KR20170095170A - Cosmetic composition comprising extract of plant belonging to the genus nynphoides for improving skin - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin improvement, containing an extract of a Nymphoides sp. plant, and specifically, to a cosmetic composition for skin improvement, containing an extract of a Nymphoides sp. plant, capable of improving skin as the extract of the Nymphoides sp. plant has effects of alleviation of skin inflammation, prevention of skin aging, improvement of skin moisturization, improvement of skin whitening, and alleviation of skin wrinkles. The cosmetic composition according to the present invention contains the extract of the Nymphoides sp. plant as an active ingredient and has such as anti-oxidation, alleviation of skin inflammation, prevention of skin aging, alleviation of skin damage caused by ultraviolet rays, improvement of skin moisturization, improvement of skin whitening, alleviation of skin wrinkles, and the like, thereby remarkably improving skin.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for improving skin comprising an extract of a plant belonging to the genus Escherichia coli,

The present invention relates to a cosmetic composition for skin improvement comprising an extract of a plant belonging to the genus Lilium, and more particularly, to a cosmetic composition for extracting an extract of a plant belonging to the genus Lilium, which is useful as an antioxidant, an anti- Which is capable of improving the skin, and having an effect of preventing the skin from being damaged by the skin.

The skin is the largest organ of the body, always in direct contact with the external environment, and acts as a protective membrane for protecting the living body from various stimuli or dry environment. It is an organ which actively produces and destroys new cells as compared with other organs. In addition, it protects the body from physical abrasion, protects the inside of the body from moisture loss, protects it from ultraviolet rays generated from the sun, and plays a very important role in controlling body temperature.

In recent years, however, environmental pollution has increased the amount of ultraviolet rays reaching the surface of the earth, increasing the number of hours of outdoor activity, and increasing threats to skin health, such as westernized lifestyles and stress. These factors can lead to skin contamination, dryness, trouble, or the skin immune system, resulting in dermatitis, roughness, and rapid aging.

As the importance of skin protection has increased, the demand for functional cosmetics has been increasing. Because of the need for too much protection action such as skin moisturizing, wrinkling, aging, inflammation and whitening, functional cosmetics having respective protective functions are individually prepared There is an inconvenience to do. If it is necessary to prepare individually, it is expensive, it is inconvenient because it is difficult to store and use, and since the number of products to be added to the skin is increased, the ingredient of the product to be applied later may not be absorbed properly. Korean Patent No. 10-1325464 discloses a patent having a combination of whitening and wrinkle-improving effects, but it has no moisturizing function as the most basic effect and can not improve the rapidly increasing skin inflammation in recent years.

Therefore, there is a need to develop new substances having various effects such as skin whitening and wrinkle improvement, skin moisturization improvement, skin inflammation improvement, and the like.

Korean Patent No. 10-1325464 (published on November 6, 2012)

Accordingly, the present inventor has found that extracts of plants belonging to a lotus flower can improve various problems of the skin while searching for a substance capable of simultaneously solving various problems of the skin, thus completing the present invention.

Accordingly, an object of the present invention is to provide a cosmetic composition for improving skin comprising an extract of a plant belonging to the genus Lilium as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for skin improvement comprising an extract of a plant of the genus Nymphoides sp. As an active ingredient.

The above-mentioned plants in the juvenile lotus are Nymphoides indica ), the Nymphoides peltata , the Nymphoides coreana ), and combinations thereof.

The skin improvement may be selected from the group consisting of antioxidation, skin inflammation improvement, skin aging prevention, skin protection from ultraviolet rays, skin moisturization improvement, skin whitening improvement, skin wrinkle improvement, and combinations thereof.

The extract may be extracted with a solvent selected from the group consisting of water, alcohol, methanol, hexane, methylene chloride, ethyl acetate, butanol, propylene glycol, butylene glycol, dipropylene glycol, chloroform, glycerin and combinations thereof.

The cosmetic composition according to the present invention has various effects such as antioxidation, improvement of skin inflammation, prevention of skin aging, protection of skin from ultraviolet rays, improvement of skin moisturizing, improvement of skin whitening, So that the skin can be remarkably improved.

