KR20180033438A - External composition for soothing effect on the skin comprising an extract of fermented wheat germ - Google Patents
External composition for soothing effect on the skin comprising an extract of fermented wheat germ Download PDFInfo
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Abstract
Description
The present invention relates to a composition for external application having excellent skin soothing effect, and more particularly to a composition for skin-soothing cosmetic composition comprising 2,6-dimethoxybenzoquinone or a wheat germ fermented product or an extract thereof, a composition for preventing or treating inflammatory skin diseases , And a quasi-quasi-product composition for skin soothing.
The skin of the human body is stimulated by various harmful substances, prolonged exposure to sunlight or cosmetic raw materials, which can cause skin problems such as skin inflammation, tautness, red spots and edema have.
In order to solve the skin problem, it is possible to consider application of an anti-inflammatory agent, but most of the conventional anti-inflammatory agents can not be used as a cosmetic raw material or have a limited use amount. In addition, steroid-based anti-inflammatory drugs are accompanied by side effects such as skin atrophy, insomnia and anxiety. Accordingly, the development of natural raw materials which are excellent in safety for skin and effective for calming skin has been actively carried out (see Korean Patent Publication Nos. 10-2011-0038546, 10-2010-0086698 and 10-2012-0015881 ).
Under these circumstances, the present inventors have intensively studied to develop a cosmetic composition for skin soothing that contains a natural substance. As a result, it has been found that a composition comprising 2,6-dimethoxybenzoquinone or a wheat germ fermented product or an extract thereof, And showed the skin soothing effect without completing the present application.
The object of the present invention is to provide a skin-soothing cosmetic composition comprising a compound of the following formula 1, or a wheat germ fermented product or an extract thereof.
[Chemical Formula 1]
Another object of the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory skin diseases, which comprises the compound of formula (I) or a fermented wheat germ or an extract thereof.
Another object of the present invention is to provide a method for preventing, ameliorating or treating an inflammatory skin disease comprising administering to a subject in need thereof a composition comprising the compound of formula 1 or a fermented wheat germ extract or an extract thereof will be.
Another object of the present invention is to provide a quasi-drug composition for skin soothing which comprises the compound of formula (1), or a wheat germ fermented product or an extract thereof.
Another object of the present invention is to provide a method of soothing the skin comprising the step of administering a composition comprising the compound of formula 1 or the fermented wheat germ extract or extract thereof to a subject in need thereof.
In one aspect to achieve the above object, the present application provides a skin-soothing cosmetic composition comprising a compound of the following formula 1, or a wheat germ fermented product or an extract thereof.
The compound which may be contained as an active ingredient in the cosmetic composition of the present application may be a compound represented by the following formula (1).
[Chemical Formula 1]
The compound of formula 1 means 2,6-dimethoxycyclohexa-2,5-diene-1,4-dione, 2,6-DMBQ, 2,6-dimethoxybenzoquinone or dimethoxybenzoquinone in the application. Specifically, the compound of Formula 1 may be in the form of a non-glycoside.
The compound of Chemical Formula 1 may be chemically or biologically synthesized or may be purchased commercially. Or from natural products known to contain the compound of the present application (for example, plants, etc.). However, the present invention is not particularly limited thereto. In the present application, the compound of formula (1) may include both physiologically acceptable salts of the compound of formula (1) as well as possible solvates and hydrates thereof which may be prepared therefrom. The compound of formula (I) may exhibit an anti-inflammatory effect and exhibit a skin soothing effect.
The wheat embryo of the present application may include, but is not limited to, the dried, ground, concentrate and mixtures thereof of the wheat embryo.
The term " fermentation " as used in this application means any activity or process involving enzymatic or metabolic degradation of an organic material with a microorganism.
The term " wheat germ fermentation product " as used in the present application means the result of enzymatic or metabolic degradation of wheat germ using microorganisms.
The wheat germ fermented product of the present application may contain the compound of formula (1) of the present application.
The wheat germ fermented product of the present application may be obtained by inoculating a microorganism into a wheat germ or a culture medium containing the same and culturing the same. Specifically, the microorganism may be at least one selected from the group consisting of yeast, lactic acid bacteria, bacteria, and fungi.
