KR101729494B1 - Cosmetic composition containing 6,7-dihydroxy bergamottin - Google Patents
Cosmetic composition containing 6,7-dihydroxy bergamottin Download PDFInfo
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- KR101729494B1 KR101729494B1 KR1020150092434A KR20150092434A KR101729494B1 KR 101729494 B1 KR101729494 B1 KR 101729494B1 KR 1020150092434 A KR1020150092434 A KR 1020150092434A KR 20150092434 A KR20150092434 A KR 20150092434A KR 101729494 B1 KR101729494 B1 KR 101729494B1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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Abstract
The present invention relates to a cosmetic composition, and more particularly, to a cosmetic composition containing 6,7-dihydroxyvergamatin, which promotes collagen production and inhibits the production of MMP-1 to improve wrinkles, Dihydroxy < / RTI > bergamotene, which inhibits the production of NO, an inflammatory factor that induces inflammation and does not have cytotoxicity, and has an anti-inflammatory effect.
Description
The present invention relates to a cosmetic composition, and more particularly, to a cosmetic composition containing 6,7-dihydroxyvergamatin, which promotes collagen production and inhibits the production of MMP-1 to improve wrinkles, Dihydroxy < / RTI > bergamotene, which inhibits the production of NO, an inflammatory factor that induces inflammation and does not have cytotoxicity, and has an anti-inflammatory effect.
Cosmetics are deeply related to our daily lives and are used by many people. The functions of cosmetics are not only clean and aesthetic but also free radicals, free radicals, free radicals, peroxides due to external factors such as pollutants, stress and ultraviolet rays, internal factors such as immune cell dysfunction, Such as protein, nucleic acid, cell membrane lipid breakage, etc. In addition, cosmetics having whitening, skin wrinkle improvement and inhibition, and ultraviolet shielding function to meet consumers' desire for high-performance cosmetics are continuously being developed. Among them, wrinkle improvement and restraining products are attracting much attention from consumers who want to keep younger.
Originally, wrinkles caused by aging can be defined as bone on skin surface due to loss of collagen and elastic fibers in skin dermal layer. MMP enzyme promotes collagen degradation. Therefore, collagen synthesis promoting substances such as retinoic acid and chlorella extract for promoting collagen production and preventing skin aging have been developed as described in the following patent documents.
<Patent Literature>
U.S. Patent No. 4,603,146 (registered on June 29, 1986) "Methods for retarding the effects of aging of the skin"
Japanese Patent No. JP10-36283 (registered on Jun. 08, 2007) "Fibroblast Growth Promoting Agent"
However, it is unstable in retinoic acid, there is a limitation in usage due to safety problems such as irritation and redness in skin application, chlorella extract and the like have insufficient effects, and skin collagen synthesis is promoted substantially, .
SUMMARY OF THE INVENTION The present invention has been made to solve the above problems,
It is an object of the present invention to provide a cosmetic composition having an effect of improving skin wrinkles.
It is another object of the present invention to provide a cosmetic composition having an anti-inflammatory effect.
In order to achieve the above object, the present invention is implemented by the following embodiments.
According to one embodiment of the present invention, the cosmetic composition according to the present invention is characterized by containing 6,7-dihydroxyvergamant as an active ingredient.
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According to another embodiment of the present invention, the cosmetic composition according to the present invention is characterized by having 6,7-dihydroxyvergamene as an active ingredient and having an effect of improving skin wrinkles.
According to another embodiment of the present invention, the cosmetic composition according to the present invention is characterized in that 6,7-dihydroxyvergamant is contained as an active ingredient and has an anti-inflammatory effect.
According to the present invention, the following effects can be obtained by this embodiment.
The present invention has the effect of increasing the synthesis of collagen and inhibiting the production of MMP-1, thereby improving the wrinkles of the skin.
The present invention also has anti-inflammatory effects by inhibiting the production of inflammatory factors found in atopic and acne diseases and the like.
