KR20210106661A - Composition for improving skin whitening and anti-wrinkle comprising Hizikia fusiformis extract as effective component - Google Patents

Abstract

The present invention relates to a composition for skin whitening and wrinkle improvement containing hizikia fusiforme extract as an active ingredient. According to the present invention, a hizikia fusiforme extract is safe for the skin due to no cytotoxicity, inhibits tyrosinase activity and melanin production, and has excellent skin whitening and wrinkle improvement effects by inhibiting elastase, collagenase activity and MMP-1 production, and thus hizikia fusiforme extract may be usefully used as a material for functional cosmetics or health functional food for skin whitening and wrinkle improvement.

Description

Composition for improving skin whitening and anti-wrinkle comprising Hizikia fusiformis extract as effective component

The present invention relates to a composition for skin whitening and anti-wrinkle improvement comprising a hibiscus extract as an active ingredient.

The skin is the first defense organ of the human body that protects our body from external stimuli, and the interest in it is increasing from an aesthetic point of view. Accordingly, research on various functional cosmetics is active, and among them, various functional cosmetics for wrinkle relief and whitening functions are being released.

Various causes of wrinkles have been reported. First, when the skin is exposed to excessive ultraviolet rays, the decomposition of collagen, which is a component of the skin, is promoted, so that the skin loses elasticity and wrinkles can be generated. In addition, when the skin becomes too dry due to excessive temperature change, humidity drop, wind, etc., the skin function as a barrier to the outside decreases, and wrinkles may occur. In addition, when the skin is exposed to reactive oxygen species or free radicals, lipid peroxide is generated by oxidation, and thus, skin constituent proteins such as collagen are deformed and wrinkles can be generated. Various cosmetic compositions are being studied to suppress the occurrence of wrinkles due to these external and internal factors.

On the other hand, when the skin is irradiated with ultraviolet rays, melanocytes in the skin are activated and melanin production is promoted by the action of tyrosinase, tyrosinase related protein 1 (TRP1) and tyrosinase related protein 2 (TRP2). The produced melanin is deposited on the skin and develops into age spots or freckles. In order to prevent this, whitening cosmetics containing various compositions are used. In addition, ultraviolet rays have been found to cause various skin disorders by promoting the oxidation of sebum and cell membranes, and in particular, more effective skin oxidation prevention measures are required due to the recent increase in ultraviolet rays due to the destruction of the ozone layer.

Ascorbic acid, phosphoric acid ester salts, hydroquinone derivatives, placental extracts, kojic acid, ellagic acid, and the like are known conventionally known ingredients for a whitening effect, and cosmetic compositions containing these ingredients are common. Recently, it has been reported that polyphenols contained in plants have a high whitening effect, and cosmetic compositions using the same have been proposed. In addition, the whitening effect of flavanones and hydroxyflavones has already been disclosed. However, these whitening ingredients are somewhat ineffective and have problems during storage, and in addition to the whitening effect, they do not sufficiently exhibit the wrinkle-relieving effect and anti-aging effect. In particular, functional cosmetics containing chemical ingredients cause skin irritation and are not suitable for sensitive skin, so there is little irritation to the skin and interest in eco-friendly, natural plant-derived functional cosmetics.

Hizikia fusiformis (Hizikia fusiformis) is a perennial seaweed that is distributed in the south-central and south-central parts of Korea, and is especially produced in Jeju Island and the southwest coast. As it has a unique taste and is rich in calcium, vitamin A and dietary fiber, it is effective for diabetes, high blood pressure, colorectal cancer, and constipation.

On the other hand, Korea Patent Application Publication No. 2013-0023326 discloses 'a cosmetic composition with excellent anti-allergic, soothing and moisturizing effects containing a seaweed extract composed of hat banyan, agar agar, and chives'. It discloses a 'cosmetic composition for wrinkle improvement comprising a Serrated Capricorn extract or a fraction thereof as an active ingredient. However, the 'composition for skin whitening and wrinkle improvement containing the extract of the present invention' as an active ingredient has not yet been disclosed.

