KR100536857B1 - Cosmetic composition comprising the extract of Actinidia arguta having anti-allergy and anti-inflammation activity - Google Patents

Abstract

 The present invention relates to a composition containing a wormwood extract having anti-allergic and anti-inflammatory activity, the worm extract of the present invention reduces IgE and selectively reduces Th2 cytokines in OVA sensitized mouse models and human cell lines and serum, Food additives useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases, which have the effect of inhibiting the release of histamine from the mast cells and inhibiting the inflammatory activity. The present invention provides animal feed additives useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases and feed comprising the same, or cosmetic compositions useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases.

Description

Cosmetic composition comprising the extract of the wormwood having anti-allergic and anti-inflammatory activity

The present invention includes food additives useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases containing the extract as a active ingredient, animal feed additives useful for the prevention and improvement of allergic or non-allergic inflammatory diseases and the same The present invention relates to a feed composition or a cosmetic composition useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases.

Allergic diseases include anaphylaxis, allergic rhinitis, and asthma, atopic dermatitis, insect allergies, food allergies, drug allergies, and urticaria. Allergic diseases such as these have been developed to about 20% of the population in many countries and their prevalence is increasing (Wuthrich B., Int. Arch. Allergy Appl . Immunol ., 90 , pp3-10, 1989). Similarly to humans, atopic dermatitis, asthma, and diarrhea and vomiting due to food allergies have been reported as serious symptoms, and the incidence of the disease is also rapidly increasing. In particular, atopic dermatitis has recently been visited at animal hospitals. Accounted for 20-30% of the total (Lund et al., J. Am. Vet. Med. Assoc. , 214 (9) , pp1336-1341, 1999).

Allergic diseases are mediated by IgE, which demonstrates that type 2 T helper (Th2) cells, mast cells and eosinophils play an important role in this process (Maggi E., Immunoyrchnology, 3 , pp233-). 244, 1998; Pawankar R., Curr. Opin.Allergy Clin.Imunmun., 1 , pp3-6, 2001; Vercelli D. , Clin.Allergy Immunol ., 16 , pp179-196, 2002). IgE is one of many immunoglobulin isotypes commonly present in the serum of human or some other animal species, without helminth infection or other stimuli (Coffman RL. Et al., J. Immunol . 136) . , pp 949-64, 1986). According to the Th2 hypothesis, IgE is likely to be produced in immunological conditions predominantly humoral immunity mediated by cytokines such as Th2 cells, IL-4, IL-5 and IL-13 (Maggi E., Immunotechnology , 3 , pp 233-244, 1998). Current concepts of the pathogenesis of allergic diseases suggest that genetic and environmental factors interact with each other, thereby producing IL-4 via Stat6-mediated signaling pathways and c-Maf, GATA3, NIP45 and Activation of specific transcription factors such as NFATc and consequently results in the development of allergen-specific T helper 2 CD4 + cells. Th2 cells activated by allergens produced once secrete IL-4, IL-5 and IL-13. IL-4 and IL-5 induce the production of IgE and IgGI by B cells, stimulate the development of acidic leukocytes in the bone marrow and induce them to collect in inflamed tissues (Erb KJ., Immunol. Today, 20 , pp 317-322, 1999; Rothenberg ME., N. Engl. Med. , 338 , pp 1592-1600, 1998). IL-13 is a cytokine closely related to IL-4, which binds to the IL-4 receptor alpha chain and induces an allergic phenotype independent of IL-4, IgE or acidic leukocytes (Wills-Karp M., et al., Science, 282 , pp 2258-61, 1998; Grunig G., et al., Science, 282 , pp2261-2263, 1998).

IgE also circulates and binds to the IgE receptors of two isoforms. Affinity IgE receptors (FcεRI) are present on the surface of mast cells and basic leukocytes, and low affinity IgE receptors (FcεRII or CD23) are present on the surface of lymphocytes, acidic leukocytes, platelets and macrophages. It is believed that the most important factor governing the pathogenesis of allergic disease is the degranulation of mast cells resulting from cross-linking and allergen support of IgE receptors on mast cells. Molecules released by mast cells include histamine, heparin, proteases and free radicals and mediate various biological effects such as vasodilation, intestinal and / or bronchial smooth muscle contraction, mucus secretion and local protein secretion. Influx of leukocytes, basic leukocytes and lymphocytes occurs after 6-24 hours following the initial mediated response of mast cells. This late stage response leads to chronic inflammation of tissues constantly exposed to the antigen. Generally, for the prevention of allergic diseases, compositions such as antihistamine, steroidal or nonsteroidal anti-inflammatory drugs and leukotriene antagonists are used. Such compositions are primarily useful for symptomatic effects and do not provide the protection required for the fundamental healing of allergic diseases such as alleviating excessive humoral immunity or inhibiting IgE production. The hypothesis that reducing serum IgE levels can improve allergic symptoms has been demonstrated by clinical trials of chimeric anti-IgE antibodies (CGP-51901) and recombinant humanized monoclonal antibodies (rhuMAB-F25) (Fahy JV et al., Am. J. Respir. Crit. Care. Med., 155 , pp 1828-1834,1997). Diacyl benzimidazole homologs and bacterial polysaccharides that inhibit IgE synthesis and secretion are described in US Pat. No. 6,369,091 and US Patent Application Publication 20020041885, respectively. Some oriental medicine regimens (Yang et al., 2001) and herbs such as Siegesbeckia glabrescens (Kim HM. Et al., Phytother Res ., 15 , pp572-576, 2001) are known for their IgE-reducing activity. In addition, many herbs have been found to be a rich source of histamine release inhibitors or anti-inflammatory drugs.

As the culture of mankind developed and the economic margins increased, food brought about quantitative increase and qualitative improvement, and the demand for food with convenient convenience increased. As a result, foods such as mass-produced and consumed processed foods, instant foods, etc. are more likely to come to the table, and new kinds of foods are pouring out every day, and this phenomenon is accompanied by the use of food additives.

Food additives (or food additives) refer to materials used by food additives, mixing, infiltration or other methods for the manufacture, processing or preservation of food. There are two types of food additives, natural and synthetic, which are usually classified according to their function and use. Currently, there are more than 370 chemical synthetic products and 50 natural additives that are approved as food additives in Korea.They are mainly preservatives, fungicides, antioxidants, colorants, colorants, bleaches, seasonings, sweeteners, flavoring agents, swelling agents, reinforcing agents, They are classified into improvers, emulsifiers, thickeners (hofues) and stabilizers, coatings, gum base agents, antifoams, solvents, mold release agents, insect repellents, quality improvers and other food additives.

