KR102085052B1 - Composition for anti-inflammatory or moisturizing on skin comprising plant extract - Google Patents

Abstract

The present invention relates to a skin anti-inflammatory or skin moisturizing composition comprising, in one aspect, a feverfew extract, a nectar extract, and a cucumber extract as an active ingredient, the composition using an active ingredient derived from a natural plant. It shows an excellent skin anti-inflammatory effect in a state where the risk of side effects is low and safety is ensured, and the expression of skin moisturizing factor is markedly promoted, thereby showing an excellent skin moisturizing effect. The composition of the present invention can be used in cosmetics, external skin preparations, or hygiene products.

Description

Skin anti-inflammatory or skin moisturizing composition comprising a plant extract {COMPOSITION FOR ANTI-INFLAMMATORY OR MOISTURIZING ON SKIN COMPRISING PLANT EXTRACT}

The present invention relates to a skin anti-inflammatory or skin moisturizing composition comprising a plant extract, specifically feverfew, lamiaceae, and / or cucumber extract.

The skin functions to protect the body from external stimuli and maintains homeostasis of the human body by inhibiting evaporation of moisture from inside the skin. Since the skin is exposed to the outside, inflammation may be caused by external factors or stimuli, and in order to keep the skin healthy, the moisture of the skin should be kept constant. The skin may include the scalp and hair, and these organs may also be inflamed by external stimuli and there is a need for constant moisturization.

Inflammation is a phenomenon aimed at minimizing and restoring a response to a cause or stimulus that causes damage to tissue or cells. Factors that cause skin irritation include physical causes such as trauma, ultraviolet rays, burns, high temperature, and radiation, chemical factors, immunological factors caused by antibody responses, complex factors caused by environmental contaminants, and blood vessel or hormonal imbalances. .

Elimination of the cause of inflammation or anti-inflammatory action to reduce the biological response and symptoms can minimize or prevent this inflammation. Among the substances used to treat skin inflammation, nonsteroidal compounds include frupnamic acid, ibuprofen, benzidamine, and indomethacin, and arachidonic acid, an essential fatty acid, reacts with cyclooxygenase and lipoxygenase. To produce inflammatory cytotoxins (leukotriene, prostaglandin), which can block the biosynthetic inflammation that reacts with cyclooxygenase, but does not block the inflammation that occurs when it reacts with lipoxygenase. There is a problem that the effect is poor. In addition, as a steroid-based anti-inflammatory agent, prednisolone, dexamethasone, etc. are effective, but may cause various side effects in the human body. If steroid is applied to the skin for a long time, the skin at the site of application may have more hairs, shrink the skin, less skin pigment, bacterial infections, acne, thin skin, and blood vessels. There is a case. In severe cases, hormonal systemic symptoms may occur, and if the prescription (coating) is stopped, the symptoms become more severe, and thus the use of the steroid may not be stopped. In addition, conventional anti-inflammatory ingredients such as aspirin and indomethacin have been widely recognized for their effects, but side effects such as gastrointestinal disorders and antivascular vasodilation have been known. The scope and level of its use is limited due to legal issues such as stability issues and restrictions on use.

On the other hand, the moisturizing function is a basic property to keep the skin healthy, moisturizing should be properly maintained to properly exhibit the immune function and protective function of the skin itself. The most commonly used moisturizing ingredients are polyhydric alcohols or polysaccharides.However, since the hydroxyl group is hydrophilic, it acts as a mechanism to keep moisture from the outside by absorbing moisture from the outside when applied to the skin. There is a disadvantage, it also makes the use of sticky feeling, and if the product contains a large amount, it also affects the stability, and expensive polysaccharides reduce the price competitiveness.

Korean Patent Registration No. 10-0656287 Korea Patent Registration No. 10-1575722

The purpose of the present invention is to protect the skin and keep it healthy by exhibiting a skin moisturizing effect through promoting the expression of the skin anti-inflammatory action through the inhibition of inflammatory mediators and promoting the expression of water supply factors without causing any side effects. It is providing the composition for anti-inflammatory or skin moisturizing.

In order to achieve the above object, the present invention provides a skin anti-inflammatory or skin moisturizing composition comprising at least one of feverfew extract, nectar extract, and cucumber extract as an active ingredient.

In addition, in one aspect of the present invention, the skin anti-inflammatory or skin moisturizing composition may be used in cosmetics, external preparations for skin, or hygiene products.

Skin anti-inflammatory composition according to an aspect of the present invention, using an active ingredient derived from a natural plant exhibits a remarkably excellent skin anti-inflammatory effect in a state where the risk of side effects and safety is guaranteed as compared to conventional inflammatory therapeutics.

In addition, the composition for moisturizing the skin according to an aspect of the present invention significantly promotes the expression of factors related to skin moisturizing, provides an excellent and lasting moisturizing effect, and ensures stability when using the product and produces a product having excellent price competitiveness. Indicates.

