KR102107196B1 - Cosmetic composition containing adoxa moschatellina as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition containing yeonbokcho extract as an active ingredient and having at least one effect of skin wrinkle improvement, skin whitening, antioxidant, moisturizing and skin elasticity improvement effects.

Description

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

The present invention relates to a cosmetic composition containing yeonbokcho extract as an active ingredient, specifically, the cosmetic composition has an 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. Consumers who feel the skin is sensitive are on the rise. In the consciousness survey of general consumers, more than 30-40% of respondents answered that they are sensitive skin, and this tends to increase. Moreover, the recent cosmetic products tend to focus on functional cosmetics (Cosmeceutical) that can give a real effect to the skin, and 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 from fine dust, fumes, and ultraviolet rays (UV) becomes serious, and the amount of ultraviolet radiation increases due to the destruction of the ozone layer, drying of the living space, personal allergies, stress, and chemical harmful substances. Due to the increase, the skin is easily damaged, and as a result, various irritants are generated, so that the skin is easily reddened or sensitive, and severely causes skin trouble.

In general, various raw materials are used in the process of making cosmetics, and typically, from 10 to 50 kinds of raw materials are used. Although these ingredients have been proven to be safe through skin irritation tests, they can cause irritation of the skin such as inflammation, itching and allergies by being applied to the product. Skin irritations that may occur due to the use of cosmetics include irritation and erythema and edema, as well as irritation such as itching, stinging and burning. Contact dermatitis can be classified into irritating 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, many cosmetics using natural products have been developed to reduce skin irritation caused by various chemical substances. Natural ingredients have relatively few side effects on skin, and recently, as the consumer's response to cosmetics using natural ingredients has increased, the development value of cosmetics has increased.

Yeonbokcho used in the present invention is scientifically Adoxa Called moschatellina L., it is a perennial plant belonging to the dicotyledonous plant river. The flower color is yellow and green, inhabiting the temperate and cold regions of the northern hemisphere, and is characterized by having a growing environment that grows to 8 to 17 cm in height at the foot of the mountain. In addition, yeonbokcho is known to contain various flavonoids.

Recently, strong antioxidant effects of flavonoids and polyphenols abundant in plant extracts such as lotus root have been studied. In addition, these vegetable flavonoid derivative components are used for various purposes such as edible, antioxidant, skin whitening, and are also used in the fields of medicines, cosmetics, and functional foods in Korea.

Under this background, the present inventors have completed the present invention by preparing an extract of lotus root that is excellent in alleviating irritation while studying a cosmetic composition 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 caused by various raw materials contained in cosmetics.

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

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

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

The present inventors sought to secure a material that is stable to relieve skin irritation and secures safety that does not cause skin problems such as irritation and seizures when applied to the skin.

Preferably, the yeonbokcho extract has a skin wrinkle improving effect, skin whitening effect, antioxidant effect, skin moisturizing effect and skin elasticity improving effect.

In addition, preferably, the yeonbokcho extract is characterized in that it contains 0.0001 ~ 30.0% by weight relative to the total weight of the cosmetic composition.

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

The Yeonbokcho extract according to the present invention has an inhibitory effect on MMP-1 production (Experimental Example 1), an effect of enhancing collagen synthesis (Experimental Example 2), an inhibitory effect on melanin production (Experimental Example 3), an antioxidant effect (Experimental Example 4), and inflammatory cytos It has an effect of inhibiting kinase expression (Experimental Example 5), a moisturizing effect (Experimental Example 6), a skin elasticity improving effect (Experimental Example 7), and a skin wrinkle improvement effect (Experimental Example 8).

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

All technical terms used in the present invention, unless defined otherwise, have the following definitions and conform to the meaning as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although a preferred method or sample is described herein, similar or equivalent ones are included in the scope of the present invention.

The term "about" refers to 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, for reference amount, level, value, number, frequency, percent, dimension, size, amount, weight or length It means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by 4, 3, 2 or 1%.

