KR20130008761A - Composition for anti-allergy and/or anti-atopy comprising extract of ecklonia cava - Google Patents

Abstract

PURPOSE: An anti-allergic and/or anti-atopic composition containing an Ecklonia cava extract is provided to maximize safety and anti-allergic effect with low cytotoxicity. CONSTITUTION: An anti-allergic or anti-atopic composition contains effective amount of water extract of Ecklonia cava leaves as an active ingredient, prepared by extracting Ecklonia cava leaves with water of 40 deg.C. The Ecklonia cava leaves are dried by far infrared ray at 100 deg.C. for 1 hour. An anti-allergic or anti-atopic functional food composition contains the water extract of Ecklonia cava leaves. The Ecklonia cava extract for the food composition is 20-90% of high concentrate liquid, powder, or granule. [Reference numerals] (1,7) Naturally drying for 48 hours; (2) Hot wind drying(50°C) for 4 hours; (3) Hot wind drying(100°C) for 2 hours; (4) Far infrared ray drying(50°C) for 3 hours; (5) Far infrared ray drying(100°C) for 1 hours; (6) Compression molding at 100°C; (8) Hot wind drying(100°C) for 24 hours; (9) Far infrared ray drying(100°C) for 4 hours; (AA) 40°C water extract of Ecklonia cava; (BB) Leaves; (CC) Stems; (DD) Ecklonia cava(ug/ml); (EE) Ecklonia cava

Description

Composition for anti-allergy and / or anti-atopy comprising extract of Ecklonia cava

The present invention relates to anti-allergic and / or anti-atopic compositions comprising Ecklonia cava extract.

WHO has already included allergy disease as the seventh most problematic disease in the 21st century. Allergic diseases include atopic dermatitis, asthma and rhinitis. Allergies are so close to incurable diseases in modern medicine that there is no clear cure to date. At present, antihistamines, which are not fundamental treatments for treating allergic diseases, have excellent effects but have many side effects.

Symptoms of atopy and allergies have been steadily increasing in the 21st century, accompanied by industrial development, westernization and eastern eating, serious environmental pollution, and new diseases such as sick house syndrome. The trend is serious not only in Korea, but also in Western societies, and even in developing countries where development is underway, so these treatments are expected to generate global market demand.

In addition, the demand for the development of safe and effective treatments for young children, the main patient group, and anti-allergic drugs should be safer and more effective than others because the main patients are young children. Rather than development, it is more appropriate to materialize therapeutic resources from previously known safe herbs and edible plants.

Recently, the number of patients suffering from allergic diseases due to environmental pollution and changes in the residential environment is increasing. According to the National Health Insurance Corporation, the number of patients treated with atopic dermatitis, allergic rhinitis and asthma increased by more than 20% from 5.45 million in 2002 to 6.66 million in 2006.

These diseases are increasing incidence of infants and the elderly who are vulnerable to disease, and children under 9 years of age have more patients than other age groups. In the case of allergic rhinitis patients in 2007, 21.2% of the patients under 9 years old accounted for 51.2% of patients with atopic dermatitis and 41.5% of patients with asthma. The social cost is 4.1 trillion won and the burden of disease is high, accounting for 0.52% of GDP.

IgE is an important component involved in allergic symptoms. In particular, IL-4 is essential for IgE production, which is more productive in Th2 cells than Th1 cells and is associated with allergic expression (Beatti PE, Lewis-Jones MS (2006), Br J Dermatol 155 (1): 145-151).

In addition, IgE binds to the FcR1 receptor, which is expressed on mast cells or basophils, to form a cross-linked structure with antigens (Hyeon-Chul Lee, et al. (2003), Immunology and Allergology, Chonnam National University). Publishing department). When IgE and allergens that bind to FcR1 on mast cells or basophils form a crosslinked structure, allergens are released (degranulated), including histamine (Spergel JM, Paller AS (2003), J Allergy Clin Immunol 112). S118-127).

The mechanism of allergy caused by immunological causative agents is that the causative antigens that enter the body bind to the high-affinity IgE receptors on the surface of mast and basophil cells. In addition, degranulation of these cells occurs, and it is explained by the sudden secretion of many chemical mediators such as histamine and trypsinase (Falcone FH, Haas H, Gibbs BF. (2000)). , Blood. 96: 402838). The mechanism of the occurrence of allergens whose non-immune causative agent is the causative factor or the cause is not clear, but the final response of the body by various chemical factors is considered to be similar. It should also be included in the scope of care to provide guidance for preventing future recurrence.

Since allergic diseases are accompanied by frequent relapses, the development of therapeutic agents is required. Allergies and atopic diseases are caused by a combination of seasonal, environmental, and temporal factors, and many of them are repeatedly aggravated or cured due to unavoidable circumstances or patterns of life. Minor Clinical Features in Patients with Atopic Dermatitis (1994): Frequency of Secondary Skin Symptoms in Pediatric Atopic Dermatitis, Korean Dermatological Association: 32 (5): 866 ~ 89.

Serious atopic dermatitis suffers from social problems that cause social problems. In fact, cured atopy patients have a great impact not only on the individual patients but also on the society in order to improve their quality of life. Therefore, there is an urgent need to develop a powerful therapeutic agent that eliminates the underlying cause rather than a popular therapeutic agent to alleviate the symptoms.

Existing allergy treatments include steroids, immunosuppressants, antihistamines, antibiotics, and skin moisturizers, but they have many side effects and are not a fundamental treatment, but are for the treatment of temporary lesions.

Steroid is a hormone secreted by the adrenal cortex. It is involved in carbohydrate, protein and fat metabolism and mineral metabolism such as sodium, potassium, calcium and has anti-inflammatory function. In most cases, steroids are used to clinically suppress inflammatory and immune responses. However, if steroids are used in excess of the dosage, they become depleted by depletion of steroid-coupled receptors, and patients are ineffective, so they are applied more frequently or with higher intensity. The drug does not appear until the steroid-binding receptors are regenerated. It is a problem for patients and therapists that these steroids deteriorate the skin to peroxidation, irritating and exacerbating the skin and making dermatitis worse.

Immunosuppressants are drugs that contain cyclosporine A, tacrolimus, and ascomycin, which are components that inhibit the activity of a series of cells related to dermatitis and inhibit the production of cytokine related to inflammation. Prototypes based on A and tacrolimus and Elidel based on pimecrolimus, an ascomycin derivative developed by Novartis, Switzerland. In addition to immunosuppressants, a number of adjuvant inhibitors such as Mycophenolate Mofetil (Roche), Rapamycine (Wyeth), Basiliximab (Novartis), Daclizumab (Roche), and Azathioprine (GSK) are available on the market. As drugs used for the purpose of suppression, the immunosuppressive factors that they suppress are immune factors that prevent cancer or infectious diseases in the body, and abuse of this drug may cause problems such as carcinogenesis and infection. Therefore, the US FDA warns against the carcinogenicity of these drugs, recommends that they be used only in intractable cases where steroids do not work.