Figure 1 shows ABTS radical scavenging activity by concentration in Examples 1 and 3, and vitamin C is a positive control.
Fig. 2 shows the inhibitory activity of Xanthine Oxidase (XO) according to the concentrations of Examples 1 and 3, and vitamin C is a positive control.
Fig. 3 shows the SOD-like action performance of each of the concentrations of Examples 1 to 4 (meaning Example 2, Example 1, Example 4 and Example 3 according to lightness (darkness to lightness)).
Figure 4 shows the DPPH radical scavenging activity by concentration of Examples 1 and 3, and vitamin C is a positive control.
Figure 5 shows the inhibitory effect of elastase according to the concentration of Examples 1 and 3, and EGCG (Epigallocatechin-3-gallate) is a positive control.
FIG. 6 shows the cytoprotective effect against ultraviolet rays of Examples 3 and 4, and EGCG (Epigallocatechin-3-gallate) is a positive control group (Blank: control group without irradiation of ultraviolet ray A or EGCG- UVA: irradiation of ultraviolet A, negative control without the example or EGCG treatment).
Fig. 7 shows the ability to inhibit nitric oxide production by concentration in Examples 1 and 3, and vitamin C is a positive control.
FIG. 8 shows the inhibitory effect of PGE2 (prostaglandin E2) expression by concentration in Examples 1 and 3, and PDTC (Pyrrolidine dithiocarbamate) is a positive control.
Figure 9 shows the inhibition of tyrosinase by concentration in Examples 1 and 3, and vitamin C is a positive control.
FIG. 10 shows the results of quantitative analysis of electrophoresis and tyrosinase protein expression by Western blot analysis of inhibition of tyrosinase protein expression in Examples 1 and 3 (Nor:? -MSH and Example / Cont: < / RTI > alpha-MSH only, negative control not treated in the Examples).
FIG. 11 is a graph showing the effect of Examples 1 and 3 on the expression of AQP3 by concentration (Nor: control group without any treatment / Cont: treatment with only TNF-? And INF-? ).

The present invention provides a cosmetic composition for skin improvement comprising an extract of a plant of the genus Nymphoides sp. As an active ingredient.

The above-mentioned plants belonging to the myrtle lotus may be all the plants belonging to the genus Lilium , including, but not limited to, Nymphoides indica ), Nymphoides peltata , Nymphoides coreana , and combinations thereof. In addition, the cosmetic composition according to the present invention can be used as an effective ingredient to provide an antioxidant, a skin anti-inflammation, a skin anti-aging, a skin protection against ultraviolet rays, a skin moisturization improvement, a skin whitening improvement, A synergistic effect can be obtained.

Nymphoides indica ) is spread in all directions in the mud and has one to three leaves at the end of long petiole. The leaf is floating on the water and is centrifugal type, one side is deeply cracked and the edge is flat. Flowers bloom on 8 woers, white on the center, yellow in the center, and about 10 are wrapped in the bottom of the petiole. The lotus lotus is called the gold medal of the gold silver lily, and it has been traditionally used for the treatment of diseases such as gogal and tobacco by the efficacy of ginseng and persimmon.

Nymphoides peltata are rootstocks that grow sideways in the subterranean soil, and the stem has a thread-like shape. Leaves are long petiole, floated on water, wide elliptical, 5 ~ 10 cm in diameter, some split in two, but some are attached. Yellow flowers bloom in July and August. Two to three peduncles come out from the leaf axil opposite to the umbeliform inflorescence, and 2 to 3 flowers are poured on the water.

Nymphoides coreana ) is a perennial aquatic plant that grows in water and clay, has long and thin stems, and 1-2 leaves are floating on the water. Leaves are heart-shaped in the ovary or 2 ~ 6cm in diameter and are deeply cracked. The edges are flat and the petiole is 1 ~ 10cm long. Flowers bloom in June ~ July, white, blooming period, and pedicel is 1 ~ 3cm long. Calyx is wide lanceolate, pointed end, 3 ~ 4mm long, and corolla divided into 4 ~ 5 pieces. In addition, there are short hairs on the edge, 5 stamens, and the fruit is oval with 4 ~ 5mm in length, and the seeds are smooth and glossy.

The extract of the plants of the present invention is not limited to water, ethanol, methanol, hexane, methylene chloride, ethyl acetate, butanol, propylene glycol, butylene glycol, dipropylene glycol, chloroform, glycerin and combinations thereof And may be extracted with a solvent selected from the group consisting of ethanol, methanol or a mixture thereof.

In addition, the extract of the plant of the present invention is not limited thereto, but may be extracted at one or more sites selected from the group including flowers, leaves, stems, fruits, seeds, roots and outposts of plants, Can be extracted from the outpost.

The cosmetic composition according to the present invention can improve the skin. The skin can be improved by the antioxidation, improvement of skin inflammation, prevention of skin aging, protection of skin from ultraviolet rays, improvement of skin moisturizing, improvement of skin whitening, improvement of skin wrinkle, Can be selected from the group.

