KR102172913B1 - Cosmetic composition containing adoxa moschatellina as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a lotus leaf extract as an active ingredient and having one or more of the effects of improving skin wrinkles, whitening skin, antioxidant, moisturizing and improving skin elasticity.

Description

Cosmetic composition containing lotus leaf extract as an active ingredient {COSMETIC COMPOSITION CONTAINING ADOXA MOSCHATELLINA AS ACTIVE INGREDIENT}

The present invention relates to a cosmetic composition containing a lotus leaf extract as an active ingredient, specifically, the cosmetic composition has the effect of improving skin wrinkles, skin whitening, antioxidant, moisturizing and skin elasticity.

In general, many products related to skin contain substances that can cause irritation or inflammation, so more than 10% of people who use personal care products complain of side effects to the skin. There is a growing trend of consumers who feel that their skin is sensitive. In the survey on consciousness of general consumers, more than 30-40% of the respondents answered that they were sensitive skin, and this tends to increase continuously. Moreover, as recent cosmetics are focused on functional cosmetics that can give real effects to the skin, skin side effects due to the use of personal care products such as cosmetics are expected to increase gradually. In particular, air pollution such as ozone, a secondary product caused by fine dust, smoke and ultraviolet rays (UV), becomes serious, and the amount of ultraviolet irradiation increases due to the destruction of the ozone layer, drying of living spaces, personal allergies, stress, and chemical harmful substances. The skin is easily damaged due to the increase, etc., and various irritants are generated due to this, causing the skin to become easily reddened or sensitive, and seriously, it causes skin trouble.

In general, a variety of raw materials are used in the process of making cosmetics, and typically, from 10 to 50 raw materials are used. These ingredients have been proven to be safe through skin irritation tests, but when applied to products, they can cause irritation such as inflammation, itchiness, and allergies to the skin. Skin irritation that may be caused by the use of cosmetics includes irritation such as itching, stinging, and burning, and erythema and edema. Contact dermatitis can be broadly classified into irritant contact dermatitis, allergic contact dermatitis, sensory irritation and phototoxic dermatitis, but it is difficult to clearly distinguish them.

In order to solve these various problems, cosmetics using natural products have been developed in recent years to reduce skin irritation caused by various chemical substances. Natural ingredients have relatively few side effects on the skin, and as consumers' response to cosmetics using natural ingredients increases in recent years, the value of development as a cosmetic ingredient is further increasing.

The lotus leaf used in the present invention has the scientific name Adoxa It is called moschatellina L. and is a perennial plant belonging to the dicotyledonous plant class. The flower color is yellow and green, and it lives in the temperate and cold regions of the northern hemisphere and is characterized by a growing environment that grows up to 8~17cm in height in the humid place at the foot of the mountain. In addition, lotus leaf is known to contain various flavonoids.

Recently, the strong antioxidant effect of the flavonoids and polyphenols contained in plant extracts such as lotus leaf has been studied. In addition, these plant flavonoid derivatives are used for various purposes such as edible, antioxidant, skin whitening, etc., and are also used in fields such as pharmaceuticals, cosmetics, and functional foods in Korea.

Under this background, the present inventors have completed the present invention by preparing a lotus bok choy extract excellent in alleviating irritation while studying cosmetic compositions originating from natural products.

The main object of the present invention is to provide an excellent new material capable of alleviating skin irritation caused by external irritation and skin problems that may be caused by various raw materials contained in cosmetics.

Another object of the present invention is to provide a cosmetic composition having various physiologically active effects such as antioxidant effect, collagen biosynthesis effect, moisturizing effect, skin wrinkle improvement effect, and skin elasticity improvement effect.

Another object of the present invention is to provide a cosmetic method using a cosmetic composition containing the lotus leaf extract.

Therefore, the object of the present invention as described above is achieved by a cosmetic composition containing the lotus bokcho extract as an active ingredient.

In order to alleviate skin irritation, the present inventors have attempted to secure a material that is stable and does not cause skin problems such as irritation and seizures when applied to the skin.

Preferably, the lotus leaf extract has a skin wrinkle improvement effect, skin whitening effect, antioxidant effect, skin moisturizing effect and skin elasticity improvement effect.