On the other hand, functional food refers to foods developed so that the primary function of the food, the nutritional function of the primary function, the sensory function of the secondary function in addition to the third function of the bioregulatory function. Bioregulatory functions that foods can exhibit include anticancer, lowering blood pressure, lowering cholesterol, inhibiting blood clots, preventing diabetes, and inhibiting aging. Functional foods directly contribute to improving the quality of life, reducing medical expenses of individuals and nations by preventing chronic diseases through bioregulatory functions, and forming a rich welfare state by implementing a healthy society. Therefore, in order to impart such functionality to food, a material having a specific function may be used in the food as a food additive.

Feed is a substance that supplies organic or inorganic nutrients necessary to maintain livestock and pets' lives and to produce milk, meat, eggs and hides. Good feed is rich in nutrients and livestock It is free from harmful ingredients, has high yields, is always readily available, always fresh and has a high digestibility, and can be classified into various types according to nutritional value, main ingredient, distribution, moisture content, blending state and processing type. .

Feed additives are edible additives used for the purpose of preventing diseases, supplementing deficiencies, improving feed efficiency and promoting growth by adding to feeds. Vitamins, furovitamins, antibiotics, antibacterial agents, antioxidants, antifungal agents, enzymes There are probiotics, amino acids and trace minerals. In general terms, a feed additive is used to enhance the effectiveness of feed by adding or supplementing small amounts to livestock feed, which mainly supplies energy and protein, in order to maximize livestock and animal production. Thus, it is distinguished from animal medicine for the purpose of prevention and treatment of diseases.

Cosmetics are articles used in other similar ways that are applied or sprayed to clean or beautify the human body and should not cause changes or malfunctions in the human body. Cosmetic raw materials have changed little by little from the past to the present, at the request of each era. In the past, many synthetic chemicals were used, but recently, many cosmetic products using natural products have appeared. However, preservatives, surfactants, pigments, UV absorbers, and other ingredients that are still essential for cosmetics may irritate skin cells, cause allergic reactions, and eventually cause skin irritation. This can be In addition, fatty acid, high alcohol, protein, etc. in sebum, sweat, and cosmetic components discharged from the body are decomposed into highly toxic substances by dermal fungi present on the skin, which may cause skin inflammation. It is well known that the sun's ultraviolet rays can also cause skin irritation. In particular, all preservatives are manufactured through chemical synthesis to perform the function of preservatives, but skin allergy due to the high irritation and toxicity of the skin even with the use of small amounts (Andrea Counti et al ., Contact Dematitis , 37 , pp35-36, 1997 ), There is a problem of skin edema, and various side effects may occur by each component such as perfume (Maibach HI, Contact Dermatitis , 6 , pp369-404, 1980). In addition, recently, as the demand for functional cosmetics increases, various cosmetic products including various functional ingredients have been developed even though there are some side effects on the skin. The symptoms of cosmetic side effects can be divided into three categories: cosmetic allergy, cosmetic irritant dermatitis, and existing dermatitis.

Therefore, to prevent the skin allergy and inflammation by maximizing the effect of each of the ingredients and minimizing side effects is a very important problem in cosmetics, it is necessary to develop various anti-allergic and anti-inflammatory substances.

Actinidia arguta , the main component of the composition used in the present invention, belongs to Actinidiaceae and is native to Siberia, northern China, and Korea. More than 30 species of similar species have been reported, and the representative ones of which are A. kolomikta , A. polygama , A. rupa , etc., commonly known as kiwi ( A. chinensis or A. deliciosa ) It is used as a herbal medicine called Mifudo. It contains the fruit of the plant of perilla and related plant, which is cold and nontoxic, and has been prescribed for the treatment of liver and gastrointestinal diseases or urinary stones (Shindong medicinal herbs) Drug database, Institute of Natural Products, Seoul National University, Dongbang Media Co., Ltd., 1999), has not been used for the prevention and treatment of allergic diseases and non-allergic inflammatory diseases.

Korean Patent Registration No. 344147 discloses a health beverage mainly based on Tadasu sap and a manufacturing method thereof, and Korean Patent Registration No. 134024 discloses a Health Drink mainly based on a tuna and its manufacturing method. Patent Application No. 1996-10396 discloses cotton wormwood extract, its liver function protector, and its use as an anticancer agent. Domestic Patent Application No. 2001-17524 discloses a beverage for restoring liver function containing wormwood and various herbal extracts. The manufacturing method is disclosed. However, there have been no reports of anti-allergic effects or inhibition of inflammatory responses.

Therefore, in the present invention, in order to determine the anti-allergic and anti-inflammatory activity of the Darae extract, the test for the lowering of serum levels of Th2 cytokine and IgE in human serum, the inhibitory rate of histamine release from mouse peritoneal mast cells and the anti-inflammatory effect on edema As a result of in vitro and in vivo experiments, it was confirmed that the Darae extract according to the present invention lowered the serum levels of Th2 cytokine and IgE, thereby inhibiting the release of histamine from mast cells and inducing the inhibition of the inflammatory response. The invention has been completed.

The present invention lowers the serum levels of Th2 cytokine and IgE containing Tara extract as an active ingredient to prevent and improve allergic or non-allergic inflammatory diseases that inhibit the release of histamine from mast cells and inhibit the inflammatory response. Provided are useful food additives, animal feed additives useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases and feed comprising the same, or cosmetic compositions useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases do.

In order to achieve the above object, the present invention provides a food additive useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases containing the crude extract or non-polar solvent soluble extract as an active ingredient.

The present invention also provides an animal feed additive useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases containing crude crude extract or non-polar solvent soluble extract as an active ingredient, and feed comprising the same.

The present invention also provides a cosmetic composition useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases containing the crude extract or non-polar solvent soluble extract as an active ingredient.

The stalks include Actinidia arguta , A. kolomikta , A. polygama or cognate plants, and include the fruit, stem or root part of the stalk tree.

The crude crude extract includes water, a lower alcohol having a carbon number of C ₁ to C ₄ and a mixed solvent thereof, preferably water or an extract soluble in 50% ethanol, and the nonpolar solvent soluble extract includes hexane, ethyl acetate, Non-polar solvents such as dichloromethane, preferably extracts soluble in ethyl acetate.

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

The non-allergic inflammatory diseases include general inflammatory diseases, and include various dermatitis, systemic lupus erythematosus, retinitis, gastritis, hepatitis, enteritis, pancreatitis, nephritis and the like.

On the other hand, the crude crude extract or non-polar solvent soluble extract is selected in the range of 0.01 to about 90 parts by weight per 100 parts by weight of the composition of the present invention.

The method for separating the crude extract or non-polar solvent soluble extract of the present invention is as follows, the present invention will be described in detail below.