The present invention was derived from the above needs, and as a result of the present inventors preparing a hot water extract of Hibiscus and testing its function, the hot water extract of Hibiscus inhibits tyrosinase activity and melanin synthesis, and elastase, cola By confirming that genase activity and MMP-1 production were inhibited, the present invention was completed.

In order to solve the above problems, the present invention provides a cosmetic composition for skin whitening or wrinkle improvement containing an extract of Hizikia fusiformis as an active ingredient.

In addition, the present invention provides a health functional food composition for skin whitening or wrinkle improvement containing an extract of Hizikia fusiformis as an active ingredient.

The extract of the present invention has no cytotoxicity, so it is safe for the skin, inhibits tyrosinase activity and melanin production, and has skin whitening and wrinkle improvement effects by inhibiting elastase, collagenase activity and MMP-1 production. Because it is excellent, it may be usefully used as a material for functional cosmetics or health functional food for skin whitening and wrinkle improvement.

1 is a result of measuring cell viability when B16F10 cells are treated with a hot water extract of the present invention. C: untreated control.
2 is a result confirming the melanin production inhibitory effect of the hot water extract of the present invention. α-MSH: melanogenesis inducer.
3 is a result of measuring cell viability when CCD-986SK cells are treated with a hot water extract of the present invention. C: untreated control.
4 is a result confirming the inhibitory effect of MMP-1 (Matrix metalloproteinase-1) production of the hot water extract of the present invention. C: untreated control.

In order to achieve the object of the present invention, the present invention provides a cosmetic composition for skin whitening or wrinkle improvement containing an extract of Hizikia fusiformis as an active ingredient.

In the cosmetic composition for skin whitening or wrinkle improvement of the present invention, the extract _{may be prepared by extracting water, a C 1} to C ₄ lower alcohol or a mixture thereof with a solvent, and preferably by extracting with water. , but not limited thereto.

In addition, the cosmetic composition is a skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, eye cream, moisture cream, hand cream, essence, nourishing essence, pack , cleansing foam, cleansing water, cleansing lotion, cleansing cream, body lotion, body cleanser, soap and powder may be prepared in any one formulation selected from the group consisting of, but is not limited thereto. The composition of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation, and the types and amounts of these components can be easily selected by those skilled in the art.

The cosmetic composition of the present invention may contain one or more ingredients for skin whitening or wrinkle improvement that exhibit the same or similar function in addition to the above active ingredients. Skin whitening Ingredients for skin whitening or wrinkle improvement include kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, cycloalkanone, methylenedioxyphenyl alkanol, 2,7-di plant extracts such as nitroindazole or tendril extract, rice extract, and licorice extract, but are not limited thereto.

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

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

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative or ethoxylated glycerol fatty acid ester and the like can be used.

The cosmetic composition of the present invention may further contain excipients including fungicides, hydrotropes, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, and the like.

The present invention also provides a health functional food composition for skin whitening or wrinkle improvement containing an extract of Hizikia fusiformis as an active ingredient.

In the health functional food composition for skin whitening or wrinkle improvement of the present invention, the extract of hibiscus is the same as described above.

The health functional food composition for skin whitening or wrinkle improvement of the present invention may be prepared in one formulation selected from the group consisting of powders, granules, pills, tablets, capsules, candy, syrups and beverages, but is not limited thereto. Preferably, the health functional food for skin whitening or wrinkle improvement has the advantage of having a more excellent effect by ingesting it in the form of an inner beauty food. The inner beauty food is a food called 'eating cosmetics or beauty food', which refers to food that absorbs various ingredients good for the skin into the body and changes the skin constitution to make it healthy. As you choose, you can choose and consume inner beauty foods that are suitable for you by considering your skin condition and lifestyle. More preferably, when mixing the cosmetic containing the extract and the inner beauty food containing the extract, compared to using only cosmetics, tyrosinase activity and melanin synthesis are inhibited, and elastase and collagenase activity And the effect of inhibiting the production of MMP-1 is significantly increased, so it has the advantage of being able to see the skin whitening and wrinkle improvement effect more effectively.

