KR102236071B1 - Composition for Improving Skin Trouble Using an Extract of Centella asiatica or Paeonia lactiflora Cultured with Magma Seawater - Google Patents

Abstract

According to the present invention, disclosed is a skin trouble composition using a centella asiatica extract and a paeonia lactiflora extract grown in lava seawater, which have anti-inflammatory activity, keratinocyte extracellular membrane formation promoting activity, and hyaluronic acid production promoting activity.

Description

Composition for Improving Skin Trouble Using an Extract of Centella asiatica or Paeonia lactiflora Cultured with Magma Seawater}

The present invention relates to a composition for improving skin troubles using extracts of centella asiatica and peony ( Paeonia lactiflora) grown using lava seawater.

Inflammation is a local protective reaction in the living body in response to physical trauma, infection by harmful chemicals, bacteria, fungi, viruses, or pathological conditions caused by irritants in metabolites in the living body. Inflammation is triggered by various inflammatory mediators produced from damaged tissue and migrating cells. During an inflammatory reaction, plasma accumulates in the inflamed area, diluting the toxicity secreted by bacteria, increasing blood flow, and accompanying symptoms such as erythema, pain, swelling, and fever. In a normal case, the living body neutralizes or removes pathogenic factors through an inflammatory reaction and regenerates damaged tissue to restore normal structure and function, but otherwise, it may progress to a disease state such as chronic inflammation.

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

Although various biochemical phenomena are involved in the inflammatory response, it is known that macrophages in particular play an important role in the inflammatory response by producing nitric oxide (NO) and various inflammatory cytokines by chemical stimulation (Ito T., et al. 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, eNOS (endothelial NOS) present in the vascular endothelial system are always expressed in the body at a certain level, and a small amount of monoxide is produced by them. Nitrogen (NO) plays an important role in maintaining homeostasis of the human body 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, produces excessive NO, and nitrogen oxide generated excessively by iNOS reacts with superoxide to form peroxynitrite. It acts as a potent oxidant, damaging cells, and is involved 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).

_{Meanwhile, cyclooxygenase (COX) synthesizes the intermediates PGG 2} and PGH ₂ from arachidonic acid with the function of COX and hydroperoxidase (HOX), and these compounds PGE ₂ and PGF ₂ , PGD ₂ , prostacyclin and thromboxane A ₂ (thromboxane A2, TxA2) are produced, and there are two types of isoforms in COX. COX-1 is always expressed in most tissues to synthesize prostaglandins (PGs), which are required for cytoprotective action, whereas COX-2 rapidly induces its expression during inflammatory reactions _{and generates PGE 2} to induce an inflammatory reaction. Plays an important role (Weisz A., Biochem. J., 316:209-215, 1996; (Miller MJ et al., Mediators of inflammation, 4:387-396, 1995: Appleton L. et al., Adv) Pharmacol., 35:27-28, 1996).

The expression of iNOS and COX-2, which induces overproduction of NO and PGs, is regulated by the nuclear transcription factor NF-κB. NF-κB is a nuclear protein of the Rel gene family. In the cytoplasm, it is combined with I-κB and exists in an inactive form. However, when I-κB kinase is activated by a certain factor, it undergoes phosphorylation. It is activated by the fall of I-κB. NF-κB, which is composed of a heterodimer of p50 and p65, is activated and then moves to the nucleus to induce gene expression of iNOS and COX-2, which induces an inflammatory response (Oh, GT et al., Atherosclerosis, 159). (1):17-26, 2001).

Bacterial endotoxins such as LPS (lipopolysaccharide) activate the transcription factor NF-κB by binding to TLR4 (toll-like receptor 4), and induce the expression of iNOS and COX-2 to form NO, inflammatory cytokines, PGE _2, etc. Secrete several inflammatory regulators (Chen YC, et al, Biochem. Pharmacol., 61:1417-1427, 2001; Dobrovolskaia MA, et al., J Immunol., 170:508-19, 2003; Ji Y, et al., Cell Physiol Biochem., 25:631-640, 2010). NO, Inflammatory cytokines such as TNF-α and IL-6, PGE _2, etc. are arthritis (Jang CH et al., Rheumatology, 2006, 45(6):703-710), fibromyalgia (Hernandez ME et. al., BMC Res). Notes., 2010, 3(1):156), Sjogren's syndrome (Baturone R. et. al., Scand J Rheumatol., 2009, 38(5):386-389) as an important factor inducing inflammatory response. It has been reported (Jang CH et al., Rheumatology, 45(6):703-710, 2006; Hernandez ME et. al., BMC Res. Notes., 3(1):156, 2010; Baturone R. et. al., Scand J Rheumatol., 38(5):386-389, 2009).