Crude crude extract for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases of the present invention is characterized in that the extraction with water or alcoholic aqueous solution, more specifically,

The coarse crude extract of the present invention is pulverized by drying the cotyledon or cotyledon stem, and then a lower alcohol such as methanol, ethanol and butanol in a volume of about 5 to 25 times, preferably about 10 times, the weight of the cotyledon. Or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably about 0.5 hours to 2 at an extraction temperature of 20 to 100 ° C, preferably 60 to 100 ° C with water or 50%, ethanol 1, preferably 1 to 5 times, preferably 1 to 3 times of continuous extraction after extraction by hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for 1 hour to 1 day, The filtrate was concentrated under reduced pressure at 20 to 100 ° C., preferably 50 to 70 ° C., using a rotary vacuum concentrator, and the residue was purified by vacuum freeze drying, hot air drying, or drying by spraying. Is to obtain a crude extract of Actinidia soluble in a mixed solvent, the obtained extract of Actinidia is used in the experiments was stored at -20 ℃. In addition, the dry powder of the stalk extract is used by dissolving to a certain concentration in distilled water.

In addition, the fusiform non-solvent soluble extract of the present invention, after suspending the crude extract in water, it is about 1 to 100 times, preferably about 1 to 5 times the volume of the non-polar, such as ethyl acetate, chloroform or hexane It can be obtained by adding a solvent and fractionating with a separatory funnel 1 to 10 times, preferably 1 to 5 times. It is also possible to further carry out conventional fractionation processes (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis . 3rd Ed . Pp 6-7, 1998).

In order to investigate the anti-allergic and anti-inflammatory effects of the crude extract or non-polar solvent soluble extract obtained above, the hypoallergenic effect of IgE and Th2 cytokines in OVA sensitized mouse model and human cell line and serum of mast extract (mast) In vitro and in vivo experiments on the inhibitory effect of the release of histamine and the inhibitory effect of inflammatory activity on the cells were confirmed.

Accordingly, the present invention provides a food additive useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases, containing the crude extract or non-polar solvent soluble extract obtained by the above method as an active ingredient.

Food additives containing crude crude extract or non-polar solvent soluble extract of the present invention include organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymeric phosphate), and the like. Any one or more of natural antioxidants such as phosphates, polyphenols, catechin, alpha-tocopherol, rose mary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid It may further comprise.

The crude feed extract or non-polar solvent soluble extract for food additives may be in a high concentration of 20 to 90% or in powder or granule form.

Similarly, the food additive containing the crude crude extract or non-polar solvent soluble 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 crude crude extract or non-polar solvent soluble extract in food preservatives, fungicides, antioxidants, spices, seasonings, sweeteners, flavoring agents, swelling agents, reinforcing agents, modifiers, emulsifiers, various nutrients, synthetic Flavoring agents such as flavoring agents and natural flavoring agents, coloring agents, coloring agents, neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, antifoaming agents, It provides a method of using a food additive, characterized in that it is used as an essential raw material for solvents, mold release agents, preservatives, quality improvers, glycerin, alcohols, carbonation agents used in carbonated drinks, or other food 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 food additive 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 emulsifier were mixed in a preheated mixer at 60 to 70 ° C., followed by mixing. , While maintaining the internal temperature of the mixer at 55 ° C., add 9 g of peppermint fragrance, mix for 30 minutes, extract, roll, and age 24-48 hours to cut and package the 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. Can be used to make spices, fish paste, fish, seasoned poultry, salted fish, etc., used as a dipping, spraying and addition method, and can also be immersed, sprayed and added to kimchi, radish, pickled radish, pickled vegetables.

The present invention also provides an animal feed additive useful for the prevention and improvement of an allergic disease or a non-allergic inflammatory disease, which contains the crude extract or a non-polar solvent soluble extract obtained by the above method as an active ingredient, and a feed comprising the same. .

Crude crude extract or non-polar solvent soluble extract for the animal feed additives may be 20 to 90% high concentrate or powder or granule form.

The crude feed extract or non-polar solvent soluble extract for animal feed additives includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates or polyphosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polymeric phosphate). It further contains any one or more of natural antioxidants such as phenol, catechin, alpha-tocopherol, rose mary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid can do.

Animal feed additives containing crude crude extract or non-polar solvent soluble extract of the present invention and feed comprising the same are amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal, enzymes, live microbial preparations, etc. Various adjuvants such as cereals, for example crushed 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.

Crude crude extract or non-polar solvent soluble extract for the animal feed additives may be administered to the animal alone in combination with other feed additives in an edible carrier.

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

When the crude feed or non-polar solvent soluble extract for the animal feed additive is administered separately from the animal feed, the dosage form of the extract, as is well known in the art, may be used in combination with a non-toxic pharmaceutically acceptable edible carrier for immediate release or It may be prepared in a sustained release formulation. 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. When 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 improving and preventing other allergic and nonallergic inflammatory diseases.

In addition, the animal feed in which the crude crude extract or non-polar solvent soluble extract is included as an animal feed additive may be any protein-containing organic cereal meal conventionally used to meet the dietary needs of animals. Such protein-containing flours typically consist mainly 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 animal feed blending method including the extract according to the present invention, the crude extract or non-polar solvent soluble 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, it is preferable to add water to the animal feed and then go through the usual pelletization, expansion, or extrusion process to prevent separation during storage. Excess water may be removed to dryness.

The present invention provides a cosmetic composition useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases, which comprises the crude extract or non-polar solvent soluble extract obtained by the above method as an active ingredient.

Accordingly, the present invention is to prevent and improve allergic skin diseases or non-allergic skin inflammatory diseases, the amount of crude extract or non-polar solvent soluble extract having 0.01 to 30% by weight, preferably 0.01 to 5.0% by weight of the total cosmetic composition The present invention provides a cosmetic composition useful in the present invention, and is added to a cleansing, face wash, soap, shampoo, rinse and treatment to provide a cosmetic composition useful for the prevention and improvement of allergic skin diseases or non-allergic skin inflammatory diseases.

Cosmetic composition containing the crude extract or non-polar solvent soluble extract of the present invention as an active ingredient may be used in a variety of cosmetics, face wash and shampoo for the prevention and improvement effect of allergic skin diseases or non-allergic skin inflammatory 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.

Cosmetics of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.

The water-soluble vitamin may be any compound that can be incorporated into cosmetics, but preferably vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , pyridoxine, pyridoxine, vitamin B ₁₂ , pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H and the like, and salts thereof (thiamine hydrochloride, sodium ascorbate salt) and derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.) can also be used in the present invention. It is included in water soluble vitamins. 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 D ₂ , vitamin D ₃ , and vitamin E (d1-α-tocopherol, d-α-tocopherol, d-δ-tocopherol) And derivatives thereof (ascorbic palmitate, ascorbic stearate, ascorbic acid dipalmitine, dl-α-tocopherol acetate, dl-α-tocopherolvitamin E, DL-pantothenyl alcohol, D- Pantothenyl alcohol, Pantothenyl ethyl ether, etc.) are contained in the oil-soluble vitamin used by this 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 its salt can be normally purified from a mammal 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.