In addition, there is no particular limitation on the type of the food. Examples of foods to which the health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea There are drinks, alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense. The beverage includes carbonated beverages, functional ionic beverages, juices (eg, apple, pear, grape, aloe, tangerine, peach, carrot, tomato juice, etc.), sikhye, and the like.

In the case of using the hibiscus extract of the present invention as a food additive, the hibiscus extract may be added as it is or used together with other foods or food ingredients, and may be suitably used according to a conventional method. The active ingredient may be used appropriately depending on the purpose of its use (prevention or improvement). In general, in the production of food or beverage, the extract of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range that does not cause any problems in terms of safety.

The functional food of the present invention includes ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.) or sugar alcohols (eg, , xylitol, sorbitol, erythritol, etc.). As the flavoring agent, natural flavoring agents (eg, taumatine, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used. In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonated beverages It may further contain a carbonation agent, etc. used for

In addition to the above health functional food composition, various nutrients, vitamins, inhibitors, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, It may further contain a carbonation agent, etc. used in carbonated beverages. The ratio of these added ingredients is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food composition of the present invention.

Hereinafter, an embodiment of the present invention will be described in detail. However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

Preparation Example 1. Preparation of hot water extract of hibiscus

Hizikia 　 fusiformis ) was washed once in running water, washed once in distilled water, and distilled water 4 times that of 2 kg of hibiscus was added and extracted at 100 ° C for 3-4 minutes. ) was obtained. Thereafter, the Brix (%) was measured to confirm the sugar content (0.4 Brix), and lyophilized to be used in the following Examples.

Example 1. Measurement of total polyphenol content and DPPH radical scavenging activity

Total polyphenol content was determined using the Folin-Denis method. The hot water extract and phenol solution (Folin-ciocalteu's phenol reagent) were mixed 1:1 and reacted for 3 minutes, and then 10% (w/v) Na ₂ CO ₃ was slowly added. After reacting this for 1 hour in the dark, it was measured by measuring the absorbance at 725 nm. At this time, the standard calibration curve was obtained by measuring gallic acid at a certain concentration (0 to 500 μg/ml) in the same manner as above, and substituting the absorbance of the sample into the standard calibration curve to determine the total polyphenol content in mg/g indicated as As a result, a total polyphenol content of 17.1 mg/g was measured in the hot water extract of hibiscus (Table 1).

In addition, DPPH radical scavenging activity was measured by reacting hot water extract with 0.2 mM DPPH (1,1-diphenyl-2-picrylhydrazyl) 1:1 in a dark room for 30 minutes and then absorbance at 528 nm. . At this time, the absorbance was measured in the same manner as the sample using L-Ascorbic acid (250 μM), which has been widely used as an antioxidant, as a positive control for activity comparison. Antioxidant activity using DPPH was expressed as a percentage (%) of the absorbance difference between the sample-added group and the non-additive group. The formula for calculating the DPPH radical scavenging activity is as follows. As a result, it was confirmed that the DPPH radical scavenging activity of the hot water extract of hibiscus was 72.7% (Table 2).

DPPH radical scavenging activity (%) = [1- (sample absorbance ₅₂₈ nm / control absorbance ₅₂₈ nm )] × 100

Determination of total polyphenol content sample Total polyphenol content (mg/g) Chickpea hot water extract 17.1

Determination of DPPH radical scavenging activity sample treatment concentration DPPH radical scavenging activity Chickpea hot water extract 1 mg/ml 72.7% L-Ascorbic Acid 250 μM 83.0%

Example 2. Skin whitening and cytotoxicity

One) in vitro Tyrosinase inhibitory activity

Tyrosinase inhibitory activity was measured using a Tyrosinase Inhibitor Assay Kit (ab204715, abcam) after reacting with hot water extract of each concentration, and then absorbance was measured at 510 nm, and Kojic acid (0.75 mM) was used as a positive control. The formula for obtaining the tyrosinase inhibitory activity is as follows. As a result, the hot water extract of hibiscus showed a tendency to increase the tyrosinase inhibitory activity in a concentration-dependent manner (Table 3).