In addition, preferably, the lotus leaf extract is characterized in that it contains 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

In addition, preferably, the lotus leaf extract is (a) water, an anhydrous or hydrous lower alcohol having 1-4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, By an extraction method selected from the group consisting of a solvent extraction method using an extraction solvent selected from the group consisting of dichloromethane, nucleic acids, and mixtures thereof, (b) a supercritical extraction method, (c) an ultrasonic extraction method, and (d) a fermentation extraction method using microorganisms. Can be extracted.

In addition, preferably, the cosmetic composition exhibits MMP-1 production inhibitory effect, collagen biosynthesis effect, melanin production inhibitory effect, inflammatory cytokine expression inhibitory effect, antioxidant effect, moisturizing effect, skin wrinkle improvement effect, skin elasticity improvement effect. It is to provide a cosmetic composition.

In addition, preferably, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream and spray. It is characterized by having a formulation selected from the group consisting of.

The lotus bokcho extract according to the present invention has an MMP-1 production inhibitory effect (Experimental Example 1), collagen synthesis enhancement effect (Experimental Example 2), melanin production inhibitory effect (Experimental Example 3), antioxidant effect (Experimental Example 4), inflammatory cytotoxic It has an effect of inhibiting kine expression (Experimental Example 5), a moisturizing effect (Experimental Example 6), an effect of improving skin elasticity (Experimental Example 7), and an effect of improving skin wrinkles (Experimental Example 8).

In addition, according to the present invention, in order to maximize the efficacy of the lotus leaf extract, the effect of inhibiting MMP-1 production, promoting collagen synthesis, inhibiting melanin production, improving skin elasticity, etc. through ultrasonic extraction, supercritical extraction, and fermentation process. It further promotes.

All technical terms used in the present invention, unless otherwise defined, have the following definitions and correspond to the meanings as commonly understood by one of ordinary skill in the relevant fields of the present invention. In addition, although preferred methods or samples are described in the present specification, those similar or equivalent are included in the scope of the present invention.

The term “about” refers to 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, or 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, or relative to a reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length. It means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length varying by 4, 3, 2 or 1%.

Throughout this specification, unless otherwise required by the context, the terms “comprise” and “comprising” include the steps or components presented, or groups of steps or components, but any other step or component, or It is to be understood that a step or group of components is not excluded.

The present inventors studied the applicability as a cosmetic containing lotus bok choy extract among various natural products and measured skin wrinkle improvement effect, skin whitening effect, antioxidant effect, skin moisturizing effect, and skin elasticity improvement effect through various experiments. I found that I can expect efficacy. Therefore, the cosmetic composition of the present invention comprising the above components is suitable for a cosmetic composition for improving skin wrinkles, whitening skin, antioxidant effects, moisturizing skin, and improving skin elasticity.

The lotus leaf extract of the present invention may be prepared using a conventional solvent according to a conventional method known in the art, that is, under the conditions of a conventional temperature and pressure, preferably (a) water, 1 carbon number. -4 anhydrous or aqueous lower alcohols (e.g., methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, Extraction is performed using a solvent extraction method using a solvent selected from the group consisting of chloroform, hexane, and mixtures thereof (b) a supercritical extraction method using reduced pressure and high temperature using carbon dioxide or (c) an ultrasonic extraction method.

In the present invention, preferably, a solvent extraction method using an extraction solvent was used.

In the present invention, the lotus leaf extract is a variety of extraction solvents, for example, water, anhydrous or hydrated lower alcohols having 1 to 4 carbon atoms (eg, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene It can be obtained using one or more extraction solvents selected from the group consisting of glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof, preferably ethanol, 70 It is obtained using% (v/v) ethanol or water, and most preferably it is obtained using 70% (v/v) ethanol.

On the other hand, it will be apparent to those skilled in the art that the lotus bokcho extract of the present invention can obtain an extract exhibiting substantially the same effect even when using not only the above-described extraction solvent but also other extraction solvents.

In addition, the lotus bokcho extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract that has undergone conventional purification and fermentation processes. For example, decompression by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by extraction by ultrasonic wave, separation by using an ultrafiltration membrane having a certain molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity) Active fractions obtained through various purification and extraction methods additionally carried out, such as separation by means of (produced for separation according to sex) or by fermentation products using natural conditions or various microorganisms, are also included in the extract of the present invention.