In addition, histamine is a substance present in mast cells, basophils, and platelets. When it is secreted, it binds to the H1 and H2 receptors distributed in subcutaneous blood vessels, causing pruritus, which is the main pain of allergy symptoms. Complete blocking of the H1 receptor does not clear the itch and it is known that H1 and H2 blockers are more effective than H1 blockers alone. However, even if the anti-histamine improves pruritus, it is difficult to cure if it leads to a vicious cycle of pruritus-scratch-allergy-pruritus, and it has not yet been proved to be effective for allergy.

And most atopic patients are frequently infected with Staphylococcus bacteria, which in turn produce IgE specific to the bacteria, causing further itching. Antibiotics of the erythromycin family are used for systemic treatment and usually can be effective in about 5 days.However, long-term use of antibiotics increases the resistance of the infected bacteria, so it is recommended to use only in emergency situations where the infection should not spread anymore. It is becoming.

In addition, atopic patients with significantly lower skin moisturizing and regenerating ability than healthy people should protect their damaged skin by applying appropriate moisturizers to dry, keratinized skin. Moisturizers are external preparations applied to the skin to maintain or preserve the moisture present in the skin. Commercially available products for moisturizing atopy skin include oil 100%, skin lotion, milk lotion and cream, and the components contained in these products include oils, humectants, plant extracts, vitamins, ceramides, and the like. There are no side effects to the moisturizer, but problems such as skin hypersensitivity due to the excipient used in the moisturizer are caused.

In recent years, marine resources such as seaweed have been well received by consumers in the well-being era, and functional drinks developed using them are sufficient to meet consumer needs. In particular, the natural products produced by marine organisms have a lot of chemical structural peculiarities, so the research for the development of anti-allergic substances from seaweed among these bioactive substances is insufficient.

Ecklonia cava is a perennial algae of Laminariales and Seaweeds (Alariaceae), widely distributed in the southern part of the East Sea of Korea, Jeju Island, and Japan. Stems are cylindrical, base is root-shaped, grows up to 1-2 m in length. It is mainly used to feed abalone and turban shell, and is also used as a major raw material for making alginic acid and potassium iodine. Ecklonia has been used as a local food for a long time.

The study of formulating functional beverages using seaweeds that can be used as foods with excellent allergy suppression, easy to secure ingredients, and contributing to the income improvement of seaweed-harvesting farmers, will contribute greatly to economic revival and improvement of local independence between regions. It seems to be.

    In particular, no papers or literatures have been reported that search for anti-allergic / antiatopic effects by drying and extracting Ecklonia cava.

In this regard, as a prior patent, Korean Patent Publication No. 10-2010-0057431 relates to an extract for treating allergy to mimosa and a pharmaceutical composition for treating allergy comprising the same as an active ingredient, in order to confirm the usefulness of the extract of the mimosa. (1) the effects of mimosa extracts on the motility of mast cells by stem cell factor, (2) the toxic effects of mimosa extracts on cells, and (3) inflammatory cytokines in EoL-1 cells (human eosinophilic leukemic cells) The inhibitory effect of mimosa extracts on the caine and chemokine MCP-1 (monocyte chemoattractant protein-1) and IL-6 (interleukin-6) and (4) the effects of mimosa extract on asthma-induced mice were identified and from these It is described that it can be effectively used to improve or treat atopy and / or asma.

As another prior patent in this regard, Korean Patent Publication No. 10-2005-0084725 relates to a composition for the prevention and treatment of inflammatory or allergic diseases, including Jeonho extract, Jeonho extract is responsible for various inflammatory and allergic diseases. Inhibits the production of PAF and inhibits the activity of cyclooxygenase-2 and lipoxygenase (5-LO) to prevent the production of eicosanoids and release of histamine from mast cells Since it inhibits, it is described that it can be usefully used as a medicine or health functional food for the prevention and treatment of various inflammatory or allergic diseases.

The present invention is to solve the above problems and the object of the present invention is to provide a high functional food with less side effects and excellent anti-allergic / anti-atopic effect.

Another object of the present invention is to provide a method for producing a high functional food with less side effects and excellent anti-allergic / antiatopic effect.

In order to achieve the above object, the present invention provides an anti-allergic or anti-atopic composition containing an effective amount of Ecklonia cava leaf extract, extracted from 40 ℃ water of Ecklonia cava as an active ingredient.

In one embodiment of the present invention, the Ecklonia cava leaves are preferably far-infrared dried at 100 ℃ for 1 hour, but is not limited thereto.

In another embodiment of the present invention, the Ecklonia cava leaf extract preferably contains 0.01 to about 90% more preferably 1 to 5% of the total composition, but is not limited thereto.

In another embodiment of the present invention, the effective amount of the Ecklonia cava leaf extract is preferably 5-25 µg / ml in consideration of cytotoxicity and effects, but is not limited thereto.

In another aspect, the present invention provides an anti-allergic or anti-atopic functional food composition containing an effective amount of Ecklonia cava leaf extract, which is extracted with 40 ° C water.

In another aspect, the present invention provides a method for producing an anti-allergic or anti-atopic composition comprising the step of obtaining the Ecklonia cava leaf extract by extracting the Ecklonia cava leaves at 100 ° C. for 1 hour to far-infrared dry water.

The allergic diseases include hypersensitivity, allergic rhinitis, allergic dermatitis, allergic asthma, allergic conjunctivitis, atopic dermatitis, insect allergy, food allergy, drug allergy or hives.

Accordingly, the present invention provides a food composition useful for the prevention and improvement of allergic diseases containing the Ecklonia cava extract as an active ingredient.

The food composition containing the Ecklonia cava extract of the present invention includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid and malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polyphosphate), polyphenols, and Carter. It may further include any one or more of natural antioxidants such as catechin, alpha-tocopherol, rose mary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid and the like. .

Ecklonia cava extract for food compositions is 20 to 90% of a high concentration or may be in the form of powder or granules.

Likewise, the food composition containing the Ecklonia cava extract of the present invention may further include any one or more of lactose casein, dextrin, glucose, sugar, sorbitol.