Throughout this specification, the terms “comprises” and “comprising”, unless the context requires otherwise, include the steps or components presented, or groups of steps or components, but any other steps or components, or It should be understood that it implies that a group of steps or components is not excluded.

The present inventors researched the applicability as a cosmetic composition containing lotus root extract among various natural products and conducted various experiments to measure skin wrinkle improvement effect, skin whitening effect, antioxidant effect, skin moisturizing effect and skin elasticity improvement effect as cosmetic products. It was discovered that efficacy can be expected. Therefore, the cosmetic composition of the present invention composed of the above components is suitable for a cosmetic composition for improving skin wrinkles, skin whitening effect, antioxidant effect, skin moisturizing effect and skin elasticity improving effect.

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

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

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

On the other hand, it is apparent to those skilled in the art that the extract of yeonbokcho of the present invention can obtain an extract exhibiting substantially the same effect by using not only the aforementioned extraction solvent, but also other extraction solvents.

In addition, the yeonbokcho extract of the present invention includes not only extracts by the above-described extraction solvent, but also extracts that have undergone conventional purification and fermentation processes. For example, reduced pressure by carbon dioxide, extraction by supercritical extraction method by high temperature, extraction by ultrasonic extraction method, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity) Active fractions obtained through various purification and extraction methods additionally performed, such as separation by sex) or extraction by fermentation products using natural 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 cold filtration at room temperature, filtered by heating, and further obtained by concentrating or freeze-drying the solvent under reduced pressure.

The decompression method by carbon dioxide and the extraction method by the supercritical extraction method by high temperature refer to a supercritical fluid extraction method. In general, a supercritical fluid is a liquid that a gas has when it reaches a critical point under high temperature and high pressure conditions, and It has the properties of a gas and has a chemically similar polarity to a non-polar solvent, and due to these properties, supercritical fluids are used for the extraction of fat-soluble materials (J. Chromatogr. A. 1998; 479: 200-205).

Carbon dioxide becomes a supercritical fluid having both liquid and gaseous properties through the process of pressure and temperature reaching a critical point through the operation of a supercritical fluid device, and as a result, solubility in fat-soluble solute increases. When supercritical carbon dioxide passes through an extraction container containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from supercritical carbon dioxide. After extracting the fat-soluble substance, passing the supercritical carbon dioxide containing a small amount of co-solvent again through the sample remaining in the extraction container may allow components not extracted with pure supercritical carbon dioxide to be extracted.

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

The co-solvent 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.

Most of the extracted sample contains carbon dioxide, and since carbon dioxide volatilizes 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 by a vacuum evaporator.

In addition, the ultrasonic extraction method is an extraction method using energy generated by ultrasonic vibration, and ultrasonic waves can destroy the insoluble solvent contained in a sample in a water-soluble solvent, and are located around due to the high local temperature generated at this time. Since the kinetic energy of the reactant particles is increased, sufficient energy necessary for the reaction is obtained, and high pressure is induced by the impact of ultrasonic energy, thereby increasing the mixing effect of the substance and the solvent contained in the sample, thereby increasing 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 is vacuum filtered to recover the filtrate, and then removed with a vacuum evaporator, and an extract can be obtained through a conventional extract preparation method of freeze drying.

Yeonbokcho extract according to the present invention also includes an extract that has undergone a fermentation process, and the fermentation extract of Yeonbokcho can be prepared as follows. After finely crushing yeonbokcho about 100 to 500 mesh, 1 to 50 g / L of a conventional microbial 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 cultured under normal microbial culture conditions of 30 ~ 37 ℃. The pH is 5 to 7 and cultured for about 5 to 10 days under aerobic or normal anaerobic conditions. It can then be obtained through aging and filtration.