In addition, the present invention not only increases the synthesis of collagen but also inhibits the production of inflammatory factors.
Fig. 1 is a graph showing the evaluation of collagen production efficacy according to Example 1. Fig.
2 is a graph showing the evaluation of the inhibitory effect of MMP-1 production according to Example 1. Fig.
3 is a graph showing the evaluation of the inhibitory effect on nitric oxide formation according to Example 2. Fig.
Hereinafter, the cosmetic composition containing 6,7-dihydroxyvergamantin according to the present invention will be described in detail with reference to the accompanying drawings. Unless defined otherwise, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs and, if conflict with the meaning of the terms used herein, It follows the definition used in the specification. Further, the detailed description of known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted. Throughout the specification, when an element is referred to as "including " an element, it is understood that the element may include other elements as well, without departing from the other elements unless specifically stated otherwise.
The cosmetic composition according to one embodiment of the present invention comprises 6,7-dihydroxyvergamantin. The 6,7-dihydroxy bergamotin has a molecular formula of C 21 H 24 O 6 and has a structure represented by the following formula (1), and the CAS Registry Number is 145414-76-2. It is a substance that can be obtained. However, the wrinkle improvement and anti-inflammatory effect on 6,7-dihydroxyvergamethin were not known at all.
The cosmetic composition according to the present invention contains 6,7-dihydroxyvergamethin to increase the synthesis of collagen to improve the wrinkles of the skin and to inhibit the production of inflammatory factors such as atopic and acne.
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The cosmetic composition of the present invention may further comprise a composition selected from the group consisting of water-soluble vitamins, oiliness vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.
As the water-soluble vitamin, any of vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C and vitamin H can be used, And salts thereof (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium ascorbic acid-2-phosphate etc.) can also be used as water-soluble vitamins . The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzymatic method, or a chemical synthesis method.
As the above-mentioned useful vitamins, any vitamins A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) , And derivatives thereof (such as palmitic acid ascorbin, stearic acid ascorbic acid, dipalmitic acid ascorbic acid, dl-alpha tocopheryl acetate, dl-alpha tocopherol nicotinic acid vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, Ethyl ether, etc.) and the like are also included in the usable vitamins used in the present invention. The usable vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzyme or a chemical synthesis method.
The polymeric peptide may be any compound as long as it can be compounded in a cosmetic, and preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like. The polymeric peptide can be obtained by a conventional method such as purification from a culture broth of a microorganism, an enzymatic method, or a chemical synthesis method, or purified from natural products such as ducks such as pigs and cows and silk fiber of silkworms .
The polymeric polysaccharide may be any compound as long as it can be compounded in cosmetics, and examples thereof include hydroxyethylcellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be usually purified from mammals or fish.
The sphingogly lipid may be any as long as it can be compounded in cosmetics, and preferable examples thereof include ceramides, phytosphingosine and sphingoglycolipids. The sphingogly lipids can be purified from ordinary mammals, fish, shellfish, yeast or plants by a conventional method or can be obtained by chemical synthesis.
The seaweed extract may be any of those which can be compounded in cosmetics, but preferably include algae extract, red tiger extract, green algae extract and the like, and furthermore, coloring guanine, arginic acid, sodium alginate , Potassium alginate, and the like are also included in the seaweed extract used in the present invention. The seaweed extract can be obtained from seaweed by a conventional method.
The cosmetic composition of the present invention may contain other ingredients usually added to the cosmetic composition. For example, it is possible to use various additives such as a preservative, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a plant extract, a pH adjuster, Cold agents, antiperspirants, purified water, and the like.