The yeast may be, for example, Saccharomyces cerevisiae , S. ellipsoideus , S. coreanus , Saccharomyces cylindusensis (" Saccharomyces cerevisiae " S. carlsbergensis), Saccharomyces access to my Paz Astoria Augustine (S. pastorianus), Saccharomyces access to my lactis (S. lactis), Saccharomyces access to my ruksi (S. rouxii), to access Shijo Saccharomyces pombe Mai (Schizosaccharomyces pombe ), Zygosaccharomyces major , Hansenula anomala , Brettanomyces bruxellensis , B. custersianus , Dekkera , Dekkera anomala , Streptomyces olivochromogenes , or S. griseus , but the wheat germ fermented product of the present application or Is not limited insofar as the extract of the present invention can exhibit a skin soothing effect for the purpose of the present application. For example, the yeast may be Saccharomyces cerevisiae.
The lactic acid bacteria may be, for example, Bifidobacterium sp . ), Lactobacillus genus (Lactobacillus sp.), In Lactococcus (Lactococcus sp . ), Peddie Oh caucus in (Pediococcus sp.), May be a microorganism Streptococcus genus (Streptococcus. Sp), or the current Kono Stock in (Leuconostoc sp.), Filed a wheat germ fermented or its extract of the present application And the like. For example, the lactic acid bacteria is Bifidobacterium bipyridinium bushes (B. bifidum), Bifidobacterium breather probe (B. breve), Bifidobacterium ronggeom (B. longum), Bifidobacterium Annie Marlies (B. animalis), Bifidobacterium lactis (B. lactis), Lactobacillus ash's also a filler (L. acidophilus), Casey Lactobacillus (L. casei), Lactobacillus joined Lee (L. gasseri), Lactobacillus del Brewer L. delbrueckii spp . L. bulgaricus , L. helveticus , L. fermentum , L. paracasei , L. vaginalis , and the like), Lactobacillus spp . plantarum , L. reuteri , L. rhamnosus , L. salivarius , L. lactis , Pediococcus sereviiae (Lactobacillus spp. ), Lactobacillus sp. P. cerevisiae), Peddie Oh Caucus rakti City Sidi Ke (P. acidilactici), Streptococcus lactis (S. lactis), Streptococcus thermostat Phillies (S. thermophiles), Streptococcus Crescent Morris (S. cremoris), or current It may be L. mesenteroides . In the present application, lactic acid bacteria may be used in the same sense as lactic acid bacteria.
The fungus used in the fermentation may include, but is not limited to, mushrooms. Examples of the mushrooms include shiitake mushroom, oyster mushroom, mushroom mushroom, mushroom mushroom, mushroom mushroom, mushroom mushroom, Ginger mushroom, or mushroom, but is not limited thereto as long as the wheat germ fermented product or extract thereof of the present application can exhibit a skin soothing effect for the purpose of the present application.
The amount of inoculation, culture temperature, culture humidity, and incubation time of the microorganism used to produce the wheat germ fermented product of the present application can be appropriately selected by those skilled in the art in consideration of the kind of microorganism used for fermentation. As a non-limiting example, the cultivation of the present application can be carried out at a temperature of 20 ° C to 40 ° C for 12 to 60 hours. Specifically, the cultivation of the present application is carried out at a temperature of 25 ° C to 35 ° C, 28 ° C to 32 ° C or 30 ° C and / or 12 hours to 50 hours, 20 hours to 50 hours, 30 hours to 50 hours, 38 hours to 42 hours or 40 hours.
As used herein, the term "extract of wheat germ fermented product" refers to a substance from which microorganisms have been removed from a wheat germ fermentation product.
The elimination of the present application may include any method for removing microorganisms (for example, yeast cells) contained in microbial fermentations known in the art. Specifically, the extract of the wheat germ fermented product of the present application may be a supernatant obtained by centrifuging the wheat germ fermented product of the present application. More specifically, the centrifugation may be carried out at a rotational speed of 6,000 rpm to 10,000 rpm, 7000 rpm to 9000 rpm, 7500 rpm to 8500 rpm, 7800 rpm to 8200 rpm or 8000 rpm and / 5 minutes to 40 minutes, 5 minutes to 30 minutes, 10 minutes to 120 minutes, 10 minutes to 90 minutes, 10 minutes to 60 minutes, 10 minutes to 40 minutes, 15 to 30 minutes, 15 to 120 minutes, 15 to 90 minutes, 15 to 60 minutes, 15 to 40 minutes, 15 to 30 minutes, 15 to 25 minutes, or 20 minutes.