In addition, the present invention provides a cosmetic composition, characterized in that the extract is obtained by concentrating under reduced pressure or freeze-drying a liquid product obtained by cooling, heating and filtration at room temperature, and an additional solvent.

The extraction method by the supercritical extraction method by the decompression by carbon dioxide and the high temperature refers to a supercritical fluid extraction method.In general, the supercritical fluid is a liquid and a liquid that is possessed when a gas reaches a critical point under high temperature and high pressure conditions. It has gaseous properties and chemically has a polarity similar to that of non-polar solvents. Due to this property, supercritical fluids are used for extraction of oil-soluble substances (J. Chromatogr. A. 1998;479:200-205).

Carbon dioxide becomes a supercritical fluid having both liquid and gaseous properties as the pressure and temperature reach a critical point due to the operation of the supercritical fluid device, and as a result, the solubility of the oil-soluble solute increases. When the supercritical carbon dioxide passes through an extraction container containing a certain amount of a sample, the fat-soluble substance contained in the sample is extracted and released into the supercritical carbon dioxide. After extracting the fat-soluble substance, if the sample remaining in the extraction container is passed through the supercritical carbon dioxide containing a small amount of co-solvent again, components that were not extracted with pure supercritical carbon dioxide can be extracted.

The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract an active ingredient by using a supercritical carbon dioxide or a mixed fluid in which a co-solvent is additionally mixed with carbon dioxide.

As such a co-solvent, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane, and diethyl ether may be used.

Most of the extracted samples contain carbon dioxide, and since carbon dioxide is volatilized into the air at room temperature, the extract obtained by the above method can be used as a cosmetic composition, and the co-solvent can be removed with a vacuum evaporator.

In addition, the ultrasonic extraction method is an extraction method that uses energy generated by ultrasonic vibration, and the ultrasonic waves can destroy the insoluble solvent contained in the sample in the aqueous solvent, and due to the high local temperature generated at this time, Since the kinetic energy of the reactant particles is increased, sufficient energy required for the reaction is obtained, and high pressure is induced by the impact effect of ultrasonic energy to enhance the mixing effect of the material contained in the sample and the solvent, thereby increasing the extraction efficiency.

The extraction solvent that can be used in the ultrasonic extraction method may be one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane, and diethyl ether. The extracted sample may be vacuum filtered to recover the filtrate, then removed with a vacuum evaporator, and freeze-dried to obtain an extract through a conventional extract preparation method.

The lotus leaf extract according to the present invention also includes an extract that has undergone a fermentation process, and the fermented extract of lotus leaf can be prepared as follows. After finely crushing the lotus plant to about 100 to 500 mesh, 1 to 50 g/L of conventional microorganism culture solution is added, and microorganisms such as yeast strains or lactic acid bacteria are added in an amount of 10,000 to 100,000 cfu/L. The culture temperature is 30~37℃ under normal microbial culture conditions. The pH is 5-7, and the culture is carried out for about 5-10 days under aerobic or usually anaerobic conditions. Then, it can be obtained through aging and filtration.

According to the present invention, the lotus leaf extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, and more preferably, the lotus leaf extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition. It is characterized. When the content of the extract is less than 0.0001% by weight, the skin improvement effect does not appear, and when it exceeds 30.0% by weight, the degree of increase in the skin improvement effect due to the increase in content is insignificant. Not even an enemy.

In the present invention, the lotus leaf extract may be obtained by extracting from various organs or parts of lotus plant (eg, leaves, flowers, roots, fruits, etc.), and preferably may be an extract obtained from outpost.

The cosmetic composition of the present invention comprising a lotus leaf extract having various functions has excellent collagen synthesis enhancing effect, MMP-1 production inhibition effect, inflammatory cytokine expression suppression effect, antioxidant effect, skin wrinkle improvement effect, skin elasticity improvement effect, It is characterized by having skin whitening effect, melanin production inhibitory effect, moisturizing effect, and skin aging prevention effect.

Ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the active ingredients as an active ingredient. For example, conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances It may include an adjuvant, and a carrier.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , Oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream and spray may be formulated, but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

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

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol aliphatic ester, polyethyleneglycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspending agents such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like can be used.