Examples of the above-mentioned fat-soluble ingredients include ester-based fats, hydrocarbon-based fats, silicon-based fats, fluorine-based fats, animal fats and plant fats and the like. Examples of the ester-based oil include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, But are not limited to, ethylbutyl, butylbutyl, ethylbutylbutyl, butylbutyl, isobutylbutyl, isobutylbutyl, isobutylisobutyl, isobutylisobutyl, isobutylisobutyl, Trimethylol propane, triisostearic acid trimethylol propane, tetra-2-ethylhexanoic acid pentaerythritol tri (2-ethylhexanoic acid) But are not limited to, tallow, caprylic estersyl, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, myristine sasyl, stearyl stearate, decyl oleate, Examples of the isocyanates include isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, Ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol , Propylene glycol dicaprylylate, dicapric acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated acid glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyl Dodecylsuccinic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, Polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, octyldecyl lactate, octyldecyl isostearate, polyglycerin oleic acid ester, But are not limited to, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, Propyl stearate, propyl stearate, propyl stearate, stearyl stearate, stearyl stearate, propyl stearate, stearyl stearate, Stearyl hydroxystearate, stearyl 12-stearoyl hydroxystearate, 12-stearoyl hydroxystearic acid, Allo one hydroxy stearic acid and the like esters such as cetearyl source. Examples of the hydrocarbonaceous oil include hydrocarbons such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and vaseline. Examples of the silicone-based oil include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethyl polysiloxane, decamethyl polysiloxane, dodecamethyl cyclosiloxane, dimethyl siloxane methyl cetyloxysiloxane copolymer, dimethyl siloxane methyl stearoxysiloxane copolymer, Alkyl-modified silicone oil, and amino-modified silicone oil. Examples of the fluorine-based oil include perfluoropolyether and the like. Examples of the animal or plant oil include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, The present invention relates to a process for producing a starch-containing starch, which is selected from the group consisting of seed oil, cottonseed oil, palm oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, Animal or vegetable fats such as oil, can-pick wax, carnauba wax, liquid lanolin, and hardened castor oil.
Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and oil-soluble polymers. Examples of the water-soluble low molecular weight moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B Propylene glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt and the like. Examples of the lipid-soluble low-molecular moisturizing agent include cholesterol, cholesterol ester and the like. Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid salt, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, . Examples of the oil-soluble polymer include polyvinylpyrrolidone / eicosene copolymer, polyvinylpyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.
Examples of the molient agent include long chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid and lanolin fatty acid cholesteryl ester.
Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants. Examples of the nonionic surfactant include self emulsifying monostearic acid glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene), sorbitan fatty acid ester, POE sorbit fatty acid ester POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POEPOP (polyoxyethylene polyoxypropylene) copolymer, POEPOP alkyl ether, polyether modified silicone, lauric acid alkanolamide , Alkylamine oxides, hydrogenated soybean phospholipids, and the like. Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl ginseng salt, alkyl Amide phosphate, alkyloyl taurine salt, N-acyl amino acid salt, POE alkyl ether carboxylate, alkylsulfosuccinate, sodium alkylsulfoacetate, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester and the like . Examples of the cationic surfactant include alkyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium bromide, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium chloride, behenyl trimethyl ammonium bromide, Benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like. Examples of the amphoteric surfactant include carboxybetaine, amidebetaine, sulfobetaine, hydroxysulfobetaine, amidosulfobetaine, phosphobetaine, aminocarboxylate, imidazoline derivative, amideamine, etc. , And the like.
Examples of the organic and inorganic pigments include silicic acid, anhydrous silicic acid, magnesium silicate, talc, sericite, mica, kaolin, spinach, clay, bentonite, titanium coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, Inorganic pigments such as aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium oxide, But are not limited to, polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, Silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and composite pigments of inorganic pigments and organic pigments thereof.
Examples of the organic powder include metal soaps such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethylmethacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride, and the like.