The extract of the wheat germ fermented product of the present application may contain the compound of formula (1) of the present application, though not particularly limited thereto.
The extract of the wheat germ fermented product of the present application includes the extract of the wheat germ fermented product of the present application and an extract of all the formulations which can be formed by using an extract such as a diluted solution, a concentrate, a dried product, a controlled preparation, a purified product or a mixture thereof. Specifically, the extract of the wheat germ fermented product of the present application may be a dried product, more specifically, a lyophilized product.
In addition, the extract of the wheat germ fermented product of the present application can be obtained from the wheat germ fermented product by any method as long as it has a skin soothing effect or an effect of preventing, improving or treating inflammatory skin diseases, A method such as a cold extraction method in which water is dipped in a solvent and extracted at a room temperature of 10 to 25 캜, a heating extraction method in which the extraction is performed by heating at 40 to 100 캜, an ultrasonic extraction method in which ultrasonic waves are applied to the extraction method and a reflux extraction method using a reflux condenser Can be used. These methods may be performed alone or in combination of two or more methods.
The kind of the extraction solvent used in the extraction of the present application is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the extraction solvent of the present application include water, a group consisting of an alcohol having 1 to 4 carbon atoms, hexane, ethyl acetate, chloroform, dichloromethane and a mixed solvent thereof . In particular, the extraction solvent of the present application may be hydrothermal.
The extract of the present application may comprise fractions thereof. The term " fraction " as used herein means a product obtained by performing a fractionation to separate a specific component or a specific component group from a mixture containing various components.
The fractionation method for obtaining the fraction of the present application is not particularly limited and may be carried out according to a method commonly used in the art. A solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity ), A combination thereof, and the like. The kind of the fraction solvent used for obtaining the fraction of the present application is not particularly limited and any solvent known in the art can be used. Non-limiting examples of the fraction solvent of the present application include polar solvents such as water and alcohols having 1 to 6 carbon atoms; And non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of one or more.
The wheat germ fermented product or extract thereof of the present application may be used in an amount of from 0.001% by weight to 10% by weight based on the total weight of the composition of the present application, specifically 0.01% by weight to 7% by weight, 0.01% by weight to 5% From 0.01% to 3%, from 0.01% to 2%, from 0.01% to 1%, from 0.05% to 10%, from 0.05% to 7%, from 0.05% to 5% % To 3 wt%, 0.05 wt% to 2 wt%, 0.05 wt% to 1 wt%, 0.1 wt% to 10 wt%, 0.1 wt% to 7 wt%, 0.1 wt% to 5 wt%, 0.1 wt% 0.5 to 3 wt%, 0.1 to 2 wt%, 0.1 to 1 wt%, 0.5 to 10 wt%, 0.5 wt% to 7 wt%, 0.5 wt% to 5 wt%, 0.5 wt% to 3 wt% %, 0.5 wt% to 2 wt%, 0.5 wt% to 1 wt%, 0.8 wt% to 10 wt%, 0.8 wt% to 7 wt%, 0.8 wt% to 5 wt%, 0.8 wt% 3% by weight, and 0.8 can be included in a weight% to 2 weight%, 0.8 weight% to 1% by weight or 1% by weight.
The wheat germ fermented product or the extract thereof of the present application may be contained in the composition of the present application at 0.1 μg / ml to 400 μg / ml. Specifically, the wheat germ fermented product or the extract thereof of the present application may be added to the composition of the present invention in an amount of 25 μg / ml to 400 μg / ml, 50 μg / ml to 400 μg / ml, 100 μg / ml to 400 μg / / ml to 400 [mu] g / ml.
The compound represented by Formula 1 of the present application may be used in an amount of 0.00001 wt% to 0.1 wt% based on the total weight of the composition, specifically 0.00001 wt% to 0.01 wt%, 0.00001 wt% to 0.001 wt%, 0.00005 wt% 0.001 to 0.001 wt.%, 0.00005 to 0.001 wt.%, 0.0001 to 0.01 wt.%, 0.0001 to 0.001 wt.%, 0.0001 to 0.001 wt.%, 0.0005 to 1 wt.% 0.01% by weight, 0.0005% by weight to 0.001% by weight, 0.0005% by weight to 0.001% by weight, or 0.0007% by weight.
