KR20160004593A - Cosmetic Composition Comprising Extract of Pittosporum tobira as Active Ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of Pittosporum tobira as an active ingredient, comprising 0.0001-30.0 wt% of an extract of Pittosporum tobira as an active ingredient of total weight of the cosmetic composition. Provided in the present invention is a cosmetic composition having an MMP-1 generation inhibiting effect, a bio-collagenesis effect, a melanin generation inhibiting effect, a preservative effect, an anti-oxidation effect, a skin wrinkle reduction effect, a skin elasticity improving and moisturizing effects, and skin anti-aging effect by comprising the extract of Pittosporum tobira.

Description

[0002] Cosmetic Composition Comprising Extract of Pittosporum to Bira as Active Ingredient [

The present invention relates to a cosmetic composition comprising a walnut extract as an active ingredient.

A variety of physical and chemical changes occur in human skin during the aging process. The cause of this phenomenon is divided into intrinsic aging and photo-aging. Aging can be caused by activation of free radicals due to ultraviolet rays, stress, disease states, environmental factors, wounds, and aging. When such conditions become severe, the antioxidant defense network existing in the living body is destroyed, Thereby promoting adult diseases and aging.

Specifically, lipids, proteins, polysaccharides and nucleic acids, which are major components of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. In particular, the oxidation of proteins can cause collagen (collagen), hyaluronic acid, elastin, proteoglycan, fibronectin, and the like that are severely hyperinflammatory reactions and skin elasticity If this gets worse, DNA mutations can lead to mutations, cancer, and immune deficiency.

Therefore, it is possible to protect the cell membrane by eliminating the free radicals generated by the free radicals, ultraviolet irradiation, and inflammation reactions that occur during metabolism of the body. In addition, already damaged cells can be regenerated by active metabolism to proliferate the cells so that the skin can quickly recover and maintain healthy skin.

Aging involves not only these free radicals, but also enzyme called matrix metalloproteinase (MMP), a collagenase. That is, synthesis and degradation of extracellular matrix such as collagen in vivo are appropriately controlled, but collagen synthesis is reduced as aging progresses, and expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, is promoted, And the wrinkles are formed. In addition, these degrading enzymes may be activated by ultraviolet irradiation.

Therefore, there is a demand for development of a substance capable of modulating MMP expression induced in the cell or inhibiting its activity. Until now, the raw materials used as cosmetic materials mostly inhibited only the enzyme activity. Therefore, we tried to control the amount and activity of MMP expression induced by intracellular natural aging and photoaging.

On the other hand, skin changes due to pigment deposition occur. The factors that determine skin color are basically the partial or total pigmentation such as stain, freckle and tanning due to ultraviolet exposure, and the distribution of acne and scars and keratin Blood circulation, stress, and health. The most important factor among these factors is pigmentation.

Melanin, melanoid, carotene and hemoglobin are the most important factors affecting skin color. Among them, melanin is the most important factor affecting the biosynthesis of melanin.

Melanin absorbs or scatters ultraviolet light and plays a major role in preventing damage to the skin from ultraviolet rays. It does not have a specific maximum absorption wavelength and absorbs light in all areas. In addition, it has excellent ability to remove reactive oxygen species, sometimes melanin itself generates active oxygen, and other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself shows free radical properties Pray.

The biosynthesis of melanin begins with the conversion of tyrosine, an amino acid, into melanosomes of melanocyte-forming melanosomes by tyrosinase and conversion to dihydroxy phenylalanine, followed by a series of oxidation processes to form pheomelanin, eumelanin.

This biosynthesis process is carried out in a special form of melanocyte in the brown intracellular organelle. The melanomas, including melanin granules, migrate from the nucleus to the tip of the dendritic process, and migrate into the cytoplasm by phagocytosis of keratinocytes. It accumulates around the nucleus.