According to the present invention, the yeonbokcho extract is contained 0.0001 ~ 30.0% by weight relative to the total weight of the cosmetic composition, more preferably, the yeonbokcho extract is contained 0.01 to 10% by weight relative to the total weight of the cosmetic composition It is characterized by. When the content of the extract is less than 0.0001% by weight, the skin improvement effect is not exhibited, and when it exceeds 30.0% by weight, the degree of improvement in the skin improvement effect on the increase in content is negligible, and there is a problem in the safety and stability of the formulation and is economical. Not even enemies

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

The cosmetic composition of the present invention comprising the lotus extract having various functions has excellent collagen synthesis promoting effect, MMP-1 production inhibitory effect, inflammatory cytokine expression inhibitory effect, antioxidant effect, skin wrinkle improvement effect, skin elasticity improvement effect, It has a skin whitening effect, a melanin production inhibitory effect, a moisturizing effect, and an anti-aging effect.

Ingredients included in the cosmetic composition of the present invention, as an active ingredient, may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances Adjuvants, and carriers.

The cosmetic composition of the present invention can 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, but may be formulated, but is not limited thereto. More specifically, it can be prepared in the form of a flexible lotion, nutrition lotion, nutrition 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, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. Can be.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or 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-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester.

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, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

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, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether.

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

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it may be wiped, peeled off, or washed with water after being applied to the skin. As a specific example, the soap is liquid soap, powdered soap, solid soap and oil soap, and the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, a cleansing towel and a cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream , Cleansing lotion, cleansing water and cleansing gel, but are not limited thereto.

Meanwhile, 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 a 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 combination 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 may be varied according to a user's skin condition or taste.

When performing a cosmetic method using a cosmetic composition comprising the yeonbokcho extract of the present invention, MMP-1 production inhibitory effect, collagen synthesis promoting 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 intended to illustrate the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Preparation Example 1: Preparation of yeonbokcho solvent extract

After dividing, the outpost of the shaded yeonbokcho was refluxed 3 times for 5 hours with 70% (V / V) ethanol aqueous solution, cooled, and then filtered with Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure at 50 ° C or lower and freeze-dried.

Preparation Examples 2 to 3: Preparation of supercritical fluid extract and ultrasonic extract

The supercritical extract (Preparation Example 2) is extracted by slicing and shading the outpost of yeonbok bokcho using a conventional supercritical extraction method (extraction using ethanol as a cosolvent in a supercritical state under a pressure of about 300 atm at a temperature of about 60 ° C). ). 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.

Preparation Example 4: Preparation of yeonbokcho fermentation extract

After naturally cooling the extract obtained in Preparation Example 1 to room temperature, the culture ph was adjusted to 4.0 to 5.0 to 5.0 to 7.0, and the pre-cultured Saccharomyces cerevisiae was 10 ⁷ to 10 ⁸ CFU / After adding to a concentration of Ml, the cells were cultured at a temperature of 30 ° C to 50 ° C for 20 hours to 110 hours. In order to end the fermentation, heat was applied at 80 ° C for 15 minutes to induce yeast loss. The precipitate was removed using a centrifuge and supernatant was collected or filtered supernatant was used.

Experimental Example  One: Bokbokcho  Of extract MMP -1 suppression effect

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

Human normal skin cells, fibroblasts (Korea Cell Line Bank, Korea), were inoculated into 48-well microplates (Nunc, Denmark) to be 1 × 10 ⁶ cells per well, and conditions of DMEM medium (Sigma, United States) and 37 ° C. After incubation for 24 hours, serum-free DMEM medium to which the yeonbokcho extracts of Preparation Examples 1 to 4 were added and serum-free DMEM medium containing no bokbokcho extract as a control were further cultured for 48 hours.