Tyrosinase inhibitory activity (%) = [1- (sample absorbance _{510 nm} / absorbance without addition _{510 nm} )] × 100

Measurement of tyrosinase inhibitory activity sample treatment concentration Tyrosinase inhibitory activity (%) Chickpea hot water extract 100 μg/ml 14.4 200 μg/ml 36.5 500 μg/ml 52.8 kojic acid 0.75 mM 80.6

2) Cytotoxicity

In order to measure the effect of hot water extract of hibiscus on B16F10 melanoma cells, MTT (3-(4,5-dimethylthaiazol-2-yl)-2,5-diphenyltetrazolium bromide) analysis was performed using B16F10 melanoma cells. . B16F10 cells were used with DMEM (Dulbeco's Modified Eagle's Media) containing 10% fetal bovine serum (FBS), 1% penicillin and 1% streptomycin, _{and cultured at 5% CO 2} and 37°C conditions. The medium was exchanged every 2 days and subcultured by treatment with trypsin-EDTA at 80% confluent status. After culturing B16F10 cells (1×10 ³ cells/well) in a 96-well plate for 24 hours, 50, 100, and 200 μg/ml of hot water extract of hibiscus were treated, respectively, and 2 mg/ml of MTT solution was added after 72 hours. and reacted for 4 hours. Then, after removing the MTT solution, DMSO (dimethyl sulfoxide) was added and the reaction was carried out in the dark for 30 minutes, and absorbance was measured at 540 nm. Cell viability (cell viability) was calculated by comparing the value of the control group that was not treated with the hot water extract of hibiscus. As a result, it was confirmed that there was no cytotoxicity in all the treatment groups of the hot water extract of hibiscus (Fig. 1).

3) Inhibition of melanin production

Using B16F10 melanoma cells, the effect of hot water extract of hibiscus on melanin synthesis was confirmed. Specifically, B16F10 cells were inoculated into a 6-well plate at 5×10 ⁴ cells/well and cultured for 24 hours, and then melanocyte-stimulating hormone (α-MSH) and hot water extract ( 50, 75 and 100 μg/ml) were treated simultaneously for 72 hours. After 72 hours, the medium was removed, washed twice with phosphate buffer (phosphatate buffer saline, PBS), and 200 μl of 1N NaOH solution containing 10% DMSO was added to the pellet obtained by centrifuging the adhered cells and 80 After dissolution by standing at ℃ for 1 hour, absorbance was measured at 475 nm. As a result, as shown in FIG. 2 , it was confirmed that the melanin content increased by α-MSH treatment was reduced by the hot water extract of hibiscus, and the melanin content was further decreased as the treatment concentration of the hot water extract of hibiscus increased. showed a trend.

Example 3. Skin Wrinkle Generation and Cytotoxicity

One) in vitro Inhibition of elastase activity

It was confirmed whether the hot water extract of hibiscus inhibited the activity of elastase, which is the main cause of skin wrinkles. Specifically, 100mM Tris-HCl buffer (pH 8.0), hibiscus hot water extract, and 2.9mM AAAPVN (N-succinyl-(L-Ala)3-p-nitroanilide) were added to a 96-well plate and left for 10 minutes. A solution of pancreatic elastase (porcine pancreas elastase, 0.2 unit) was added and reacted at 25° C. for 10 minutes, and then absorbance was measured at 410 nm. As a control, distilled water was added instead of the hot water extract of hibiscus to measure the enzyme activity, and ursolic acid (0.5 mM) was used as a positive control for activity comparison. The formula for determining the inhibitory activity of elastase is as follows. As a result, as shown in Table 4, it was confirmed that the hot water extract of hibiscus inhibited the elastase activity in a concentration-dependent manner.