Figure 1 is a graph showing the degree of inhibition of expression for TNF-α (tissue necrosis factor) of the embodiment according to an aspect of the present invention in the inflammatory response induced by UVB (UVB, Ultraviolet B).
Figure 2 is a graph showing the degree of inhibition of expression for NF-κB (cell nuclear transcription factor) of the embodiment according to an aspect of the present invention in the inflammatory response induced by UVB.
Figure 3 is a graph showing the degree of inhibition of expression for TNF-α of the embodiment according to an aspect of the present invention in the inflammatory response induced by hydrogen peroxide.
Figure 4 is a graph showing the degree of inhibition of expression for NF-κB of the embodiment according to an aspect of the present invention in the inflammatory response induced by hydrogen peroxide solution.
Figure 5 is a graph showing the expression promoting degree for hyaluronan synthase-3 (HAS-3), which is a skin moisturizing factor of an embodiment of the present invention.
Figure 6 is a graph showing the expression promoting degree for the skin moisturizing factor aquaporin of the embodiment according to an aspect of the present invention.

In one aspect of the invention, the "skin" is an organ covering the exterior of the organism, consisting of the epidermis, dermis and subcutaneous fat layer from the outside of the organ and covering the scalp and hair as well as tissue covering the outside of the face or the entire body. It is the broadest concept to include.

In one aspect of the invention, an “extract” includes all materials obtained from natural products and extracts materials obtained from natural products, regardless of the method of carrying out the extraction, the extraction solvent, the extracted components or the form of the extract. It is a broad concept that includes all materials that can be obtained after processing or processing in other ways.

In one aspect of the invention, "fiberfew extract," "honey extract," "cucumber extract" is to include all the materials obtained by performing the extraction of each plant as a raw material, for example the process of performing the extraction It is possible to treat plants with heat, acids, bases, enzymes, and the like, and this is a broad concept that includes all materials obtained by extracting and then processing the materials obtained from plants.

In one aspect of the present invention, “ Tanacetum parthenium ,” “ Prunella vulgaris ,” and “ Sanguisorba officinalis ” are the young plants, leaves, flowers, stems, bark, fruit, At least one selected from the group consisting of seed, root and combinations thereof.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention may be directed to a skin anti-inflammatory composition comprising one or more of feverfew extract, nectar extract, and cucumber extract as an active ingredient.

In one aspect, the present invention may be directed to a skin moisturizing composition comprising at least one of feverfew extract, nectar extract, and cucumber extract as an active ingredient.

Feverfew ( Tanacetum parthenium ) is a perennial herb that is distributed from South and Eastern Europe to Asia and is also called summer white or feverfew. The generic name Tanacetum comes from the Greek 'athanasia' and is called 'feverfew' because of its heat-reducing effect. The components of feverfew discovered to date include beta-carotene, parthenolide, calcium, iron, magnesium, manganese, phosphorus, potassium, selenium, zinc, vitamins B1, B2, B3, C , Sesquiterpene lactone, volatile oils, tannins, monoterpins and the like.

In one aspect of the invention, the feverfew extract may be one or more extracts selected from the group consisting of feverfew herb leaves, flowers, stems, roots, and combinations thereof.

Prunella vulgaris ( Prunella vulgaris ) is a perennial herb, also called hyacinth and honey bat, widely distributed in Korea, China, and Japan. As a wheat plant, it used to pick flowers and suck honey. The entire herbaceous herb contains ursolic acid, caffeic acid, rosemary acid, carotenoids, tannins, organic acids, and the like, and saponins, triterpenes, flavonoids, sterols, and fatty acids are constituents of flowers.

In one aspect of the invention, the nectar extract may be one or more extracts selected from the group consisting of young shoots, leaves, flowers, stems, and combinations thereof of nectar herb.

Cucumber ( Sanguisorba officinalis ) is a perennial herb belonging to the family Rosaceae, and in oriental medicine, cucumber roots are called fat milk. Cucumber roots contain about 17% of tannins and saponins, as well as gallic acid, ursolic acid, ellagic acid, quercetin glycosides, kaempferol glycosides, and leucoanthocyanidins. (leucoanthocyanidin) and the like are known.

In one aspect of the invention, cucumber extract may be root extract of cucumber herbal.

In one aspect of the invention, the total content of the extract relative to the total volume of the composition is 0.0001 to 20% by volume, 0.001 to 15% by volume, 0.05 to 10% by volume, 0.1 to 5% by volume, 0.5 to 2% by volume, 0.8 to It may be 1.2% by volume. In addition, the total content of the extract relative to the total volume of the composition may be, for example, 1.0% by volume. If the total content of the extract is too small, it may not exhibit the skin anti-inflammatory or skin moisturizing effect to achieve the object of the present invention, and if excessively excessive, it may affect the formulation used.

In one aspect of the invention, the extract is water; Organic solvents; And it may be extracted with one or more selected from the group consisting of a mixture thereof.

In one aspect of the invention, the water comprises distilled or purified water, the organic solvent is C1 ~ C6 lower alcohol, acetone, ether, propanediol, ethyl acetate, diethyl ether, ethyl methyl ketone, chloroform, hexane, methylene One or more selected from the group consisting of chloride, ethyl acetate, butylene glycol, hexanediol and propylene glycol, but is not limited thereto.