In addition to the said essential component, you may mix | blend the cosmetics of this invention with the other components normally mix | blended with cosmetics as needed.

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, cooling agents, 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 phthalic acid, neopentyl glycol dicapric acid, neopentyl glycol dioctanoate, glyceryl tricaprylate, glyceryl tritridecyl, glyceryl triisopalmitine, glyceryl triisostearate, octyl dodecyl neopentane 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.

Perfluoro polyether etc. are mentioned as fluorine-based fats and oils.

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.

Water-soluble low molecular humectants include 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, hydroxy stearic acid cholesterol, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl esters, 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 the carboxybetaine type, the amide betain type, the sulfobetain type, the hydroxysulfobetain type, the amide sulfobetain type, the phosphobetaine type, the aminocarboxylate type, the imidazoline derivative type, and the amideamine type. An amphoteric surfactant etc. are mentioned.

Organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, 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; Amide sulfonic acid polyvalent metal salts, such as N-lauroyl-taurine calcium and N-palmitoyl-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-hydric When the like can be mentioned 5-methylphenyl) benzotriazole.

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 0.01-3 weight% compounding.

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 crude extracts or soluble extracts of non-polar solvents, which are essential ingredients (as illustrated above. Compounding ingredients, etc. which may be added in addition to the above) can be obtained by following a known method, for example, according to the method described in the "Dermal Application Formulation Manual" Matsumoto Michio Supervision Edition (Issuyoshi 1985).

In addition, the present invention provides a cosmetic additive for the prevention and improvement effect of allergic skin disease or non-allergic skin inflammatory disease containing the crude extract or non-polar solvent soluble extract as an active ingredient.

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

Crude crude extract or non-polar solvent soluble extract of the present invention can be used with confidence even for long-term use for prevention purposes because there is little toxicity and side effects.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Example 1 Preparation of Crude Crude Extracts

(1-1) Botanical hot water extraction method using water solvent

In order to obtain perilla crude extract (hereinafter referred to as perennial hot water extract), first, dried persimmon ( Mifudo , Actinidia arguta: hereinafter, perilla ), A. kolomikta , perilla perilla ( A. polygama ) and stalk stem ( backlight ) were purchased and used in the experiment. 100 g of pulverized stalk and stalk stem were added to 1 liter of distilled water and stirred well, followed by extraction under reflux for 3 hours at an extraction temperature maintaining 90 to 95 ° C., and then the filtrate was separated and the crude extract was decompressed to 55 to 65 ° C. After concentrating, lyophilization was performed to obtain 15.6 g, 16.2 g and 17.0 g of the crude water composition, namely, the extracts of the wild boar, and the wild boar, respectively, and 10.4 g of the wild boar stem water extract.

(1-2) Botanical hot water extraction method using water-alcohol mixed solvent

To obtain crude crude extract (hereinafter, referred to as ethanol hot water extract), 30%, 50%, and 70% aqueous ethanol solution 1 in 100 g of pulverized tuna (Mifudo, Actinidia arguta ) as in Example 1-1 was used. After adding ℓ and stirring well, the mixture was refluxed for 3 hours at an extraction temperature maintained at 70 to 90 ° C., and the filtrate was separated. The herbal extracts were concentrated under reduced pressure at 55 to 65 ° C., and then lyophilized to obtain a herbal composition. , 30%, 50% and 70% ethanol hot water extracts were obtained as powder X 11 g-13 g, respectively.

Example 2 Preparation of Soluble Extract of Polar and Non-Polar Solvents

Tara (Mifudo, Actinidia arguta ) hydrothermal extract prepared in Example (1-1) was fractionated using an organic solvent.

(2-1) Chloroform Soluble Fraction Separation

After dissolving 5 g of Tadasu hot water extract (or crude extract) of Example (1-1) in 50 ml of distilled water, and then 50 ml of chloroform and separating it into a separatory funnel and then vigorously mixing the chloroform insoluble layer (aqueous layer) and The chloroform soluble layer was separated. Then, 50 ml of chloroform was added to the chloroform insoluble layer (aqueous layer), followed by mixing and fractionation to collect a chloroform insoluble fraction and a chloroform soluble fraction.

(2-2) Separation of ethyl acetate soluble fraction

After adding 50 ml of ethyl acetate to the chloroform insoluble fraction (aqueous layer) of Example (2-1), the ethyl acetate soluble fraction and the insoluble fraction were separated, and further 50 ml of ethyl acetate was added to the ethyl acetate insoluble fraction (aqueous layer). After mixing, the mixture was partitioned to collect ethyl acetate soluble fraction and insoluble fraction (aqueous layer).

The chloroform soluble fraction, ethyl acetate soluble fraction, and the remaining aqueous layer were concentrated under reduced pressure, and then lyophilized to obtain 0.34 g of the chloroform fraction, 0.05 g of the petroleum acetate fraction, and 4.61 g of the petroleum water fraction.

Example 3 Botanical Extract Fractions by Silica Gel Column Chromatography

Silica gel column chromatography (Daisogel IR-60-W-40: 63 mm) was performed on the soluble ethyl acetate soluble extract of Example (2-2), and the developing solvent was a chloroform: methanol: water mixed solvent ([ 1] 90: 11: 1, [2] 60: 10: 1, [3] 60: 20: 2) and methanol [4] were used, which was carried out at 300 mL per hour to obtain 4 additional fractions.

Each fraction obtained from 2,784 mg of the ethyl acetate fraction was [1] 2,381 mg, [2] 135 mg, [3] 148 mg, [4] 98 mg.

TLC (TLC plate: Merck, developing solvent: chloroform: methanol: water = 9: 5: 1) was performed using the above-mentioned hot water extract, ethyl acetate soluble extract and each fraction (see FIG. 1A). In FIG. 1a, lane 1 is a stratum hydrothermal extract, lane 2 is a stratum ethyl acetate fraction, lane 3 is a stratum extract silica gel fraction [4], lane 4 is a stratum extract silica gel fraction [3], and lane 5 is a stratum extract silica gel fraction [ 2], lane 6 is the Tsu extract silica gel fraction [1].

In addition, 2D-TLC was performed on the silica extract fractions [1] and [2]. As the first developing solvent, chloroform: methanol: water = 9: 5: 1 and the second developing solvent were performed using chloroform: acetone: water = 3: 8: 0.5 (see FIGS. 1B and 1C).

