KR20200093347A - Composition for Anti-inflammation and Atopy Dermatitis Using an Extract of Barly Nuruk - Google Patents

Abstract

Disclosed are an anti-inflammatory composition using a barley yeast extract and a composition for alleviating atopic dermatitis. The anti-inflammatory composition shows concentration-dependent NO production inhibitory activities on a mouse macrophage cell line (RAW 264.7 cells) stimulated with lipopolysaccharide (LPS), and also shows inhibitory activities on the expression of MDC and TARC in HaCaT cells, a human keratinocyte line treated with INF-γ and TNF-α.

Description

Composition for anti-inflammation and Atopy Dermatitis Using an Extract of Barly Nuruk}

The present invention relates to an anti-inflammatory composition and a composition for improving atopic dermatitis using an extract of barley yeast extract (that is, an archaea fermentation of barley).

Inflammation is a local biological defense response that appears in response to physical trauma, harmful chemicals, infections with bacteria, fungi, viruses, or pathological conditions caused by irritating substances in metabolites in vivo. Inflammation is triggered by a variety of inflammatory mediators produced from damaged tissue and migrating cells. During an inflammatory reaction, plasma accumulates in the inflammatory site, diluting the toxicity secreted by bacteria, increasing blood flow, and accompanying symptoms such as erythema, pain, edema, and fever. In the normal case, the living body neutralizes or eliminates the onset factors through the inflammatory reaction and regenerates the upper tissue to restore normal structure and function, but in other cases, it progresses to a disease state such as chronic inflammation.

With the recent development of molecular biology, many studies have been conducted on the inflammatory response at the molecular level.

Although various biochemical phenomena are involved in the inflammatory reaction, macrophages are known to play an important role in the inflammatory reaction by producing nitric oxide (NO) and various inflammatory cytokines by chemical stimulation (Ito T., et. al., Curr Drug Traget Inflamm Allergy, 2(3):257-265, 2003). Nitric oxide is synthesized from L-arginine by the action of nitric oxide synthase (NOS), and NOS has several isoforms. BNOS (brain NOS) present in the brain, nNOS (neuronal NOS) present in the nervous system, and eNOS (endothelial NOS) present in the vascular endothelial system are always expressed at a certain level in the body, and monoxide produced in small amounts by these Nitrogen (NO) plays an important role in maintaining homeostasis by performing various physiological reactions related to blood pressure regulation, neurotransmission, learning, and memory. On the other hand, iNOS (induced NOS), whose expression is induced by a certain stimulus, generates excessive NO, and nitric oxide generated excessively by iNOS reacts with superoxide to form peroxynitrite. It acts as a powerful oxidizing agent and damages cells, thereby engaging in various pathological processes including inflammation and cancer (Gupta SC et al., Exp Biol Med., 236:658-671, 2011; Riehemann et al., FEBS Lett., 442:89-94, 1999; Stamler et al., Science, 258:1898-1902, 1992).

The results of these studies suggest that drugs that inhibit NO production or inhibit the expression of iNOS have potential as effective anti-inflammatory agents (Karin M. et al., Cold Spring Harb Perspect Biol., 1, pp1-14, 2009).

Atopic dermatitis is a recurrent chronic skin disease, which is common in infants and toddlers, but persists even in adults or may develop into adulthood, and its prevalence is increasing (Kor J Pharmacogn., 43:59-65, 2012) ). Atopic dermatitis is characterized by symptoms such as pruritus, dryness of the skin, hyperactivity around the hair follicles, stomatitis, eczema, and eczema bile lesions (Korean J Invest Dermatol., 14:67-72, 2007).

The cause of atopic dermatitis has not been clearly identified, but abnormal increase in IgE, decrease in the number and function of T cells that play a pivotal role in cellular immunity, infiltration of monocytes and macrophages, increase in the number of mast cells and eosinophils, Immunological factors such as increased numbers of CD4+ T lymphocytes have been reported (J Invest Dermatol., 96:523-526, 1991; J Invest Dermatol., 97:389-394, 1991; Immunol., 11:81- 88, 1999; Curr Drug Targets Inflamm Allergy., 2:199-120, 2003; J Allergy Clin Immunol., 107:871-877, 2001; Adv Immunol., 78:57, 2001; International Immunology, 14(7) :767-773, 2002; Pediatr Allergy Immunol., 19:605-613, 2008), especially the Th1/Th2 imbalance due to the increased number of Th2 cells compared to Th1 cells is known as an important factor (Kor J Pharmacogn, 43: 59-65, 2012).