Plant extracts from the myrtle lotus extract can remove the ABTS radical, inhibit the activity of xanthine oxidase, and have an SOD-like activity, which can be antioxidant. Aerobic organisms use oxygen to conduct energy metabolism. When oxygen in the body is subjected to various physical, chemical and biological stresses, it is oxidized to O ₂ ^- (superoxide), NO (nitric oxide), NO ₂ It is converted into harmful active oxygen species such as hydroxyl, ROO · proxyl, RO · alkoxyl, NO ₃ · peroxynitrite and radicals such as ABTS ⁺ Cause. These reactive oxygen species attack the unsaturated fatty acid which is a constituent of the cell membrane and cause the peroxidation reaction, and it is called antioxidation action to prevent such peroxidation reaction. As means for protecting the living body from the radicals described above, antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSHpx), vitamin E, glutathione And the like to protect the living body from oxidative damage by eliminating free radicals. In addition, Xanthine oxidase (XO) is an enzyme that oxidizes purines hypoxanthine and xanthine to form uric acid, which is a cause of diseases known as gout. Gout is an inflammation accompanied by pain due to the deposition of uric acid at the joints. Therefore, inhibition of xanthine oxidase calms gout (Chiang et al ., Journal of Enzyme Inhibition , 8, pp 61-71, 1994). Xanthine oxidase is a biological source of oxygen-derived free radicals that act on oxidative damage in living cells. These various oxidative impairments cause many pathologic diseases including inflammation, arteriosclerosis, cancer and aging (Cos P et al ., Journal of Natural Products , 61, pp71-76, 1998). Inhibition of xanthine oxidase has been shown to be useful in the treatment of gout and other xanthine oxidase-induced diseases (Goodman Gilman A et al ., Pharmacological Basis of Therapeutics (eighth edition), pp 674-681, 1990).

Plant extracts from the myrtle lotus suppress the expression of nitrogen monoxide and prostaglandin E2. Inflammation is a mechanism to repair and regenerate the injured area when an invasion of biological or tissue causes physical changes such as physical action, chemical substances, bacterial infection, etc. Nitric oxide plays an important role in biological defense in the early stage of infection And regulates acute or chronic inflammatory responses, and prostaglandin E2 is an inflammatory mediator. Therefore, inhibiting the production of nitrogen monoxide or prostaglandin E2 can alleviate the inflammatory response.

Plant extracts of the myrrh lotus suppresses the activity of elastase, protects the skin from ultraviolet rays, and can prevent skin aging and wrinkles and improve wrinkles due to these actions.

Wrinkles are formed and accelerated by muscle movements in a particular direction and long-term accumulation of these movements, and are further promoted by a variety of factors such as aging, environmental effects, especially exposure to ultraviolet light. Elastin is an elastic fiber with rubber-like elasticity, which is easily deformed by a small force and returns to its original shape when the force is removed. Therefore, elastin is very important material for skin elasticity and wrinkle improvement. Therefore, inhibiting the activity of elastase degrading allostatin can improve skin elasticity and wrinkles.

Plant extracts from mature lotus plants increase the expression of aquaporin. Aqua purin is a membrane protein and is known to play a role as a water channel (Proc. Nadl. Acad Sci USA 91: 6255 (1994).) The first aquaporin was correctly identified by Peter Achre in 1991, The aquaporin family has been known for many years, but among them, aquaporin-3 is the most abundant in the aqueduct It has been known that it plays an important role, and studies on substances that activate the aquaporin-3 are active, because substances that activate the aquaporin-3 are associated with the moisturizing function of the skin (Journal of Investigative Dermatology 131: 865 , 2011).

Plant extracts of the myrtle lotus suppress the activity and expression of tyrosinase. The mechanism of melanogenesis is tyrosine, a tyrosine enzyme, which produces DOPAquinone, which undergoes autoxidation and enzymatic reaction from dopaquinone to produce melanin, a copolymer. The resulting melanin is transferred to the keratinocyte through the bag of melanomas, and then keratinocytes are exposed to the surface of the skin through a keratinization process for 28 days. However, in this process, melanin is produced in excess by the factors promoting melanin production, and pigmentation phenomenon occurs when melanin is not completely eliminated by keratinization. Therefore, in order to prevent the pigmentation phenomenon, it can be suppressed by controlling some processes in the melanin production process. Important enzymes involved in the biosynthesis of melanin include tyrosinase, tyrosinase related pro- tein 1 (TRP1), and dopachrome tautomerase, but enzymes that play a crucial role in melanin synthesis Is a tyrosinase.