In addition, the present invention is a food additive containing the Ecklonia cava extract in food preservatives, fungicides, antioxidants, spices, seasonings, sweeteners, flavors, swelling agents, reinforcing agents, modifiers, emulsifiers, various nutrients, synthetic flavors and natural flavors Flavoring agents, coloring agents, coloring agents, neutralizing agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, antifoaming agents, solvents, mold release agents, preservatives, It provides a method of using a food additive, characterized in that it is used as an essential raw material of the quality improving agent, glycerin, alcohol, carbonation agent used in carbonated drinks, or other food manufacturing additives and food ingredients (additives). In this case, the food additive may be added to the food by immersing, spraying or mixing the food, and the ratio of such additive is not so important, but it is generally selected from 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention. .

Foods are fruits, vegetables, dried or cut products of fruits or vegetables, fruit juices, vegetable juices, mixed juices of these, chips, noodles, livestock processed foods, fish processed foods, dairy products, fermented milk foods, legumes foods, grain foods , Microbial fermented food, bakery, condiments, meat processing, acidic beverages, licorice, herbs or any one or more.

Each of these will be described below.

In the preparation of gum, 0.5 g or 1 g of the extract of the present invention, 90 g of gum base, 200 g of sorbitol, 1 g of softener, 3 g of xylitol and 12 g of an emulsifier were mixed in a preheated mixer at 60 to 70 ° C., followed by mixing. 9 g of peppermint flavor is added while maintaining the temperature inside the mixer at 55 ° C., then mixed for 30 minutes, extracted, rolled, aged for 24-48 hours, and cut and packaged to prepare a gum.

In addition, when kneading in the manufacturing process of confectionery baking, it is added and used in sauces, sauces, and also can be used as a functional or palatable food by adding powder or liquid to fermented milk or yogurt. The herbal extract powder can be directly added to kimchi, salted fish, or vegetable processing to be used for the purpose of preserving taste, anti-oxidation and discoloration.

In meat processing, it is used by dipping or spraying for the purpose of final sanitary agent and microbial growth suppression in ham, sausage, etc., by mixing garlic, green onion, ginger, herb, licorice, etc. It can be used to make spices, fish paste, fish, seasoned fish, salted fish, etc., as a dipping, spraying, and adding method, and can also be immersed, sprayed, and added to kimchi, radish, diced vegetables, and pickled vegetables.

The present invention also provides an animal feed additive useful for the prevention and improvement of allergic diseases containing the Ecklonia cava extract obtained by the above method as an active ingredient and a feed comprising the same.

Ecklonia cava extract for animal feed additives may be 20 to 90% high concentrate or powder or granule form.

The Ecklonia cava extract for animal feed additives includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polymeric phosphate), polyphenols, and caterine (catechin). ), Alpha-tocopherol, rose mary extract (rose mary extract), vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid and the like may further include any one or more than one.

Animal feed additives containing Ecklonia cava extract of the present invention and feed containing them are various supplements such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal, enzymes, living microbial preparations, grains, For example milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed, such as rape, soybeans and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugars and dairy products, for example, a dry ingredient consisting of various powdered milk and whey powder, and a drying additive, all mixed together with a liquid component, and a component which becomes a liquid after heating, that is, a lipid, for example, an animal liquefied by heating. In addition to the main components such as fats and vegetable fats can be used with substances such as nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents and the like.

The Ecklonia cava extract for animal feed additives may be administered to animals alone in combination with other feed additives in an edible carrier.

In addition, the Ecklonia cava extract for animal feed additives can be easily administered as a top dressing or directly mixed with animal feed or separately from the feed, in a separate oral formulation, by injection or transdermal or in combination with other ingredients. Typically, single or divided daily dosages can be used as is well known in the art.

When the Ecklonia cava extract for animal feed additives is administered separately from the animal feed, the dosage form of the extract, as is well known in the art, may be prepared in immediate release or sustained release formulation in combination with a non-toxic pharmaceutically acceptable edible carrier. Can be. Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the dosage form of the extract may be tablets, capsules, powders, torokies or sugar-containing tablets or top dressings in microdisperse form. If a liquid carrier is used, it may be in the form of soft gelatin capsules or syrups or liquid suspensions, emulsions or solutions. In addition, the dosage form may contain auxiliaries such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like. They may also contain substances useful for the improvement and prevention of other allergic diseases.

In addition, the animal feed, wherein the Ecklonia cava extract is included as an animal feed additive, may be any protein-containing organic cereal meal commonly used to meet the dietary needs of animals. Such protein-containing flours are typically mainly composed of corn, soy flour, or corn / bean flour mixtures.

The animal feed additive may be used by adding to the animal feed by dipping, spraying or mixing.

The invention is applicable to a number of animal diets, including mammals, poultry and fish. More specifically, the diet is commercially important mammals such as pigs, cows, sheep, goats, laboratory rodents (rats, mice, hamsters and gerbils), furry animals (eg, mink and fox), and Zoo animals (eg monkeys and tailless monkeys), as well as livestock (eg cats and dogs). Commercially important poultry typically include chickens, turkeys, ducks, geese, pheasants and quails. Commercially raised fish, such as trout, may also be included.

In the method of formulating animal feed comprising the extract according to the present invention, the Ecklonia cava extract is incorporated into the animal feed in an amount of about 5 g to 100 g per kg of feed on a dry weight basis.

In addition, the feed mixture is thoroughly mixed and then a viscous granulated or granular material is obtained depending on the degree of grinding of the components. It is fed as a mash or formed into the desired discrete shape for further processing and packaging. At this time, in order to prevent separation during storage, it is preferable to add water to the animal feed, followed by a conventional pelleting, expansion, or extrusion process. Excess water may be removed to dryness.

The present invention provides a cosmetic composition useful for the prevention and improvement of allergic skin diseases containing the Ecklonia cava extract obtained by the above method as an active ingredient.

Accordingly, the present invention provides a cosmetic composition useful for the prevention and amelioration of allergic skin diseases, wherein the amount of Ecklonia cava extract has 0.01 to 30% by weight, preferably 0.01 to 5.0% by weight of the total cosmetic composition, and includes cleansing, face wash, and soap. In addition to shampoos, rinses and treatments, the present invention provides a cosmetic composition useful for the prevention and improvement of allergic skin diseases.

Cosmetic composition containing the Ecklonia cava extract of the present invention as an active ingredient may be used in a variety of cosmetics, face wash and shampoo for preventing and improving allergic skin diseases. Examples of products to which the cosmetic composition can be added include cosmetics such as various creams, lotions, and skins, and shampoos, rinses, cleansing agents, face washes, soaps, treatments, and cosmetic liquids.