In one aspect of the invention, the extract may be extracted with a mixed solvent of purified water, 1,3-butylene glycol and 1,2-hexanediol. For example, the mixed solvent includes 60 to 75 mass% of purified water, 24 to 35 mass% of 1,3-butylene glycol and 1 to 5 mass% of 1,2-hexanediol based on the total mass of the mixed solvent. It may be. In addition, the content of the purified water contained in the mixed solvent may be 60 to 75 mass%, 63 to 73 mass%, or 65 to 70 mass% with respect to the total mass of the mixed solvent; The content of 1,3-butylene glycol may be 20 to 40 mass%, 25 to 35 mass%, or 28 to 32 mass% with respect to the total mass of the mixed solvent; The content of 1,2-hexanediol may be 0.1 to 7% by mass, 1 to 5% by mass, or 2 to 4% by mass relative to the total mass of the mixed solvent, but is not limited thereto.

In one aspect of the present invention, the extract can be extracted using, for example, hot water extraction, heat extraction, cold extraction, reflux extraction, reflux cooling extraction, or ultrasonic extraction and the like, which will be apparent to those skilled in the art. Is not limited. Extraction of the extract may be carried out at room temperature, but for efficient extraction may be extracted at the conditions of about 40 to 120 ℃, 80 to 110 ℃, or 90 to 100 ℃, specifically at a heating condition such as a temperature of about 95 ℃ Can be extracted, but is not limited thereto. Extraction time may be performed for about 1 to 4 hours, specifically about 1.5 to 2.5 hours, but is not limited thereto. The extraction time may vary depending on conditions such as extraction solvent and extraction temperature. For example, when the extraction solvent is water, it may be carried out at 90 to 100 ℃, 65 to 70 ℃ if ethanol. The extraction may be extracted one or more times in order to increase the extraction yield of the active ingredient, for example, may be used by extracting the combined successive extraction 1 to 5 times.

Inflammatory reactions in the skin include physiological phenomena such as vasodilation, changes in blood flow, and infiltration of leukocytes in tissues, resulting in an increase in vascular permeability, as well as symptoms of redness, fever, swelling, and pain. And macrophages, etc. are concentrated to the site of inflammation may cause erythema. Various biochemical phenomena are involved in this inflammatory response. Nitric oxide synthase (NOS) induced by inflammatory cytokines such as TNF-α is known as an important mediator of the inflammatory response.

The composition according to one aspect of the present invention inhibits the secretion of TNF-α, a cytokine involved in the inflammatory response, and inhibits the expression of NF-κB, thereby exhibiting an anti-inflammatory activity.

On the other hand, the moisture of healthy human skin supports the elasticity and flexibility of the skin, and the proper moisture content can be maintained to exhibit the protective function and immune function of the skin itself. If the moisture in the stratum corneum is less than 10%, the skin is known to dry out and cause skin trouble. Aquaporin is a membrane protein that regulates the entry and exit of water molecules in cell membranes and increases water retention in the skin. In addition, along with proteins such as collagen and elastin, hyaluronic acid, a major component of the extracellular matrix, is a polysaccharide glycosaminoglycan (GAG) that functions as a moisture adsorbent that allows the skin to contain sufficient moisture. To perform. Increasing the amount of HAS-3, an enzyme that synthesizes hyaluronic acid, promotes the synthesis of hyaluronic acid, thereby improving skin moisture retention and moisturizing retention.

The composition according to one aspect of the present invention promotes the synthesis of aquaporin and increases the HAS-3, thereby improving the moisture retention of the skin and exhibiting a moisturizing action of the skin, thereby exhibiting the effect of protecting and maintaining the skin.

In one aspect of the present invention, the composition may include a combination of two or more of feverfew extract, nectar extract and cucumber extract as an active ingredient. The composition according to one aspect of the present invention exhibits a remarkably superior skin anti-inflammatory effect and skin moisturizing effect compared to the case comprising each of them by a combination of two or more of feverfew extract, lamiaceae extract and cucumber extract.

In one aspect of the present invention, the volume ratio of feverfew extract: Lamiaceae extract: Cucumber extract in the composition may be an integer ratio of 1 ~ 9: 1 ~ 9: 1 ~ 9. Specifically, the volume ratio of Feverfew extract: Lavender extract: Cucumber extract in the composition is 1-5: 1-5: 1-5, 1-3: 1-3: 1-3, 1-2.7: 1-2.7: It can be 1 to 2.7, 1 to 2.3: 1 to 2.3: 1 to 2.3, or 1 to 2: 1 to 2: 1 to 2. In addition, the volume ratio of the Feverfew extract: Lavender extract: Cucumber extract in the composition may be about 2: 1: 2, for example.

In one aspect of the present invention, the composition is a conventional and known components that can be obviously applied to those skilled in the art within the range that does not impair the effects of the present invention, antioxidants, preservatives, absorbents, exfoliants, thickeners, It may further comprise one or more of a sunscreen, emulsifiers, surfactants, fragrances, deodorants, stabilizers, skin protectants, and pH adjusting agents.