When the formulation of the present invention is a powder or 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, additionally chlorofluorohydrocarbon, propane / May contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a surfactant containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

When the cosmetic composition of the present invention is a soap, a cleansing formulation containing a surfactant, or a cleansing formulation containing no surfactant, it may be applied to the skin and then wiped off, removed, or washed with water. As a specific example, the soap is liquid soap, powder soap, solid soap and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel and cleansing pack, and the cleansing formulation without surfactant is a cleansing cream , Cleansing lotion, cleansing water, and cleansing gel, but is not limited thereto.

On the other hand, the cosmetic method of the present invention refers to all cosmetic methods using the cosmetic composition of the present invention. That is, all methods known in the art using the cosmetic composition belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in duplicate, or may be used in conjunction with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention may be used according to a conventional method of use, and the number of times of use thereof may be varied according to a user's skin condition or taste.

When performing the cosmetic method using the cosmetic composition containing the lotus leaf extract of the present invention, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, melanin production inhibitory effect, inflammatory cytokine expression inhibitory effect, antioxidant effect, moisturizing effect, elasticity Improvement effect and skin wrinkle improvement effect can be obtained.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Manufacturing example One: Lotus leaf Solvent extract preparation

Shredded and shaded outpost of lotus leaf was extracted with reflux 3 times for 5 hours each with 70% (V/V) ethanol aqueous solution, cooled, and then filtered through Whatman #4 filter paper. The filtered extract was concentrated under reduced pressure at 50° C. or lower and freeze-dried.

Manufacturing example 2 to 3: Lotus leaf Supercritical Fluid extract and ultrasonic extract manufacturing

Chopped and shaded outpost of lotus leaf is extracted with a conventional supercritical extraction method (extracted using ethanol as a co-solvent in a supercritical state under a pressure of about 300 atmospheres at a temperature of about 60°C) and a supercritical extract (Preparation Example 2) ). An extract (Preparation Example 3) was obtained through extraction using ultrasonic waves (extracted at 30° C. for about 2 hours at an intensity of 25 KHz using a Supersonic ultrasonic device). However, in the case of Preparation Example 3, ethanol was used as a co-solvent.

Manufacturing example 4: Lotus leaf Ferment extract manufacturing

After naturally cooling the extract obtained in Preparation Example 1 to room temperature, the pH of the culture solution was adjusted to 4.0 to 5.0 to 5.0 to 7.0, and previously cultured Saccharomyces cerevisiae was added to 10 ⁷ to 10 ⁸ CFU/ After addition to the concentration of Ml, it was incubated for 20 to 110 hours at a temperature of 30°C to 50°C. To end fermentation, heat was applied at 80° C. for 15 minutes to induce loss of yeast function. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.

Experimental example One. Lotus leaf Extract of MMP -1 production inhibition effect

In Experimental Example 1, the effect of inhibiting the production of MMP-1, a collagen-degrading enzyme, was measured for the extracts of lotus bok choy obtained in Preparation Examples 1 to 4.

Fibroblasts (Korea Cell Line Bank, Republic of Korea), which are human normal skin cells, were inoculated into 48-well microplates (Nunc, Denmark) to 1×10 ⁶ cells per well, and DMEM medium (Sigma, United States of America) and 37°C conditions After culturing at for 24 hours, a serum-free DMEM medium to which the lotus bok choy extract of Preparation Examples 1 to 4 was added and a serum-free DMEM medium containing no lotus bok choy extract as a control were further cultured for 48 hours.

After incubation, the supernatant of each well was collected and the amount of newly synthesized MMP-1 (ng/ml) was measured using an MMP-1 assay kit (Amersham, United States of America), and MMP-1 was generated according to Equation 1 below. The inhibition rate (%) was calculated, and the results are shown in Table 1.