The cosmetic composition according to the present invention comprises an extract of a plant belonging to the genus Lily of the Lotus as an active ingredient and is used in combination with a dermatologically acceptable excipient in a basic cosmetic composition (such as a lotion, cream, essence, cleansing foam and cleansing water, (Shampoos, rinses, hair conditioners, hair gels) and soaps, for example, in the form of a cosmetic composition (oil, oil), a color cosmetic composition (foundation, lipstick, mascara, makeup base)

Such excipients include, but are not limited to, emollients, skin penetration enhancers, colorants, perfumes, emulsifiers, thickeners and solvents. In addition, it may further contain flavors, pigments, bactericides, antioxidants, preservatives, moisturizers and the like, and may include thickeners, inorganic salts and synthetic polymeric substances for the purpose of improving physical properties. For example, when the cleanser and the soap are prepared using the cosmetic composition of the present invention, the present invention can be easily produced by adding the present biarylamide derivative to a common cleanser and a soap base. In the case of producing a cream, the cream can be prepared by adding a bivalent amide derivative of the present invention to a cream base of a typical underwater type (O / W). A synthetic or natural material such as a flavor, a chelating agent, a coloring matter, an antioxidant, an antiseptic, and a protein, a mineral, and a vitamin for the purpose of improving a physical property may be further added.

The content of the plant extract in the present invention is not limited thereto, but is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight based on the total weight of the composition. If the content is less than 0.001% by weight, the desired effect of the present invention can not be expected. If the content is more than 10% by weight, safety or formability may be difficult.

The formulations of the cosmetic composition of the present invention may be in the form of a cosmetic or dermatological composition such as a skin lotion, a skin softener, a skin toner, a convergent lotion, a softening longevity, a nutritional lotion, an astringent, a lotion, a milk lotion, a moisturizing lotion, Nourishing Cream, Moisture Cream, Hand Cream, Essence, Nutrition Essence, Pack, Soap, Shampoo, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion, Body Cleanser, Treatment, Serum, Milk, Press Powder, Loose Powder, Eye Shadow Or the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.

< Example  Manufacturing>

Example  1: Preparation of Ethanol Extract of Young Lotus Flower

Nymphoides indica ( Nymphoides indica ) In 50 g of the outpour , 500 ml of 70% ethanol was added. After dipping at room temperature for 24 hours, the extract was filtered with filter paper. The filtered filtrate was volatilized through a concentrator and then lyophilized to prepare an ethanol extract of Mulberry lotus flower.

Example  2: Preparation of Methanol Extract of Young Lotus Flower

A methanol extract of Mulberry lotus was prepared in the same manner as in Example 1, except that 80% methanol was used as the extraction solvent.

Example  3: Yellow lotus lotus  Preparation of ethanol extract

An ethanol extract of Yellow Lily lotus was prepared in the same manner as in Example 1, except that Nymphoides peltata was used.

Example  4 : Yellow lotus lotus  Preparation of methanol extract

A methanol extract of Yellow Lily lotus methanol was prepared in the same manner as in Example 3, except that 80% methanol was used as the extraction solvent.

< Experimental Example  : Verification of skin improvement effect>

Experimental Example  1. Validation of antioxidant activity

<1-1> ABTS  Radical Scatters  Measure

The antioxidant capacity of ABTS radical was measured by ABTS + cation decolorization assay [40]. After incubation at room temperature for 24 hrs, ABTS + · was formed and diluted with ethanol, and the sample was added to ABTS + · 100uL. And the absorbance was measured at 732 nm after standing for 1 minute. The results of the other experimental groups were relatively calculated with the elimination ability of the group treated with vitamin C at a concentration of 1000 占 퐂 / ml as 100%, and the results are shown in Fig.

In FIG. 1, the ABTS radical scavenging activity of Examples 1 and 3 was increased in a concentration-dependent manner, and it was found that the ABTS radical scavenging activity was similar to that of vitamin C at the concentration of 1000 μg / ml.

<1-2> xanthine of oxidase (XO, xanthine oxidase)  activation Low performance

The superoxide radical scavenging activity of xanthine oxidase was analyzed by NBT (nitro-blue tetrazolium) reduction method. 0.2 ml of xanthine (2 mM) was added to 0.1 ml of each sample solution and 0.6 ml of 0.1 M potassium phosphate buffer (pH 7.5), 0.1 ml of xanthine oxidase (0.2 U / ml) was added, and the mixture was reacted at 37 ° C for 15 minutes After 1 ml of 1N HCl was added to terminate the reaction, the amount of uric acid produced in the reaction solution was measured at 292 nm. Xanthine oxidase inhibitory activity was expressed by the absorbance reduction ratio of the sample solution and the non-additive solution. On the other hand, vitamin C was prepared at the same concentrations as in the examples and the absorbance was measured by the same method. The inhibition of NBT reduction by concentration-based samples was calculated as follows.