In one aspect of the invention, the antioxidant is tocopheryl acetate, tocopherol, dibutylhydroxytoluene (BHT), acetyl cysteine, ascorbic acid polypeptide, erythorbic acid, diamyl within the range that does not impair the effects of the present invention At least one selected from the group consisting of hydroquinone and ascorbyl palmitate. Antioxidants are conventionally used content may be included, for example, but not limited to 0.01 to 2% by weight relative to the total weight of the composition.

In one aspect of the invention, the preservative is composed of methylparaben, propylparaben, phenoxyethanol, potassium sorbate, dehydroacetic acid, pentanediol and sorbic acid within the scope of not impairing the effects of the present invention. It may be one or more selected from the group. Preservatives are conventionally used content may be included, for example, but not limited to 0.01 to 2% by weight relative to the total weight of the composition.

In one aspect of the invention, the absorbent may be one or more selected from the group consisting of aluminum starch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrin, talc, and zeolite. Absorbents are conventionally used, for example, but may be included, but is not limited to, 0.01 to 2% by weight relative to the total weight of the composition.

In one aspect of the invention, the exfoliating agent may be one or more selected from the group consisting of alpha-hydroxy acid, beta-hydroxy acid, lactic acid, glycolic acid, salicylic acid and salts thereof. Exfoliants are conventionally used amounts, for example, may be included in 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the thickeners are carboxylic acid polymers (eg acrylic acid), paraffins, crosslinked polyacrylate polymers, polyacrylamide polymers (eg polyacrylamides, or isoparaffins), poly At least one selected from the group consisting of saccharides (eg cellulose), and gums (eg, agar, algin, ammonium alginate, or gelatin). Thickeners are conventionally used amounts, for example, may be included in 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the sunscreen preparation is para-aminobenzoic acid (PABA), glyceryl PABA, octyldimethyl PABA, butyl PABA, cinnamate, anthranilate, salicylate, kaolin, talc, petrolatum, and metals It may be at least one selected from the group consisting of oxides. Sunscreen preparations are conventionally used content may be included, for example, 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the emulsifier is beeswax, cetearyl glucoside, carboxyvinyl polymer, polysorbate 20, polysorbate 60, dodecylbenzenesulfonate, sorbitan within the range that does not impair the effects of the present invention At least one selected from the group consisting of fatty acid esters, glycerin monostearate, stearic acid, palmitic acid, stearyl alcohol, behenyl alcohol, and lecithin. Such emulsifiers may be included in 0.01 to 4% by weight based on the total weight of the composition, but is not limited thereto.

In one aspect of the present invention, the surfactant is sorbitan sesquioleate, disodium coco ampoacetate, sodium coco ampoacetate, fiji-12 nonylphenyl ether, and sebum within a range that does not impair the effects of the present invention. At least one selected from the group consisting of -120 methylglucose dioleate, and potassium cocoylglycinate. The surfactant may be included in 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the fragrance may be one or more selected from the group consisting of herbs, ylang-ylang essential oil, orange essential oil, cananga essential oil, cinnamon essential oil, chamomile flower extract, lemon glass essential oil, lemon essential oil, and rosemary essential oil. Perfume may be included in the conventionally used content, for example, 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the deodorant may be one or more selected from the group consisting of potassium permanganate, bleach powder, cypress extract, bamboo extract, green tea extract, pine needle extract, and lemon juice. Deodorant is a conventionally used content and may include, for example, 0.01 to 2% by weight relative to the total weight of the composition, but is not limited thereto.

In one aspect of the invention, the stabilizer may be one or more selected from the group consisting of sodium sulfite, ascorbic acid, carotenoids, retinol, thioglycerol, and cysteine. Stabilizers are conventionally used content may be included, for example, but not limited to 0.01 to 2% by weight relative to the total weight of the composition.

 In one aspect of the invention, the skin protectant is caprylic / capric triglyceride, cetearyl alcohol, squalane, allantoin, betaglucan, hyaluronic acid extract, rosemary extract, tocopheryl acetate, colostrum, allantoin, and At least one selected from the group consisting of D-panthenol. Skin protectants can be included, but are not limited to, amounts typically used, for example, from 0.01 to 2% by weight relative to the total weight of the composition.

In one aspect of the invention, the pH adjusting agent is citric acid, sodium citrate, malic acid, sodium malate, fmaric acid, sodium pmarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate. At least one selected from the group consisting of triethanolamine and calcium carbonate. pH adjusters are conventionally used content, for example, may be included, but is not limited to 0.01 to 2% by weight relative to the total weight of the composition.

In one aspect of the invention, the composition can be used in cosmetics, external preparations for skin, or hygiene products.

In one aspect of the invention, the cosmetic is a cosmetic, such as a lotion, essence, sun cream, sun lotion, creams; Makeup cosmetics such as powdered powder, solid white powder, cream white powder, foundation, lip balm, eyebrow pencil, unflavored powder, etc .; Hair care cosmetics such as hair lotions, hair tonics, liquid and oil-based hair dressings, hair creams, hair treatment creams, shampoos, hair sprays, and the like; Facial care cosmetics such as packs; Or body care cosmetics such as soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

In one aspect of the invention, the external preparation for the skin may be any one selected from the group consisting of ointments, lotions, sprays, patches, creams, powders, suspensions, and gels.