These findings suggest that either inhibit NO production or Drugs that inhibit the production of inflammatory cytokines such as TNF-α and IL-6, and drugs that inhibit the expression of iNOS or COX-2 suggest that they have potential as effective anti-inflammatory agents (Karin M. et al., Cold Spring Harb Perspect Biol., 1, pp1-14, 2009).

Non-steroidal anti-inflammatory drugs (NSAIDS), which are currently widely used as anti-inflammatory drugs, are known to cause serious side effects such as gastrointestinal disorders, liver disorders, and renal disorders (Rainsford KD., Subcell biochem., 42:3). -27, 2007; Guruprasad P. Aithal., Rheumatology., 7:139-150, 2011; Praveen PN Rao et al., Pharmaceuticals., 3:1530-1549, 2010).

Therefore, it can be said that it is still necessary to develop a new drug that has anti-inflammatory activity, has few side effects, and is effective.

On the other hand, the skin accounts for about 16% of the human body, and since it is in direct contact with the external environment, it plays an important role in protecting the human body from external harmful factors such as temperature, humidity, and ultraviolet rays. The skin is composed of the epidermis, the dermis, and the subcutaneous tissue under the keratin. The epidermis is a thin protective layer composed of keratinocytes, mainly composed of keratin, immune cells such as melanocytes, langerhans cells, etc., and Merkel's copuscles that produce and secrete melanin. It prevents external irritation and invasion of pathogens, regulates body temperature, and maintains moisture and lipid components.

The dermis is the connective tissue under the epidermis and is mostly composed of a network of macromolecules called extracellular matrix. This extracellular stroma is made from fibroblasts and consists of fibrous proteins such as collagen and elastin and polysaccharides such as hyaluronic acid.

Hyaluronic acid is mainly synthesized by hyaluronic acid synthase in keratinocytes and fibroblasts, and is a polysaccharide with a molecular weight of 200,000 to 400,000 accumulated in the extracellular matrix. It is very strong and acts to prevent water evaporation from the epidermis. Three types of HAS genes known so far, HAS-1, HAS-2, and HAS-3, have been reported, and the defects in the moisturizing barrier due to their decreased expression are epidermal atrophy, wrinkle formation, skin moisture reduction, elasticity decrease, etc. It has been reported to cause skin aging (J Eur Acad Dermatol Venereol. 19, 308-318, 2005; Int J Dermatol. 33, 119-122, 1994; Kor J Herbology. 22,213-218, 2007; Skin Pharmacol Appl Skin Physiol. 12, 276-283; 1999).

The present invention has been completed based on the anti-inflammatory activity of centella asiatica cultivated in desalted lava seawater, peony extract, keratin extracellular membrane formation promoting activity, hyaluronic acid production promoting activity with strong water retention ability, and the like.

It is an object of the present invention to provide a composition for improving skin problems using centella asiatica and peony extract grown in desalted lava seawater.

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

As confirmed in the Examples and Experimental Examples below, the present inventors believe that Centella asiatica extract and Paeonia lactiflora extract grown using desalted lava seawater are P.acne or LPS (lipopolysaccharide), which are acne bacteria. While it was confirmed that the inflammatory response induced by this was suppressed, it was confirmed that it promotes the formation of the keratinocyte outer membrane and the production of hyaluronic acid in the keratinocyte HaCaT cells. The keratin extracellular membrane forms the stratum corneum together with the corneocytes and plays an important role in the skin barrier function. Specifically, it provides an environment in which the skin can maintain normal biological functions by suppressing the loss of moisture and electrolytes through the skin. It plays a role in preventing the invasion of harmful factors from the external environment through the skin (Kor. J. Aesthet. Cosmetol., Vol. 13 No. 6, 713-720, December 2015).