Sample name Treatment concentration MMP-1 production inhibition rate (%) Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Manufacturing Example 4 Lotus leaf extract 20ppm 24.37 22.17 21.52 23.41 50ppm 35.93 32.25 33.65 34.84 100ppm 49.74 40.81 42.50 45.58 500ppm 75.96 72.23 73.58 74.25 1000ppm 84.52 80.19 81.35 83.21 TGF-β (200nM) 76.4

As shown in the results of Table 1, it can be seen that the lotus bokcho extract of the present invention inhibits the production of MMP-1 in a concentration-dependent manner (inhibits MMP-1 activity). Even considering the difference in the concentration (200 nM) of TGF-β, a substance known to have an inhibitory effect on the production of MMP-1, and the concentration (500 ppm) of the lotus leaf extract of the present invention, the above The degree of inhibition of MMP-1 indicates that the extract of the present invention has an inhibitory effect on MMP-1 production.

Experimental example 2. Lotus leaf Collagen synthesis enhancement effect of extract

Fibroblasts, which are human normal epithelial cells, were inoculated into 48-well microplates at 1 x 10 ⁶ cells per well, and cultured in DMEM medium for 24 hours. Serum-free DMEM medium to which the lotus bok vine extract prepared according to Preparation Examples 1 to 4 was added and a serum-free DMEM medium containing no lotus bok vine extract as a control were further cultured for 48 hours. After incubation, the supernatant of each well was collected and the amount of procollagen type I C-peptide (PICP) was measured and converted into ng/ml using a collagen kit (Takara, Japan), and the amount of collagen synthesized thereby Was measured. Collagen biosynthesis increase rate (%) was calculated according to Equation 2 below, and the results are shown in Table 2.

Sample name Treatment concentration Collagen biosynthesis rate (%) Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Manufacturing Example 4 Lotus leaf extract 2.5ppm 12.8 11.9 12.0 12.3 5ppm 26.8 26.9 25.8 27.6 10ppm 45.4 41.6 42.8 44.2 25ppm 97.1 100.9 99.8 95.9 50ppm 136.4 125.5 128.3 130.2 TGF-β (200nM) 137.8

According to Table 2, it was confirmed that the collagen biosynthesis rate of the purified lotus leaf extract prepared according to Preparation Examples 1 to 4 of the present invention was increased in a concentration-dependent manner. In particular, it can be seen that the effect of the extract prepared by Preparation Example 1 is better than the extract of Preparation Examples 2 to 4.

Experimental example 3: Lotus leaf Extract of B16F1 Inhibitory effect of melanogenesis using melanocytes

In this Experimental Example 3, in order to confirm the whitening effect of the extracts obtained in Preparation Examples 1 to 4, the whitening effect was determined by looking at the degree of inhibition of melanin production on B16F1 melanin-forming cells. The B16F1 melanin-forming cells used in Experimental Example 3 are a cell strain derived from a mouse, and are cells that secrete a black pigment called melanin. During the artificial culture of these cells, the samples were treated to evaluate the degree of reduction in melanin black pigment. The B16F1 melanin-forming cells used in this experimental example were pre-sold from ATCC (American Type Culture Collection) and used.

The measurement of the inhibitory effect of melanin biosynthesis of B16F1 melanogenic cells was carried out as follows. B16F1 melanin-forming cells were dispensed into a 6-well plate at a concentration of 2×10 ⁵ per well, and after attaching the cells, the extracts of Lotus Botanical Plants according to Preparation Examples 1 to 4 were treated at a concentration that did not cause toxicity, and cultured for 72 hours. After 72 hours of culture, the cells were removed with trypsin-EDTA, the number of cells was measured, and then the cells were recovered by centrifugation. The quantification of intracellular melanin was performed by slightly modifying the method of Rotan (Cancer Res., 40: 3345-3350, 1980). After washing the cell pellet with PBS once, 1 ml of a homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After dissolving the extracted melanin by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), the absorbance of melanin was measured at 405 nm with a microplate reader, and then the melanin was quantified. The melanin production inhibition rate (%) was measured. The melanin production inhibition rate (%) of B16F1 melanin-forming cells was calculated by the following equation (3), and the results are shown in Table 3.