In addition, the compound represented by the formula (1) of the present application may be contained in the composition of the present application at 15 μg / ml to 120 μg / ml. Specifically, the wheat germ fermented product or the extract of the present application may be produced by adding the wheat germ fermented product or the extract thereof of the present application to the composition of the present application in an amount of 30 to 120 μg / ml or 60 μg / ml To 120 μg / ml.
The term "skin soothing" in this application means to alleviate and calm the erythema or irritated skin area. For example, the skin sedation may include improvement or treatment of inflammation, reduction of transdermal water loss and / or reduction of redness, as compared with the case of not treating the composition according to the present invention, but is not particularly limited thereto.
As long as the skin is soothed by the cosmetic composition in the present application, the cause of skin sedation is not limited. Specifically, the skin irritation can be improved by the composition of the present application to calm the skin.
The cosmetic composition of the present invention can be used in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation, And may be formulated and provided in a selected form.
The cosmetic composition of the present application may further comprise a cosmetically acceptable carrier.
The type of cosmetically acceptable carrier of the present application is not particularly limited so long as it does not impair the activity and properties of the present cosmetic composition, and any cosmetically acceptable carrier conventionally used in the art can be used. Non-limiting examples of the cosmetically acceptable carrier include saline, sterilized water, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and the like. These may be used alone or in combination of two or more. The cosmetically acceptable carrier of the present application may comprise a non-naturally occuring carrier.
The cosmetically acceptable carrier of the present application varies according to the formulation of the cosmetic composition.
When the formulation of the cosmetic composition of the present application is a paste, a cream or a gel, it is preferable to use an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, And the like may be used, but the present invention is not limited thereto.
When the formulation of the cosmetic composition of the present application is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or the like may be used as a carrier component. But are not limited to, propellants such as hydrofluorocarbons, propane / butane or dimethyl ether.
When the formulation of the cosmetic composition of the present application is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Benzoate, propylene glycol, 1,3-butyl glycol oil and the like can be used. Particularly, it is possible to use a cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan Of fatty acid esters may be used, but are not limited thereto.
When the formulation of the cosmetic composition of the present application is in the form of a suspension, the carrier component may be a suspending agent such as water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, but are not limited thereto.
When the formulation of the cosmetic composition of the present application is a soap, it is preferable to use, as a carrier component, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, an isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, May be used, but is not limited thereto.
In the case where the formulation of the cosmetic composition of the present application is a pack, a pack off pack containing polyvinyl alcohol or the like, a wash off containing a pigment such as kaolin, talc, zinc oxide, or titanium dioxide, Pack, or mask sheet pack, and is not particularly limited thereto.
The cosmetic composition of the present application further comprises a component included in a conventional skin external agent component such as water, a surfactant, a humectant, a lower alcohol having 1 to 6 carbon atoms, a chelating agent, a bactericide, an antioxidant, an antiseptic, .
In another aspect, the present invention provides a pharmaceutical composition for preventing or treating inflammatory skin diseases, which comprises the compound of formula (1) or a fermented wheat germ or an extract thereof.
The term "inflammatory skin disease" in the present application collectively refers to a skin disease in which inflammation is the main lesion, and the inflammatory skin disease is seborrheic dermatitis, contact dermatitis, systemic lupus erythematosis, acne, eczema, But are not limited to, those selected from the group consisting of hypersplenic lupus, chronic simple hypoxic, cholestatic, deprived dermatitis and sunlight dermatitis.
The extracts of the compounds of formula 1, wheat germ, wheat germ fermented product and wheat germ fermented product are as described above.
The term "prophylactic" in this application refers to any act that inhibits or delays the onset of inflammatory skin disease by administration of the composition according to the present invention.
The term "treatment" in the present application means the administration of a pharmaceutical composition comprising the compound of formula 1, the wheat germ fermented product or the wheat germ fermented product of the present application to all the actions for improving or alleviating symptoms of inflammatory skin diseases do.
In the present application, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier.
The term "pharmaceutically acceptable carrier" as used herein refers to a carrier or diluent which does not irritate the organism and does not inhibit the prophylactic or therapeutic activity and properties of the pharmaceutical composition of the present application. Examples of the pharmaceutical carrier which is acceptable for the composition to be formulated into a liquid solution include sterilization and sterilization which are suitable for a living body and include saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol And one or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.
In addition, the pharmaceutically acceptable carrier of the present application may comprise a non-naturally occuring carrier.
The pharmaceutical compositions of the present application may be administered in single or multiple doses in a pharmaceutically effective amount.