The cosmetic of the present invention includes a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts. The water-soluble vitamins may be any compound that can be incorporated into cosmetics, but preferably vitamins. B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H and the like, and their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) Derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-phosphate magnesium salt, etc.) are also included in the water-soluble vitamins usable in the present invention. The water-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from microorganism culture, enzyme or chemical synthesis.

The oil-soluble vitamin may be any compound that can be formulated in cosmetics, but preferably vitamin A, carotene, vitamin D2, vitamin D3, and vitamin E (d1-α-tocopherol, d-α-tocopherol, d-δ-tocopherol) and the like. Derivatives thereof (ascorbic palmitate, ascorbic stearate, ascorbic acid dipalmitate, dl-α-tocopherol acetate, dl-α-tocopherolvitamin E, DL-pantothenyl alcohol, D-pantothenyl nicotinic acid) Alcohols, pantotenylethyl ether, etc.) are also included in the oil-soluble vitamins used in the present invention.

Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification of microorganism culture, enzyme or chemical synthesis.

The polymer peptide may be any compound as long as it can be incorporated into cosmetics. Preferably, collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like can be given. Polymeric peptides can be purified and obtained by conventional methods such as purification from microbial cultures, enzymatic methods or chemical synthesis methods, or can be purified and used from natural products such as dermis and pig silk such as pigs and cattle.

The polymer polysaccharide may be any compound as long as it can be blended into cosmetics. Preferably, hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.) may be mentioned. For example, chondroitin sulfate or a salt thereof can be usually purified from mammals or fish.

The sphingolipid may be any compound as long as it can be blended into cosmetics. Preferably, ceramide, pit spingosine, sphingolipid lipid and the like can be given. Sphingo lipids can usually be purified from mammals, fish, shellfish, yeasts or plants by conventional methods or obtained by chemical synthesis.

The seaweed extract may be any compound as long as it can be blended into cosmetics. Preferably, the seaweed extract may include brown algae extract, red algae extract, green algae extract, and the like. Also, calginine, arginic acid, sodium arginate, Potassium arginate and the like are also included in the seaweed extract used in the present invention. Seaweed extract can be obtained by purification from seaweed by conventional methods.

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients.

Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cold depressants, restriction agents, purified water and the like.

Examples of the fat or oil component include ester fats, hydrocarbon fats, silicone fats, fluorine fats, animal fats, and vegetable fats and oils.

As ester fats and oils, glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl mystinate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl acid isocetyl, isostyl acid isostearyl, isostaryl palmitate, octylate acid octyldodecyl, isostearic acid isetyl, diethyl sebacate, adipine Acid isopropyl, isoalkyl neopentane, tri (capryl, capric acid) glyceryl, trimethyl ethyl trimethylol propane, triisostearic acid trimethylol propane, tetra 2-ethylhexanoic penta erythritol Cetyl caprylate, lauric acid decyl, hexyl laurate, decyl myristin, myristin acid myristyl, myritic acid cetyl, stearyl stearate, decyl oleate, rininooleic acid , Isostearyl laurate, isotridecyl myristin, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecate oleate, octylate decyl oleate, octyl dodecyl linoleate, isopropyl isopropyl acid, 2 -Cetostearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicapric acid, propylene glycol dicacapate, capric acid Propylene glycol, dicapric acid neopentyl glycol, dioctanoate neopentyl glycol, tricaprylic acid glyceryl, triundecyl glyceryl, triisopalmitinate glyceryl, triisostearate glyceryl, neopentane dodecyl Isostearyl octanoate, octyl isononate, hexyl decyl neodecanoate, octyl dodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, Sothete octylate, polyglycerol oleate, polyglycerine isostearate, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myritic lactate, octyl lactate, octyl lactate Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipic acid, diisopropyl sebacinate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, physteryl isostearate, phytic oleate, 12-Steloylhydroxystearate isocetyl, 12-Steloylhydroxystearate stearyl, 12-stealo Esters such as monohydroxystearic acid isostearyl; and the like.

Examples of the hydrocarbon-based oils and fats include hydrocarbon oils such as squalene, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and vaseline.

Examples of the silicone-based oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane and methylcetyloxysiloxane copolymer, dimethylsiloxane and methyl steoxysiloxane copolymer and alkyl. Modified silicone oil, amino modified silicone oil and the like.

Examples of the fluorine-based oil include perfluoropolyether and the like.

Animal or vegetable fats and oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, soybean oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil, Cottonseed oil, palm oil, coukin nut oil, wheat germ oil, rice germ oil, shea butter, moon dog colostrum oil, marker demy nuts nut, meadow home oil, egg yolk oil, Uji, horse oil, mink oil, orange rape oil, jojoba oil, Animal or plant fats and oils, such as candeler wax, carnava wax, liquid lanolin, and hardened castor oil.

Examples of the moisturizing agent include a water-soluble low molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.

As the water-soluble low molecular moisturizer, serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol (polymerization degree n = 2 or more), polypropylene glycol (Polymerization degree n = 2 or more), polyglycerol (polymerization degree n = 2 or more), lactic acid, lactic acid salt, etc. are mentioned.

Examples of the fat-soluble low molecular humectants include cholesterol and cholesterol esters.

Examples of the water-soluble polymer include carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water soluble chitin, chitosan, and dextrin. Can be.

Examples of the fat-soluble polymers include polyvinylpyrrolidone-eicosene copolymers, polyvinylpyrrolidone-hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, polymer silicones, and the like.

Examples of the emollient include long-chain acyl glutamic acid cholesteryl esters, hydroxystearic acid cholesterol, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

As surfactant, nonionic surfactant, anionic surfactant, cationic surfactant, amphoteric surfactant, etc. are mentioned.

Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerol fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbitan fatty acid esters, POE Glycerin Fatty Acid Ester, POE Alkyl Ether, POE Fatty Acid Ester, POE Cured Castor Oil, POE Castor Oil, POE · POP (Polyoxyethylene Polyoxypropylene) Copolymer, POE / POP Alkyl Ether, Polyether Modified Silicone, Laurin Acid Alkanolamide, alkylamine oxide, hydrogenated soybean phospholipid, etc. are mentioned.

As anionic surfactant, fatty acid soap, (alpha)-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphorus salt, alkylamide Phosphates, alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfo acetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. have.

As cationic surfactant, alkyl trimethylammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium bromide, behenyl trimethyl ammonium chloride, chloride Benzalkonium, diethylaminoethyl stearate, dimethylaminopropyl stearate, lanolin derivatives, quaternary ammonium salts, and the like.