A: The amount of melanin in the well to which no sample was added

B: The amount of melanin in the well to which the sample was added

Sample name Treatment concentration (%) Melanin production inhibitory effect (%) Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Manufacturing Example 4 Lotus leaf
extract 0.001 28.27 23.85 24.83 27.32 0.005 42.45 38.12 39.71 40.45 0.01 57.43 53.70 55.32 56.67 0.05 82.61 76.82 79.19 80.26 Arbutin 200ppm 75.12

As can be seen from the results of Table 3, it was found that the purified lotus leaf extract prepared by Preparation Examples 1 to 4 of the present invention greatly inhibited melanin production in B16F1 melanogenic cells. In particular, it can be seen that the effect of the extract prepared by Preparation Example 1 is better than that of the extracts of Preparation Examples 2 to 4.

Experimental example 4: Lotus leaf Extract of DPPH Radical Scavenging ability analysis

In this Experimental Example 4, the antioxidant effect (free radical scavenging rate) was measured using the DPPH method in order to measure the antioxidant effect of the lotus bok vine extract obtained in Preparation Examples 1 to 4. The DPPH method uses a free radical called DPPH(2,2-Di(4-tert-octylphenyl)-1-picrylhydrazyl) free radical to measure the antioxidant effect by reducing power. The degree of decrease in absorbance due to reduction of DPPH by the test substance (perilla extract) was compared with the absorbance of the blank test solution, and the free radical scavenging rate (%) at a wavelength of 560 nm was measured. The reagent used was 2,2-Di(4-tert-octylphenyl)-1-picrylhydrazyl free radical (Aldrich Chem. Co., MW=618.76) 0.1 mM solution, 61.88 mg dissolved in methanol to make 100 ml.

As a measurement method,

① Add 0.15 ml of sample solution to 0.15 ml of 0.1 mM DPPH solution to a 96-well plate, stir quickly, and start incubation for 10 minutes at 25°C.

② After that, measure the absorbance St at 560nm.

③ A blank of the sample solution is operated in the same manner as when measuring the absorbance St, except that methanol is used instead of the 0.1mM DPPH solution, and the absorbance So is measured.

④ In the blank test, the absorbance Bt is measured in the same manner as the absorbance St, except that distilled water is used instead of the sample solution.

⑤ The absorbance Bo was measured in the same manner as when measuring the absorbance St, except that methanol was used instead of the 0.1mM DPPH solution and distilled water was used instead of the sample solution.

The result of the free radical elimination rate (%) was calculated by Equation 4, and the results are shown in Table 4 below.

St: Absorbance at 514 nm after free radical elimination of sample solution

Bt: Absorbance at 514 nm after free radical scavenging of blank test solution

So: Absorbance at 514 nm before reaction when free radicals are not added to the sample solution

Bo: Absorbance at 514 nm before reaction when free radicals are not added to the blank test solution

Sample name SC ₅₀ (%) Manufacturing Example 1 0.023 Manufacturing Example 2 0.030 Manufacturing Example 3 0.032 Manufacturing Example 4 0.035

As a result of checking SC ₅₀ , which is the concentration of the sample required to eliminate 50% of DPPH free radicals, Preparation Examples 1 to 4 showed antioxidant effects, and in particular, the antioxidant effect of Preparation Example 1 was excellent.

Experimental example 5: Lotus leaf Inhibitory Effect of Extracts on Inflammatory Cytokine Expression by UV Irradiation

This Experimental Example 5 is for evaluating the effect of inhibiting the expression of inflammatory cytokines expressed by ultraviolet irradiation of the extracts of Lotus Botanic herb obtained in Preparation Examples 1 to 4, and fibroblasts isolated from human epidermal tissue were 24- Each of 5 X 10 ^{4 was} put in a well test plate and attached for 24 hours.

Each well was washed once with PBS, and 500 μl of PBS was added to each well. After irradiating the fibroblasts with UV rays of 10 mJ/cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), remove PBS and remove the cell culture medium Medium) was added 350 μl.

Here, the lotus bokcho extract to be evaluated was treated and incubated for 5 hours. By taking 150 µl of the culture supernatant and quantifying IL-1α, the effect of inhibiting the expression of inflammatory cytokines of the lotus leaf extract was determined. The amount of IL-1α was quantified using an Enzyme-linked Immunosorbent Assay, and the inhibition rate (%) of expression of inflammatory cytokines (IL-1α) was calculated by Equation 5 below.