Examples of the ultraviolet absorber include ultraviolet absorbers such as p-aminobenzoic acid, ethyl p-aminobenzoate, amyl paranobenzoate, octyl p-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, benzyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, Benzyl, 2-ethoxyethyl methoxycinnamate, octyl methoxycinnamate, mono-2-ethylhexane glyceryl dipyrromethoxycinnamate, isopropyl methoxycinnamate, diisopropyl-diisopropyl cinnamate ester mixture, Dihydroxymethoxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid and its salt, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzo Phenanone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino-p- (carbo-2'-ethylhexyl- 1,3,5-triazine, 2- (2- And the like can be de-hydroxy-5-methylphenyl) benzotriazole.
Examples of the bactericide include antibiotics such as hinokitiol, trichloroacid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, No. 301, mononitro and eicoll sodium, undecylenic acid, and the like.
Examples of the antioxidant include butylhydroxyanisole, gallic acid propyl, and eicosorbic acid.
Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogenphosphate and the like.
Examples of the alcohol include higher alcohols such as cetyl alcohol.
In addition, any of the above components may be blended within the range not impairing the object and effect of the present invention, but is preferably 0.01 to 5% by weight based on the total weight, More preferably 0.01 to 3% by weight.
The cosmetic composition of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.
The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients conventionally used in cosmetic compositions in addition to the above-mentioned compounds, for example, conventional additives such as stabilizers, solubilizers, vitamins, And a carrier.
The cosmetic composition of the present invention can be prepared in any formulations conventionally produced in the art, and examples thereof include emulsions, creams, foundations, lotions, essences, and hair cosmetic compositions. Specifically, the cosmetic composition of the present invention can be used as a skin lotion, a skin softener, a moisturizing lotion, a nutritional lotion, a massage cream, a nutritional cream, a hand cream, a moisturizing cream, an essence, a pack, a soap, a lotion, a milk lotion, Cleansing foams, body cleansers, astringents, and spray formulations. When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component . 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 the case of a spray, in particular, / Propane or dimethyl ether. In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters. When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used. When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.
Hereinafter, the present invention will be described in more detail with reference to Examples. However, these are only for the purpose of illustrating the present invention in more detail, but the scope of the present invention is not limited thereto.
≪ Example 1 > Evaluation of skin wrinkle improving effect
1) Cell Culture (Cell Culture)
Human fibroblast dermal cell HDFn (ATCC-CRL-2076) was cultured in DMEM / F12 (3: 1) medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin incubated in the ℃, 5% CO 2 conditions, and the mixture was sub-cultured by trypsinization.
2) Quantitative detection of
The HDFn was inoculated in a 96-well plate at 1 × 10 5 cells / ml, and then stabilized for 24 hours at 37 ° C. and 5% CO 2 . Thereafter, the medium was replaced with DMEM / F12 (3: 1) medium not containing FBS, and a sample (6,7-dihydroxyvergamotin) was added thereto in concentration and cultured for 48 hours. TGF-β was used as a positive control. Subsequently, the supernatant of the medium was collected and the amount of procollagen was measured using a Procollagen type 1 C-peptide (PIP) EIA kit. The results are shown in FIG.
3) Quantification of MMP-1 activity
The HDFn was inoculated in a 96-well plate at 1 × 10 5 cells / ml, and then stabilized for 24 hours at 37 ° C. and 5% CO 2 . Thereafter, the medium was replaced with DMEM / F12 (3: 1) medium not containing FBS, pretreated for 30 minutes, removed again and TNF-alpha (10 ng / ml) Lt; / RTI > Then, the supernatant of the medium was collected and the amount of MMP-1 was measured with Amercham MMP-1 and Biotrack ELISA system (GE healthcare). The results are shown in FIG.
4) Evaluation of experimental results
1, the amount of pro-collagen type-1 biosynthesis in human dermal fibroblast cell (HDFn) was measured. As a result, the amount of pro-collagen increased with increasing concentration of 6,7-dihydroxy Bergamottin from 12.5 to 100 μM . FIG. 2 also shows that MMP-1 production decreases as the concentration of 6,7-dihydroxy Bergamottin increases from 12.5 to 100 μM. Therefore, it was confirmed that the 6,7-dihydroxy Bergamottin used in the present invention had an excellent effect for improving the wrinkles of the skin.