In consideration of the experimental results as described above, in one aspect, the present invention is for anti-inflammatory (inflammatory) containing as an active ingredient a peony extract cultivated using desalted lava seawater or centella asiatica extract cultivated using desalted lava seawater. For inhibiting reaction or for inhibiting inflammatory skin irritation) composition, and in another aspect, a skin barrier comprising as an active ingredient a peony extract grown using desalted lava seawater or centellar extract grown using desalted lava seawater It can be understood as a composition for enhancing function, and in another aspect, it can be understood as a composition for skin moisturizing comprising a peony extract grown using desalted lava seawater or centellar extract grown using desalted lava seawater as an active ingredient.

In the present specification, "the centella asiatica grown using desalted lava seawater" refers to centella asiatica obtained through field cultivation by supplying desalted lava seawater during cultivation. The supply cycle of desalted lava seawater may be 1 to 5 days, preferably 1 to 3 days, and the cultivation period is 15 to 30 days based on the time of sowing or transplanting seedlings grown 1 to 2 days after sowing. It may be, and preferably it may be 20 to 30 days. The supply method of desalted lava seawater may be supplied by a sprinkling method or a spray method. During cultivation, appropriate fertilizer can be supplied with desalted lava seawater.

In addition, in the present specification, "peony grown using desalted lava seawater" refers to a product obtained by supplying desalted lava seawater during cultivation and field cultivation, as in centella cultivated using desalted lava seawater. The supply cycle of the desalted lava seawater, the cultivation period, and the supply method of the desalted lava seawater are as described in relation to the peony grown using the desalted lava seawater.

In addition, in the present specification, "extract" refers to an extract obtained by leaching the object to be extracted using water (including hot water), ethanol, or a mixed solvent thereof. Preferably it refers to a hot water (especially 50 ~ 90 ℃) extract, a mixed solvent extract of water and ethanol, particularly 70% ethanol extract. The extract may be a concentrated liquid extract or a solid extract from which the extraction solvent has been removed by a method such as extraction stock solution, freeze drying, vacuum drying, hot air drying, spray drying, or the like.

In addition, in the present specification, "lava seawater" is defined in Korean Patent No. 0853244 (application number: 10-2008-0027861) in which the name of the invention is "a method for preparing a mineral composition and a mineral composition obtained by the method" It has the same meaning as "Salt underground water existing in the eastern part of Jeju Island" as described. Specifically, "Jeju-do eastern region" means Gujwa-eup, Seongsan-eup, and Pyoseon-myeon, Jeju-do, and "Saltji sewage" means groundwater containing more than a certain amount of salt. Low-salt groundwater (electrical conductivity of 1,700㎲/cm) based on the salinity concentration used in the study on the hydrogeological properties of the eastern part of Jeju Island (II), 2003, Kyungwon University; ~17,350 µs/cm) and brine groundwater (electrical conductivity of 17,350 µs/cm or more) can be understood as a meaning, but preferably the electrical conductivity is 17,350 µs/cm or more or the concentration of salt is 30 ‰ (permilliage ) It can be understood to mean brine groundwater having a salinity above (usually the salt concentration of seawater is 32 to 35‰).

In addition, in the present specification, "desalted lava seawater" refers to a result obtained by desalting lava seawater. Desalted lava seawater can be understood as mineral water from which salt has been removed. Desalination of lava seawater may be performed by applying any technique known in the art, such as a reverse osmosis method, an ion exchange resin method, an electrodialysis method, and the like.

In addition, in the present specification, the term "active ingredient" refers to an ingredient that can exhibit a desired activity alone or exhibit activity with a carrier that is not itself active.

The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit anti-inflammatory activity, moisturizing activity, skin barrier function strengthening activity, etc., and a typical effective amount is based on the total weight of the composition. When it will be determined within the range of 0.001% to 15% by weight. Herein, the term "effective amount" refers to the intended dermatology, such as moisturizing effect, skin barrier function strengthening activity, etc., when the composition of the present invention is administered to a mammal, preferably a human, during the administration period as suggested by a medical or skin expert It refers to the amount of the active ingredient contained in the composition of the present invention that can exhibit a pharmacological effect. Such effective amounts can be determined empirically within the range of ordinary skill in the art.