The immune system, which is a defense mechanism against external invasion of the human body, is mainly focused on the activation of T cells. Cytokines such as IL-2, IFN-γ, and TNF produced by Th1 cells induce differentiation of Th1 cells and inhibit proliferation and differentiation of Th2 cells, whereas IL-4, IL-5, IL produced by Th2 cells Cytokines such as -6, IL-10, IL-13 induce the proliferation and differentiation of Th2 cells and inhibit the differentiation of Th1 cells (J Am Acad Dermatol., 44, S1-S12, 2001). Under normal conditions, the immune response is maintained by balancing the interaction between Th1 cells and Th2 cells, but in atopic dermatitis, the conversion of T cells to Th2 cells is promoted, and increased Th2 cells secrete a large amount of cytokines to generate IgE And induces hypersensitivity reactions by inducing mast cell and eosinophil differentiation (J Clin Invest. 113(5):651-7, 2004; J Allergy Clin Immunol. 2003;112(2) : 252-62; Kor J Pharmacogn, 43:59-65, 2012).

Chemokine (chemokine) is a low-molecular protein that represents chemoattractants, and is divided into four types, C, CC, CXC, and CX3C, depending on its structure and function. Recently, these chemokines are immune cell development and maturation, T cell differentiation. And migration, and Th1/Th2 balance control.

Among various chemokines, THRC (Thymus & activation-regulated chemokine) and MDC (macrophage-derived chemokine) are typical CC chemokines acting on Th2 cells, acting on CCR4 (CC chemokine receptor 4) of Th2 cells to Th2 cells as inflammatory lesions Induces migration and infiltration (J Clin Invest. 113(5):651-7, 2004; Curr Allergy Asthma Rep. 5(4):284-90, 2005)

Recently, there have been reports of a significant increase in serum concentrations of TARC and MDC in patients with atopic dermatitis (J Allergy Clin Immunol., 113(2):334-340), and reports of increased skin expression of TARC and MDC in animal models of atopic dermatitis There is a report that the serum concentrations of MDC and TARC decrease when Cyclosporin A or corticosteroid used as a therapeutic agent for atopic dermatitis is administered to patients with atopic dermatitis (J Dermatol Sci., 34:201-208, 2004). In addition, in the in vitro experiment, when INF-γ or TNF-α was treated with human keratinocyte cell line HaCaT cells, MDC and TARC were expressed in large quantities, and it was suggested that substances capable of inhibiting such expression could be used as atopic dermatitis therapeutics. Yes (Int Immunol., 14(7):767-773, 2002).

Currently, as a therapeutic agent for atopic dermatitis, steroids that suppress the inflammatory response and cytokine production are mainly used, but when used for a long time, the use of nonsteroidal drugs has increased recently due to various side effects such as atrophy or delay in growth of the skin. . However, nonsteroidal drugs also have various side effects such as erythema, itching, swelling, swelling, and lichenification, and weakened immunity, making it difficult to treat fundamental atopic dermatitis (Arellano FM et al., J Invest Dermatol. 2007 Apr ;127(4):808-16.). Therefore, there is a need for research to find substances with excellent therapeutic effects from relatively safe natural products.

The present invention discloses anti-inflammatory and anti-atopic activity of barley yeast extract.

An object of the present invention is to provide an anti-inflammatory composition using barley yeast extract and a composition for improving atopic dermatitis.

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

The present invention shows concentration-dependent NO production inhibitory activity in a mouse macrophage cell line (RAW 264.7 cells) in which barley yeast extract and its fractions are stimulated with lipopolysaccharide (LPS), and human keratinocyte cell line treated with INF-γ and TNF-α It was completed by confirming that it inhibits the expression of MDC and TARC in the phosphorus HaCaT cells.

Considering the above, the present invention, in one aspect, can be identified as an anti-inflammatory composition comprising barley yeast extract as an active ingredient, and in another aspect, a composition for improving atopic dermatitis comprising barley yeast extract as an active ingredient (Or a composition for suppressing skin hypersensitivity immune response).