* B (Blank): Absorbance of reaction solution containing only 0.8 mM xanthine in 0.1 mM buffer (pH 8.0) and 69 mM SDS only

* C (control): absorbance of the reaction solution to which 0.48 mM NBT was added instead of the sample (example, vitamin C)

* S (Sample): absorbance of the reaction solution to which Example 1, Example 3, or vitamin C was added

The measured absorbance is shown in Fig. Examples 1 and 3 all exhibited better performance than vitamin C, and in particular, the XO inhibition of Example 2 was significantly superior to that of Example 1 and vitamin C.

<1-3> Similar to SOD Bow performance

20 μl of each sample solution having concentrations of 1 μg / ml, 10 μg / ml, and 100 μg / ml of each of Examples 1 to 4 was administered to a 20 μl sample of SOD assay kit-WST After adding 200 μl of working solution (WST working solution) and stirring, 20 μl of enzyme working solution was added and mixed. The mixture was incubated at 37 ° C. for 20 minutes in an incubator, and absorbance at 450 nm Were measured. Superoxide dismutase (100 units) was used as an experimental control. Superoxide dismutase-like activity was expressed by the absorbance reduction rate of the sample solution added to the superoxide dismutase treatment group. The measurement results are shown in Fig.

In FIG. 3, it was confirmed that the SOD-like activity was dependent on the concentration of the plant extract in the seed lotus flower. Especially, treatment with 100 ug / ml showed almost the same effect as that of SOD treatment. The extracts of the mature lotus showed higher SOD activity than those of the mature mature lotus extract.

<1-4> DPPH  Radical Scatters

100 μl of 5 × 10 ^-4 M DPPH (2,2-Diphenyl-1-picrylhydrazyl) was added to 100 μl of each concentration solution of Examples 1 and 3, stirred for 1 minute, and reacted in a 25 ° C. incubator for 30 minutes And the absorbance was measured at 517 nm using a microplate counter. Vitamin C was used as a positive control, and DPPH radical scavenging activity was expressed as a reduction rate of absorbance based on the absorbance of the control without the sample (example, vitamin C). The results are shown in Fig.

Experimental Example  2. Wrinkle improvement and ultraviolet ray damage repair activity verification

&Lt; 2-1 > Elastase ( elastase ) Activity inhibition test

The effect of inhibiting the activity of elastase which promotes wrinkle formation by decomposing elastin involved in dermal tissue flexibility was tested and tested by the following method. The amount of p-nitroanilide produced from the substrate at 37 ° C for 20 minutes was measured at 445 nm using N-succinyl- (L-Ala) ₃ -p-nitroanilide as a substrate. 0.5 ml of porcine pancreas elastase (2.5 U / ml) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added to the test tube, and 50 mM tris- N-succinyl- (L-Ala) _3- p-nitroanilide (0.5 mg / ml) dissolved in HCl buffer (pH 8.6) was added and reacted for 20 minutes. Elastase inhibitory activity was expressed as the absorbance reduction ratio of the sample solution and the non - added sample. FIG. 5 shows the inhibition efficiency (%) of the elastase according to the concentration of the positive control and the positive control, as compared with the case where the wrinkle improving raw material was not added as the negative control.

In FIG. 5, it can be seen that Examples 1 and 3 inhibit elastase activity in a concentration-dependent manner.

<2-2> Cytoprotective effect on ultraviolet rays

Human fibroblasts were cultured in a CO ₂ incubator at 37 ° C. Thereafter, the culture medium was removed, washed with HBSS (Hank's Balanced Salt Solution), and then irradiated with ultraviolet ray A by using a UV irradiator system. Then, And cultured under the same culture conditions for 21 hours. After the incubation, the MTT 3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide solution reagent was added to the wells and incubated at 37 ° C. for 3 hours. After the culture was removed, DMSO (dimethyl sulfoxide) was added, and the resultant was shaken appropriately. The absorbance was measured using a microplate reader. For comparison of the test results, a negative control group irradiated only with ultraviolet rays and a positive control group treated with 10 uM of EGCG (Epigallocatechin-3-gallate) were used without the examples. The absorbance measurement results are shown in Fig.

All of Examples 3 and 4 showed a cytoprotective effect against ultraviolet rays, and in particular, Example 4 showed excellent effects similar to those of positive control.