In one aspect of the invention, the hygiene article may be any one selected from the group consisting of wet wipes, tissues, diapers, sanitary napkins, sanitary wipes, wet wipes, cleaning wipes, facial wipes, skin care wipes, and antiperspirant wipes.

Hereinafter, the configuration and effects of the present specification will be described in more detail with reference to Examples and Test Examples. However, these examples and test examples are provided only for the purpose of illustration to help the understanding of the present specification, the scope and scope of the present specification is not limited by the following examples.

[Examples 1-4, a control example, and a comparative example]

(1) A mixture consisting of leaves, flowers, stems, and roots of Feverfew Herb was extracted in a weight ratio of 1:20 (67 mass% of purified water, 30 mass% of 1,3-butylene glycol, and 1,2-hexane). 3% by weight of diol) was extracted for 2 hours at 95 ℃ to remove the residual solvent to prepare a feverfew extract.

(2) Instead of the feverfew, a mixture consisting of young shoots, leaves, flowers, and stems of Lamiaceae herb was extracted by the same method, and Lamiaceae extract was prepared by extracting the roots of Cucumber herb by the same method. .

(3) Each of the extracts thus prepared was diluted to 10% by volume with purified water, and then diluted again with the following media to the contents of Table 1 to prepare the compositions of Examples 1 to 4.

Medium: RPMI Medium 1640 (Fetal Bovine Serum), 100 IU / mL penicillin and 100 μg / mL streptomycin added (Test Example 1);

Fibroblast line medium with low serum growth supplement (add 50X low serum growth component to 1X medium) and 100 units / mL streptomycin (Test Example 2)

(4) In the case of the control means that no evaluation sample (active ingredient) was added, and Comparative Examples 1 and 2 were prepared by diluting dexamethasone and hyaluronic acid with the above-described media in the amounts shown in Table 1, respectively.

Item Active ingredient Content (% by volume) Control none - Example 1 Feverfew Extract One% Example 2 Lamiaceae Extract One% Example 3 Cucumber Extract One% Example 4 Mixture of Feverfew Extract + Lamiaceae Extract + Cucumber Extract (Mixing ratio by volume 2: 1: 2) One% Comparative Example 1 Dexamethasone (Sigma, product no .: D8893) 0.065% Comparative Example 2 Hyaluronic acid 0.005%

Test Example 1 Evaluation of Skin Anti-inflammatory Effect of Plant Extracts

(1) After dispensing RAW264.7 cells (obtained from Korea Cell Line Bank) into 6-well plates at a concentration of 5 × 10 ⁵ cells / well, 10 wells of fetal bovine serum (GIBCO, Cat. No. 12483-020), RPMI Medium 1640 (GIBCO, Catalog No. 61870) with 100 IU / mL penicillin (SIGMA, Cat.No. P3032) and 100 μg / mL streptomycin (SIGMA, Cat.No. S9137). Medium was added and cultured for 2 days at 37 ° C. under 5% CO ₂ (the same conditions for cell culture). The cultured RAW264.7 cells were washed with PBS and treated with each well of Examples 1 to 4, Comparative Example 1 and Control Example and incubated for 3 days at 37 ℃, 5% CO ₂ .

(2) Each cell was treated with UVB or hydrogen peroxide as follows to induce cell damage.

-Cell damage by UVB treatment

Remove the medium of the cells cultured in (1), wash the cells with PBS and induce cell damage by irradiating UVB of 40 mJ / cm ² for 2 hours in UVP Crosslinkers (UVP, United Kingdom) After replacing the medium and incubated for 1 day.

-Cell damage by hydrogen peroxide

After removing the culture medium of the cells cultured in (1) and washing the cells with PBS, the cells were treated with hydrogen peroxide (DAEJUNG, Korea) at a concentration of 500 μM and cultured for 1 day.

(3) In order to analyze anti-inflammatory genes from the cultured cells, RNA was extracted and quantified in the following manner and cDNA was synthesized to perform RT-PCR (Reverse transcription-polymerase chain reaction).

① Remove the medium of the cells incubated with the sample in (2) and washed the cells with 1X PBS.

② 1X PBS was removed, and the triazole reagent (Invitrogen, USA) was added to each well of the cells incubated in the plate evenly to the cells and allowed to stand at room temperature for 3-5 minutes.

③ Carefully scrape the cells in each well with a scraper, collect in microtubes, add 40 μL of chloroform and mix.

④ After standing for 2 to 3 minutes at room temperature, centrifuged for 15 minutes at 12,000 rpm, 4 ℃.

⑤ After centrifugation, the transparent supernatant was transferred to a new tube, 100 μL of isopropyl alcohol was added and carefully inverted and left at room temperature for 10 minutes, followed by centrifugation at 12,000 rpm and 4 ° C. for 10 minutes.

⑥ After discarding the supernatant and washing the pellet with 200 μL of 75 vol% ethanol diluted with purified water treated with DEPC (diethyl pyrocarbonate), the supernatant was discarded and the RNA pellet was dried at room temperature.