Examples of amphoteric surfactants include carboxybetaine type, amide betatype, sulfobetaine type, hydroxysulfobetaine type, amide sulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type and amideamine type. An amphoteric surfactant etc. are mentioned.

Organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengal, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, carbon black and composites thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene, styrene copolymer, Organic pigments such as silk powder, cellulose, CI pigment yellow, CI pigment orange, and composite pigments of these inorganic pigments and organic pigments;

As organic powder, Metal soaps, such as a calcium stearate; Alkyl phosphate metal salts such as sodium cetylinate, zinc lauryl acid and calcium laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-β-alanine calcium, N-lauroyl-β-alanine zinc and N-lauroylglycine calcium; N-lauroyl-taurine calcium, N-palmitoyl- Amide sulfonic acid polyvalent metal salts, such as taurine calcium; N-acyl basic amino acids, such as N (epsilon) -lauroyl-L- lysine, N (epsilon) -palmitolysine, N (alpha)-partoylol nitin, N (alpha)-lauroyl arginine, and N (alpha) -cured fatty acid acyl arginine; N-acyl polypeptides, such as N-lauroyl glycyl glycine; α-amino fatty acids such as α-aminocaprylic acid and α-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride and the like.

Examples of the ultraviolet absorber include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoic acid, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate and cinnamic acid benzyl , Paramethoxy cinnamic acid 2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono 2-ethylhexane glyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl diisopropyl cinnamic acid ester mixture, urokanoic acid Ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone, tetra Hydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy) -1,3 , 5-triazine, 2- (2-hydroxy 5-methylphenyl) benzotriazole and the like.

As fungicides, hinokithiol, trichloric acid, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, ginxitlionone, benzalkonium chloride, photosensitive Sodium No. 301, sodium mononitroguicol, undecylenic acid, and the like.

Examples of the antioxidant include butylhydroxyanisole, propyl gallic acid, and erythorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fmaric acid, sodium pmarate, succinic acid, sodium succinate, sodium hydroxide, sodium dihydrogen phosphate, and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, the compounding component which may be added other than this is not limited to this, Moreover, Although all said components can be mix | blended within the range which does not impair the objective and effect of this invention, Preferably it is 0.01-5 weight% with respect to gross weight, More Preferably it is mix | blended with 0.01-3 weight%.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.

Examples of the form of the cosmetic composition are not particularly limited, and examples thereof include milky lotion, cream, lotion, pack, foundation, lotion, beauty liquid, hair cosmetic, and soap.

Specific examples of the cosmetic of the present invention face wash, face wash foam, cleansing cream, cleansing milk, cleansing lotion, massage cream, cold cream, moisturizing cream, emulsion, lotion, pack, aftershave cream, sunburn prevention cream, suntan oil, soap , Body shampoo, hair shampoo, hair rinse, hair treatment, wool, hair care, hair cream, hair liquid, set lotion, hair spray, hair bridge, coloring, color spray, permanent wave liquid, press powder, loose powder Eye shadows, hand creams, lipsticks, and the like.

The cosmetic of the present invention is an ingredient selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extracts in addition to the ecstasy extract, which is an essential ingredient (combination may be added in addition to those exemplified above as necessary). Components, etc.) can be obtained according to a known method, for example, according to the method described in the "transdermal application formulation development manual" Matsumoto Michio supervision first edition (Seishi-ken, 1985).

In addition, the present invention provides a cosmetic additive for the prevention and improvement effect of allergic skin disease containing the Ecklonia cava extract as an active ingredient.

The cosmetic additives can be used in addition to the production process or finished products of existing cosmetics and cleansing agents, and has the function of preventing and improving allergic skin diseases. It can also be used in creams, lotions, massage packs, systemic cleansers, soaps, shampoos, and other care preparations.

Ecklonia cava extract of the present invention has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of prevention.

Hereinafter, the present invention will be described.

Extract processing RBL -2 H3  Impact on Cell Viability

MTT assay was performed to investigate the effect of red ginseng and algae extracts on cell viability in RBL-2H3 cells. MTT assay is a water-insoluble MTT formazan (3- (4,5-dime thylthiazol-2-yl) -2,5-diphenyl -tetrazoliumbromide that has blue purple color of water-soluble MTT tetrazolium by dehydrogenase action). This test uses the ability of the mitochondria to reduce to).

As can be seen in Figure 1, each extract A: 200, B: 200, C: it was confirmed that there is no cytotoxic at the concentration below 100 (ug / ml). Since the experiment was conducted at the highest concentration without cytotoxicity (Fig. 1).

Influences of Extracts from Korean Dried Algae Extracted with Various Solvents on Cell Viability

As can be seen in Figure 2, each extract purified water: 800, alcohol: 100, ethanol: 50, methanol: 100 (ug / ml) was confirmed that there is no cytotoxicity. Since the experiment was conducted at the highest concentration without cytotoxicity (Fig. 2).

Ecklonia  40 ℃, 90 ℃ Purified water , 80% Methanol Extract and Red Ginseng Purified water  Effect of Extracts on Cell Viability

As can be seen in Figure 3, each Ecklonia cava extract 40 ℃ purified water: 25, 90 ℃ purified water: 50, methanol: 100, red ginseng purified water: 1000 (ug / ml) concentration was confirmed that there is no cytotoxicity, Eckloniax 40 Treatment of 10ug / ml purified water and 250ug / ml of red ginseng extract showed simultaneous cell growth rate of about 90%. Since the experiment was conducted at the highest concentration without cytotoxicity (Fig. 3).

Extract RBL -2 H3  cell Degranulation  Impact

When basophils and mast cells are stimulated by degranulation inducers or IgE by crosslinking of antigens, they secrete histamine through degranulation, causing immediate hypersensitivity and allergy. Hexosaminidase is also a granule component secreted from cells along with histamine and is used as a degranulation indicator for basophils and mast cells. RBL-2H3 degranulation inhibitory effect was confirmed by β-hexosaminidase assay in order to find possible anti-allergic extracts. The concentration of each extract was treated with a concentration without cytotoxicity to determine the inhibitory effect.

Seaweed Extract Processing RBL -2 H3  cell Degranulation  Impact

As can be seen in Figure 4, three kinds of seaweed extracts in RBL-2H3 cells inhibited β-hexosaminidase secretion in a concentration-dependent manner below the highest concentrations that do not exhibit cytotoxicity. Among them, the degranulation inhibitory effect was good at the lowest concentration (100 ug / ml), and there was no rejection, and the easily picked king radish was selected as a material so that it could act advantageously for sale during product development (FIG. 4).