Bo: IL-1α production in wells without UV irradiation and sample treatment

Bt: The amount of IL-1α produced in a well that was irradiated with ultraviolet rays and not treated with a sample

St: IL-1α production amount in the well treated with UV irradiation and sample

Sample name Treatment concentration Inhibition rate of inflammatory cytokine expression (%) Manufacturing Example 1 Manufacturing Example 2 Manufacturing Example 3 Manufacturing Example 4 Lotus leaf
extract 10ppm 23.61 23.39 21.13 22.82 50ppm 41.52 37.28 39.12 40.09 100ppm 59.50 54.25 56.78 54.08 500ppm 71.12 65.92 68.77 67.23 1000ppm 79.28 72.86 74.10 74.86 Indomethacin 100 ppm 59.36

According to Table 5, it was found that the solvent extract of the lotus leaf extract (Preparation Example 1) inhibited the production of IL-1α, an inflammatory cytokine by ultraviolet rays at a concentration of 1000 ppm, by 79.28%, and the ultrasonic extract (Preparation Example 3) was It can be seen that the production of IL-1α, an inflammatory cytokine caused by ultraviolet rays, was inhibited by 74.10% at a concentration of 1000 ppm. It can be seen that the lotus bokcho extract according to the present invention exhibits superior inflammatory cytokine expression inhibition rate than indomethacin, an inflammatory cytokine expression inhibitor, at the same concentration. Therefore, it can be seen that the lotus bokcho extract of the present invention effectively protects the occurrence of inflammation caused by UV irradiation even at a low concentration.

Formulation example 1 to 4: Lotus leaf Containing extract Cosmetic Preparation of the composition

Cosmetics containing lotus bok vine extract were prepared cosmetics (Formulation Examples 1 to 4) each containing the lotus bok vine extract of Preparation Examples 1 to 4. And formulation examples For comparison of efficacy, a cosmetic composition containing glycerin and 1,3-butylene glycol as a moisturizing agent instead of the lotus leaf extract (Comparative Formulation Example 1), and a cosmetic composition containing glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 2), a cosmetic composition that does not contain any extract or moisturizing agent (Comparative Formulation Example 3) is shown in Table 6 below.

division ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation 2 Comparative Formulation Example 3 A Lotus leaf extract
(Production Example 1) 5.0 - - - - Lotus leaf extract
(Production Example 2) - 5.0 - - - Lotus leaf extract
(Production Example 3) - - 5.0 - - - Lotus leaf extract
(Production Example 4) 5.0 glycerin - - - 5.0 2.0 - 1,3-butylene glycol - - - 5.0 - - Sorbitol - - - - 3.0 - EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 B Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Floating paraffin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Trioctanoine 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl Acetate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount

Experimental example 6: Lotus leaf Moisturizing effect of extract

The clinical moisturizing effect of the cosmetic compositions of Formulation Examples 1 to 4 and Comparative Formulation Examples 1 and 3 was measured as follows. Through the questionnaire survey, 60 healthy adult men and women who feel dry skin were randomly divided into 6 groups (A, B, C, D, E, and F) of 10 people, and formulation examples in groups A, B, C, and D. The cosmetic compositions of 1 to 4, a cosmetic composition containing glycerin and 1,3-butylene glycol as a moisturizing agent instead of a lotus leaf extract in the E group (Comparative Formulation Example 1), and the F group did not contain a lotus leaf extract and a moisturizing agent. The cosmetic composition (Comparative Formulation Example 3) was applied to the face twice a day for 8 weeks, respectively. The moisturizing effect was evaluated every two weeks from the start of the actual use test, and the improvement effect after the end was evaluated using Corneometer CM 820 (Corage +Khazaka, Germany). The experimental results are shown in Table 7 below.

division Before use 2 weeks after use 4 weeks after use Formulation Example 1 25.4 38.6 49.8 Formulation Example 2 21.1 36.7 45.9 Formulation Example 3 24.1 37.5 49.7 Formulation Example 4 24.0 38.2 48.1 Comparative Formulation Example 1 22.7 35.7 45.5 Comparative Formulation Example 3 21.5 22.4 23.9

As shown in Table 7 above, it can be seen that the moisturizing effect of the cosmetics containing the lotus leaf extract of the present invention (Formulation Examples 1 to 4) is excellent even compared with the cosmetics containing other general moisturizing agents (Comparative Formulation Example 1). .