⑦ Put 50 μL of DEPC treated purified water into the dried RNA pellet and suspend it and heat it for 10 minutes at 55 ~ 60 ℃.

⑧ RNA was then quantified by absorbance at 260 nm.

Formula for quantitative value: quantitative value = OD value * dilution factor * 40 μg / mL

(OD260 = 1 = 40 μg / mL)

(4) cDNA synthesis was performed using PrimeScript 1st strand cDNA Synthesis Kit (TaKaRa, Cat. No. 6110A) and the experiment was performed by the method of the kit. PCR was performed according to the method of the kit using a Taq polymerase Kit (TaKaRa, Cat.No R001A) and using a PCR equipment (Biofree, Cat.No TC-96 / G / H / (b) B) The gene was amplified with the primer of (all primers used in the following experiment was custom-made in Daejeon, Korea). The product produced by PCR was confirmed by electrophoresis on 1% agarose gel and analyzed by an image analyzer (KOREALABTECH, Bundang, Korea).

primer order Where to get TNF-α Forward: 5'-CAG AGG GAA GAG TTC CCC AG-3 '(SEQ ID NO: 1)
Reverse: 5'-CCT TGG TCT GGT AGG AGA CG-3 '(SEQ ID NO: 2) Bioneer NF-κB Forward: 5'- TCT CAG CAA TGT CAA CGA C-3 '(SEQ ID NO: 3)
Reverse: 5'- TTT ATG CCT ACA GCC TCC T-3 '(SEQ ID NO: 4) Bioneer

The results of expression levels of TNF-α and NF-κB after the treatment of Examples 1 to 4, Comparative Example 1 and the control examples are shown in FIGS. 1 to 4 and Tables 3 to 6, respectively. The expression rate of each gene was calculated using the following equation.

[Equation]

% Expression of each gene = (quantity of gene expression / control gene expression in sample treatment) X 100

Table 3 shows the results of expression of TNF-α in UVB treatment, Table 4 shows the results of expression of NF-κB in UVB treatment, Table 5 shows the results of expression of TNF-α in treatment with hydrogen peroxide and Table 6 shows the number of hydrogen peroxide The result of expression level of NF-κB upon treatment. 1 to 4 correspond to graphs of the results of Tables 3 to 6, respectively.

sample Content (% by volume) UVB Expression level of TNF-α (%) Standard Deviation Control 0 + 100.00 0.00 Example 1 One + 73.85 1.64 Example 2 One + 73.82 0.73 Example 3 One + 74.37 0.58 Example 4 One + 59.69 2.69 Comparative Example 1 0.065 + 50.80 4.92

sample Content (% by volume) UVB Expression level of NF-κB (%) Standard Deviation Control 0 + 100.00 0.00 Example 1 One + 69.95 0.93 Example 2 One + 71.19 0.10 Example 3 One + 71.45 2.01 Example 4 One + 51.25 0.27 Comparative Example 1 0.065 + 54.84 1.29

sample Content (% by volume) H ₂ O ₂ Expression level of TNF-α (%) Standard Deviation Control 0 + 100.00 0.00 Example 1 One + 70.80 0.96 Example 2 One + 71.22 1.83 Example 3 One + 71.44 1.48 Example 4 One + 46.36 1.09 Comparative Example 1 0.065 + 42.14 0.77

sample Content (% by volume) H ₂ O ₂ Expression level of NF-κB (%) Standard Deviation Control 0 + 100.00 0.00 Example 1 One + 72.43 1.02 Example 2 One + 66.10 1.98 Example 3 One + 55.88 2.83 Example 4 One + 47.89 0.41 Comparative Example 1 0.065 + 67.17 7.82

1 to 4, and Tables 3 to 6, when treated with the composition of Examples 1 to 4 according to one aspect of the present invention, the amount of expression of anti-inflammatory factors induced by damage caused by UVB or hydrogen peroxide solution of Examples 1 to 4 It can be seen that the decrease by 4.

Specifically, according to Table 3 and Figure 1 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) is about 25 to 26% TNF in the inflammatory response induced by UVB -α expression inhibitory effect, in particular, a combination of all of these compositions (Example 4) showed an expression inhibitory effect of about 40%, significantly inhibiting the expression of TNF-α compared to each of the extracts. The effect of Example 4 is similar to the inhibitory effect of the expression of dexamethasone of Comparative Example 1, especially considering that it is a natural component, the effect of Example 4 is very excellent.

According to Table 4 and Figure 2 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) is about 28 to 30% NF-κB in the inflammatory response induced by UVB In particular, the combination of these compositions (Example 4) exhibited an expression inhibitory effect of about 49%, which significantly inhibited the expression of NF-κB relative to each of the extracts. The effect of Example 4 is higher than the inhibitory effect of the expression of dexamethasone, Comparative Example 1, especially considering that it is a natural component, the effect of Example 4 is very excellent.