After incubation, the supernatant of each well was collected to measure the amount of newly synthesized MMP-1 (ng / ml) using the 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 (%) Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Yeonbokcho 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 yeonbokcho extract of the present invention inhibits the production of MMP-1 in a concentration-dependent manner (inhibits MMP-1 activity). Considering the difference between the concentration of TGF-β (200 nM), which is a substance known to have an inhibitory effect on MMP-1 production, and the concentration (500 ppm) of the Yeonbokcho extract of the present invention, it is the same as the above at the level of extracts in which various types of substances are mixed. The degree of inhibition of MMP-1 indicates that the extract of yeonbokcho of the present invention has an inhibitory effect on MMP-1 production.

Experimental Example  2: Bokbokcho  Effect of extract to enhance collagen synthesis

Human normal epithelial cells, fibroblasts, were seeded in 48-well microplates to be 1 x 10 ⁶ cells per well, and cultured in DMEM medium for 24 hours. Serum-free DMEM medium to which the yeonbokcho extracts prepared in Preparation Examples 1 to 4 were added and serum-free DMEM medium containing no bokbokcho extract as a control were further cultured for 48 hours. After cultivation, the supernatant of each well was collected, and the amount of procollagen type I C-peptide (PICP) was measured using a collagen kit (Takara, Japan), and converted to ng / ml. Was measured. The increase rate (%) of collagen biosynthesis was calculated according to Equation 2 below, and the results are shown in Table 2.

Sample name Treatment concentration Collagen biosynthesis rate (%) Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Yeonbokcho 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 yeonbokcho extracts prepared by Preparation Examples 1 to 4 of the present invention increased in a concentration-dependent manner. In particular, it can be seen that the extract prepared by Preparation Example 1 has a better effect than the extracts of Preparation Examples 2 to 4.

Experimental Example  3: Bokbokcho  Of extract B16F1  Inhibiting effect of melanin production using melanocytes

This Experimental Example 3 is to determine the whitening effect on the B16F1 melanin-forming cells to determine the whitening effect to determine the whitening effect of the yeonbokcho extract obtained in Preparation Examples 1 to 4 above. The B16F1 melanin-forming cells used in Experimental Example 3 are cell strains derived from mice, and are cells secreting a black pigment called melanin. During the artificial culture of these cells, samples were treated to compare and evaluate the degree of melanin black pigment reduction. The B16F1 melanin-forming cells used in this experimental example were used for pre-sale from the American Type Culture Collection (ATCC).

Measurement of melanin biosynthesis inhibitory effect of B16F1 melanocytes was performed as follows. B16F1 melanin-forming cells were dispensed at a concentration of 2 X 10 ⁵ per well in a 6-well plate, and the cells were attached to the cells, followed by treatment with the yeonbokcho extracts according to Preparation Examples 1 to 4 at a concentration not causing toxicity, and cultured for 72 hours. After incubation for 72 hours, the cells were separated with trypsin-EDTA, and the number of cells was measured, followed by centrifugation to recover the cells. Quantification of melanin in cells was performed by slightly modifying the method of Rotan (Cancer Res., 40: 3345-3350, 1980). After washing the cell pellet once with PBS, 1 ml of 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 adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes) to dissolve the extracted melanin, measure the absorbance of melanin at 405 nm with a microplate reader, and then quantify melanin to quantify the sample The rate of melanin production inhibition (%) was measured. The inhibition rate (%) of melanin production in B16F1 melanocytes was calculated by Equation 3 below, and the results are shown in Table 3.

A: The amount of melanin in the well without the sample added

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

Sample name Treatment concentration (%) Melanin production inhibitory effect (%) Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Bokbokcho
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 in Table 3, it was found that the purified yeonbokcho extracts prepared by Preparation Examples 1 to 4 of the present invention significantly inhibit melanin production in B16F1 melanin-forming cells. In particular, it can be confirmed that the extract produced by Preparation Example 1 is more effective than the extracts of Preparation Examples 2 to 4, among others.