Experimental Example  3. Validation of anti-inflammatory activity

<3-1> Inhibitory effect on the formation of NO (nitric oxide)

Mouse macrophage Raw 264.7 cells were seeded at a density of 5 × 10 ³ cells / ml, and 100 μl of each was dispensed into a 96-well plate and cultured in a CO ₂ incubator at 37 ° C for 24 hours. The medium was DMEM medium containing 10% bovine serum and 1% antibiotics. After replacing the cells with DMEM medium in which bovine serum and antibiotics were excluded, Examples 1 and 3 were added for each concentration, and incubated for 10 minutes under the same culture conditions. Then 1 μl of 1 μg / ml of LPS was added And cultured for 24 hours. After the incubation, the same amount of Griess reagent reagent I and II was added to the 96-well plate and incubated for 10 minutes at room temperature. The result was measured at 540 nm on a microplate reader. As a positive control, vitamin C was used, and the NO production rate (%) was measured based on the results of the negative control not treated with the examples, and the results are shown in FIG.

In FIG. 7, it can be seen that Examples 1 and 3 reduce the amount of NO produced in a concentration-dependent manner.

<3-2> PGE2  Expression inhibitory effect

PGE2 (prostaglandin E2) plays an important role in inflammation induction, confirming that Examples 1 and 3 inhibit PGE2 expression.

Human keratinocytes were adjusted to 5 × 10 ⁵ cells / well using DMEM medium supplemented with 10% FBS and then inoculated on a 6-well plate. Each of the cells was treated with each of the concentrations of Examples 1 and 3, (Lipopolysaccharide, 1 ㎍ / ㎖) was added and cultured for 18 hours. Pyrrolidine dithiocarbamate (PDTC), an antioxidant, was used as a positive control. The amount of PGE2 produced was measured using a PGE2 ELISA kit (R & D systems, Inc, USA), and the inhibition rate (%) of PGE2 expression was measured based on the amount of PGE2 produced in the negative control group Is shown in Fig.

In FIG. 8, it can be confirmed that Examples 1 and 3 suppress PGE2 expression in a concentration-dependent manner.

Experimental Example  4. Verification of whitening activity

<4-1> Tyrosinase ( Tyrosinase ) Inhibitory activity

Tyrosinase inhibitory activity was measured by modifying a method of Yagi et al. Which measured DOPA chrome produced by the action of tyrosinase by colorimetry. That is, 0.2 ml of mushroom tyrosinase (110 U / ml) was added to a mixture of 0.2 ml of a substrate solution in which 10 mM L-DOPA was dissolved in 0.5 ml of 0.175 M sodium phosphate buffer (pH 6.8) and 0.1 ml of a sample solution, And the DOPA chrome produced in the reaction solution was measured at 475 nm. Relative tyrosinase inhibitory activity of each test group was measured based on a control group in which no substance was treated, and the results are shown in FIG.

In FIG. 9, it can be seen that Examples 1 and 3 inhibit tyrosinase in a concentration-dependent manner.

<4-2> Western Western Blot  through Tyrosinase  Measurement of protein expression level

Western blot analysis was performed to examine the effect of Examples 1 and 3 on protein expression of tyrosinase mediating melanin biosynthesis. B16F1 melanoma cells were cultured in DMEM / high glucose (Dulbeco's Modified Eagle Medium Hyclone) supplemented with 10% bovine serum and 1% antibiotics, dispensed at the appropriate concentration in 96-well plates, CO ₂ for 24 hours, and then Examples 1 and 3 were cultured for 3 days in a culture medium of B16F10 melanoma cells at different concentrations. At the same time, α-MSH (melanicyte stimulating hormone) was treated at a concentration of 10 nM to induce melanin synthesis.

After completion of the incubation, the cells were recovered by adding a tyrosine-EDTA solution to recover the cells. The cells were collected from microtubes and centrifuged to remove the supernatant.

The separated pellet was washed with 1 × PBS (-) and centrifuged. The pellet was treated with RIPA buffer (Thermo, USA) and reacted at -20 ° C. for 15 minutes. After centrifugation, the supernatant was transferred to a tube and stored at -20 ° C. The protein purified by the above method was subjected to electrophoresis using 8% SDS-PAGE and transferred to a PVDF membrane. After blocking for 1 hour in a Tris buffer solution containing 5% skim milk, the cells were reacted with tyrosinase and GAPDH antibody, respectively.

The protein purified by the above method was subjected to electrophoresis using 8% SDS-PAGE and transferred to a PVDF membrane. After 5% reaction, donkey anti-goat IgG-HRP conjugated antibody was added, and Immobilon ^TM Western Chemiluminescent HRP Substrate (Millipore). The amount of expression of tyrosinase in each experimental group was compared and analyzed by using a LAS 4000 image analyzer. The results are shown in FIG.

In Fig. 10, it can be confirmed that Examples 1 and 3 inhibit tyrosinase protein expression in a concentration-dependent manner.