Experimental Example 1. Inhibition of IgE Production of U266B1 Cell Line by Thigh Extract

(1-1) Comparison of Efficacy of Botanical Extracts

U266B1 cells (lymphoblastoma cell line), a human B cell cell line producing IgE, were used to confirm the inhibitory effect of IgE production by the extract of T. U266B1 cells (American Type Culture Collection, Manassas, VA) were loaded with RPMI-1640 medium (15% FBS, 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyriuvate, 50 μg streptomycin, 100 U / ml penicillin) Incubated in a 24-well culture plate at 37 ° C., 5% CO ₂ . The prepared cells were again adjusted to a concentration of 2 × 10 ⁵ cells / well, first treated with LPS (2 μg / ml) and prepared in Examples (1-1) and (1-2). Hot water extract of T. alba, T. alba stem and 30%, 50%, 70% ethanol hot water extract were treated with 100 ㎍ / ml concentration, respectively, and the control group was treated with dexamethasone (2 μM) or medium (RPMI-1640). Treated. After culturing for 7 days under the same culture conditions, IgE concentration of the medium was measured by ELISA method (PharMingen; San Diego, CA).

In Table 1, all of the hot water extracts of the dai, dae, and tsudae showed the same inhibitory effect on IgE production as dexamethasone. In addition, Darae hot water extract using the ethanol aqueous solution showed the strongest activity in 50% ethanol aqueous solution. The hydrothermal extract of the stalk stem also showed similar efficacy as the stalk extract.

LPS Stimulation process IgE (IU / mL) - badge 187.8 ± 8.4 + badge 392.9 ± 3.4 + Tadasu hot water extract 228.8 ± 7.1 + Corvus hot water extract 233.2 ± 5.8 + Takesu hot water extract 211.7 ± 7.9 + Thigh Stem Hot Water Extract 241.5 ± 5.8 + 30% Ethanol Hot Water Extract 306.2 ± 16.5 + 50% Ethanol Hot Water Extract 201.9 ± 27.6 + 70% Ethanol Hot Water Extract 266.8 ± 17.0 + Dexamethasone 203.6 ± 30.9

(1-2) Comparison of Potency of Soluble Extract of Polar and Non-Polar Solvents

In order to compare the efficacy of inhibiting IgE production of U266B1 cells by the chloroform fraction obtained in Example 2-1, the ethyl ethyl acetate fraction of Example 2-2, and the water fraction, compared with Experimental Example (1-1) The same experiment was performed. The chloroform fraction, ethyl acetate fraction and water fraction were all treated in the U266B1 cell line at a concentration of 30 μg / ml. In Table 2, the ethyl acetate fraction of the Darae hydrothermal extract showed a stronger inhibitory effect on IgE production than dexamethasone.

LPS Stimulation process IgE (IU / mL) - badge 117.4 ± 7.6 + badge  212.5 ± 11.8 + Tara hot water extract of Example (1-1)  151.7 ± 11.9 + Chloroform fraction  206.7 ± 15.6 + Ethyl acetate fraction 123.3 ± 8.1 + Water fraction  218.9 ± 13.3 + Dexamethasone 138.9 ± 2.1

(1-3) Comparison of efficacy of extract fractions using silica gel column chromatography

In order to compare the effect of inhibiting IgE production of the U266B1 cell part by the silica gel column chromatography fractions [1] to [4] prepared in Example 3, the same experiment as in Experiment 1-1 was performed. Silica gel column chromatography fractions [1] to [4] were all treated with U266B1 cell line at a concentration of 10 μg / ml.

In Table 3, activity is shown in silica gel fractions [1] and [2].

LPS Stimulation process IgE (IU / mL) - badge  379.7 ± 13.2 + badge  540.4 ± 35.1 + Ethyl Acetate Fraction of Example (2-2) 298.1 ± 9.7 + Silica Gel Fraction 1  293.5 ± 12.5 + Silica Gel Fraction 2  307.6 ± 24.1 + Silica Gel Fraction 3  453.1 ± 17.3 + Silica Gel Fraction 4  396.9 ± 26.8 + Dexamethasone  277.6 ± 12.4

Experimental Example 2. Anti-allergic efficacy experiment of Tsurae extract in OVA-sensitized mouse model

(2-1) Preparation of Ovalbumin-sensitized mouse model

The white egg albumin sensitized mouse model is an animal experimental model that is widely used as an animal model of allergy. Six-week-old BALB / c mouse females (Seoul National University Animal Experiment Center) were allowed to acclimate to the environment for one week. At 7 weeks of age, 20 μg of egg albumin (OVA, crude grade V; Sigma) and 2.25 mg of aluminum hydroxide (ImujectAlum; Pierce) were mixed together to form a 100 μl emulsion and then placed in mice. Intraperitoneal injections and once again on day 14 for boosting.

After boosting, 30 mice were divided into three groups, which were prepared in Example (1-1), the hot water extract (300 μg / mouse / day), dexamethasone (10 μg / mouse / day), and drinking water (100 μl / mouse). / day) was orally administered daily for 11 days. On day 25 mice were sacrificed and serum and spleen were collected.

(2-2) Serum white albumin-specific IgE, IgG1, IgG2a, IgG2b concentration analysis

The concentrations of ovalbumin-specific IgE, IgG1, IgG2a, and IgG2b in the serum collected above were measured by ELISA method (PharMingen).

As a result, Table 4 shows the relative concentration of each antibody of the white albumin sensitized mouse to the concentration of each antibody of the normal mouse. In the mice taking the above-mentioned hot water extract of Example (1-1), IgE to white albumin was reduced to less than 1/3 level and IgG1, which is another allergic antibody, was also significantly reduced. Similarly, IgE and IgG1 were reduced in mice receiving dexamethasone. However, IgG2a, which is involved in cellular immunity, increased more than two-fold in the mice taking hot water extract, but not in the serum of dexamethasone-treated mice.

This suggests that the extract of Dalai Lai can fundamentally improve allergic constitution by reducing IgE causing allergy and increasing IgG2a involved in normal immune function.

Normal mouse Drinking water Tadasu hot water extract Dexamethasone IgE 1.0 ± 0.0 7.6 ± 0.2 2.2 ± 0.5 2.0 ± 0.6 IgG1 1.0 ± 0.0 5.1 ± 0.1 3.3 ± 0.3 3.1 ± 0.3 IgG2a 1.0 ± 0.1 2.1 ± 0.4 4.6 ± 0.3 1.9 ± 0.4 IgG2b 1.0 ± 0.0 1.4 ± 0.1 1.5 ± 0.2 1.2 ± 0.1

(2-3) Analysis of cytokine expression in spleen cells

Splenocytes were prepared from the spleen prepared in Experimental Example (2-1) to analyze the expression pattern of cytokines in immune cells of mice taking Tara extract.