Dried perilla extract treatment RBL -2 H3  cell Degranulation  Impact

As can be seen in Figure 5, the four extracts of Rhizome extract from RBL-2H3 cells had less degranulation inhibitory effect at or below the highest concentration of each of which did not exhibit cytotoxicity. Purified water extract had no inhibitory effect. The best inhibitory effect of 70% ethanol extract from the previous experiment was obtained from Jeju useful biomass extract bank. Therefore, it is thought that the content of the anti-granular inhibitory active ingredient is low in Wanggoe extract directly from the mountains near Konwol University, Danwol-dong, Chungju-si, Chungju (Fig. 5). Later, in the experiment, there was no objection to people and reselected easily available ecstasy as a material so that it could be advantageous for sales in product development.

Ecstatic  40 ℃ Purified water , 90 ℃ Purified water Methanol extract treatment RBL -2 H3  cell mask Impact on Granules

As can be seen in Figure 6, three Ecklonia cava extracts in RBL-2H3 cells inhibited β-hexosaminidase secretion in a concentration-dependent manner below each peak concentration, which did not exhibit cytotoxicity. Among them, the degranulation inhibitory effect was good at the smallest concentration (25ug / ml), and it was determined as an extract of 40 ° C. purified water extraction method that can be used as a food additive composition (FIG. 6).

Ecstatic  40 ℃ Purified water and  Red ginseng Purified water  Simultaneous treatment of extracts RBL -2 H3  cell Degranulation  Impact

As can be seen in Figure 7, 40 ℃ purified water Ecklonia cava extract in RBL-2H3 cells inhibited β-hexosaminidase secretion in a concentration-dependent manner. However, when the red ginseng purified water extract alone treatment and 40 ℃ purified water Ecklonia cava extract simultaneously treatment did not increase the significant degranulation inhibitory effect (Fig. 7).

 Dry in various drying ways Ecklonia  40 ℃ Purified water  Extract RBL -2 H3  Cell cell viability and Degranulation  Impact

As can be seen in Figure 8, the eccentric leaves and stems dried by various drying methods were extracted with purified water at 40 ℃ was measured for cytotoxicity, except for leaf extrusion 100 did not show cytotoxicity at 400ug / ml concentration in the leaves . But the stem showed about 20% more cytotoxicity than the leaves at the same concentration (FIG. 8).

The degranulation inhibitory effect was then measured at each highest concentration without cytotoxicity. The leaf part of Ecklonia cava showed 25% more degranulation inhibitory effect than the stem part, and among the various drying methods, the dry granules with 100 hours of far-infrared drying for 1 hour had the highest degranulation inhibition effect.

Extract RBL -2 H3  In a cell IL -4 effect on transcription, expression

Interleukin-4 (IL-4) is a cytokine produced mainly by Th2 subclass of CD4 + helper T cells. Its function induces the differentiation of Th2 cells from naïve CD4 + progenitor cells, stimulates IgE production by B cells, And inhibition of IFN-γ-dependent macrophage function.

The expression of the mRNA of IL-4 in moths and worms was confirmed at the transcriptional stage using RT-PCR. As can be seen in FIG. 10, the worms and the worms hatched the mRNA expression of IL-4 in a concentration-dependent manner (FIG. 10).

Red ginseng purified water extraction, 100 ℃ far infrared ray drying 1 hour production of IL-4 of Ecklonia cava 40 ℃ purified water extract was confirmed by ELISA.

As can be seen in Figure 11, red ginseng purified water extraction, dried Ecklonia cava 40 ℃ purified water extract dried at 100 ℃ for 1 hour did not significantly inhibit the production of IL-4 (Fig. 11).

Ecklon Feed  Manual skin hypersensitivity ( passive cutaneous anaphylaxis  ; PCA ) On the reaction spirit incense

Mediators such as histamine and heparin, which are involved in the early stage allergic reactions, may lead to increased vascular permeability, bronchial contraction, bowel hyperactivity, tissue injury, and early inflammatory responses. BALB / c mice with passive cutaneous anaphylaxis reactions were investigated for their potential as anti-allergic beverages after 5 days of free feeding with 1% and 5% dried Ecklonia cava at 100 ° C for 1 hour. Mice were used for the experiment in 5 rats in each group.

Ecklonia cava was dried at 100 ° C for 1 hour, and the secretion of allergens (histamine, heparin) was suppressed in the in vitro mast cells. Based on these results, IgE- mast cell dependent passive skin hypersensitivity (PCA) response animal model was used.

In this animal model, IgE is injected into the mouse's ear to sensitize local mast cells, and evans blue pigment due to activation of mast cells present in the ear after intravenous tail injection of antigen (DNP-HSA) and evans blue pigment from blood vessels to ear tissue. This is an experiment to measure the degree of leakage. As can be seen in Figure 12, the outflow of evans blue from the blood vessels caused by the antigen was observed to be significantly increased by the antigen stimulation, feed containing 1%, 5% Eckloniasis for 5 days free feeding group to antigen stimulation It was confirmed that the outflow of evans blue into the ear tissues significantly inhibited (FIG. 12).

As can be seen through the present invention, physiological activity from the processing step by confirming the efficacy by developing an optimal drying method and 100 ℃ purified water extraction method of 100 ℃ far infrared 1 hour to maximize the anti-allergic efficacy of Ecklonia cava extract The method of maximization was established, and it showed the possibility of developing anti-allergic active ingredient of Ecklonia cava extracted in the above manner as a new drug.

In addition, by using the leaves with less cytotoxicity and better anti-allergic effect among the leaves and stems of Ecklonia cava, we maximized the safety and anti-allergic effect of functional drinks for young children, the main patient group. By establishing an immunological research model that can be quickly identified at any time, it can be used for future research on the development of various materials using domestic natural products, and contribute to the diversification and industrialization of functional products through the development of high-efficiency, high-functional anti-allergic bioactive substances. It has an effect.

In addition, it was possible to confirm the potential for anti-allergic functional materials by mixing other components such as red ginseng in Ecklonia cava.

