that causes allergic reactions. Therefore, there is a need for the development of a composition for the treatment of atopic dermatitis, which has an effect on atopic dermatitis but has no side effects.

On the other hand, Pinus desiflora for. erecta) is widely distributed in the valleys and mountain ranges of the Kumgang mountain range along Ulbaek, Bonghwa-gun in northern Gyeongbuk, Gangneung, and Samcheok in southern Gangwon. The country's largest colony is formed. Pine trees are native to Korea, Japan, and Manchuria, and all parts of leaves, such as leaves, pine cones, pollen, rosin, and bark, have been used as sulfur plants. In particular, pine needles, which can be easily collected, have been used as folk remedies, and have been used as health foods for the treatment of adult diseases such as hypertension, diabetes, skin diseases, neuralgia, and longevity.

Geumgangsong, which is native to Uljin, produces infinite amount of thinning by-products due to continuous thinning work for the formation, farming, and management of Uljin area. Most of the by-products are used for composting and sawdust production. Although there are many reports on the functionality of pine needles at present, research on the functionality of mixtures of pine needles and pine needles, which are thinning by-products, is not actively conducted. It is possible to use it as a new source of income by developing functional foods and high value-added products through functional studies of thinning by-products with excellent economic efficiency as insoluble resources.

On the other hand, Korean Patent Application Publication No. 2017-0045085 discloses a skin improvement composition comprising pine bark extract and its use, but has not been disclosed for the anti-atopic composition containing the extract of the Geumgangsong thinning by-product of the present invention as an active ingredient. .

The present invention has been derived by the above requirements, provides an anti-atopic composition containing the extract of Geumgangsong thinning by-products as an active ingredient, the extracts of Geumsong-song thinning by-products that are a mixed extract of pine needles and pine leaves are pine leaf and Songjeol extract alone Compared to the beta-hexosa minidase (β-hexosaminidase) secretion effect compared to the remarkable synergistic effect, the present invention was completed.

In order to solve the above problems, the present invention provides a health functional food composition for the prevention or improvement of atopic dermatitis containing the extract of Geumgangsong thinning by-products as an active ingredient.

The present invention also provides a cosmetic composition for the prevention or improvement of atopic dermatitis, containing the extract of Geumgangsong thinning by-products as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of atopic dermatitis, containing the extract of Geumgangsong thinning by-products as an active ingredient.

The present invention relates to an anti-atopic composition containing an extract of Geumgangsong thinning by-products, the extract of Geumgangsong thinning by-products secretion beta-hexosaminidase (β-hexosaminidase) secretion, histamine secretion, IL by T-cell activity Since the effect of inhibiting the secretion of -4 and TNF-α, the composition containing the extract of the Geumsong thinning by-product of the present invention as an active ingredient is a health functional food for the prevention or improvement of atopic dermatitis, cosmetics for atopic dermatitis or atopic treatment Can be used.

1 is a result confirming the cytotoxicity of Kumgang pine thinning by-products, pine needles and songjeol. CON is the negative control.
Figure 2 is a result confirming the beta-hexosaminidase (β-hexosaminidase) production inhibitory ability of the Geumsong thinning by-products, pine needles and songjeol. CON is a negative control and beta-hexosaminidase production was induced using PA (100 pg / ml PHORBOL 12-MYRISTATE 13-ACETATE (PMA) and 1 mg of Calcium Ionophore A23187).
Figure 3 is the result confirming the inhibitory effect of histamine secretion by the Geumsong thinning by-products. CON is a negative control and histamine secretion was promoted using PA (100 pg / ml PHORBOL 12-MYRISTATE 13-ACETATE (PMA) and 1 μM of Calcium Ionophore A23187).
4 is a result confirming the secretion inhibitory effect of inflammatory cytokines (IL-4 and TNF-α) by the Kumgangsong thinning by-products. CON is a negative control and PA (100 pg / ml PHORBOL 12-MYRISTATE 13-ACETATE (PMA) and 1 μM of Calcium Ionophore A23187) was used to induce the expression of inflammatory cytokines.

The present invention relates to a health functional food composition for the prevention or improvement of atopic dermatitis, containing the extract of Geumgangsong thinning by-products as an active ingredient.

The kumgang pine thinning by-product is composed of a mixture of pine needles (leaf of the Geumsong pine) and songjeol (branch and stem of the Geumsong pine), preferably mixing the pine needles and Songjeol in a weight ratio of 1 to 3: 0.5 to 2, even more preferably Is mixed with pine leaves and cut flowers in a weight ratio of 2: 1, but is not limited thereto.

In a preferred embodiment of the present invention, the concentration of the extract of the Geumgangsong thinning by-product in the composition may be 100 µg / ml or more, preferably 150-250 µg / ml, more preferably 200 µg / ml, but is not limited thereto. It doesn't work.