In addition to the active ingredient, the composition of the present invention is administered or ingested through addition of similar activities such as skin wrinkle improvement activity, skin hypersensitivity reaction suppression activity, and skin protection activity (skin whitening, skin damage suppression by ultraviolet rays, skin moisturizing, etc.). In order to enhance the convenience of, the safety has already been verified in the art and may further include any compound or natural extract known to have the corresponding activity. These compounds or extracts include each country's pharmacopoeia (“Korean Pharmacopoeia” in Korea), each country's health functional food code (in Korea, it is “health functional food standards and standards” notified by the Ministry of Food and Drug Safety), and each country's functional cosmetics code (in Korea, it is notified by the Ministry of Food and Drug Safety). Compounds or extracts that have been licensed in accordance with the laws of each country (“Pharmaceutical Affairs Act” in Korea) governing the manufacture and sale of compounds or extracts and pharmaceuticals listed in the fair documents such as "Functional Cosmetics Standards and Test Methods"), health In accordance with the laws of each country governing the manufacture and sale of functional foods (in Korea, it is the 「Act on Health Functional Foods」), national laws governing the manufacture and sale of functional cosmetics, extracts or compounds with recognized functionality (in Korea, the 「Cosmetics Act」 ”), the functional compounds or extracts are included. For example, arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, and oil-soluble licorice extract (Oil Soluble Licorice(Glycyrrhiza) Extract) are mentioned as skin whitening ingredients, and retinol, retinyl palmitate as skin wrinkle improvement ingredients. , Adenosine, polyethoxylated amide, and the like, and examples of the UV protection component include dromethrizol, dromethrizoltrisiloxane, digaloyltrioleate, dimethicodiethylbenzalmalonate, and diethylhexylbutamido. And complex extracts such as triazone and pine bark extract, phosphatidylserine, fingerroot extract powder, and complex extracts such as red ginseng and cornus milk. In addition, as a moisturizing ingredient, AP collagen enzyme decomposition peptide, collactive collagen peptide, N-acetylglucosamine, konjac potato extract, complex extract such as dandelion, rice bran extract, corn germ extract, low molecular collagen peptide, extract powder, phosphatidylserine, hyaluronic acid, etc. Examples of functional ingredients for'improving irritable skin condition' include L. sakei Probio 65, oil containing gammarinolenic acid, L. plantarum CJLP133, a lactic acid bacteria derived from fruits and vegetables, and probiotics ATP. One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredient.

In a specific aspect, the composition of this invention can be grasped as a food composition.

The food composition of the present invention may be prepared in any form, such as beverages such as tea, juice, carbonated drinks, ion drinks, processed oils such as milk, yogurt, gums, rice cakes, Korean sweets, bread, snacks, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrup, gels, jelly, can be prepared in health functional food preparations such as bars.

In addition, the food composition of the present invention can be classified as a product as long as it conforms to 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 according to the Korean 「Health Functional Food Act」, or confectionery, bean, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (``Food Standards and Standards'' notified by the Ministry of Food and Drug Safety). , Special use food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to food and mixed or infiltrated in manufacturing, processing, or preserving food. Since they are consumed daily and for a long time with food, their safety must be ensured. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Code according to the laws of each country governing the manufacture and distribution of food (the “Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Standards” notified 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.These food additives are sweeteners and flavors in terms of function. It is categorized into agents, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart an appropriate sweet taste to foods, and both natural and synthetic can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners 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 can be used. Preferably, it is the case of using a natural one. In the case of using natural ones, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, you can use those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo. The natural flavoring agent may be a liquid concentrate or a solid extract. 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 calcium 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, etc. may be mentioned, 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. In addition to the purpose of enhancing taste, the acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms.

As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include a physiologically active substance or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutritional properties and ensuring stability as a food additive.

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 minerals include calcium preparations such as calcium citrate, magnesium stearate. Magnesium preparations such as, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, and zinc.

In the food composition of the present invention, the food additive described above may be included in an appropriate amount to achieve the purpose of addition according to the product type.

Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the food code of each country or the food additive code.

The composition of the present invention may be understood 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 an administration route 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 a case in which two or more formulations of drugs are combined according to the route of administration, for example, when one drug is first administered by an intravenous route and the second drug is administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopoeia of each country, including the “Korean Pharmacopoeia”.