Figure 1 is a cytotoxicity (cytotoxicity) measurement of each extract of algae in RBL-2H3 cells. Three extracts were treated for 24 hours at the indicated concentrations, followed by MTT assay.
Figure 2 is a measurement of cytotoxicity (cytotoxicity) of the extract of each extract from RBL-2H3 cells. The extracts were treated with the indicated concentrations for 24 hours and then the results of MTT assay.
Figure 3 is an illustration of the cytotoxicity (cytotoxicity) of each extract of Ecklonia cava and red ginseng in RBL-2H3 cells. The extracts were treated with the indicated concentrations for 24 hours and then the results of MTT assay.
Figure 4 shows the degranulation inhibitory effect of seaweed extract in RBL-2H3 cells. The results measured using the β-hexosaminidase assay are shown.
Figure 5 shows the degranulation inhibitory effect of the extract from the extract from Rhwang-2H3 in RBL-2H3 cells. Shows the results measured using the β-hexosaminidase assay.
Figure 6 shows the degranulation inhibitory effect of Ecklonia cava extract in RBL-2H3 cells. The results measured using β-hexosaminidase assay.
Figure 7 shows the degranulation inhibitory effect of Ecklonia cava extract and red ginseng extract in RBL-2H3 cells. The results measured using the β-hexosaminidase assay are shown.
Figure 8 is a cytotoxicity measurement of Ecklonia cava extract in RBL-2H3 cells. The extracts were treated for 24 hours at the indicated concentrations, showing the results of MTT assay.
Figure 9 shows the degranulation inhibitory effect of Ecklonia cava extract in RBL-2H3 cells. Shows the results measured using β-hexosaminidase assay.
Figure 10 is a diagram showing the inhibitory effect of IL-4 mRNA of the worms in the worms in RBL-2H3 cells. Measured using RT-PCR.
Figure 11 shows the results of measurement of IL-4 production of red ginseng, Ecklonia cava extract in RBL-2H3 cells. Measured using ELISA.
12 is a diagram showing the effect of Ecklonia cava diet on passive cutaneous anaphylaxis (PCA) response.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples. The following examples are intended to illustrate the invention and the scope of the invention is not to be construed as being limited by the following examples.

Insects used in the present invention (Sargassum thunbergii, starch, 80% methanol extraction), Hoksaeng yameri (Sargassum horneri, male starch, 80% methanol extraction), King Dwarf (Lactuca indica, starch, 70% ethanol extraction), Each extract of Ecklonia cava ( Ecklonia cava , outpost, 40 ℃, 90 ℃ purified water extract, 80% methanol extract ) was distributed by Jeju useful biological resource extract bank, and selected after anti-allergic screening, the king eel was selected from Dankook-dong, Danju-dong, Chungju-si, Chungju Extracts were obtained directly from the extract, and Ecklonia cava was distributed from Jeju useful biomass extract bank and dried after various drying methods. Methanol and ethanol were purchased from Deoksan Pharmaceutical Co., Ltd., and Alcohol Alcohol was purchased from Natural F & P Co., Ltd. Red ginseng purified water extract was distributed by Chungbuk Ginseng Farming Corporation. MEM (minimum essential medium) medium was purchased from Thermo scientific, anti-DNP-HSA IgE, DNP-HSA (dinitrophenyl-human serum albumin), and evans blue from Sigma Aldrich. QIAzol was purchased from Qiagen and chloroform, isopropanol, and DEPC (diethylpyrocarbonate) -treated water were purchased from Sigma Aldrich. DNTP mix, 10xbuffer, MgCl ₂ , DTT, and RNaseOUT used in reverse transcriptase reaction were purchased from Invitrogen and oligo dT from Bioneer. 10 × Taq buffer, dNTP mix, and EX Taq HS used in the PCR reaction were purchased from Takara. IL-4 ELISA kit was purchased from BD biosciences. Eckloniaticis 5% feed diet composition (when manufacturing 100g) was Casein: 20g, Starch: 50g, sucrose: 15g, Vit. Mix: 1g, Mineral mix 3.5g, Cellulose: 5g, DL-methionine: 0.3g, Choline barbiturate: 0.2g, DL-alpha-tocopherol: 0.12g, Corn oil: 5g, Easiness dried at 1 hour at 100 ℃ Used as.

The mice used in the experiment were supplied from Nara Biotech Co., Ltd. and sterilized rug and feed were used. In animal experiments, female mice used BALB / c species and were 6-7 weeks old and weighed 20-22 g.

Example  1: extraction and drying method

1) King Dwarf

The extract of Wanggogi extract was collected directly from the mountain near Konkuk University, Danwol-dong, Chungju-si, Chungbuk, and then pulverized and 10g was put in a Erlenmeyer flask. Repeat three times to extract the stirring. The extracted extract proceeds while storing at 4 ℃. After extraction is complete, each extract is filtered with filter paper (Whatman No. 2). After evaporating to dry at 45 ° C. under reduced pressure using a rotary evaporator, 100 ml of water is added thereto, and the concentrate is recovered. After evaporation and drying, the weight of the obtained solid was measured to be 20 mg / ml, and then the experiment was performed.

2) drying, grinding and extraction of Ecklonia cava leaves

Ecklonia cava leaf used in the present invention was removed from the stem portion was used only the leaf portion.

The leaves were used after washing 2-3 times in running water to remove water. The drying method used natural drying, hot air drying, and far-infrared drying. During drying, the leaves were chopped and dried, and the drying time was dried until the moisture content of the Ecklonia cava was about 10% after drying. Natural drying for 48 hours in the shade, hot air drying for 2 hours at 100 ℃ using a dryer (Vision Science. Bucheon, Korea), far infrared drying at 100 ℃ using a far-infrared roaster (Korea Energy Technology, Seoul, Korea) 1 hour of treatment. The dried Ecklonia cava leaves were pulverized using a grinder (Daehwa Precision. Daegu, Korea). The ground leaves were screened to 600μm or less using a test sieve (chung gye. Seoul, Korea) and used for extraction and feed preparation.

Example  2: cell culture

Rat basophil leukemia (RBL) -2H3 cells were cultured in an incubator at 37 ° C and 5% CO ₂ with MEM (Minimum Essential Medium) containing 10% FBS (fetal bovine serum) and 1% antibiotics. RBL-2H3 cells were passaged in 1x10 ⁶ cells in a cell culture dish (100π).

Example  3: Measurement of cytotoxicity MTT assay )

RBL-2H3 cells were dispensed into 96 well plates at 1 × 10 ⁴ cells / well and stabilized for 4 hours. Each sample was treated for each concentration (㎍ / ml) and then incubated for 24 hours under the same conditions. After removing the medium, 100 μl of MTT (3- (4,5-Dimethyl-thiazol-2-yl) -2,5-diphenyl tetrazolium bromide) at a concentration of 0.5 mg / ml was added to each well and incubated for 4 hours. It was. MTT was removed and 200 μl of DMSO was added, followed by stirring for 10 minutes using a stirrer to dissolve the MTT formazan produced in the cells. After stirring, the absorbance was measured at 540 nm with an ELISA reader.