The term "pharmaceutically effective amount" as used herein means an amount sufficient to prevent or treat a disease at a reasonable benefit / risk ratio applicable for medical prophylaxis or treatment, and the effective dose level will depend on the severity of the disease, Including, but not limited to, the activity, the patient's weight, health, sex, sensitivity of the patient to the drug, the time of administration of the presently used composition, the route of administration and the rate of release, Factors and factors well known in the medical arts.
In another aspect, the present application provides a method of preventing or treating inflammatory skin diseases comprising administering to a subject in need thereof a composition comprising a compound of formula (I) or a fermented wheat germ extract or extract thereof .
The compound of formula 1, the wheat germ fermented product, the extract of the wheat germ fermented product, the inflammatory skin disease, the prevention and the treatment are as described above.
In the present application, the suspected individual of the inflammatory skin disease refers to all animals including humans who have developed or can develop an inflammatory skin disease. By administering the pharmaceutical composition of the present application to a subject suspected of having an inflammatory skin disease, It can be treated efficiently.
The term "administering" as used herein means introducing the pharmaceutical composition of the present invention to an individual suspected of having an inflammatory skin disease by any appropriate method, and the composition of the present application may be administered by any conventional route ≪ / RTI > The route of administration of the composition of the present application is not particularly limited, but may be oral or parenteral administration. Specifically, it can be administered parenterally, and more specifically, it can be applied to the skin (i.e., transdermal administration). Specifically, administration of the present application can be carried out once to four times a day, twice to three times, or twice. The administration of the present application can also be carried out for a period of 4 weeks, 8 weeks, 4 to 12 weeks or 8 to 12 weeks.
In another aspect, the present application provides a quasi-drug composition for skin soothing comprising the compound of formula (I) or a fermented wheat germ or an extract thereof.
The compound of Formula 1, the wheat germ fermented product, the extract of the wheat germ fermented product, and the skin soothing are as described above.
The term "quasi-drug" as used in the present application means products that are less active than drugs, among the products used for diagnosing, treating, improving, alleviating, treating or preventing diseases of human or animal. For example, Quasi-drugs are products that are used for the purpose of treating or preventing diseases or diseases of humans or animals, products which are mild or have no direct action on the human body.
The quasi-drug composition of the present application can be manufactured in a form selected from the group consisting of body cleanser, foam, soap, mask, ointment, cream, lotion, essence and spray, but is not limited thereto.
When the compound of formula 1 or the wheat germ fermented product or the extract thereof of the present application is used as a quasi-drug additive, the extract of the compound of formula 1, the wheat germ fermented product or the wheat germ fermented product of the present application may be directly added or the other quasi-drug or quasi-drug ingredient Can be used together, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.
In another aspect, the present application provides a method of soothing the skin comprising administering to a subject in need thereof a composition comprising a compound of formula (I), or a wheat germ fermented product or extract thereof.
The compound of Formula 1, the wheat germ fermented product, the extract of the wheat germ fermented product, and the skin soothing are as described above.
2,6-DMBQ, or a composition comprising the wheat germ fermented product or the extract thereof has good cytostability and safety, has excellent anti-inflammatory effect, and also reduces transdermal water loss and skin redness, Can be used for prevention or treatment of diseases.
Fig. 1 shows the results of cytotoxicity test according to the concentration of wheat germ fermented product.
Fig. 2 shows the ability to inhibit NO production according to the concentration of wheat germ fermented product.
FIG. 3 shows the results of cell viability evaluation according to the concentration of 2,6-DMBQ.
4 shows the inhibitory effect of 2,6-DMBQ on NO production according to the concentration.
Fig. 5 shows the change in the transdermal water loss (TEWL) according to Experimental Example 4. Fig.
FIG. 6 shows the a-value change according to Experimental Example 4. FIG.
The present application will be described in detail by the following examples. However, the following examples are for illustrative purposes only, and the scope of the present application is not limited by the following examples.
Throughout the specification of the present application, the term "%" used to denote the concentration of a specific substance means that the solid / solid is (weight / weight)%, the solid / liquid is (weight / volume) , And liquid / liquid is (volume / volume)%.