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to a method known in the art together with a suitable carrier It can be prepared in a formulation such as. Examples of suitable carriers at this time include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol, etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, colorants, and diluents may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferryst, 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, polydecylene glycol, etc., and as disintegrating agents starch, methyl cellulose , Agar, bentonite, xanthan gum, starch, alginic acid, or a sodium salt thereof. 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 an injection, a transdermal administration, a nasal inhalation and a suppository according to a method known in the art together with a suitable carrier. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, triethanol amine-containing PBS (phosphate buffered saline), sterile water for injection, an isotonic solution such as 5% dextrose, etc. may be used. . When formulated as a transdermal administration, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, liniment, air roll, and the like. In the case of nasal inhalants, suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. can be used to form an aerosol spray.When formulated as a suppository, the carrier is Withepsol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like.

The specific formulation of pharmaceutical compositions is known in the art, and for example, Remington's Pharmaceutical Sciences (19th ed., 1995) may be referred to. This document is considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. May be in the /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.

The composition of the present invention can be understood as a cosmetic composition in another specific aspect. When the composition of the present invention is regarded as a cosmetic composition, the use of the present invention can be regarded as relieving inflammatory skin irritation, in particular, relieving inflammatory skin irritation caused by acne bacteria.

Even if the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified as a product for its use and legally, and specifically, functional cosmetics with uses such as improving skin troubles and improving atopic dermatitis, non-functional It may be a general cosmetic or the like. The product form can also take any product form, specifically solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, waxes. It can take the form of a product such as a foundation or spray. In a specific product form, it may be a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder formulation.

In addition to the active ingredient, the cosmetic composition of the present invention may include components commonly used in the cosmetic composition, such as stabilizers, solubilizers, surfactants, vitamins, conventional adjuvants such as pigments and perfumes, and carriers.

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

When the formulation of the present invention is a powder or 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 / May contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or 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 may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and crystallites Castle cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

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

The cosmetic composition of the present invention can be prepared according to a method for preparing a cosmetic composition commonly used in the art, except that it contains the active ingredient that exhibits skin moisturizing activity and the like.

As described above, according to the present invention, it is possible to provide a skin trouble composition using centella asiatica extract and peony extract grown in desalted lava seawater having anti-inflammatory activity, keratin extracellular membrane formation promoting activity, and hyaluronic acid production promoting activity.

The composition of the present invention may be commercialized as food or cosmetics.

1 to 4 are results of cytotoxicity evaluation for Raw264.7 cells and HaCaT cells.
5 and 6 are NO generation evaluation results.
7 to 10 are results of evaluation of IL-8 production.
11 to 14 are results of evaluation of the production of TNF-α.
15 is a result of evaluation of keratin extracellular membrane formation ability.
16 and 17 are results of evaluation of hyaluronic acid production.

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 Centella asiatica and Peony extract cultivated in fresh water or lava seawater

<Example 1> Preparation of Centella asiatica extract grown in fresh water or desalted lava seawater

After transplanting centella seedlings (Joy Plant, seedling length 100 to 120 mm) to the cultivation table where the blended soil (Nongwoo Bio Co., Ltd.) is located, vinyl mulch for rapid growth, and after installing a sunshade, 2 in the open field. Once a day, a liquid fertilizer containing fresh water (tap water) or a liquid fertilizer containing desalted lava seawater (hardness 100) was sprayed and cultivated for 2, 4 and 6 weeks. Here, the liquid fertilizer is a diluent (containing 50g of agent A and 50g of agent B based on 50L) and desalted lava seawater (hardness 100) or fresh water of a mixture of commercially available nutrient fertilizers A and B (Coseal Co., Ltd., comprehensive herbal fertilizer). Was obtained by mixing in the same amount. Ingredients in Table 1 below are the components of Koseal's comprehensive herbal fertilizer products.

Composition of nutrient fertilizers A and B ingredients Nutrient fertilizer Agent A nitrogen
Whole quantity Phosphoric acid weir boron manganese zinc calcium 13% 0% 21% 0.2% 0.07% 0.007% 14% B agent nitrogen
Whole quantity Phosphoric acid weir Goto iron 8.5% 9.5% 22% 5.5% 0.35%

Add 20 times the weight of 70% ethanol to the dried powder of cultivated centella (outpost) , extract for 24 hours , filter with filter paper, concentrate the filtrate under reduced pressure, freeze-dry, and cultivate fresh water in powder form (2 weeks cultivation) 70% ethanol Extracts and 70% ethanol extracts grown in desalted lava seawater were obtained.