Example  4: Degranulation  Measurement (β- hexosaminidase assay )

After suspending RBL-2H3 in MEM, add anti-DNP-HSA IgE (70ng / ml final concentration) and dispense 2.5x10 ⁵ cell / well in a 24 well plate. 37 ℃, 5% CO ₂ 7 hours incubation in the incubator was sensitized. The cells in each well were washed twice with Siraganian buffer (NaCl 119 mM, KCl 5 mM, Glucose 5.6 mM, MgCl ₂ (6H ₂ O) 0.4 mM, PIPES 25 mM, NaOH 40 mM, CaCl ₂ 1 mM, pH 7.2). Each well was treated with 200 μl Siraganian buffer and sample for each concentration and reacted for 30 minutes. The cells were then reacted for 15 minutes using DNP-HSA (25 ng / ml final concentration) and incubated for 10 minutes in an ice bath to terminate the reaction. Transfer 150 µl of the supernatant to the tube, centrifuge at 3000 rpm for 3 minutes, transfer 30 µl to a 96 well plate, and transfer the substrate buffer (4- p -Nitrophenyl-N-acetyl--D-glucosaminide 1 mM, 0.1 M citrate buffer, pH). 4.5) 30 μl of the solution was reacted at 37 ° C. for 1 hour, and 250 μl of stop solution (0.1M NaHCO 3) was added to each well to stop the reaction. The absorbance was measured at 405 nm using an ELISA reader. Degranulation percent (%) was calculated by the following equation as the absorbance values of the sample and the control.

degranulation percent (%) = (sample treatment group OD-blank OD) / (control OD-blank OD) x100

Example  5: IL -4 mRNA  Expression measurement ( RT - PCR )

After RBL-2H3 was suspended in MEM medium, anti-DNP-HSA IgE (70ng / ml final concentration) was added, and 1x10 ⁶ cells were dispensed into a cell culture dish (60π) and incubated for 7 hours. Each sample was treated for 30 minutes at different concentrations, and cultured under the same conditions for 1 hour after the addition of DNP-HSA (25 ng / ml final concentration). The medium was removed, washed with PBS (phosphate buffered saline) and transferred to ice to terminate the reaction. Cells were lysed with 1 ml of QIAzol, 200 μl of chloroform was added, mixed well with vortex and centrifuged at 4 ° C. and 13,000 rpm for 15 minutes. 300 μl of the supernatant was added and 500 μl of isopropanol was added and centrifuged. The supernatant was removed, washed with 1 ml of 75% ethanol, centrifuged and dried. After adding 30 μl of DEPC treated water, the concentration was measured using Nanodrop, and the RNA was used for quantification, and the result was confirmed by electrophoresis. 2 g of RNA and 1 μl oligo dT, 1 μl of 10 mM dNTP mix, 4 μl of 10 × buffer, 4 μl of 25 mM MgCl 2, 2 μl of 0.1 M DTT, 1 μl of RNase OUT 1 μl of SS Ⅲ RT was added, followed by filling up to 20 μl with water, followed by a reverse transcriptase reaction. 1 μl of β-actin primer (F / R), 2 μl of 10x EX Taq buffer, 2 μl of dNTP mix, and 0.2 μl of EX Taq HS were added to 2 μl of cDNA of each group. After mixing, PCR (polymerase chain reaction) was performed and confirmed by agarose gel electrophoresis. IL-4 primer (F / R) was performed in the same manner.

Example  6: IL -4 protein expression measurement ( ELISA )

After RBL-2H3 cells were suspended in MEM medium, anti-DNP-HSA IgE (70ng / ml final concentration) was added, and the cells were dispensed at 1x10 ⁵ cell / well in a 24 well plate and 37 ° C 5% CO ₂ 4 hours incubation in the incubator was sensitized. Four hours after DNP-HSA stimulation, cell culture supernatants were collected and subjected to ELISA for IL-4 quantification. Phosphate containing blocking solution (5% skim milk / 0.05% Tween20) in a 96 well plate (Nunc, Roskilde, Denmark) for Sandwich ELISA and rat rat anti-IL-4 (BD biosciences). Buffered saline (PBS) was added and reacted at room temperature for 2 hours. After adding the IL-4 standard and the culture supernatant and reacting for 2 hours, the reaction antibody (1: 250) + Av-HRP (1: 250) was reacted with PBS-T containing 2% skim milk for 1 hour and then tetramethylbenzidine substrate. The solution was added and reacted, and the reaction was stopped with 2N H ₂ SO ₄ , and the absorbance was measured at 450 nm with an ELISA reader.

Example  7: manual skin Anaphylaxis ( passive cutaneous anaphylaxis , PCA ) reaction

Passive cutaneous anaphylaxis reaction was caused by intravenous injection of anti-DNP IgE into the mouse ear and 24 hours after sensitization of the skin, intravenous DNP-HSA was injected intravenously into the tail. Each iris extract was applied to the ear 1 hour before administration of the antigen (DNP-HSA). Azelastine is an antihistamine that is applied to the ear in the same way as irises at 1%. DNP-HSA was diluted with phosphate-buffered saline (PBS). 0.5 ug / 30 ul of anti-DNP IgE was injected into the ears of the mouse, and 24 hours later, a solution containing 1% of 2% evans blue and 100 ug DNP-HSA was injected into the tail vein of the rat. After killing the mice, the reaction site was cut to the same size, 200 μl formamide was added, and evans blue was extracted at 64 ° C. Absorbance of evans blue was measured at 650 nm of spetrophotometer and compared with control.

Claims (7)

An anti-allergic or anti-atopic composition comprising an effective amount of Ecklonia cava leaf extract, which is extracted from Ecklonia cava leaf with 40 ° C. water. The anti-allergic or anti-atopic composition of claim 1, wherein the Ecklonia cava leaves are dried at 1 ° C. in far infrared rays. The anti-allergic or anti-atopic composition according to claim 1, wherein the Ecklonia cava leaf extract contains 1-5% of the total composition. The anti-allergic or anti-atopic composition according to claim 1, wherein the effective amount of the Ecklonia cava leaf extract is 5 to 25 µg / ml. An anti-allergic or anti-atopic functional food composition containing an effective amount of Ecklonia cava leaf extract extracted from Ecklonia cava leaf with water at 40 ° C. The anti-allergic or anti-atopic functional food composition according to claim 5, wherein the Ecklonia cava leaves are dried at 1 ° C. in far infrared rays. Method of producing an anti-allergic or anti-atopic composition comprising the step of extracting the Ecklonia cava leaves after extracting the Ecklonia cava leaves at 100 ° C. for 1 hour with far infrared rays.

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