The synthesis of melanin, the number of melanosomes, and the migration to keratinocytes in the periphery are partially affected by hormones and ultraviolet rays, and are primarily affected by genetics. In addition, it is known that interleukins, prostaglandins, histamines, and the like are involved in the expression of tyrosinase and cytokine, which is an intracellular regulator involved in the synthesis and transmission of melanin, and metal ions such as copper, zinc and iron.

(Japanese Patent Laid-open Publication No. 4-9320), hydroquinone (Japanese Patent Laid-Open No. 192062), kojic acid (Japanese Laid-Open Patent No. 56-7710), arbutin Have a tyrosinase-inhibiting activity and are used as whitening cosmetics. However, since these substances are low in stability in cosmetic formulations, their use is restricted due to decomposition and coloring, generation of odor, efficacy at a living body level, uncertainty of effects, and safety problems. The above substances have proved their tyrosinase inhibitory effect, but their effect is low in experiments similar to actual biomolecular level. Therefore, it is required to evaluate the inhibitory effect by the melanoma cell culture similar to the biological level. Hydroquinone is also defined as a carcinogenic substance and its use in cosmetics is prohibited or restricted. Kojic acid and ascorbic acid are very unstable substances. When a cosmetic containing a small amount of the above components is kept at room temperature for several weeks, browning occurs.

In order to solve these various problems, recently, many cosmetics using natural products have been developed to reduce skin irritation caused by various chemical substances and the like. Natural materials are not only less harmful to the skin, but also have increased their value as a raw material for cosmetics due to the recent popularity of cosmetics using natural materials.

In cosmetics, various components such as water, oil, surfactant, protein, and plant extract are mixed and microorganisms may multiply when the microorganisms are introduced under proper environment. Microbial contamination of cosmetics can degrade product quality and seriously harm consumers' health. In order to prevent such contamination, cosmetics use an appropriate amount of preservative such as paraoxybenzoic acid ester, phenoxyethanol, imidazolidinyl urea, and methyl isothiazolinone. However, these synthetic preservatives have been reported to cause skin irritation such as irritant contact dermatitis and allergic dermatitis accompanied by erythema, edema, swelling, and palsy, and thus there is a growing interest in natural preservatives using natural products.

As a result of studying the applicability of various natural substances not yet known to the cosmetics, the present inventors have selected extracts of Yongam and a plant of Pinus densiflora to produce extracts, which have been shown to inhibit MMP-1 production, antioxidant effect, collagen biosynthesis The present inventors have found that the efficacy can be expected as a cosmetic product because of its excellent efficacy as a result of measuring melanin production inhibitory effect, preservative effect, moisturizing effect, skin wrinkle improving effect, skin elasticity improving effect and skin antiaging effect.

It is an object of the present invention to provide a method of inhibiting MMP-1 production, collagen biosynthesis effect, melanin production inhibitory effect, antioxidative effect, preservative effect, moisturizing effect, skin wrinkle improvement effect, skin elasticity improvement effect, And to provide a cosmetic composition showing the same.

It is also an object of the present invention to provide a cosmetic composition which exhibits excellent effects by preparing a supercritical extract or an ultrasonic extract to maximize the efficacy of a walnut extract.

Accordingly, the object of the present invention as described above is achieved by a cosmetic composition comprising an extract of Fenugreek as an active ingredient.

Preferably, the cosmetic composition may be for improving skin wrinkles.

Further, preferably, the cosmetic composition may be used for skin whitening.

Further, preferably, the cosmetic composition may be for antioxidation.

Further, preferably, the cosmetic composition may be for skin moisturizing.

Preferably, a cosmetic composition comprising a walnut extract as a preservative component can be provided.

Preferably, the cosmetic composition is characterized in that it is for preventing skin aging.

Further, preferably, the walnut extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

Preferably, the walnut extract is a mixture of (a) water, an anhydrous or a lower alcohol having 1-4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, (B) a supercritical extraction method, or (c) an ultrasonic extraction method, wherein the solvent extraction method is an extraction method using an extraction solvent selected from the group consisting of methanol, hexane, and mixtures thereof.