Experimental Example  5. Measurement of moisturizing activity

Aquaporins (AQPs) are a family of transdermal proteins that form channels that facilitate diffusion of water and small molecules in the solution, such as glycerol and urea. Among them, AQP3 is an aqua glycerophorin, which plays an important role in maintaining the hydration level of tissues, such as glycerol, urea, purines and pyrimidines. Human Aquaporin 3 and Gill Blood Group ELISA kit were purchased from Mybiosource (USA) to measure the amount of AQP-3 in cell culture medium. 5 × 10 ⁵ cells were seeded on a 6-well plate and cultured for 24 hours. Then, TNF-α (10 ng / ml) and INF-γ (10 ng / ml) were pretreated and treated with the concentrations of Examples 1 and 3 for 24 hours. After culturing for 24 hours, cell culture was obtained and used for AQP-3 measurement. Measurement of AQP-3 was performed according to the protocol of Human Aquaporin 3 and Gill Blood Group ELISA kit.

In FIG. 11, it can be seen that Examples 1 and 3 increase the expression of AQP-3 in a concentration-dependent manner.

< Prescription example  >

Prescription example  One. Manufacture of softening longevity

The prescriptions of the softening longevity containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 1 below.

Specifically, sodium hyaluronate was dispersed in purified water with a propeller mixer (3000 rpm) to prepare a 1% solution. Ingredients 1 to 8, except for sodium hyaluronate, were homogenized at 500 rpm in a water-dissolving tank using a propeller mixer, completely dissolved by heating at 75 ° C, and then cooled to room temperature. Thereafter, the raw materials 9 to 11 were completely dissolved in a separate dissolution tank, and the mixture was added to the above-mentioned water dissolution tank and mixed with stirring. Each of the extracts of Examples 1 and 3 was poured therein and thoroughly stirred and mixed to prepare a softening agent.

Flexible longevity manufacturing ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 Polyoxyethylene hardened castor oil 0.5 3 Glycine 3.3 4 Dipotassium glycyrrhizate 0.1 5 1,3-butylene glycol 3.0 6 Sodium hyaruronate 0.1 7 ethanol 5.0 8 Antioxidant 0.1 9 Triethanolamine 0.1 10 EDTA 0.1 11 antiseptic Suitable amount 12 Purified water Balance

Prescription Example 2. Preparation of nutrition lotion

The prescriptions of the nutritional lotion containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 2 below.

Specifically, the carbomer was dispersed at 4000 rpm using a propeller mixer to prepare a 2% solution state. Raw materials 1 to 6 were fed into an aqua yard and stirred with a homomixer (2000 rpm) and dispersed, and the mixture was heated to 75 캜. The raw materials 7 to 14 were added to the oily melting tank and dissolved by heating to 75 ° C. Then, the oil phase dissolved in the water dissolution tank was charged and emulsified (3000 rpm / 5 minutes) and then cooled to room temperature. Each of the extracts of Examples 1 and 3 was added thereto and sufficiently stirred to prepare a nutrition lotion.

Nutritional lotion manufacturing ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 glycerin 7.0 3 Sorbitan stearate sucrose cocoate 2.0 4 Mineral oil 4.0 5 Trioctanoin 1.0 6 Stearic Acid 1.0 7 Glyceryl stearate 0.5 8 Sorbitan monostearate 1.0 9 Dimethicone 0.5 10 Antioxidant 0.3 11 Triethanolamine 0.1 12 Carbomer 0.2 13 EDTA 0.1 14 antiseptic Suitable amount 15 Purified water Balance

Prescription example  3. Manufacture of Essence

The prescriptions of the essences containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 3 below.

Specifically, sodium hyaluronate and hydroxyethylcellulose were each dispersed in purified water in a propeller mixer (2000 rpm) to prepare a solution containing 1% by weight. The carbomer was also dispersed in purified water with a propeller mixer (4000 rpm) to prepare a solution containing 2% by weight. On the other hand, raw materials 1 to 12 were added to an aqueous dissolution tank, and the mixture was stirred and dispersed by a homomixer (2000 rpm), then heated to 75 캜, and the warmed water phase was further cooled to room temperature. The ingredients 13 to 15 were completely dissolved in a separate dissolution tank, and the mixture was added to the above-mentioned water dissolution tank and mixed by stirring. Thus, an essence containing each of the extracts of Examples 1 and 3 as an active ingredient was prepared.