<Example 2> Preparation of peony extract grown in fresh water or desalted lava seawater

In the same manner as in Example 1, the peony was grown using fresh water or lava seawater (2 weeks of freshwater cultivation, 2 weeks of lava seawater, 2 weeks of lava seawater, cultivation of 6 weeks) and 70% ethanol extract for freshwater cultivation and 70% ethanol for desalted lava seawater cultivation. The extract was prepared. However, peony seedlings were purchased from Geumsan Seedling Farm, and seedlings with a length of 70-90 mm were used.

<Experimental Example> Cytotoxicity, anti-inflammatory activity, skin barrier strengthening and skin moisturizing activity test

1. Cytotoxicity test

HaCaT cells and Raw264.7 cells were aliquoted into a 96 well plate by 1.5×10 ⁴ cells/well, and Raw264.7 cells were divided by 6×10 ⁴ cells/well, and cultured for 24 hours under the culture conditions of each cell. After culturing for 24 hours, the medium was discarded, washed with PBS, and then the cells were starved using DMEM medium not containing FBS. The next day, a sample of a certain concentration was treated and cultured for 24 hours. 100ul of the WST-1 reagent diluted 10 times in the medium was added to each well and incubated for 2 hours. Absorbance was measured at 450 nm with an ELISA reader and cell viability was calculated as follows.

Cell viability (%)= (Sample treatment group/control group)×100

The cytotoxicity results for Raw264.7 cells are shown in FIGS. 1 and 2, and the cytotoxicity results for HaCaT cells are shown in FIGS. 3 and 4.

As a result of the experiment, it was confirmed that there was no cytotoxicity in Peony at 10 ug/ml and Centella asiatica 50 ug/ml in Raw264.7 cells, and it was confirmed that there was no cytotoxicity in Peony at 15 ug/ml and Centella asiatica 200 ug/ml in HaCaT cells.

The following experiment was conducted at the treatment concentration determined in consideration of these results.

2. Anti-inflammatory activity test

2.1 NO production inhibition activity test

Raw264.7 cells were aliquoted into 96 well plates by 6×10 ⁴ cells/well, and cultured for 24 hours under cell culture conditions. The next day, a sample was treated with LPS (lipopolysaccharide) (1 µg/ml) to the cells and cultured for 24 hours. After the cell culture solution and Griess reagent were mixed in a ratio of 1:1 and reacted for 10 minutes, absorbance at 540 nm was measured with an ELISA reader. The concentration of NO was determined using the sodium nitrite standard curve, and the final amount of NO was converted to the amount of NO per certain protein and compared with the negative control group.

The results are shown in FIGS. 5 and 6. In FIGS. 5 and 6, INDO (Indomethacin) was used as a positive control.

In the case of crops cultivated in lava seawater, the inhibitory activity of NO production was increased compared to crops cultivated in freshwater (general water) from centella and peony grown for 4 weeks, especially in lava seawater.

2.2 Cytokines IL-8 and TNF-α Inhibitory activity test

After dispensing HaCaT into a 96 well plate by 1.5×10 ⁴ cells/well, the cells were cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded, washed with PBS, and then the cells were starved using DMEM medium (serum free medium) not containing FBS. The next day, P.acne (for P.acne, use the supernatant after centrifuging the P.acne lysate with an OD (600nm) of 2-2.5) or LPS (1㎍/㎖ ) to the cells. Treated and incubated for 24 hours. After the experiment using IL-8 and TNF-a ELISA kit, respectively, the absorbance at 450 nm was measured with an ELISA reader. The final amounts of IL-8 and TNF-α were compared with the negative control group by converting the amount of IL-8 and TNF-a per certain protein.

The results of inhibition of IL-8 production by LPS stimulation are shown in Figs. 7 and 8, and the results of inhibition of IL-8 production by P.acne stimulation are shown in Figs. 9 and 10.

In terms of the inhibitory activity of IL-8 production by LPS stimulation, compared to crops grown with lava seawater, especially peony grown for 2 weeks with lava seawater, and crops grown with fresh water (normal water) from Centella asiatica grown for 4 weeks with lava seawater. -8 production inhibitory activity was increased.