Preferably, the cosmetic composition is used for the preparation of a cosmetic composition which has an effect of inhibiting MMP-1 production, collagen biosynthesis effect, melanin formation inhibitory effect, preservative effect, antioxidative effect, moisturizing effect, skin wrinkle improving effect, skin elasticity improving effect, And to provide a cosmetic composition showing the same.

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 Lt; RTI ID = 0.0 > of: < / RTI >

The extracts of the walnut trees according to the present invention exhibited the effects of inhibiting MMP-1 production (Experimental Example 1), collagen synthesis enhancing effect (Experimental Example 2), melanin formation inhibitory effect (Experimental Example 3), antioxidative effect (Experimental Example 4) (Experimental Example 5), preservative effect (Experimental Example 6), wrinkle improving effect (Experimental Example 7) and skin elasticity improving effect (Experimental Example 8).

Further, according to the present invention, in order to maximize the efficacy of the walnut extract, the effect of suppressing MMP-1 production, the effect of promoting collagen synthesis, the effect of inhibiting melanin production, and the skin elasticity improvement are further enhanced through ultrasonic extraction or supercritical extraction

In the cosmetic composition of the present invention, the pinna (anthocyanin, thawing powder, sea tangle, clover) used as an active ingredient is Pittosporum tobira . The fir tree grows at the foot of the beach, has no hair on the branches, and the bark is dark brown. The stem is divided into several branches in the base, and the water tube is semicircular. Leaves are alternate but they are gathered at the ends of the branches and are thick. The front surface of the leaf is dark green and has a long, long egg shape. It is 4 ~ 10cm long and 2 ~ 3cm wide. The flower of the Pinus is bisexual, and it hangs on the end of new branch in May ~ June with gunshot inflorescence. Petals, calyx, and stamen are 5 each. The fruit is a capsule, round or wide oval, about 1.2cm long, and split into three pieces in October, resulting in a red seed. It is distributed in Korea (Jeonnam, Jeonbuk, Gyeongnam), Japan, Taiwan and China.

Pinna contains essential oils and its main constituents are Limonen, Pinen and Sesquiterpen. It is known that it has the effect of lowering blood pressure, helping circulation of blood, and promoting boils. The diseases include hypertension, arteriosclerosis, bony edema and sickness. It is also used as a remedy for eczema and boils.

In the present invention, the walnut extract is obtained by extracting from various organs or parts (e.g., leaves, branches, flowers, roots, stems, etc.) of the fir tree. More preferably, an extract obtained from a plant or root can be used, and the extract obtained from a plant can be most preferably used.

In addition, the fruit extract of the present invention can be prepared by using conventional solvents known in the art, that is, under the conditions of ordinary temperature and pressure, but preferably (a) water, Propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane (such as methanol, ethanol, (B) solvent extraction using a solvent selected from the group consisting of chloroform, hexane and mixtures thereof; (b) supercritical extraction with carbon dioxide and supercritical extraction with high temperature; or (c) extraction with ultrasonic extraction.

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

The Fennel Extract may be used in a variety of extraction solvents, such as water, anhydrous or lower alcohols having 1-4 carbon atoms (such as methanol, ethanol, propanol or butanol), propylene glycol, 1,3-butylene glycol, glycerin, Can be obtained using at least one extraction solvent selected from the group consisting of acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof. Preferably ethanol, 70% (v / v) ethanol or water, and most preferably 70% (v / v) ethanol.

On the other hand, it is obvious to those skilled in the art that the extract of the present invention can be obtained not only by the above-mentioned extraction solvent but also by other extraction solvent, which exhibits substantially the same effect.