Manufacturing example One: Wheat embryo Fermented Extract preparation
50 g of wheat embryo (CJ Cheiljedang) and 500 g of water were added to the flask, followed by thorough mixing and sterilization at 121 占 폚 for 15 minutes. Then, 5% (2.5 g) of dry yeast (Baker's yeast (Angelica japonica), Angel Yeast) was inoculated on the wheat germ, and fermented at 30 DEG C for 40 hours and then incubated at 8000 rpm for 20 minutes The supernatant was taken by centrifugation. The supernatant was lyophilized and recovered to prepare an extract of the wheat germ fermented product.
In this Example, the extract of the wheat germ fermented product was named 'CJ'.
Manufacturing example 2: Wheat embryo Fermented Analysis of components of extract
The content of 2,6-dimethoxybenzoquinone (2,6-dimethoxybenzoquinone) was analyzed by high performance liquid chromatography (HPLC). The content of the extract was higher than that of wheat germ It was 700 ppm based on the lyophilized product of the fermented product extract. The 2,6-DMBQ content was analyzed as follows.
The same amount of chloroform was added to 5 g of the sample, followed by stirring for a predetermined period of time, followed by centrifugation to recover the chloroform layer. This procedure was repeated 2-3 times, then the chloroform was evaporated using a vacuum concentrator, dissolved in chloroform and analyzed using HPLC. HPLC analysis was performed on Inertsil ODS3 (GL science) column and the mobile phase was analyzed with 25% MeOH and UV detector at 290 nm. At this time, 2,6-DMBQ reagent (Sigma) dissolved in chloroform was standardized.
The 2,6-DMBQ in the following experimental examples was purchased from Sigma-Aldrich.
Experimental Example One: Wheat embryo Fermented Cytotoxicity of Extracts ( MTT assay)
Experimental Example 1-1: Cell culture
Mouse macrophage RAW 264.7 cells were cultured in a 5% CO 2 incubator using a 100 mm cell culture dish using DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin Respectively. Cells that reached confluence were maintained by subculture using a scraper.
Experimental Example 1-2: Identification of cytotoxicity
The dehydrogenase present in the mitochondria in living cells produces a chromogenic substance called formazan in tetrazolium salt (WST). Therefore, we measured the number of living cells in the extract of the wheat germ fermented product and confirmed the toxicity.
RAW 264.7 cells at a concentration of 1 x 10 4 cells / well were cultured in 96-well cell culture plates for 24 hours in DMEM supplemented with 10% FBS and 1% penicillin / streptomycin. Then, the RAW 264.7 cells were cultured for 24 hours at 25, 50, 100, 200 and 400 / / ml of the extract of the wheat germ fermented product obtained in Preparation Example 1, and then 5 mg / ml MTT solution Was added to the wells and incubated in a 5% CO 2 incubator at 37 ° C for 48 hours. Then, the MTT reagent was removed from the culture solution, and 100 μl of DMSO was added to each well, and then formazan was dissolved. After measuring the absorbance through a 540 nm ELISA detector, the cell viability was determined by the following equation (1).
[Formula 1]
Cell viability = (absorbance of cells with added sample / absorbance of untreated cells) * 100
As a result, it was confirmed that the extract of the wheat germ fermented product showed no cytotoxicity up to the concentration of 400 / / ml (Fig. 1).
Experimental Example 2: Wheat embryo Fermented Identification of anti-inflammatory efficacy of extract
RAW 264.7 macrophages were cultured in DMEM medium containing 10% FBS at 37 ° C and 5% CO 2 . Then, RAW 264.7 macrophages were divided into 5 × 10 4 cells / well in a 96-well plate. After 24 hours, the extracts of the wheat germ fermented extract were cultured at 37 ° C. in a concentration of 25%, 100, 200 and 400 μg / 2 for 1 hour. Then, 1 μg / ml Lipopolysaccharide (LPS) was treated for 24 hours each to induce NO production to induce an inflammatory reaction.
The anti-inflammatory efficacy evaluation (NO assay) was measured using an NO detection kit (intron) based on the Griess reaction. Specifically, a 1: 1 mixture of 1% sulfanilamide and 0.1% N- (1-naphthyl) ethylenediamine dihydrochloride (N- (1-naphthyl) ethylenediamine dihydrochloride) The supernatant of the 96-well plate treated with the extract of the wheat germ fermentation product and the reagent were mixed at a ratio of 1: 1 and reacted at room temperature. Thereafter, NO production was measured at 540 nm through an ELISA reader. NO production was calculated by using a standard calibration curve using 1M NaNO 2 (Sodium Nitrite).