Experimental Example 4: Analysis of DPPH radical scavenging activity of Yeonbokcho extract

In Experimental Example 4, the antioxidant effect (free radical scavenging rate) was measured using the DPPH method to measure the antioxidant effect of the Yeonbokcho extracts obtained in Preparation Examples 1 to 4. The DPPH method measures the antioxidant effect by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The free radical scavenging rate (%) is measured at a wavelength of 560 nm by comparing the degree of decrease in absorbance due to the reduction of DPPH by the test substance (yeonbokcho extract). The reagent used was a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76). 61.88 mg was 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 in a 96-well plate, stir quickly, and start incubation for 10 minutes at 25 ° C.

② After that, absorbance St at 560 nm is measured.

③ Blank of the sample solution is measured in the same manner as when measuring the absorbance St, except that methanol is used instead of the 0.1 mM DPPH solution to measure the absorbance So.

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

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

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

St: absorbance at 514 nm after free radical scavenging of sample solution

Bt: Absorbance at 514 nm after free radical scavenging of the 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 (%) Preparation Example 1 0.023 Preparation Example 2 0.030 Preparation Example 3 0.032 Preparation Example 4 0.035

As a result of confirming SC ₅₀ , which is the concentration of the sample required to eliminate DPPH free radicals by 50%, Preparation Examples 1 to 4 showed an antioxidant effect, and particularly, the antioxidant effect of Preparation Example 1 was excellent.

Experimental Example  5: Bokbokcho  Inhibitory effect of inflammatory cytokine expression by UV irradiation of extract

This Experimental Example 5 is for evaluating the inhibitory effect of inflammatory cytokine expression expressed by ultraviolet irradiation of the lotus root extracts obtained in Preparation Examples 1 to 4, and the fibroblasts isolated from human epidermal tissues are 24- 5 x 10 ⁴ pieces were placed in well test plates and attached for 24 hours.

Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of UV light using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then PBS was removed and FBS was not added to the cell culture medium (DMEM. Unused medium) 350 μl was added.

After treating the extract of yeonbokcho to be evaluated, it was cultured for 5 hours. By taking 150 µl of the culture supernatant and quantifying IL-1α, the effect of suppressing the inflammatory cytokine expression of the lotus root extract was determined. The amount of IL-1α was quantified using an enzyme immunoassay (Enzyme-linked Immunosorbent Assay), and the expression inhibition rate (%) of inflammatory cytokines (IL-1α) was calculated by Equation 5 below.

Bo: IL-1α production in wells that were not sampled without UV irradiation

Bt: IL-1α production in wells that were not irradiated with ultraviolet light and treated with samples

St: IL-1α production in wells treated with UV irradiation and samples

Sample name Treatment concentration Inhibition rate of inflammatory cytokine expression (%) Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Bokbokcho
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 Yeonbokcho extract (Preparation Example 1) inhibited the production of inflammatory cytokines IL-1α by UV at a concentration of 1000 ppm by 79.28%, and the ultrasonic extract (Preparation Example 3) It can be seen that the production of IL-1α, an inflammatory cytokine caused by ultraviolet light, at a concentration of 1000 ppm is inhibited by 74.10%. It can be seen that the Yeonbokcho extract according to the present invention exhibits a superior inflammatory cytokine expression inhibition rate than indomethacin, an inflammatory cytokine expression inhibitor, at the same concentration. Therefore, it can be seen that the yeonbokcho extract of the present invention effectively protects inflammation caused by ultraviolet irradiation even at a low concentration.

Formulation example  1 to 4: Bokbokcho  Containing extracts Cosmetic  Preparation of the composition

Cosmetics containing the yeonbokcho extract were prepared cosmetics (formulation examples 1 to 4) each containing the yeonbokcho extract of Preparation Examples 1 to 4. And for comparison of formulation example efficacy, a cosmetic composition comprising glycerin and 1,3-butylene glycol as a moisturizer instead of lotus root extract (Comparative Formulation Example 1), a cosmetic composition comprising glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 2), a cosmetic composition (Comparative Formulation Example 3) that does not contain any extract or moisturizing agent 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 Yeonbokcho extract
(Production Example 1) 5.0 - - - - Yeonbokcho extract
(Production Example 2) - 5.0 - - - Yeonbokcho extract
(Production Example 3)  - - 5.0  -  -  - Yeonbokcho 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 Microcrystal 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 Trioctanoin 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 Proper Proper Proper Proper Proper Proper Proper