In addition, the fruit of the present invention includes not only the extract obtained by the above-mentioned extraction solvent, but also the extract obtained through a conventional purification process. For example, decompression by carbon dioxide, extraction by supercritical extraction with high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cutoff value, various chromatography (size, charge, hydrophobic or affinity And the active fraction obtained through various purification and extraction methods, such as the extract by fermentation products using natural or various microorganisms, may be included in the extract of the present invention.

Further, the present invention provides a cosmetic composition characterized in that the above extract is obtained by liquid-freezing obtained by cold-pressing and heating filtration at room temperature, and further by concentrating or lyophilizing the solvent under reduced pressure.

The supercritical fluid extraction refers to supercritical fluid extraction. Generally, the supercritical fluid is extracted from the liquid and the liquid when the gas reaches the critical point at high temperature and high pressure. (J. Chromatogr. A. 1998; 479: 200-205). In addition, it has been reported that the supercritical fluid has a polarity similar to that of a nonpolar solvent.

Carbon dioxide is a supercritical fluid with both liquid and gaseous nature, with the operation of supercritical fluid equipment reaching its critical pressure and temperature, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the lipophilic substance contained in the sample is extracted into supercritical carbon dioxide.

When the supernatant carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction vessel after extracting the lipid-soluble substance, components that were not extracted by 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 mixed fluid in which a cosolvent is further mixed with supercritical carbon dioxide or carbon dioxide.

These co-solvents may be used alone or in admixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether.

Most of the extracted samples contain carbon dioxide. Since the carbon dioxide is volatilized into air at room temperature, the extract obtained by the above method can be used as a cosmetic composition, and the cosolvent can be removed by a reduced pressure evaporator.

In addition, the ultrasonic extraction method is an extraction method using energy generated by ultrasonic vibration. Ultrasonic waves can destroy an insoluble solvent contained in a sample in a water-soluble solvent. Due to the high local temperature generated at this time, Since the kinetic energy of the particles is increased, sufficient energy required for the reaction is obtained. By inducing the high pressure by the shock effect of the ultrasonic energy, the mixing efficiency of the substance contained in the sample and the solvent is increased, thereby increasing the extraction efficiency.

As the extraction solvent which can be used for the ultrasonic extraction method, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether can be used. The extracted sample is recovered by vacuum filtration, and the filtrate is recovered, and the extract is removed by a vacuum evaporator and freeze-dried to obtain an extract.

According to the present invention, the walnut 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 walnut extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition do. When the content of the extract is less than 0.0001% by weight, the effect of improving the skin is not exhibited. When the content of the extract is more than 30.0% by weight, the degree of improvement of the skin against the increase of the content is insignificant, It is not even.

Therefore, the cosmetic composition of the present invention including the fruit extract having the various functions of the present invention is excellent in collagen synthesis promoting effect, MMP-1 production inhibiting effect, antioxidant effect, skin wrinkle improving effect, skin elasticity improving effect, skin whitening effect, An antiseptic effect, a moisturizing effect, and an anti-aging effect on the skin.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the above-mentioned effective ingredients, and may contain conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, Supplements, and carriers.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powdered foundation, emulsion foundation, wax foundation, pack, massage cream and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

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

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

In the case where 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. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

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 applied to the skin and then wiped off, washed or rinsed with water. 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, a cleansing water, a cleansing towel and a cleansing pack, , Cleansing lotion, cleansing water and cleansing gel, but is not limited thereto.

On the other hand, the cosmetic process of the present invention refers to all cosmetic processes 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 combination, or may be used in combination with other cosmetic compositions other than the present invention. The cosmetic composition according to the present invention may be used according to a conventional method of use, and may be used in a number of times depending on the skin condition or taste of the user.

By carrying out the cosmetic method using the cosmetic composition comprising the fruit extract of the present invention, it is possible to provide a cosmetic composition containing the extract of the present invention, which is effective for inhibiting MMP-1 production, enhancing collagen synthesis, inhibiting melanin production, antioxidant effect, preservative effect, moisturizing effect, Effect can be obtained.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Manufacturing example  One: Wicker  Extract preparation

After shredding, the shrubs of the shrubs were washed three times with a 70% (v / v) aqueous ethanol solution for 5 hours, cooled, and filtered through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or lower.