First, each spleen was put together and then broken down into homogenous state for homogenization. Rinse the RPMI-1640 medium, filter out the large mass with a nylon mesh (60 μm pore size), centrifuge at 1500 rpm for 5 minutes to separate only the precipitated cells and add to RPMI 1640 (FBS 10% added) medium. . The spleen cells thus prepared were seeded in 24 well-plates to a final concentration of 5 × 10 ⁶ cells / ml / well. After adding white albumin (100 ㎍ / ml) to the culture, the cells were incubated in a carbon dioxide (CO ₂ ) incubator for 3 days at 37 ° C. After the incubation was completed, the culture medium was collected and an allergic cytokine (IL-4) by ELISA method. , IL-5, IL-12, interferon-gamma) were measured.

As a result, as shown in Table 5, the splenocytes of the mice taking the water extract of the above-mentioned Example (1-1) reduced the allergy-induced Th2 cytokines IL-4 and IL-5 and allergic The inhibitory Th1 cytokines IL-12 and interferon-gamma were significantly increased. Dexamethasone has been shown to inhibit both Th1 and Th2 cytokines.

Through these results, it was confirmed that dexamethasone inhibits the overall immunity, while the taraxa extracts can help to prevent and improve allergic diseases by selectively inhibiting allergic Th2 cytokines.

Normal mouse OVA sensitized mouse Drinking water Tadasu hot water extract Dexamethasone IL-4   21.6 ± 11.8  159.5 ± 15.7   76.5 ± 10.0 23.1 ± 8.7 IL-5   0.5 ± 0.8 2573.6 ± 42.0 1638.3 ± 33.3 884.1 ± 80.7 IL-12 1626.0 ± 58.5  906.2 ± 66.0 1321.2 ± 92.4 297.7 ± 16.4 Interferon-gamma   14.1 ± 19.6 1016.0 ± 25.6 1688.2 ± 15.8 470.5 ± 38.6 Unit: pg / ml

Experimental Example 3 Regulation of Th1 / Th2 Cytokines in Human PBMC

To investigate the regulating ability of Th1 / Th2 cytokines in human PBMCs of T. aeruginosa extracts, human peripheral blood mononuclear cells (PBMCs) were prepared using picol hypaque from total blood from allergic patients with high basal serum levels of IgE. Incubated in RPMI medium with 10% FBS. PBMCs were treated with 100 μg / ml of Tara hot water extract prepared in Example (1-1) and 5 μg / ml of plant hemagglutinin (phytohemagglutinin (PHA)). Thereafter, after 48 hours of incubation at 37 ° C. with 5% CO _{2, the} levels of IL-5, IL-13, IL-10 and interferon-gamma in the cell culture supernatant were measured using ELISA.

As a result, as shown in Table 6 below, the water extract of Darae significantly lowered the serum levels of Th2 cytokines. The levels of IL-5 and IL-13 were lowered by 52% and 47%, respectively. In contrast, serum levels of interferon-gamma, or Th1 cytokine, increased by 3.2 times.

Through these results, it can be seen that the extract of the wormwood can increase the production of Th1 cytokines, and at the same time reduce the production of Th2 cytokines. It has been previously reported that downregulation of Th2 cytokines may contribute to the relief of IgE synthesis and allergic inflammation.

PHA Stimulation process IL-5 IL-13 IL-10 Interferon-gamma - - 92.2 ± 6.4 0 ± 0 0 ± 0 53.3 ± 30.2 + badge 518.1 ± 120.3 667.8 ± 46.5 480.3 ± 15.3 334.1 ± 277.7 + Tadasu hot water extract 248.5 ± 62.0 355.7 ± 93.1 570.2 ± 56.3  1067 ± 345.1 Unit: pg / ml

Experimental Example 4. Reduction of IgE Level in Human Serum

In order to observe whether the extract of the wormwood reduced IgE levels in human serum, two allergic patients (patients K and E with allergic rhinitis and allergic dermatitis, respectively) 1g) was taken orally daily for 42 days, and the IgE serum levels of the patients were measured by ELISA every two weeks.

As a result, the serum levels of IgE in the two patients continued to decrease, reducing by more than two-thirds after 42 days, and improving the symptoms of dermatitis in patient E during the trial (see Table 7 below). .

1 day 14 days 28 days 42 days Allergic Patient K 950.8 IU / mL 855.6 IU / mL 679.1 IU / mL 266.1 IU / mL Allergic Patient E 278.3 IU / ml 236.0 IU / mL 189.2 IU / ml 95.0 IU / ml

Experimental Example 5 Inhibition of Histamine Release from Mouse Peritoneal Mast Cells

Release of histamine from mast cells is one of the major causes of allergic reactions. Therefore, the effect on histamine release from mast cells was observed.

Each mouse was anesthetized with ether and injected into the peritoneal cavity with 20 ml of thyroid buffer B (Tyrode buffer B, NaCl, glucose, NaHCO ₃ , KCl, NaH ₂ PO ₄ ) containing 0.1% gelatin. Then gently massage the abdomen for about 90 seconds. After carefully opening the peritoneal cavity, the solution containing the peritoneal cells was aspirated with a Paster pipette. Peritoneal cells were then precipitated at 150 xg for 10 minutes at room temperature and resuspended in thyroid buffer B. Mast cells were isolated from major components of mouse peritoneal cells by Yurt's method (Yurt RW. Et al., J. Biol. Chem ., 252 , pp518-521, 1977). Peritoneal cells suspended in 1 ml of thyroid buffer B were packed with 0.225 g / ml methizamide (metrizamide, density 1.120 g / ml) and centrifuged at room temperature for 15 minutes at 400 x g. Cells remaining at the buffer-methrazimide interface were aspirated and discarded. The cells in the pellet were resuspended in 1 ml of thyroid buffer A containing calcium after washing. Peritoneal cells suspended in 1 ml of thyroid buffer B were filled with 2 ml of metrizamite at 0.225 g / ml (density 1.120 g / ml) and centrifuged at room temperature for 15 minutes at 400 x g. The cells in the pellet were resuspended in 1 ml of thyroid buffer A containing calcium after washing.

Mast cells were added in an amount of 2 × 10 ⁵ cells / well to 0.4 ml of culture per well of 24-well culture plate. Cells were incubated overnight at 37 ° C. and sensitized to 0.5 μg / ml anti-DNP24-BSA IgE. After the cells with IgE sense drawing, removal of the medium, and the cells are washed twice with 0.5 ㎖ PIPES buffer solution, PIPES buffer solution (control) 200 ㎕, cromolyn (10 ^-4 M) or the embodiment (1 -1) Tara hot water extract (100 ㎍ / ㎖) was added and incubated for 10 minutes at 37 ℃. Mast cells were sensitized with the antigen DNP24-BSA 20 ng / ml for 30 minutes and histamine released into the medium was measured by ELISA (ALerCHEK).