In a preferred embodiment of the present invention, the extract of the Kumgangsong Gimbal by-product is extracted using water, a lower alcohol of C ₁ ~ C ₄ or a mixture thereof as a solvent, more preferably extracted using an alcohol as a solvent But is not limited thereto.

The health functional food composition for preventing or improving atopic dermatitis of the present invention may be prepared in any one formulation selected from granules, pills, tablets, capsules, candy, syrups and beverages, but is not limited thereto. The dietary supplement composition is not particularly limited as long as it can be ingested to prevent or improve atopic dermatitis.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used with other foods or food ingredients, and may be appropriately used according to a conventional method. The active ingredient may be appropriately used depending on the purpose of use (prevention or improvement). However, in the case of long-term intake for health control purposes, it can be used in an amount in a range without any problem in terms of safety. There is no particular limitation on the kind of food. Examples of foods to which the health functional food composition may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products, including ice cream, various soups, drinks, tea Drinks, alcoholic beverages, vitamin complexes, and the like, and include all health foods in the conventional sense.

In addition, the nutraceutical composition of the present invention may be prepared as food, in particular functional food. Functional foods of the present invention include ingredients that are commonly added in food production, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when prepared with a drink, natural carbohydrates or flavors may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates can be monosaccharides (e.g. glucose, fructose, etc.), disaccharides (e.g. maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g. dextrins, cyclodextrins, etc.) or sugar alcohols (e.g. , Xylitol, sorbitol, erythritol and the like). The flavourant may be a natural flavourant (eg, taumartin, stevia extract, etc.) and a synthetic flavourant (eg, saccharin, aspartame, etc.). Various nutritional supplements, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid The carbonation agent etc. which are used for a drink can be contained further.

The present invention also provides a cosmetic composition for the prevention or improvement of atopic dermatitis, containing the extract of Geumgangsong thinning by-products as an active ingredient.

In the cosmetic composition for improving atopic dermatitis of the present invention, the cosmetic composition for improving atopic dermatitis is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, It is preferably one of the formulations selected from emulsions, foundations, wax foundations, and sprays, and more preferably skin ointments, creams, softeners, nutrients, packs, essences, hair tonics, shampoos, rinses, hair conditioners, Hair Treatment, Gel, Skin Lotion, Skin Softener, Skin Toner, Astringent, Lotion, Milk Lotion, Moisture Lotion, Nutrition Lotion, Massage Cream, Nutrition Cream, Eye Cream, Moisture Cream, Hand Cream, Foundation, Nutrition Essence, Sunscreen , Soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser You can have a single dosage form slow, but not limited to this. The composition of each of these formulations may contain a variety of bases and additives necessary for the formulation of the formulation and are suitable, and the types and amounts of these components can be readily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal carriers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc acid may be used as carrier components. Can be. 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, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether. When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used. When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of atopic dermatitis, containing the extract of Geumgangsong thinning by-products as an active ingredient.

The pharmaceutical composition for preventing or treating atopic dermatitis of the present invention may further include a suitable carrier, excipient or diluent commonly used in the preparation of the pharmaceutical composition. Pharmaceutical dosage forms of the compositions according to the invention may be used alone or in combination with other pharmaceutically active compounds as well as in suitable combinations.

The pharmaceutical compositions according to the invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, external preparations, suppositories, and injections, respectively, according to conventional methods. . Carriers, excipients and diluents that may be included in the pharmaceutical composition comprising the extract of the Kumgangsong thinning by-products include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin Compounds or mixtures, including calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Can be mentioned. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, It is prepared by mixing gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, syrups, and the like, which are commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. The pharmaceutical composition of the present invention can be administered to various mammals such as rats, mice, livestock, humans, and the like. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. These preparations and examples are only intended to explain the present invention more specifically, it is apparent to those skilled in the art that the scope of the present invention is not limited thereto.

Production Example  1. Preparation of Extract of Geumgangsong Gimbal By-products

Kumgang pine thinning by-products were used by mixing the leaves and songjeol harvested in the fall of 2015 in a weight ratio of 2: 1. The collected thinning by-products were crushed after washing and drying. Thereafter, 10 parts by weight of 70% (v / v) ethanol was added per 1 part by weight of the sample, followed by stirring extraction at room temperature for 10 hours. After stirring and extraction, the supernatant and the precipitate were separated by Whatman No. 1 filter paper, and concentrated and lyophilized, and then stored as a sample in the refrigerator. The extracts of pine needles or songjeol extracts of Geumgangsong were also extracted using the same method as that of the extract of Geumsong Songgae byproduct.

Example  1. Confirmation of cell viability

Rat Basophil Leukemia (RBL-2H3) cells from ATCC (American Type Culture Collection) were cultured in DMEM medium. In order to determine the concentration range to be used for the experiment, MTT (3- (4,5-dimethylthiazol-2-yl) using the method of Mosmann (1983) was treated by concentration of the Geumsong thinning by-products, pine needles and pine extract of Preparation Example 1 -2,5-diphenyltet razolium bromide) analysis was performed.