Elastase inhibitory activity (%) = [1- (B-C) / (A-D)] × 100

A: Absorbance after reaction by adding distilled water instead of sample and adding enzyme

B: Absorbance of sample after reaction by adding enzyme

C: Absorbance of sample after reaction by adding distilled water instead of enzyme

D: Absorbance after reaction by adding distilled water instead of sample and enzyme

Measurement of elastase inhibitory activity sample treatment concentration Elastase inhibitory activity (%) Chickpea hot water extract 100 μg/ml 7.0 200 μg/ml 9.2 500 μg/ml 19.1 Ursolic acid 0.5 mM 54.1

2) in vitro Inhibition of collagenase activity

The decomposition of collagen, which constitutes the skin cell connective tissue, directly affects the decrease in elasticity of the connective tissue and the generation of wrinkles. Therefore, by measuring the collagenase inhibitory activity of the hot water extract of hibiscus, it was attempted to confirm the skin wrinkle improvement effect. Collagenase inhibitory activity was determined by 0.3 mg/mL of 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-Agr (4-phenylazovenzyloxycarbonyl-Pro-Leu-Gly-Pro-Arg) and hot water extract (100 , 200 and 500 μg/ml) were added with 0.2 mg/mL of collagenase, left at 25° C. for 20 minutes, and then 6% citric acid was added to stop the reaction. Then, after adding 100% ethyl acetate, the supernatant was taken and absorbance was measured at 320 nm to calculate collagenase inhibitory activity, and EGCG (epigallocatechin gallate, 2 mM) was used as a positive control for activity comparison. . As a result, it was confirmed that the collagenase inhibitory activity of the hot water extract of H. shiitake was increased in a concentration-dependent manner (Table 5).

Measurement of collagenase inhibitory activity sample treatment concentration Collagenase inhibitory activity (%) Chickpea hot water extract 100 μg/ml 19.2 200 μg/ml 27.1 500 μg/ml 27.2 EGCG 2 mM 54.1

3) Cytotoxicity

MTT analysis was performed using CCD-986SK melanoma cells. For CCD-986SK cells, DMEM (Dulbeco's Modified Eagle's Media) containing 10% fetal bovine serum (FBS), 1% penicillin and 1% streptomycin was used, _{and cultured at 5% CO 2} and 37°C conditions. The medium was exchanged every 2 days and subcultured by treatment with trypsin-EDTA at 80% confluent status. After culturing CCD-986SK cells (1×10 ³ cells/well) in a 96-well plate for 24 hours, 50, 100, and 200 μg/ml hot water extract of shiitake shiitake extract were treated, respectively, and 2 mg/ml MTT solution after 24 hours. was added and reacted for 4 hours. After the removal of the MTT solution, DMSO was added, and the absorbance was measured at 540 nm after reacting in the dark for 30 minutes. Cell viability (cell viability) was calculated by comparing the value of the control group that was not treated with the hot water extract of hibiscus. As a result, it was confirmed that there was no cytotoxicity when 50, 100 and 200 μg/ml of hot water extract of hibiscus was treated (FIG. 3).

4) Inhibition of MMP-1 production

By analyzing the change in the expression level of matrix metalloproteinases (MMPs), which has an important effect on the reduction of collagen, it was confirmed whether or not the wrinkle-improving effect of hot water shiitake extract. CCD-986SK cells were inoculated in a 48-well plate to 5×10 ⁴ cells per well and cultured for 24 hours. , MMP-1 content was measured using the Human MMP-1 ELISA Kit (Sigma-Aldrich) according to the manufacturer's protocol. As a result, it was confirmed that the MMP-1 content was reduced by about 36-52% in the hot water extract treated group compared to the untreated control group (C) (FIG. 4).

Claims (5)

Hizikia 　 fusiformis ) A cosmetic composition for skin whitening or wrinkle improvement containing an extract as an active ingredient. The cosmetic composition for skin whitening or wrinkle improvement according to claim 1, wherein the extract is prepared by extracting water, C ₁ to C ₄ lower alcohol or a mixture thereof with a solvent. According to claim 1, wherein the cosmetic composition is skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, eye cream, moisture cream, hand cream, essence, Cosmetics for skin whitening or wrinkle improvement, characterized in that it is prepared in any one formulation selected from the group consisting of nutritional essence, pack, cleansing foam, cleansing water, cleansing lotion, cleansing cream, body lotion, body cleanser, soap and powder composition. A health functional food composition for skin whitening or wrinkle improvement containing an extract of Hizikia fusiformis as an active ingredient. The health functional food composition for skin whitening or wrinkle improvement according to claim 4, wherein the composition is an inner beauty food.

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