Manufacturing example  2 And 3 : Supercritical  Manufacture of fluid extracts and ultrasonic extracts

The outpost of a shrine The supercritical extract was obtained by extracting with a conventional supercritical extraction method (extracting with ethanol as a co-solvent in a supercritical state under a pressure of about 300 atm at a temperature of about 60 DEG C). The extract was obtained by extraction using a supersonic wave (a supersonic ultrasonic device at 25 KHz at 30 캜 for about 2 hours). In Production Example 3, ethanol was used as a co-solvent.

Experimental Example  One. Wicker  Extract MMP -1 production inhibitory effect

In Experimental Example 1, the effect of inhibiting the production of MMP-1, a collagenase, was evaluated by the extracts of the wince obtained in Preparation Examples 1, 2 and 3.

Human normal skin cells were inoculated into a 48-well microplate (Nunc, Denmark) at a concentration of 1 × 10 ⁶ cells per well and cultured in DMEM medium (Sigma, USA) and 37 ° C For 24 hours, and then cultured in serum-free DMEM medium supplemented with the fruit extracts of Preparation Examples 1, 2 and 3 and in a serum-free DMEM medium containing no fennel extract for 48 hours as a control.

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

Name of sample Treatment concentration Inhibition rate of MMP-1 production (%) Production Example 1 Production Example 2 Production Example 3 Wicker
extract 10ppm 21.79 23.57 24.82 50 ppm 35.53 36.95 37.95 100ppm 50.94 53.81 55.14 500ppm 75.45 77.53 78.25 1000ppm 86.52 88.79 90.15 TGF-beta 200 nM 76.8

As shown in the results of Table 1, it was confirmed that the fruit extract of the present invention inhibits the production of MMP-1 (inhibits MMP-1 activity) in a concentration-dependent manner. Even considering the difference between the concentration of TGF-β (200 nM), a substance known to have an inhibitory effect on MMP-1 production, and the concentration of the extract of winky leaf extract (500 ppm) of the present invention, Showed that the wince extract of the present invention has an inhibitory effect on the production of MMP-1.

Experimental Example  2. Wicker  Enhancement of collagen synthesis of extract

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. The cells were further cultured for 48 hours in serum-free DMEM medium supplemented with the fruit extract prepared in Preparation Examples 1, 2, and 3 and in serum-free DMEM medium containing no wormwood extract as a control group. After the incubation, 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 into ng / Respectively. Collagen biosynthesis increase rate (%) was calculated according to the following formula (2), and the results are shown in Table 2.

Name of sample Treatment concentration Collagen biosynthesis rate (%) Production Example 1 Production Example 2 Production Example 3 Wicker
extract 2.5 ppm 11.2 13.7 12.9 5 ppm 20.4 21.9 20.8 10ppm 41.5 46.5 45.1 25 ppm 96.8 104.1 98.8 50 ppm 151.4 167.5 163.1 TGF-beta 200 nM 165.5

According to Table 2, it was confirmed that the collagen biosynthesis rate of the purified walnut extract prepared in Production Examples 1, 2, and 3 of the present invention increased in a concentration-dependent manner. In particular, it was confirmed that the extract prepared in Preparation Example 2 was more effective than the extracts prepared in Preparation Example 1 and Preparation Example 3.

Experimental Example  3: Wicker  Inhibitory Effect of Extracts on Melanin Production by B16F1 Melanocytes

Experimental Example 3 was conducted to examine the whitening effect of the extract of the wince obtained in the above Preparation Examples 1, 2, and 3 and to determine the inhibitory effect of melanin formation on B16F1 melanin-forming cells. The B16F1 melanin-forming cell used in Experimental Example 3 is a cell strain derived from a mouse, and is a cell that secretes a melanin pigment called melanin.