As a result, inhibition of NO production was observed in a concentration dependent manner. In particular, 45.9% inhibition activity of 200 g / ml of the extract of wheat germ fermentation product and 99.1% inhibition activity of 400. The IC 50 value, which is the concentration at which NO production is inhibited by 50%, is 215.5 ㎍ / ml, and is calculated by using the trend line of data about 50% in the inhibition results by concentration.
Experimental Example 3: 2 , 6- DMBQ's Identify cytotoxicity and anti-inflammatory efficacy
Experimental Example 3-1: Cell culture
The procedure of Experimental Example 1-1 was repeated.
Experimental Example 3-2: Cytotoxicity check
RAW 264.7 cells at a concentration of 1 × 10 4 cells / well were cultured in a 96-well cell culture plate for 24 hours at 37 ° C. and 5% CO 2 in DMEM culture medium supplemented with 10% FBS and 1% penicillin / streptomycin. Then 2,6-DMBQ (Sigma) in the concentrations (15 μg / ml, 30 μg / ml, 60 μg / ml and 120 μg / ml) 37 ℃, 5
As a result, 2,6-DMBQ did not show cytotoxicity up to a concentration of 120 μg / ml (FIG. 3).
Experimental Example 3-3: Anti-inflammatory efficacy confirmation
RAW 264.7 cells were plated on 60 mm plates at a density of 2 x 10 cells / dish and adhered to the plates for 24 hours. (15 μg / ml, 30 μg / ml, 60 μg / ml and 120 μg / ml) containing 1 μg / ml of LPS and the test substance, 2,6-DMBQ The medium was treated and cultured for 24 hours to obtain a culture solution. Thereafter, the amount of NO produced in each culture was measured in the same manner as in Experimental Example 2.
As a result, it was found that 2,6-DMBQ also had an ability to inhibit NO production in a concentration-dependent manner (FIG. 4).
Experimental Example 4: Wheat embryo Fermented Extract Skin soothing Check efficacy
A variety of methods have been used to measure the sedative effects of skin irritation reactions. The most sensitive and accurate method is to measure trans-Epidermal Water Loss (TEWL) and skin redness (a-value). Specifically, TEWL is known to continuously increase for 2 or 3 days after patch removal when 24 hour patches of 1 to 2% sodium lauryl sulfate (SLS) are applied. During this period, TEWL and a-values are known to be reduced in the increase of TEWL or decrease in TEWL and a-value when applying sedative external preparations. The skin soothing effect of the extract of the wheat germ fermented product was measured using the TEWL measurement as described below.
1) Day 0 (Day 0)
The subject's arm was cleaned using a standard detergent, and then the water was removed. Then, two portions were separated at a
Thereafter, 1% laurylsulfate was patched using an 8 mm finn chamber.
2) One day (Day-1)
After 24 hours of patch application, the patches were removed, washed and dried with water, and TEWL 1 and a 1 - values were measured at each site.
Thereafter, a test site and a control site were randomly assigned to two sites, and a patch containing 15 占 퐇 of 1% wheat germ fermented product was attached to the test site for 24 hours. The control site contained an extract of the wheat germ fermentation product Patches were attached.
3) Day 2 (Day 2)
TEWL 2 and a 2 - values were measured at the test site after the patch was removed, rinsed with water and dried, and the measurement was performed 5 times.
Thereafter, a patch containing 15 μl of 1% wheat germ fermented extract was attached to the test site for 24 hours, and no treatment was performed on the control site.
4) Three days (Day-3)
The patches were removed, washed with water and dried. TEWL 3 and a 3 - values were measured at the test site and the control site, and the measurement was performed 5 times.
Day-1
Day-1
*: p < 0.05, **: p < 0.01, ***: p < 0.001
The TEWL and a-values measured in each step are shown in Tables 1 and 2, respectively.
Table 1 shows statistical significance between test site and control site measurements before and after patch attachment (Day-0) with 1% SLS solution to confirm homogeneity between test site and control site The level of homology between the test site and the control site before the sample treatment can be confirmed.
Table 2 shows the effect on the TEWL and a-value by attaching the patch containing the extract of the wheat germ fermentation product to the test site where the TEWL and the a-value were increased by attaching the patch containing 1% SLS solution for 24 hours to 48 hours . As a result, the increased TEWL and a-value decreased to a statistically significant level (p <0.05) after 24 hours and 48 hours compared to the control site, confirming that the extract of the wheat germ fermented product had a skin soothing effect.