Histamine release inhibition was calculated by the following equation:

% Inhibition = 100 × (Histamine-released during PIPES buffer treatment-Histamine-released during drug treatment) / (Histamine-released during PIPES buffer treatment) Histamine released in culture)

As a result, Table 8 shows that the hot water extract inhibited histamine release from 100 μg / ml to approximately 44%, and was as effective as chromoline used as a positive control.

process Cromoline (Control) Tadasu hot water extract % Inhibition 52 ± 13 44 ± 10

Experimental Example 6. Anti-inflammatory effect on arachidonic acid induced mouse edema

In order to determine the anti-inflammatory effect of the arachidonic acid-induced edema of the mouse ear of Tara extract, the following experiment was performed.

First, mice (8-week-old male BALB / c) were allowed to freely drink water, fasted for 18 hours, and 15 were divided into three groups. Inflammation was induced by topical application of arachidonic acid (0.5 mg / 20 μL acetone) in the right ear of each mouse. The left ear was used as a negative control and was administered excipient (20 μl acetone). Tara hot water extract (200 mg / kg in water) administration group of Example (1-1) was orally administered 1 hour before the application of arachidonic acid. The positive control group was administered indomethacin (10 mg / kg orally) 1 hour before arachidonic acid application. Inflammation was followed for 1 hour, after which the animals were sacrificed. From each ear a 6 mm diameter disk piece was obtained and weighed. The edema index of the control mice that did not receive any treatment was 7.2 ± 1.1 mg. However, when the experimental animals were treated with taraxacum extract, the edema index decreased by 62.5% to 2.7 ± 0.8 mg. Positive control ears treated with indomethacin had a decrease in edema index by 81.9% compared to untreated ears (see Table 9 below).

* As a result, it was confirmed that the taraxacum extract has anti-inflammatory activity comparable to indomethacin in this experimental model.

process Edema Index (mg) % Inhibition Control 7.2 ± 1.1 - Tadasu hot water extract 2.7 ± 0.8 62.5 Indomethacin 1.3 ± 0.3 81.9

Experimental Example 7. Atopic Dermatitis Mouse Model Experiment

In order to confirm anti-allergic efficacy in animals with allergic diseases, the following experiments were conducted using Nc / Nga mice, which are widely used as animal models of atopic dermatitis. On the other hand, Nc / Nga mice, which have been used in many atopic dermatitis animal experiments, have low production of interferon gamma, which suppresses Th1 immunity, and overproduces Th2 cytokines and IgE, resulting in natural conditions from 8 weeks of age. As a mouse model in which dermatitis develops, it is known as a model suitable for the study of human atopic dermatitis (Vestergaar CH. Et al., J. Clin. Invest ., 104 , pp 1097-1105, 1999).

First, 15 7-week-old Nc / Nga female mice (SLC; Shizuoka, Japan) were divided into three groups for the experiment, and then adapted to the new environment for one week. From 8 weeks of age to 16 weeks of age, the treated group was orally administered with Tadasu hot water extract (300 ㎍ / mouse / day) prepared in Example (1-1), and the control group was treated with dexamethasone (10 ㎍ / mouse / day) and drinking water ( 100 μl / mouse / day) was orally administered. In order to compare the progression of dermatitis symptoms at 12 and 14 weeks, the number of scratches of the mice was measured (scratch time per 20 minutes). At 16 weeks, IgE, IgG1, and IgG2a in blood were sacrificed using the ELISA method. It was. In addition, in order to compare the production of Th1 / Th2 cytokines, splenocytes were prepared in a general manner from each mouse, and then transferred to a 24-well culture plate (5 × 10 ⁶ cells / ml / well). After incubation for 3 days with ConA (1 ㎍ / ㎖), IL-4, IL-5, IL-12 and interferon gamma in the culture was quantified by ELISA method.

As a result of comparing the degree of scraping of mice at 12 and 14 weeks, it was found that the extract of Darae inhibited the symptoms of itching of the mouse as much as the steroidal anti-inflammatory dexamethasone (FIG. 2A: observation at 12 weeks, FIG. 2B: 14 Weekly observations).

In addition, Table 10 below shows that the concentration of IgE in the blood of the mice administered with the hot water extract was significantly reduced. In Table 11 below, IL-4 and IL-5 production of splenocytes prepared from mice treated with Tara extract significantly decreased, and interferon gamma and IL-12 production increased markedly.

In the above experiments, the extract of Darae showed similar effect to dexamethasone, improved itch symptoms, decreased IgE concentration in blood, and inhibited Th2 cytokine production of splenocytes, but unlike dexamethasone, Th1 cytokines, namely IL- 12 and production of interferon gamma significantly increased.

Through these results, it was confirmed that unlike the steroidal drug, which is a strong immunosuppressive agent, the extract of Darae extract can improve and prevent allergic symptoms by inhibiting Th2 immunity and increasing Th1 immunity.

IgE IgG1 IgG2a Drinking water 1572.9 ± 77.4 242.2 ± 14.2 177.5 ± 17.2 Tadasu hot water extract  699.0 ± 348.5 263.4 ± 21.1 263.5 ± 16.2 Dexamethasone 130.0 ± 55.3 247.3 ± 10.8 148.0 ± 14.8 Unit: pg / ml

IL-4 IL-5 IL-12 Interferon gamma Drinking water 151.9 ± 2.2 778.3 ± 34.1 1925.7 ± 134.5 10407.5 ± 130.8 Tadasu hot water extract 24.1 ± 5.2 248.0 ± 17.8 2346.2 ± 98.4 15847.9 ± 1693.1 Dexamethasone 42.7 ± 19.4 646.5 ± 51.7 201.2 ± 12.1 10096.4 ± 192.4 Unit: ng / ml

Experimental Example 8. Toxicity Test

Mice were used to carry out repeat toxicity test of the extracts. Female Balb / c mice were divided into two groups of ten rats each day and the dose of 150 mg / kg body weight per day was administered for 4 weeks, and the control group was administered water. Signs of toxicity were identified for 4 weeks through changes in body weight, hematological analysis, and biopsy.

As a result of the experiment, the extract of Dalai was not observed at 150 mg / kg of the dose. I could confirm it.

(1) Body weight and visual observation: No abnormal weight change was observed.

(2) Hematology test: any abnormalities in the number of WBCs, lymphocytes, monocytes, neutrophils, eosinophils, basophils, RBCs, hemoglobin, platelets No signs were found.