During the artificial culture of these cells, samples were treated to compare the degree of reduction of melanin pigment. The B16F1 melanin-forming cells used in this experiment were purchased from the American Type Culture Collection (ATCC).

Melanin biosynthesis inhibitory effect of B16F1 melanin-forming cells was measured as follows.

B16F1 melanocyte-forming cells were seeded at a concentration of 2 × 10 ⁵ per 6-well plate, and the cells were adhered and treated for 72 hours with treatment of the fruit extracts according to Preparations 1 to 3 at a concentration not causing toxicity. After incubation for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Lotan (Cancer Res., 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 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 centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin and the absorbance of melanin was measured at 405 nm with a microplate reader. The inhibition rate (%) of melanin formation was measured. Melanin formation inhibition rate (%) of B16F1 melanin-forming cells was calculated by the following equation (3), and the results are shown in Table 3. [

A: Amount of melanin in wells to which no sample was added

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

Name of sample Treatment concentration (%) Melanin inhibition rate (%) Production Example 1 Production Example 2 Production Example 3
Fruit Extract 0.001 20.17 19.15 22.13 0.005 38.55 39.12 40.11 0.01 53.53 54.72 54.52 0.05 73.96 75.11 76.42 Arbutin 200ppm 68.25

As can be seen from the results in Table 3, it was found that the extract of the wince obtained by Preparation Examples 1, 2 and 3 significantly inhibited melanogenesis in B16F1 melanin-forming cells.

Experimental Example  4: Wicker  Extract DPPH Radical Scatters  analysis

In Experimental Example 4, the antioxidative effect (free radical scavenging ratio) was measured by the DPPH method in order to measure the antioxidative effect of the walnut extract obtained in Production Examples 1, 2, and 3. The DPPH method measures the antioxidative effect by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The degree of decrease in absorbance of DPPH is reduced by the test substance (elderberry extract) to the absorbance of the blank test solution and the free radical scavenging ratio (%) is measured at a wavelength of 560 nm. As a reagent used, 61.88 mg of a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) was dissolved in methanol to make 100 ml.

As a measuring method

(1) Add 0.15 ml of the sample solution to 0.15 ml of 0.1 mM DPPH solution in a 96-well plate, stir rapidly, and start culturing at 25 ° C for 10 minutes.

② Measure the absorbance St at 560 nm thereafter.

(3) The blank of the sample solution is measured by the same procedure as that for measuring the absorbance St except that methanol is used instead of the 0.1 mM DPPH solution.

(4) The absorbance Bt is measured by operating in the same manner as in the measurement of the absorbance St except that distilled water is used instead of the sample solution.

(5) Blank of the blank test is carried out in the same manner as in the measurement of 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, and the absorbance Bo is measured.

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

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

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

So: the absorbance at 514 nm before the reaction in the absence of free radicals in the sample solution

Bo: absorbance at 514 nm before the reaction in the absence of the free radical of the blank test solution

Name of sample SC ₅₀ (%) Production Example 1 0.050 Production Example 2 0.035 Production Example 3 0.045

SC ₅₀ , which is the concentration of the sample required for 50% removal of the DPPH free radical, was confirmed, and the production examples 1, 2 and 3 showed antioxidative effect.

Formulation Example  One: Wicker  Extract-containing Cosmetics  Preparation of composition

The cosmetic compositions containing the walnut extracts (Examples 1, 2 and 3) each containing the extract of the walnut trees of Preparation Examples 1 to 3 were prepared. (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as moisturizers (Comparative Formulation Example 1), a cosmetic composition containing glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 2), an extract (Comparative Formulation Example 3) which does not contain both the moisturizing agent and the moisturizing agent are shown in Table 5 below.

division ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 compare
Formulation Example 1 compare
Formulation Example 2 compare
Formulation Example 3 A