Prescription example : Wheat embryo Fermented Softening longevity containing extract Prescription example
Hereinafter, a cosmetic comprising the extract of the fermented product of 2,6-DMBQ or wheat germ according to the present application is prepared and presented based on the results of the above Experimental Example. Examples of the formulation of softening water in cosmetics containing wheat germ fermented product are as follows.
Prescription example 1: Lotion
Among the cosmetics containing the extract of the wheat germ fermented product, the prescription examples of the lotion are as follows.
Prescription example 2: essence
Examples of prescription of essence in cosmetics containing wheat germ fermented product are as follows.
Prescription example 3: Cream
Examples of the formulations of cream in cosmetics containing wheat germ fermented product are as follows.
From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present application is to be interpreted as being within the scope of the present application, all changes or modifications derived from the meaning and scope of the appended claims and from their equivalents rather than the detailed description.
Claims (5)
[Chemical Formula 1]
[Chemical Formula 1]
[Chemical Formula 1]
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20200102061A (en) | 2019-02-21 | 2020-08-31 | 주식회사 유진바이오텍 | Composition for improving acne and trichophytia contating wheat germ extract |
| KR20210050617A (en) * | 2019-10-28 | 2021-05-10 | 주식회사 유진바이오텍 | A composition for improving acne skin containing wheat germ extract and natural extract as active ingredients. |
| KR20210066497A (en) * | 2019-11-28 | 2021-06-07 | 주식회사 래디안 | Cosmetic composition for hair care with extraction of wheat germ |
| KR20210092350A (en) * | 2020-01-15 | 2021-07-26 | 주식회사 유진바이오텍 | A composition for improving skin having wheat germ |
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| CN110613664A (en) * | 2019-10-28 | 2019-12-27 | 海南女人春天美容有限公司 | Soothing massage cream and preparation method thereof |
| CN114276958B (en) * | 2021-12-20 | 2023-06-27 | 无锡弘焕微生态科技有限公司 | Preparation method and application of triple probiotics fermentation compound with anti-inflammatory effect |
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| JP2005015450A (en) * | 2003-06-30 | 2005-01-20 | Kanebo Cosmetics Inc | Skin cosmetic |
| JP2005330273A (en) * | 2004-04-21 | 2005-12-02 | Aol Corporation | Free radical-scavenging composition in living body |
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| JPH08337536A (en) * | 1995-06-14 | 1996-12-24 | Asahi Breweries Ltd | Agent for antiactive oxygen, antiactive oxygen agent, cosmetic, food and medicine containing the same as active ingredient |
| FR2834718B1 (en) * | 2002-01-15 | 2004-12-24 | Cognis France Sa | COSMETIC AND / OR PHARMACEUTICAL ACTIVE SUBSTANCES |
| HUP0202638A3 (en) * | 2002-08-09 | 2007-08-28 | Hidvegi Mate Dr | Use of fermented wheat-germ extract for preparation of antiphlogistic compositions |
| JPWO2005077349A1 (en) * | 2004-02-17 | 2007-10-18 | 大塚製薬株式会社 | Human β-defensin production promoter |
| US20150182442A1 (en) * | 2013-12-31 | 2015-07-02 | Shang Chen Mei Bang (Beijing) Biotechnology Co., Ltd. | Traditional chinese medicine composition that has the effect of soothing and cleaning face |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2005015450A (en) * | 2003-06-30 | 2005-01-20 | Kanebo Cosmetics Inc | Skin cosmetic |
| JP2005330273A (en) * | 2004-04-21 | 2005-12-02 | Aol Corporation | Free radical-scavenging composition in living body |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200102061A (en) | 2019-02-21 | 2020-08-31 | 주식회사 유진바이오텍 | Composition for improving acne and trichophytia contating wheat germ extract |
| KR20210050617A (en) * | 2019-10-28 | 2021-05-10 | 주식회사 유진바이오텍 | A composition for improving acne skin containing wheat germ extract and natural extract as active ingredients. |
| KR20210066497A (en) * | 2019-11-28 | 2021-06-07 | 주식회사 래디안 | Cosmetic composition for hair care with extraction of wheat germ |
| KR20210092350A (en) * | 2020-01-15 | 2021-07-26 | 주식회사 유진바이오텍 | A composition for improving skin having wheat germ |
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| KR101955111B1 (en) | 2019-03-07 |
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