Experimental Example  6: Bokbokcho  Moisturizing effect of extract

The clinical moisturizing effect was measured as follows for the cosmetic compositions of Formulation Examples 1 to 4 and Comparative Formulation Examples 1 and 3. After the survey was conducted, 60 healthy men and women who felt dry skin were randomly divided into 6 groups (A, B, C, D, E, and F) of 10 people, and the formulation examples were in A, B, C, and D groups. Cosmetic compositions of 1 to 4, E group is a cosmetic composition comprising glycerin and 1,3-butylene glycol as a moisturizing agent instead of yeonbokcho extract (Comparative Formulation Example 1), F group does not contain bokbokcho extract and moisturizing agent The non-cosmetic composition (Comparative Form 3) was applied twice a day to the face for 8 weeks. The moisturizing effect was evaluated by using Corneometer CM 820 (Corage + Khazaka, Germany) for the improvement effect after every two weeks from the start of the practical use test and after completion. 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, the moisturizing effect of the cosmetic composition containing the Yeonbokcho extract of the present invention (Formulation Examples 1 to 4) is excellent even when compared to a cosmetic composition containing other moisturizing agents (Comparative Formulation Example 1). .

Experimental Example  7: Measure skin elasticity improvement effect

In order to investigate the effect of improving the skin elasticity of the cosmetic compositions according to Formulation Examples 1 to 4 containing the extract of lotus root vinegar, this experimental example 7 was measured for 60 women with decreased elasticity in their 30s and 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 the A, B, C, and D groups, and the moisturizing agent was used instead of the lotus root extract in the E group. As a cosmetic composition comprising glycerin and 1,3-butylene glycol (Comparative Formulation Example 1), the group F contains a cosmetic composition (Comparative Formulation Example 3) containing no lotus extract and moisturizer on the face every morning and evening After applying 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 the R2 (biological elasticity), which indicates skin elasticity, was measured before and after application to evaluate the improvement (%). . The results are shown in Table 8 below, and the results are average values 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, it was found that Formulation Examples 1 to 4 containing the extract of Yeonbokcho were significantly improved in 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 order to investigate the effect of improving the skin elasticity of the cosmetic compositions according to Formulation Examples 1 to 4 containing the extract of lotus root vinegar, this experimental example 8 was measured for 60 men and women with reduced elasticity in their 30s and 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 the A, B, C, and D groups, and the moisturizing agent was used instead of the lotus extract in the E group. As a cosmetic composition comprising glycerin and 1,3-butylene glycol (Comparative Formulation Example 1), group F contains a cosmetic composition (Comparative Formulation Example 3) that does not contain yeonbokcho extract and a moisturizing agent every morning and evening on the face. After applying 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 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 the yeonbokcho extract have a very excellent wrinkle improvement effect on the skin compared to Comparative Formulation Examples 1 and 3, and among them, especially formulation example 1 is another formulation It turns out that it shows a high effect compared with an example.

Since the specific parts of the present invention have been described in detail above, it is clear that for those skilled in the art, this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (3)

Cosmetic composition for moisturizing the skin, characterized in that it contains an extract of bokbokcho ( Adoxa moschatellina L.) as an active ingredient, and the bokbokcho extract is contained in an amount of 0.0001 to 30.0% by weight relative to the total weight of the cosmetic composition. The method of claim 1, wherein the yeonbokcho extract is (a) water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, Solvent extraction method using an extraction solvent selected from the group consisting of dichloromethane, nucleic acid and mixtures thereof, (b) supercritical extraction method, (c) ultrasonic extraction method and (d) fermentation extraction method using microorganisms. Extracted, cosmetic composition for moisturizing the skin. delete