In the present specification, "barley yeast" means an archaea fermentation product of barley (seed). Specifically, it was obtained by inoculating and culturing Bacillus subtilus by adding water to the barley powder or finely cut water to be fermented, and the culture temperature was in the temperature range of 15°C to 50°C, especially in the temperature range of 25°C to 45°C. It can be done. When the culture temperature is lower than the above range, the fermentation rate may be slow and unwanted fermentation products may be generated, and when the culture temperature is higher than the above range, the fermentation rate may be lowered or the desired fermentation product may be generated similarly to the death of fermenting microorganisms. Can. Cultivation is desirable to continue until the barley component is sufficiently fermented and decomposed by the archaea, such culturing time will be determined within the ordinary ability of those skilled in the art depending on the culture temperature, the amount of fermentation raw materials, the amount of microorganisms to be inoculated, and the like. Such a fermentation product is used as it is, concentrated under reduced pressure and/or lyophilized to be used in a liquid or powder form, or alcohol, acetone, ethyl acetate, chloroform, methylene chloride, water or a mixture of 1 to 4 carbon atoms such as methanol and ethanol. It can be used by extraction with a solvent.

In addition, in the present specification, the term "barley yeast extract" refers to water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane, ether, chloroform, and ethyl acetate. Supercriticals such as butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or extracts obtained by leaching using mixed solvents thereof, carbon dioxide, pentane, etc. It refers to the extract obtained by using an extraction solvent or a fraction obtained by fractionating the extract, and the extraction method is arbitrary, such as cold acupuncture, reflux, warming, ultrasonic radiation, supercritical extraction in consideration of the polarity, extraction degree, and storage degree of the active substance. Method can be applied. In the case of fractionated extracts, the fractions obtained by suspending the extract in a specific solvent and mixing and policing with a solvent having a different polarity, adsorb the crude extract on a column filled with silica gel, etc., and then add a hydrophobic solvent, a hydrophilic solvent or a mixed solvent thereof. It means that it contains the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, an extract obtained using water, ethanol or a mixed solvent thereof as an extraction solvent, more preferably an extract obtained using a mixed solvent of water and ethanol as an extraction solvent and an extract obtained using a mixed solvent of water and ethanol Means ethyl acetate fraction and butanol fraction obtained by suspending in water and sequentially fractionating with ethyl acetate and butanol. Here, "sequential fractionation" means that the water layer used for fractionation is continuously fractionated even when fractionated with a fractional solvent in the next step. In addition, the ethyl acetate fraction means a fraction obtained by suspending the mixed solvent extract of ethanol and water in water and mixing and policing with ethyl acetate, and the butanol fraction is similarly suspended in the mixed solvent extract of ethanol and water in water, and with butanol. It is meant to include fractions obtained by mixing and policing.

In addition, in the present specification, "active ingredient" refers to a component that can exhibit a desired activity alone or itself can exhibit activity together with an inactive carrier.

Also, as used herein, "anti-inflammatory" is meant to include the improvement (reduction of symptoms), treatment, suppression or delay of the onset of such diseases as defined below.

In addition, in the present specification, the "inflammatory disease" refers to an inflammatory reaction characterized by a local or systemic bio-defense response to an external infection or autoimmunity such as external physical and chemical stimulation or infection of bacteria, fungi, viruses, and various allergens. It can be defined as the pathological symptom it causes. These inflammatory reactions include activation of various inflammatory mediators and enzymes associated with immune cells (eg iNOS, COX-2, etc.), secretion of inflammatory mediators (eg, secretion of NO, TNF-α, IL-6, etc.), body fluid infiltration, It involves a series of complex physiological reactions, such as cell migration and tissue destruction, and is manifested externally by symptoms such as erythema, pain, edema, fever, and a decrease or loss of certain functions in the body. The inflammatory disease may be acute, chronic, ulcerative, allergic or necrotic, so as long as any disease is included in the definition of such inflammatory disease, it is acute, chronic, ulcerative, allergic, or Necrotic or not. Specifically, the inflammatory diseases include asthma, allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, pulmonary fibroids , Irritable bowel syndrome, inflammatory pain, migraine, headache, back pain, fibromyalgia, fascia disease, viral infections (such as type C infection), bacterial infections, fungal infections, burns, wounds caused by surgical or dental surgery, pro Stargladin E hypersyndrome, atherosclerosis, gout, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, irisitis, scleritis, uveitis, dermatitis (including atopic dermatitis), eczema, multiple sclerosis, etc. Will be included.

In addition, in the present specification, “atopic dermatitis” is defined to include all diseases classified as atopic dermatitis in the art, accompanied by skin hypersensitivity immune responses, regardless of the direct or indirect cause of the occurrence. Atopic dermatitis is generally classified into infantile atopic dermatitis, pediatric atopic dermatitis, adult atopic dermatitis, and maternal atopic dermatitis depending on the time of its onset or the subject of the invention, wherein atopic dermatitis includes all these types of atopic dermatitis. .