Experimental Example 7: Measurement of skin elasticity improvement effect

This Experimental Example 7 was measured on 60 women in their 30s to 40s with reduced elasticity in order to find out the effect of improving skin elasticity of the cosmetic compositions according to Formulation Examples 1 to 4 containing lotus bok choy extract. After dividing into 6 groups (A, B, C, D, E, and F) of 10 people, the cosmetic compositions of Formulation Examples 1 to 4 were added to groups A, B, C, and D, and moisturizers instead of lotus leaf extract in group E. As a cosmetic composition containing glycerin and 1,3-butylene glycol (Comparative Formulation Example 1), a cosmetic composition (Comparative Formulation Example 3) containing no moisturizing agent and lotus leaf extract in Group F was applied to the face every morning and evening. After being applied twice for 12 weeks, skin elasticity was measured after 12 weeks. The elasticity of the facial area was measured using a cutometer SEM575 (MPA 580, Courage+Khazaka GmbH, Germany), and R2 (biological elasticity), representing skin elasticity, was measured before and after application to evaluate its improvement (%). . The results are shown in Table 8 below, and the results are an average of the degree of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 28.1 Formulation Example 2 23.5 Formulation Example 3 25.8 Formulation Example 4 26.5 Comparative Formulation Example 1 5.7 Comparative Formulation Example 3 2.1

As shown in Table 8, Formulation Examples 1 to 4 containing Lotus Bokcho extract were found to have very improved skin elasticity compared to Comparative Formulation Examples 1 and 3, and among them, Formulation Example 1 in particular compared to other formulation examples. , It can be seen that it shows a high degree of improvement.

Experimental Example 8: Measurement of skin wrinkle improvement effect

In this Experimental Example 8, in order to examine the effect of improving skin elasticity of the cosmetic compositions according to Formulation Examples 1 to 4 containing lotus bok choy extract, it was measured on 60 men and women with reduced elasticity in their 30s to 40s. After dividing into 6 groups (A, B, C, D, E, and F) of 10 people, the cosmetic compositions of Formulation Examples 1 to 4 were added to groups A, B, C, and D, and moisturizers instead of lotus leaf extract in group E. As a cosmetic composition containing glycerin and 1,3-butylene glycol (Comparative Formulation Example 1), a cosmetic composition (Comparative Formulation Example 3) containing no moisturizing agent and lotus leaf extract in Group F was applied to the face every morning and evening. After being applied twice for 12 weeks, skin elasticity was measured after 12 weeks. The wrinkle improvement effect of each subject was evaluated through visual observation. The experimental results are as shown in Table 9 below.

Sample Wrinkle improvement effect Effective rate Great slightly none Formulation Example 1 48 12 3 97.8 Formulation Example 2 20 47 3 95.3 Formulation Example 3 19 48 3 95.7 Formulation Example 4 29 37 4 94.2 Comparative Formulation Example 1 2 11 57 18.6 Comparative Formulation Example 3 One 6 63 10.0

As shown in Table 9, it was found that formulation examples 1 to 4 containing lotus bok choy extract had very excellent skin wrinkle improvement effect compared to Comparative Formulation Examples 1 and 3, and among them, Formulation Example 1 was a different formulation Compared to the example, it can be seen that the effect is high.

As described above, specific parts of the present invention have been described in detail, and it is obvious that these specific techniques are only preferred embodiments for those of ordinary skill in the art, and the scope of the present invention is not limited thereto. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (8)

As a cosmetic composition for skin whitening containing lotus bok choy extract as an active ingredient,
The lotus leaf extract is (a) water, anhydrous or hydrous lower alcohol having 1-4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, dichloromethane, nucleic acids, and these It is characterized in that it is extracted by an extraction method selected from the group consisting of a solvent extraction method using an extraction solvent selected from the group consisting of a mixture of, (b) a supercritical extraction method, (c) an ultrasonic extraction method, and (d) a fermentation extraction method using microorganisms. , Cosmetic composition for skin whitening. The method of claim 1,
The lotus leaf extract is a cosmetic composition for skin whitening, characterized in that it contains 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. delete delete delete delete delete delete