In terms of inhibitory activity of IL-8 production by P.acne stimulation, IL-8 production inhibitory activity compared to crops grown in lava seawater, especially peony grown for 4 weeks in lava seawater, and crops grown in freshwater (normal water) from Centella asiatica. Increased.

The results of inhibiting the production of TNF-α by LPS stimulation are shown in Fig. 11 and Fig. 12, and the results of suppression of TNF-α production by P.acne stimulation are shown in Figs. 13 and 14.

In terms of the inhibitory activity of TNF-α production by LPS stimulation, TNF-α compared to crops grown in lava seawater, especially peony grown for 6 weeks in lava seawater, and crops grown in freshwater (normal water) from Centella asiatica grown for 4 weeks in lava seawater. -α production inhibitory activity was increased.

In terms of inhibiting the production of TNF-α by P.acne stimulation, crops cultivated in lava seawater, especially peony cultivated in lava seawater for 6 weeks, crops cultivated in freshwater (general water) from centella asiatica grown for 4 weeks in lava seawater. In contrast, the inhibitory activity of TNF-α production was increased.

3. Skin barrier strengthening test-Cornified envelope (CE) formation ability test

After dispensing HaCaT cells into 6 well plates by 2.5×10 ⁵ cells/well, they were cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded, washed with PBS, and cultured using DMEM medium not containing calcium. After cultivation, a sample of a certain concentration was treated and cultured. The cells were washed with PBS and lysed using 0.1M Tris buffer containing 2% sodium dodecyl sulphate (SDS) and 20 mM dithiothreitol (DTT). After boiling at 100° C. for 5 minutes, absorbance was measured at 340 nm with an ELISA reader.

The results are shown in FIG. 15.

In terms of CE formation ability, CE formation ability was increased compared to crops grown with lava seawater, especially peony grown for 4 weeks with lava seawater, and crops grown with fresh water (normal water) from Centella asiatica.

4. Skin moisturizing activity test

After dispensing HaCaT cells into a 96 well plate by 1.5×10 ⁴ cells/well, they were cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded, washed with PBS, and then the cells were starved using DMEM medium not containing FBS. The next day, a sample of a certain concentration was treated and cultured. After the experiment using a hyaluronic acid ELISA kit, the absorbance was measured at 450 nm with an ELISA reader. The final amount of hyaluronic acid was converted to the amount of hyaluronic acid per certain protein and compared with the negative control group.

The results are shown in Figs. 16 and 17.

In terms of the hyaluronic acid production activity, the hyaluronic acid production activity was increased compared to the crops grown with lava seawater, especially the peony grown for 2 weeks with lava seawater, and the crops grown with fresh water (normal water) from Centella asiatica grown for 4 weeks.

Claims (10)

A mixed solvent extract of water and ethanol from Centella asiatica cultivated using desalted lava seawater or a mixed solvent extract of water and ethanol from a peony extract cultivated using desalted lava seawater as an active ingredient,
The peony is an anti-inflammatory composition, characterized in that obtained by cultivation in desalted lava seawater for 4 to 6 weeks.
The method of claim 1,
The composition, characterized in that the mixed solvent of water and ethanol is 70% ethanol.
The method according to claim 1 or 2,
The composition is a composition, characterized in that the food composition.
The method according to claim 1 or 2,
The composition is a cosmetic composition,
The anti-inflammatory composition, characterized in that the relief of inflammatory skin irritation.
The method of claim 4,
The composition, characterized in that the inflammatory skin irritation is an inflammatory skin irritation caused by acne bacteria.
As an active ingredient, a mixed solvent extract of centella oil and ethanol cultivated using desalted lava seawater or a mixed solvent extract of water and ethanol cultivated using desalted lava seawater,
The peony is a composition for skin moisturizing, characterized in that obtained by cultivation in desalted lava seawater for two weeks.
The method of claim 6,
The composition, characterized in that the mixed solvent of water and ethanol is 70% ethanol.
The method according to claim 6 or 7,
The composition is a composition, characterized in that the food composition.
The method according to claim 6 or 7,
The composition is a composition, characterized in that the cosmetic composition.
It contains as an active ingredient a mixed solvent extract of water and ethanol of peony grown using desalted lava seawater,
The peony is characterized in that obtained by cultivation for 4 weeks in desalted lava seawater
Cosmetic composition for enhancing skin barrier function.

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