In addition, in this specification, "improvement" is meant to include treatment, prevention and improvement (reduction of symptoms) of atopic dermatitis.

The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit the activity of inflammatory disease, atopic dermatitis treatment, prevention, etc. intended to be treated according to the use, formulation, formulation purpose, etc. , A typical effective amount will be determined within the range of 0.001% to 15% by weight based on the total weight of the composition. Herein, the term "effective amount" means intended medical and pharmacological effects, such as an anti-inflammatory effect and an atopic dermatitis-improving effect, when the composition of the present invention is administered to a mammal, preferably a person, to which the object is applied, according to the recommendation of a medical expert or the like. It refers to the amount of the active ingredient contained in the composition of the present invention, which can exhibit an effect. Such effective amount can be determined empirically within the range of ordinary skill in the art.

The composition of the present invention, in addition to the active ingredient, for the increase or reinforcement of anti-inflammatory effect, atopic dermatitis improvement effect or skin protection activity (suppression of skin damage caused by ultraviolet rays, skin moisturization, etc.) In order to enhance the convenience of, any compound or natural extract already known in the art for safety and having known activity may be further included.

These compounds or extracts include compounds or extracts, medicines, etc. that are listed in the Pharmacopoeia of each country ("Korea Republic of Korea Pharmacopoeia"), and health functional food exhibitions of each country ("Korea Food and Drug Administration""Health functional food standards and standards") Laws governing the manufacture and sale of compounds or extracts and health functional foods that have been granted an item in accordance with the laws of each country governing the manufacture and sale of foods (the "Pharmaceutical Law" in Korea). ”) includes compounds or extracts whose functionality is recognized. For example, Enterococcus faecalis heat-treated dry powder, Guava leaf extract, etc., which has been individually recognized for its function of'reducing hypersensitivity immune response' in accordance with the Korean Act on Health Functional Food, complexes, sage extract, leaflet extract, and picopreto powder Complex, L. sakei Probio 65, individually recognized for its functionality through'improvement of hypersensitivity skin', PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol), oil- and fat-derived lactic acid bacteria Phosphorus L.plantarum CJLP133, Probiotic ATP, etc. may correspond to these compounds or extracts.

One or more of these compounds or natural extracts may be included in the composition of the present invention together with their active ingredients.

In the specific aspect, the composition of the present invention can be grasped as a food composition. When the composition of the present invention is identified as a food composition, its use may be understood as inhibiting skin hypersensitivity or suppressing hypersensitivity reactions.

The food composition of the present invention can be produced in any form, such as tea, juice, carbonated beverages, beverages such as ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, bread, cookies, noodles, etc. Food, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, etc. can be prepared as health functional food formulations.

In addition, the food composition of the present invention can be classified into any product as long as it complies with the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food pursuant to the Korean Act on Health Functional Food, or a confectionary, soybean, tea, or beverage according to each food type in the Food Fair of the Korean Food Sanitation Act (「Food Standards and Standards」) , Special-purpose food.

The food composition of the present invention may include a food additive in addition to the active ingredient. Food additives can be generally understood as substances added to foods to be mixed or infiltrated in manufacturing, processing, or preserving foods, and their safety must be ensured because they are consumed daily and for a long time with foods. Food additives in accordance with national laws governing the manufacture and distribution of food ("Food Sanitation Act" in Korea) are limited in terms of ingredients or functions in terms of food additives with guaranteed safety. In the Korean Food Additives Fair (「Food Additive Standards and Standards」 published by the Ministry of Food and Drug Safety), food additives are classified into chemical synthetic products, natural additives, and mixed preparations in terms of ingredients, and these food additives are sweeteners and flavors in terms of functionality. It is divided into agents, preservatives, emulsifiers, acidulants, and thickeners.

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

Flavoring agents can be used to enhance taste or aroma, and both natural and synthetic ones can be used. Preferably, it is the case of using a natural thing. In addition to flavor, when using natural ones, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, or the like, or may be obtained from green tea leaves, perilla, large leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. Natural flavors may be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, and the like.

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

As a thickener, a suspending agent, sedimentation agent, gel forming agent, swelling agent, etc. may be used.

The food composition of the present invention may include, in addition to the food additives as described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functional and nutritional properties, and having stability as food additives.