According to Table 5 and Figure 3 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) is about 28 to 29% TNF- in the inflammatory response induced by hydrogen peroxide α expression inhibitory effect, in particular, a combination of all of these compositions (Example 4) showed an expression inhibitory effect of about 53%, significantly inhibiting the expression of TNF-α compared to each of the extracts. The effect of Example 4 is similar to the inhibitory effect of the expression of dexamethasone of Comparative Example 1, especially considering that it is a natural component, the effect of Example 4 is very excellent.

According to Table 6 and Figure 4 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) is about NF- of about 27 to 44% in the inflammatory response induced by hydrogen peroxide κB expression inhibitory effect was shown, and in particular, the composition combining all of them (Example 4) showed an expression inhibitory effect of about 52%, significantly inhibiting the expression of NF-κB relative to each of the extracts. The effect of Example 4 is higher than the inhibitory effect of the expression of dexamethasone, Comparative Example 1, especially considering that it is a natural component, the effect of Example 4 is very excellent.

Through the above results it can be confirmed that the composition according to one aspect of the present invention shows a significant skin anti-inflammatory effect.

Test Example 2 Evaluation of Skin Moisturizing Effect of Plant Extract

(1) After dispensing HDFa cells (ThermoFisher SCIENTIFIC), which is a human dermal fibroblast, into each well at a concentration of 5 × 10 ⁵ cells / well in a 6-well plate, low serum growth components (LSGS, Gibco) BRL, Grand Island, NY, USA) (adding 50X of low serum growth to the medium to 1X) and 100 units / mL of streptomycin (Sigma-Aldrich Co., St. Louis, MO, USA) Fibroblast line basal medium (Medium 106, Gibco, product number: M-106-500) was added to each well and incubated at 37 ° C., 5% CO _{2 for} 2 days.

(2) The cultured HDFa was washed with PBS and treated with Examples 1 to 4, Comparative Example 2 and Control with medium replacement and incubated under 37 ° C., 5% CO _{2 for} 3 days.

(3) According to the method of Test Example 1 (3) and (4) in the cultured cells as described above, using the primers of Table 7 below to measure the expression level of HAS-3 (Hyalruronan synthase 3) and aquaporin It was.

primer order Where to get HAS-3 Forward: 5'- GGA AAG CTT GGC ATG TAC CGC AAC AG-3 '(SEQ ID NO: 5)
Reverse: 5'- AGA GGA GGG AGT AGA GGG AC-3 '(SEQ ID NO: 6) Bioneer Aquaporin Forward: 5'- ACC CTC ATC CTG GTG ATG TTT G-3 '(SEQ ID NO: 7)
Reverse: 5'- TCT GCT CCT TGT GCT TCA CAT-3 '(SEQ ID NO: 8) Bioneer

And the expression rate of each gene calculated using the above equation is shown in Figure 5, Figure 6, Table 8, Table 9, respectively. Table 8 shows the results of gene expression of HAS-3, and Table 9 shows the results of gene expression of aquaporin. 5 and 7 graphically show the results of Tables 8 and 9, respectively.

sample Content (% by volume) Expression level of HAS-3 (%) Standard Deviation Control 0 100.00 0.00 Example 1 One 111.72 3.15 Example 2 One 117.21 1.45 Example 3 One 128.63 0.31 Example 4 One 133.85 3.14 Comparative Example 2 0.005 150.31 3.15

sample Content (% by volume) Expression level of aquaporin (%) Standard Deviation Control 0 100.00 0.00 Example 1 One 112.01 3.53 Example 2 One 128.65 3.46 Example 3 One 123.43 0.40 Example 4 One 150.91 5.05 Comparative Example 2 0.005 147.87 2.27

According to the results of Table 8, Table 9, 5 and 6 it can be seen that when treated with the composition of Examples 1 to 4 according to one aspect of the present invention promotes the expression of factors related to moisturizing.

Specifically, according to Table 8 and Figure 5 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) the effect of promoting the expression of HAS-3 gene about 12 to 28% In particular, a combination of all of these compositions (Example 4) showed a gene expression promoting effect of about 34%, showing an effect of significantly promoting the expression of the HAS-3 gene compared to each of the extracts.

In addition, according to Table 9 and Figure 6 Feverfew extract (Example 1), Lamiaceae extract (Example 2) and Cucumber extract (Example 3) has an effect of promoting the expression of aquaporin gene about 12 to 28% In particular, the combination of all of these compositions (Example 4) exhibited a gene expression promoting effect of about 51%, showing an effect of significantly promoting the expression of the aquaporin gene compared to each of the extracts. The result of Example 4 is higher than that of the hyaluronic acid of Comparative Example 2, especially considering that it is a natural component, the effect of Example 4 is very excellent.

Through these results it can be confirmed that the composition according to one aspect of the present invention shows a remarkably excellent moisturizing effect.

Therefore, according to the results of Test Examples 1 and 2 it can be seen that the composition according to one aspect of the present invention has a remarkably excellent skin anti-inflammatory and skin moisturizing effect, thereby protecting and maintaining the skin healthy.

Examples of the formulation of the composition according to an aspect of the present invention will be described below, but can be applied to various other formulations, which are intended to explain in detail only, not intended to limit the present specification.