RBL-2H3 cells were incubated in 96 well plates with 100 μl of DMEM medium for one day, and then treated with samples of Preparation Example 1 at various concentrations (50, 100 and 200 μg / ml) for 24 hours. 10 μl of 5 mg / ml MTT solution was added to each well and incubated for 3 hours to induce a reduction reaction. 100 μl of dimethyl sulfoxide (DMSO) solution was added to complete purple formazan crystals. Dissolved. The degree of color development was measured by absorbance at 570 nm using a spectrophotometer. The cytotoxicity was calculated based on the relative cell survival rate of the sample treated group based on the 100% survival rate of the cell-free group.

As a result, as shown in Figure 1 was confirmed that the cytotoxicity by the Geumsong thinning by-products, pine needles and songjeol up to 200㎍ / ㎖.

Example  2. Beta-Keumsong Gimbal By-products Hexosaminidase (β- hexosaminidase Expression Inhibition  Confirm

RBL-2H3 cells were dispensed with 3 × 10 ⁵ cells per well in 24 well plates and incubated for 24 hours. Then, PA (PHORBOL 12-MYRISTATE 13-ACETATE (PMA) 100pg / ml and Calcium Ionophore A23187 1μM) was treated to induce beta-hexosaminidase production, and the sample of Preparation Example 1 was prepared at each concentration ( 50, 100 and 200 ㎍ / ㎖) was reacted for 1 hour and then stopped for 10 minutes on ice, then 1mM 4-Nitrophenyl N-acetyl-β-D-glucosaminide (0.05M citrate buffer, pH 4.5 1 hour after the reaction with the substrate was added to the stop reagent (0.1M Carbonate buffer) and the absorbance was measured at 405nm.

As a result, as shown in FIG. 2, the beta-hexosaminidase secretion was further reduced compared to the treatment of pine needles and pine needles when 200 μg / ml of Geumsong thinning by-products were treated, respectively. Has a synergistic effect on atopy treatment.

Example  3. Confirmation of the Histamine Inhibitory Effect of Gumgangsong Gimbal By-products

RBL-2H3 cells were incubated for 24 hours after dispensing 3 × 10 ⁵ cells per well into 24 well plates in order to confirm the effect of the Geumsong thinning by-product extract on histamine secreted by degranulation. Subsequently, PA (100 pg / ml PHORBOL 12-MYRISTATE 13-ACETATE (PMA) and Calcium Ionophore A23187 1 μM) were treated. By incubating for 1 hour to release histamine. The reaction was allowed to stand for 10 minutes on ice, and the histamine concentration was measured using a histamine measuring kit (Histamine ELISA kit, abcam) using the cultured supernatant.

As a result, as shown in Figure 3 it was confirmed that the concentration of histamine released to the extracellular that was increased by PA decreases when treated by Geumsong thinning by-products.

Example  4. Confirmation of Inhibitory Inflammatory Cytokine Secretion by Geumgangsong Ginseng

In order to measure the concentration of IL-4 and TNF-α secreted from mast cells, RBL-2H3 cells were dispensed into 24 well plates and incubated for 24 hours. Subsequently, PA (100 pg / ml PHORBOL 12-MYRISTATE 13-ACETATE (PMA) and Calcium Ionophore A23187 1 μM) were treated, and the samples of Preparation Example 1 were treated together at respective concentrations (50, 100 and 200 μg / ml). 1 hour of incubation induced expression of IL-4 and TNF-α. After completion of the reaction, the mixture was allowed to stand on ice for 10 minutes, and the cultured supernatant was centrifuged (1200 rpm, 5 minutes), and then the content was measured using IL-4 and TNF-α kit (ELISA kit, R & D). .

As a result, as shown in Figure 4, the treatment of the extracts of the Geumsong thinning by-products was confirmed that the secretion of IL-4 and TNF-α was inhibited, especially when the extracts of the Geumsong thinning by-products were treated at a concentration of 200㎍ / ㎖ It was confirmed that the secretion inhibitory effect is excellent.

Claims (6)

A pharmaceutical composition for the prevention or treatment of atopic dermatitis, which contains 200 µg / ml of the extract of the Geum pine pine pine and Songjeol mixture mixed at a weight ratio of 1 to 3: 0.5 to 2 as an active ingredient. delete delete The pharmaceutical composition for preventing or treating atopic dermatitis according to claim 1, wherein the extract inhibits the expression of beta-hexosaminidase and histamine. A cosmetic composition for the prevention or improvement of atopic dermatitis, which contains 200 µg / ml of the extract of the Geum pine pine pine and Songjeol mixture mixed at a weight ratio of 1 to 3: 0.5 to 2 as an active ingredient. A dietary supplement for the prevention or improvement of atopic dermatitis, which contains 200 µg / ml of the extracts of the Geumsong pine needles and Songjeol mixtures mixed at a weight ratio of 1 to 3: 0.5 to 2 as an active ingredient.

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