Examples of such bioactive substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoyl thiamine, and the like, and calcium preparations such as calcium citrate and magnesium stearate as minerals Magnesium preparations such as iron, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, and the like.

The food composition of the present invention may include the food additives as described above in an appropriate amount to achieve the purpose of addition according to the product type.

With regard to other food additives that may be included in the food composition of the present invention, it is possible to refer to national foods or food additives.

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

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

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically refer to the pharmacopeia of each country, including "Korea Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers And the like. At this time, examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropyl methylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, green Serol, etc. are mentioned. In the case of formulation, if necessary, suitable binders, lubricants, disintegrants, colorants, diluents, etc. may be included. Suitable binders include starch, magnesium aluminum silicate, starch ferist, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and lubricants include oleic acid Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydetylene glycol, and the like, and starch and methyl cellulose as disintegrants , Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Moreover, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine etc. are mentioned as a diluent.

When the pharmaceutical composition of the present invention is prepared in a parenteral dosage form, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories according to methods known in the art with suitable carriers. When formulated as an injectable agent, an aqueous isotonic solution or suspension may be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanol amine, sterile water for injection, or 5% dextrose may be used. . When formulated as a transdermal dosage form, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pasta, linen agents, aerosols, and the like. For nasal inhalants, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. can be formulated in the form of an aerosol spray using a suitable propellant. witepsol), tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like.

Regarding the specific formulation of the pharmaceutical composition, it is known in the art, and for example, see Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. The above documents are regarded as part of this specification.

Preferred dosages of the pharmaceutical compositions of the present invention range from 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g per day, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. /kg range. Administration can be made once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.

In another specific aspect, the composition of the present invention can be identified as a cosmetic composition. When the composition of the present invention is identified as a cosmetic composition, its use is to relieve inflammatory skin irritation, relieve skin irritation immune response, or suppress skin troubles such as erythema, dryness, lichenification, dullness, dryness, and bleeding caused by skin irritation or abnormal immunity. It can be understood for such purposes.

Even if the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in accordance with its use, and in particular, functional cosmetics having non-functionality such as improvement of skin trouble, atopic dermatitis, etc. It may be a general cosmetic. In the form of a product, any type of product can be taken. Specifically, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, and waxes It can take the form of products such as foundations and sprays. In a specific product form, it may be flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.

The cosmetic composition of the present invention may include, in addition to its active ingredients, ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, pigments and conventional auxiliary agents such as pigments, and carriers.

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

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane or dimethyl ether.

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

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspending agents such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition of the present invention can be prepared according to the manufacturing method of a cosmetic composition that is conventionally performed in the art, except that it contains an active ingredient showing anti-inflammatory activity or atopic dermatitis-improving activity.

As described above, according to the present invention, it is possible to provide an anti-inflammatory composition using barley yeast extract and a composition for improving atopic dermatitis.

The anti-inflammatory composition of the present invention and the composition for improving atopic dermatitis may be commercialized as food, cosmetics, and drugs for anti-inflammatory or atopic dermatitis improvement.

1 to 5 are experimental results for NO production inhibitory activity and cytotoxicity of barley yeast extract and fractions.
6 is an experimental result for the MDC and TARC production inhibitory activity of barley yeast extract and fractions.

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

<Example> Preparation of barley yeast extract and fraction

<Example 1> Preparation of extract

20% by weight of 30%, 50%, 70% ethanol was added to barley yeast powder, stirred at room temperature for 24 hours, filtered, and the filtrate was concentrated under reduced pressure and lyophilized to extract powdery (30% extract: BN- 30, 50% extract: BN-50, 70% extract: BN-70). The extract thus obtained was used in the following experiment.

Barley yeast was obtained by adding 100 parts by weight of purified water to the barley cut water, mixing it uniformly, and inoculating Bacillus subtilus prepared in nutrient broth medium with 2% (v/w) and fermenting at 30±10°C for 50 days.

<Example 2> Preparation of fractions

The 50% extract was suspended in 1 L of distilled water, and the same amount of ethyl acetate was added thereto, followed by fractionation twice. After the layers were separated, the ethyl acetate layer was recovered and concentrated under reduced pressure. After adding 1 L of butanol to the water layer, it was fractionated twice. After the layers were separated, the butanol layer and the water layer were recovered and concentrated under reduced pressure, and the obtained fraction was used in the following experiment (ethyl acetate fraction: BN-Ea, butanol fraction: BN-Bu).