Essence manufacturing ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 glycerin 5.0 3 1,3-butylene glycol 2.0 4 Polyethylene glycol 2.0 5 Carbomer 1.0 6 Sodium hyaruronate 0.1 7 Glycine 3.0 8 Polyacrylamide 2.0 9 Hydroxyethylcellulose 0.2 10 ethanol 3.0 11 Antioxidant 0.3 12 Triethanolamine 0.1 13 Polyoxyethylene hardened castor oil 1.0 14 EDTA 0.1 15 antiseptic Suitable amount 16 Purified water Balance

Prescription example  4. Manufacture of nutritional cream

The nutritional creams containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 4 below.

Specifically, raw materials 1 to 8 were fed into an aeration dissolving tank, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 캜. The raw materials 9 to 16 were added to a separate oily melting tank and dissolved by heating to 80 DEG C, and the oil phase dissolved in the water dissolution tank was introduced to emulsify (3000 rpm / 10 minutes) and then cooled to room temperature. Each of the extracts of Examples 1 and 3 was poured therein and thoroughly stirred and mixed to prepare a nutritive cream.

Nutritional cream composition ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 1,3-butylene glycol 3.0 3 glycerin 3.0 4 Hydrogenated lecithin 1.0 5 Octyldodecanol 3.0 6 Trioctanoin 2.0 7 Stearic Acid 1.5 8 Cetostearyl alcohol 2.0 9 Polysorbate 60 1.5 10 Sorbitan sesquioleate 2.0 11 Dimethicone 3.0 12 Antioxidant 0.3 13 Xanthan gum 0.2 14 Triethanolamine 0.1 15 EDTA 0.1 16 antiseptic Suitable amount 17 Purified water Balance

Prescription example  5. Manufacture of pack

The prescription of the pack containing each extract of Examples 1 and 3 as an active ingredient is shown in Table 5 below.

Specifically, raw materials 1 to 8 were completely dissolved in a separate dissolution tank and stirred and mixed. Each of the extracts of Examples 1 and 3 was added thereto, and the mixture was thoroughly stirred to prepare a pack.

Composition of pack ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 glycerin 7.0 3 1,3-butylene glycol 3.0 4 Squalene 3.0 5 Dimethicone 3.0 6 Sodium hyaluronate 0.1 7 Glycine 2.0 8 Polyacrylamide 5.0 9 Antioxidant 0.3 10 Triethanolamine 0.1 11 EDTA 0.1 12 antiseptic Suitable amount 13 Purified water Balance

Prescription example  6. Manufacture of ointment

The ointments containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 6 below.

Composition of ointment ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 glycerin 8.0 3 1,3-butylene glycol 4.0 4 Liquid paraffin 15.0 5 Beta Glucan 7.0 6 Carbomer 0.1 7 Caprylic / capric triglyceride 3.0 8 Squalane 1.0 9 Cetylaryl glucoside 1.5 10 Sorbitan stearate 0.4 11 Cetearyl alcohol 1.0 12 antiseptic Suitable amount 13 incense Suitable amount 14 Pigment Suitable amount 15 Purified water Balance

Prescription example  7. Patchy  Produce

The prescriptions of the patches containing the extracts of Examples 1 and 3 as active ingredients are shown in Table 7 below.

Composition of patch ingredient Content (% by weight) One The extracts of Examples 1 and 3 0.01 2 Hexylene glycol 20 3 Diethyleneamine 0.7 4 Polyacrylic acid 1.0 5 Sodium sulphate 0.1 6 Polyoxyethylene lauryl ether 1.0 7 Polyhydroxyethylene cetyl stearyl ether 1.0 8 Viscous paraffin oil 2.5 9 Caprylic acid ester / capric acid ester 2.5 10 Polyethylene glycol-400 3.0 11 Purified water Balance

It will be apparent to those skilled in the art that various modifications and equivalent arrangements may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, it is to be understood that the present invention is not limited to the above-described embodiments. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims. It is also to be understood that the invention includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.

Claims (4)

A cosmetic composition for improving skin comprising an extract of a plant of the genus Nymphoides sp. As an active ingredient.
The method according to claim 1,
The above-mentioned plants in the juvenile lotus are Nymphoides indica ), the Nymphoides peltata , the Nymphoides coreana ), and a combination thereof. The cosmetic composition for skin improvement according to claim 1,
The method according to claim 1,
Characterized in that said skin improvement is one selected from the group consisting of antioxidation, skin inflammation improvement, skin aging prevention, skin protection from ultraviolet rays, skin moisturization improvement, skin whitening improvement, skin wrinkle improvement, Cosmetic composition.
The method according to claim 1,
The extract is extracted with a solvent selected from the group consisting of water, ethanol, methanol, hexane, methylene chloride, ethyl acetate, butanol, propylene glycol, butylene glycol, dipropylene glycol, chloroform, glycerin and combinations thereof Wherein the cosmetic composition is a cosmetic composition for improving skin.

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