Fruit Extract
(Production Example 1) 5.0 - - - - Fruit Extract
(Production Example 2) - 5.0 - - - Fruit Extract
(Production Example 3) - - 5.0 -  -  - glycerin - - - 5.0 - - 1,3-butylene glycol - - - 5.0 - - Adenosine - - - - 3.0 - EDTA-2Na 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 B









Cetostearyl alcohol 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 Microcrystalline 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 Liquid paraffin 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 Polysorbate 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 Tocopheryl acetate 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 BHT 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  5: Wicker  Moisturizing effect of extract

The clinical moisturizing effects of the cosmetic compositions of Formulation Examples 1, 2 and 3 were measured as follows. A, B, and C groups were divided into 5 groups (A, B, C, D, and E) by randomly dividing 25 healthy adults (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as moisturizing agents, a cosmetic composition which does not contain a fruit extract and a moisturizer, and a cosmetic composition Example 3) were applied to the face for 8 weeks twice a day, respectively. The moisturizing effect was evaluated by Corneometer CM 820 (Corage + Khazaka, Germany) for every 2 weeks and after the start of the test. The experimental results are shown in Table 6 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 22.6 36.9 45.3 Formulation Example 2 23.3 37.8 48.7 Formulation Example 3 24.2 39.5 48.9 Comparative Formulation Example 1 24.5 37.5 45.5 Comparative Formulation Example 3 23.1 24.5 23.8

As shown in Table 6, it was confirmed that the moisturizing effect of the cosmetic compositions (Formulation Examples 1, 2 and 3) containing the fruit extract of the present invention was excellent in comparison with the cosmetic compositions containing other moisturizers (Comparative Formulation Example 1) there was.

Experimental Example  6: Wicker  Preservative effect of extract

In order to examine the preservative effect of the walnut extract, the cosmetic compositions of Formulation Examples 4 to 11 containing the walnut extracts of Preparation Examples 1, 2 and 3 at various ratios in the composition of the following Table 7, the comparative formulation containing purified water instead of the walnut extract The cosmetic of Comparative Formulation Example 5 containing methylparaben and imidazolidinyl urethane as a preservative instead of the cosmetic and the fir tree extract of Comparative Example 4 was prepared.

ingredient Formulation Example Comparative Formulation Example 4 5 6 7 8 9 10 11 4 5 Content (unit:% by weight) Fruit Extract
(Production Example 1) 0.5 1.0 2.0 - - - - - - Fruit Extract
(Production Example 2) - - - 0.5 1.0 2.0 - - - - Fruit Extract
(Production Example 3) 0.5 1.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Butylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Propylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Wax 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Macadamia nut oil 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Spices a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Methylparaben - - - - - - - - - 0.2 Imidazolidinyl
Urea - - - - - - - - - 0.2 Purified water To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100

The cosmetic effects of the formulation examples and comparative formulations of Table 7 were used to measure the preservative effect on bacteria and fungi.

First, in the case of bacteria, the cosmetic composition of Formulation Example 4-11 and the cosmetic composition of Comparative Formulation Examples 4 and 5 were mixed with 20-30 g of Escherichia coli ; ATCC 8739), Pseudomonas aeruginosa aeruginosa ; ATCC9027) and Staphylococcus aureus (ATCC6538) were added and mixed at an initial concentration of 10 ⁷ cfu / g per sample. 1 g of each cosmetic was taken at 1, 7, 14, 21 and 28 days intervals while culturing them in a thermostat at 30-32 ° C for 4 weeks, and the number of viable cells was measured. The measurement results are shown in Table 8 below.

Next, in the case of mold, Candida albicans ( Candida albicans ; ATCC 10231), Penicillium citrinum ; KCTC2123) and Aspergillus niger ; ATCC16404) was added at a concentration of 10 ⁷ cfu / g per sample and incubated at 25 ° C. in a thermostatic chamber to observe the presence or absence of swelling and the presence of hyphae and spore on the surface of the sample at intervals of 7 days. 8.