<Experimental Example> Anti-inflammatory activity experiment and atopic dermatitis improvement activity experiment

<Experimental Example 1> Anti-inflammatory activity experiment

For the analysis of anti-inflammatory activity, RAW 264.7 cells (murine macrophage cell line) are pre-sold from ATCC (American Type Culture Collection, VA, USA) and have 100 units/mL penicillin-streptomycin and 10% fetal bovine serum (FBS). The culture was performed in a 37°C, 5% CO ₂ incubator using the contained DMEM medium (Gibco, Grand Island, NY, USA), and subculture was performed once every 2-3 days.

In vitro anti-inflammatory activity evaluation, RAW 264.7 cells were divided into 3.0×10 ⁵ cells/mL, incubated for 20 hours at 37°C 5% CO ₂ incubator conditions, and each sample was treated by concentration, and LPS (0.1 μg/ mL) and incubated for 24 hours. The amount of NO (nitric oxide) generated was measured in the form of NO2-(nitrite) present in the cell culture using Griess reagent.

For cytotoxicity evaluation, an MTT assay was used. MTT was added to the cell culture medium to measure the amount of formazan precipitate formed by reacting with living cells. Cytotoxicity of the sample against RAW 264.7 cells was evaluated compared to the control.

The results are shown in FIGS. 1 to 5. All extract samples (30%, 50% and 70% extracts and ethyl acetate fractions and butanol fractions) showed concentration-dependent NO production inhibitory activity, and cytotoxicity was clearly observed in ethyl acetate fractions only at a concentration of 1000 μg/mL. The remaining extract and fraction samples showed no particular cytotoxicity at all treatment concentrations.

<Experiment 2> Experiment for improving atopic dermatitis

To analyze the efficacy of inhibiting expression of atopy-related factors, HaCaT cells were cultured in a 6-well plate at 1×10 ⁶ cells/well, followed by concentration of IFN-γ (10 ng/ml) and TNF-α (10 ng/ml) and samples. Treated separately and cultured for 18 hours. Next, the cultured HaCaT cells were recovered, washed twice with PBS, and total RNA was separated using trizol (Invitrogen), and cDNA was synthesized. Using the synthesized cDNA as a template, RT-PCR kit (Bioneer, Korea) was used to perform 94°C 5 minutes, [94°C 30 seconds, 60°C 30 seconds, 72°C 2 minutes] × 35 cycles, 72°C 7 minutes. Did. The primers used in the analysis are shown in Table 1, and each MDC and TARC was analyzed and the expression of each gene was confirmed by using the control gene GAPDH for quantitative analysis.

The results are shown in Figure 6 and Table 2 below. Referring to FIG. 6 and Table 2, the 50% extract showed distinct MDC and TARC expression inhibitory activity at a concentration of 100 μg/mL, and the ethyl acetate fraction showed distinct inhibitory activity at all treatment concentrations. Dexamethasone was used as a positive control.

Claims (10)

An anti-inflammatory composition comprising an extract of Bacillus subtilis Bacillus fermentation as an active ingredient.
According to claim 1,
The barley yeast extract is a composition characterized in that any one of the following (a) to (c) extract:
(a) water, ethanol or mixed solvent extracts of barley yeast,
(b) the ethyl acetate fraction obtained when these mixed solvent extracts are fractionated with water and ethyl acetate, and
(c) The butanol fraction obtained when the mixed solvent extract is fractionated with water and butanol.
The method according to claim 1 or 2,
The composition is an anti-inflammatory composition, characterized in that the pharmaceutical composition.
The method according to claim 1 or 2,
The composition is an anti-inflammatory composition, characterized in that the food composition.
The method according to claim 1 or 2,
The composition is an anti-inflammatory composition, characterized in that the cosmetic composition.
A composition for improving atopic dermatitis comprising barley yeast extract as an active ingredient.
The method of claim 6,
The barley yeast extract is a composition characterized in that any one of the following (a) to (c) extract:
(a) water, ethanol or mixed solvent extracts of barley yeast,
(b) the ethyl acetate fraction obtained when these mixed solvent extracts are fractionated with water and ethyl acetate, and
(c) The butanol fraction obtained when the mixed solvent extract is fractionated with water and butanol.
The method of claim 6 or 7,
The composition is a composition for improving atopic dermatitis, characterized in that the pharmaceutical composition.
The method of claim 6 or 7,
The composition is a composition for improving atopic dermatitis, characterized in that the food composition.
The method of claim 6 or 7,
The composition is a composition for improving atopic dermatitis, characterized in that the cosmetic composition.

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