(3) Serum biochemical test: Biochemical test using serum AST, ALT, LDH, bilirubin, creatinine, glucose, cholesterol, cholesterol, minerals, albumin, BUN Did not show any signs of abnormality in levels of lipase, amylase.

(4) Biopsy: No abnormalities found in the tissues of the kidneys, spleen, liver and thymus.

The food additives, feed additives and cosmetic compositions of the present invention can be used in the preparation examples below, the following production examples are merely to illustrate the invention, thereby not limiting the contents of the present invention.

Preparation Example 1 Preparation of Candy

Tara hot water extract of Example 1-1 .............. 1 g

Maltitol ............... 55 g

Reduced syrup ......... 44.5 g

Oligosaccharides ....................... 0.2 g

Citric Acid ......................... 0.1 g

By combining the above raw materials can be prepared candy effective in preventing hair loss by the conventional candy manufacturing method in the art.

Preparation Example 2 Preparation of Beverage

Tara hot water extract of Example 1-1 ............ 1000 mg

Citric acid ..................... 1000 mg

Oligosaccharide ......................... 100 g

Plum concentrate ............... 2 g

Taurine .................................... 1 g

Purified water is added ............... 900 ml total

According to the conventional beverage production method, the above components are mixed, then stirred and heated at 85 ° C. for about 1 hour, the resulting solution is filtered, sterilized in a sterilized 2 L container, sealed and sterilized and then refrigerated and then stored in a beverage composition. Used for manufacturing.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

Preparation Example 3 Soap Preparation

Tara hot water extract of Example 1-1 ........ 1-30 (%)

Maintenance ...

Sodium Hydroxide ..................

Sodium Chloride ...

Spices .......................................

The whole amount was 100 with purified water, and soap was prepared by the above blending ratio.

Preparation Example 4 Lotion Preparation 1

Tara hot water extract of Example 1-1 ............... 3.00 (%)

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Water Soluble Collagen (1% Aqueous Solution) ................. 1.00

Sodium Citrate ............... 0.10

Citric Acid ........................... 0.05

Licorice Extract ......... 0.20

1,3-butylene glycol ............ 3.00

The whole amount was 100 with purified water, and a lotion was prepared by the above blending ratio.

Preparation Example 5 Lotion Preparation 2

Tara hot water extract of Example 1-1 ... 3.00 (%)

L-ascorbic acid-2-phosphate magnesium salt .............. 1.00

Water Soluble Collagen (1% Solution) .................. 1.00

Sodium Citrate ............... 0.10

Citric Acid ... 0.05

1,3-butylene glycol ...... 3.00

The whole amount was 100 with purified water, and a lotion was prepared at the above compounding ratio.

Preparation Example 6 Lotion Preparation 3

Tara hot water extract of Example 1-1 .... 4.00 (%)

Brown algae extract .................... 1.00

Sodium Citrate ............... 0.10

Citric Acid ... 0.05

1,3-butylene glycol ...... 3.00

The whole amount was 100 with purified water, and a lotion was prepared at the above blending ratio (%).

Preparation Example 7 Cream Preparation

Tara hot water extract of Example 1-1 ................. 1.00 (%)

Polyethylene Glycol Monostearate ............. 2.00

Self-emulsifying glycerin monostearate ............ 5.00

Cetyl Alcohol ... 4.00

Squalene ............... 6.00

Glyceryl tri2-ethylhexanoate ... 6.00

Sphingose Sugar Lipids ......................... 1.00

1.3-Butylene Glycol ... 7.00

The whole amount was 100 with purified water, and a cream was prepared at the above compounding ratio.

Preparation Example 8 Pack Preparation

Tara hot water extract of Example 1-1 .... 5.00 (%)

Polyvinyl Alcohol ............... 13.00

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Lauroylhydroxyproline ........................ 1.00

Water Soluble Collagen (1% Aqueous Solution) .... 2.00

1,3-butylene glycol ...... 3.00

Ethanol ......................................... 5.00

The whole amount was 100 with purified water, and a pack was prepared at the above compounding ratio.

Preparation Example 9 Preparation of Serum

Tara hot water extract of Example 1-1 .... 2.00 (%)

Hydroxyethylene Cellulose (2% Aqueous Solution) ......... 12.00

Xanthan gum (2% aqueous solution) ............. 2.00

1,3-butylene glycol ...... 6.00

Concentrated glycerin ... 4.00

Sodium hyaluronate (1% aqueous solution) ................. 5.00

The whole amount was 100 with purified water, and a cosmetic liquid was prepared at the blending ratio (%).

Crude crude extract or non-polar solvent soluble extract of the present invention lowers serum levels of Th2 cytokine and IgE to inhibit the release of histamine from mast cells and inhibits the inflammatory response, thereby causing hypersensitivity, allergic rhinitis, asthma, atopic dermatitis, food It can be used as a food additive useful for the prevention and amelioration of allergic diseases such as allergy and urticaria and non-allergic inflammatory diseases. Thus, when added to the food, it can eliminate the odor and add a unique flavor to enhance the taste and aroma of the food. In addition, it can be used in animal feed additives useful for the prevention and improvement of allergic diseases or non-allergic inflammatory diseases and feed containing the same, and in various cosmetics such as creams, skins, lotions, shampoos, rinses, treatments, soaps, etc. It can be used as an anti-allergic and anti-inflammatory substance.

Figures 1a to 1c is a TLC picture of the silica gel fraction, Figure 1a is a diagram showing the TLC results of the extract and fractions, Figure 1b is a 2D-TLC analysis picture of silica gel fraction 1, Figure 1c is a 2D of silica gel fraction 2 -Degree of TLC analysis result,

Figures 2a to 2b relates to the results of the skin itching observed after administration of the wormwood extract to atopic dermatitis Nc / Nga mice, Figure 2a observed at 12 weeks of administration of the water extract to atopic dermatitis Nc / Nga mice FIG. 2B is a diagram illustrating skin itch symptoms observed at the 14th week of the administration of sputum hot water extract in atopic dermatitis Nc / Nga mice. FIG.

Claims (4)

A cosmetic composition useful for the prevention and improvement of allergic skin diseases containing water, ethanol or a mixed solvent soluble extract of Actinidia arguta fruit as an active ingredient. The cosmetic composition according to claim 1, wherein the composition is a skin, lotion, cream, essence, nutrient, lotion, pack, soap, shampoo, rinse, cleansing, systemic cleanser, face wash, treatment, essence. Additives for the prevention and improvement of allergic skin diseases containing water, ethanol or a mixed solvent soluble extract of Actinidia arguta fruit as an active ingredient. 4. The cosmetic additive according to claim 3, wherein the additive is usable in creams, lotions, massage packs, systemic cleaners, soaps, shampoos and other care preparations.