<Example 2> Evaluation of anti-inflammatory effect
1) Cell Culture (Cell Culture)
RAW 264.7 cells were cultured in a DMEM medium supplemented with 10% FBS and 100 units / ml penicillin-streptomycin in a incubator maintained at 37 ° C and 5% CO 2 and subcultured three times a day.
2) Evaluation of cytotoxicity in vitro
6,7-dihydroxybigamotin was used for MTT assay. RAW264.7 cells were co-cultured with LPS (1 μg / mL) and a sample (6,7-dihydroxy Bergamottin) for 24 hours. After the incubation, 10 μL of MTT solution was added and incubated for 2 hours. The supernatant was removed, and DMSO was added. After shaking well using a plate mixer, absorbance was measured at 540 nm. The cytotoxicity was evaluated in comparison with the control group treated with the stimulant.
3) Perform NO assay
The NO assay was performed to investigate the effect of nitric oxide production, one of the active oxygen species and known to play an important role in inflammation induction. RAW264.7 cells were adjusted to 1.5 × 10 5 cells / mL using DMEM medium, and then seeded in 96-well plates. A new medium containing 6,7-dihydroxy Bergamottin and LPS (1 μg / mL) And cultured for 24 hours. The amount of NO produced was measured in the form of NO 2 - present in cell culture medium using Griess reagent. 100 μL of the cell culture supernatant and 100 μL of the glycated reagent were mixed and reacted for 10 minutes, and the absorbance was measured at 540 nm using an ELISA reader. Standard concentration curves were obtained by serial dilution of sodium nitrite (NaNO 2 ) (1-100 μM).
4) Evaluation of experimental results
In the RAW 264.7 cells, no cytotoxicity was observed for the sample 6,7-dihydroxy Bergamottin. FIG. 3 shows that the NO production was decreased as the concentration of 6,7-dihydroxy Bergamottin increased from 50 to 200 μM there was. Therefore, it was confirmed that the 6,7-dihydroxy Bergamottin used in the present invention had an anti-inflammatory effect.
The above results show that 6,7-dihydroxy Bergamottin inhibits the production of NO, which is an inflammatory factor that induces collagen production, suppresses the production of MMP-1 and has a wrinkle-reducing effect and also causes atopy and acne It has anti-inflammatory effect because it does not have cytotoxicity.
While the present invention has been described in connection with what is presently considered to be practical exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, Should be interpreted as belonging to the scope.
Claims (4)
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100510809B1 (en) | 1997-08-19 | 2005-08-31 | 워너-램버트 캄파니 엘엘씨 | Compositions Containing Bergamottin for Increasing the Oral Bioavailability of Pharmaceutical Agents |
| KR101593624B1 (en) | 2009-07-31 | 2016-02-15 | (주)아모레퍼시픽 | Composition for reducing irritation and inflammation in skin |
| JP2016220576A (en) | 2015-05-28 | 2016-12-28 | 池田食研株式会社 | Citrus extract preparation method |
-
2015
- 2015-06-29 KR KR1020150092434A patent/KR101729494B1/en active IP Right Grant
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100510809B1 (en) | 1997-08-19 | 2005-08-31 | 워너-램버트 캄파니 엘엘씨 | Compositions Containing Bergamottin for Increasing the Oral Bioavailability of Pharmaceutical Agents |
| KR101593624B1 (en) | 2009-07-31 | 2016-02-15 | (주)아모레퍼시픽 | Composition for reducing irritation and inflammation in skin |
| JP2016220576A (en) | 2015-05-28 | 2016-12-28 | 池田食研株式会社 | Citrus extract preparation method |
Non-Patent Citations (1)
| Title |
|---|
| 21170_IL799 ANNEX - Furocoumarins in finished cosmetic products |
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