Formulation Example 1 Flexible Cosmetic (Skin Lotion)

According to the composition described in Table 10 below to prepare a flexible longevity in a conventional manner.

Compounding ingredients Content (% by volume) One or more of feverfew extract, nectar extract and cucumber extract 1.0 Carboxy Vinyl Polymer 0.1 Fiji-12 nonylphenyl ether 0.2 Polysorbate 60 0.4 Triethanolamine 0.1 Preservatives, Spices Quantity Purified water Remaining amount

Formulation Example 2 Nutrients (Milk Lotion)

Nutrients were prepared in a conventional manner according to the composition shown in Table 11.

Compounding ingredients Content (% by volume) One or more of feverfew extract, nectar extract and cucumber extract 1.0 Carboxy Vinyl Polymer 0.1 Beeswax 4.0 Polysorbate 60 1.5 Caprylic / Capric Triglycerides 2.0 Squalane 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 1.0 Triethanolamine 0.2 Preservatives, spices Quantity Purified water Remaining amount

Formulation Example 3 Nutrition Cream

Nutritional cream was prepared in a conventional manner according to the composition shown in Table 12.

Compounding ingredients Content (% by volume) One or more of feverfew extract, nectar extract and cucumber extract 1.0 Beta Glucan 2.0 Caprylic / Capric Triglycerides 2.0 Squalane 2.0 Cetearyl Glucoside 1.5 Polysorbate 60 1.2 Triethanolamine 0.1 Preservatives, spices Quantity Purified water Remaining amount

Formulation Example 4 Massage Cream

To prepare a massage cream in a conventional manner according to the composition described in Table 13.

Compounding ingredients Content (% by volume) One or more of feverfew extract, nectar extract and cucumber extract 1.0 Beta Glucan 2.0 Caprylic / Capric Triglycerides 2.0 Beeswax 4.0 Cetearyl Glucoside 1.5 Sorbitan sesquioleate 0.9 paraffin 2.0 Preservatives, spices Quantity Purified water Remaining amount

[Formulation Example 5] Pack

To prepare a pack in a conventional manner according to the composition described in Table 14.

Compounding ingredients Content (% by volume) One or more of feverfew extract, nectar extract and cucumber extract 1.0 Hyaluronic acid extract 2.0 Beta Glucan 2.0 Allantoin 0.1 Polysorbate 60 1.2 Preservatives, spices Quantity Purified water Remaining amount

As described above in detail a specific part of the present specification, it is apparent to those skilled in the art that the specific description is merely a preferred embodiment, and the scope of the present specification is not limited thereto. Therefore, the substantial scope of the present specification will be defined by the appended claims and equivalents thereof.

<110> Yuhan-Kimberly, Limited <120> COMPOSITION FOR ANTI-INFLAMMATORY OR MOISTURIZING ON SKIN COMPRISING          PLANT EXTARCT <130> KC161016 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of TNF-alpha <400> 1 cagagggaag agttccccag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of TNF-alpha <400> 2 ccttggtctg gtaggagacg 20 <210> 3 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of NF-kappa B <400> 3 tctcagcaat gtcaacgac 19 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of NF-kappa B <400> 4 tttatgccta cagcctcct 19 <210> 5 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of HAS-3 <400> 5 ggaaagcttg gcatgtaccg caacag 26 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of HAS-3 <400> 6 agaggaggga gtagagggac 20 <210> 7 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of Aquaporin <400> 7 accctcatcc tggtgatgtt tg 22 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of Aquaporin <400> 8 tctgctcctt gtgcttcaca t 21

Claims (11)

A composition for skin anti-inflammatory or skin moisturizing, comprising feverfew extract, nectar extract and cucumber extract as an active ingredient, used in cosmetics or hygiene products. The method of claim 1,
The total content of the extract is 0.0001 to 20% by volume relative to the total volume of the composition for skin anti-inflammatory or skin moisturizing composition. The method of claim 1,
The feverfew extract is one or more extracts selected from the group consisting of feverfew herbal leaves, flowers, stems, roots, and combinations thereof. The method of claim 1,
The nectar extract is a skin anti-inflammatory or skin moisturizing composition is one or more extracts selected from the group consisting of young shoots, leaves, flowers, stems and combinations of nectar herb. The method of claim 1,
The cucumber extract is a skin anti-inflammatory or skin moisturizing composition which is a root extract of cucumber herb. The method of claim 1,
The composition may further comprise at least one of an antioxidant, a preservative, an absorbent, an exfoliant, a thickener, a sunscreen, an emulsifier, a surfactant, a perfume, a deodorant, a stabilizer, a skin protectant, and a pH adjusting agent. . The method of claim 1,
The cosmetic is a composition for anti-inflammatory or skin moisturizing any one selected from the group consisting of basic cosmetics, makeup cosmetics, hair care cosmetics, facial care cosmetics, and body care cosmetics. The method of claim 1,
The hygiene article is any one selected from the group consisting of wet wipes, tissues, diapers, sanitary napkins, sanitary wipes, wet wipes, cleaning wipes, facial wipes, skin care wipes, and antiperspirant wipes. delete delete delete

Images