Cosmetics Bacteria (cfu / g) mold* Initial number of bacteria Day 1 Day 7 Day 14 Day 21 Day 28 Formulation Example 4 1 x 10 ⁷ 820000 39000 5100 500 <100 - Formulation Example 5 1 x 10 ⁷ 710000 15000 1000 250 <100 - Formulation Example 6 1 x 10 ⁷ 320000 4000 400 <100 <100 - Formulation Example 7 1 x 10 ⁷ 740000 21000 3100 400 <100 - Formulation Example 8 1 x 10 ⁷ 670000 11000 800 150 <100 Formulation Example 9 1 x 10 ⁷ 290000 3500 200 <100 <100 Formulation Example 10 1 x 10 ⁷ 700000 26000 2200 400 <100 - Formulation Example 11 1 x 10 ⁷ 660000 9100 700 300 <100 - Comparative Formulation Example 4 1 x 10 ⁷ 8500000 8200000 6900000 6000000 5800000 +++ Comparative Formulation Example 5 1 x 10 ⁷ 370000 5300 450 <100 <100 -

-: No spawning and mycelium spawning for 8 weeks and good

+: Molding on the wall or lid within 4 weeks

++: mending within 4 weeks and mold on part of the surface

+++: Mildew and mold on the entire surface within 4 weeks

As can be seen from the above Table 8, the cosmetic composition of Comparative Example 4, which did not use an antiseptic, showed molds on the whole surface and on the surface within 4 weeks, and bacterial bacteria were remarkably increased even after 1 day. However, the cosmetic compositions of Formulation Examples 4 to 11 containing the fruit extract of the present invention exhibited a concentration-dependent preservative effect on bacteria and fungi, and after about 28 days, 2.0% (w / v) The preservation force similar to or better than Comparative Formulation Example 5 in which the preservative methylparaben and imidazolidinyl urea were mixed was secured. When the content of the preservative was judged to be 2.0% or more, It was found that a far superior buoyant force was obtained as compared to Comparative Formulation Example 5 in which methylparaben, which is methylparaben, and imidazolidinyl urea were mixed. In particular, it was confirmed that the wince extract prepared in Preparation Examples 2 and 3 was more excellent in flotation power than Preparation Example 1.

Experimental Example  7. Wicker  Effect of extracts on skin wrinkles

Clinical trials of the skin wrinkle-improving effect of the cosmetic composition containing the fruit extract of the present invention were carried out. Clinical trials were evaluated through actual use tests of each cosmetic product.

That is, the clinical test was conducted in the same manner as in Formulation Example 2 containing the fruit extract of Preparation Example 2 of Table 5 and the cosmetic composition of Comparative Formulation Example 1 containing glycerin and 1,3-butylene glycol as moisturizing agents instead of the fruit extract, Comparative Example 2 containing adenosine 3% (v / v) in Daesan was used to confirm and compare the effect of improving the skin wrinkles caused by the use of cosmetics in humans.

The experiment group of 90 women (30 to 50 years old) was divided into three groups: a group treated with cosmetic composition (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as humectants, (Comparative Formulation Example 2) treated with 3% (v / v) (Formulation Example 2), and the test group of Formulation Example 2 containing the walnut extract. The wrinkle- And the degree of wrinkle improvement was confirmed. The results of the wrinkle-improving effect based on this evaluation are shown in Table 9 below.

The effect of improving wrinkles after the completion of the experiment was evaluated by visual examination and instrument measurement (cutometer SEM 575, C + K Electronic Co., Germany) of a skilled physician before and after 3 months of use. The experimental results are shown in Table 9 below.

Wrinkle-improving effect (visual evaluation and device evaluation) of a cosmetic composition containing a walnut extract Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 2
(Fruit extract) 21 6 3 90.0 Comparative Formulation Example 1
(Moisturizer) 4 6 20 33.3 Comparative Formulation Example 2
(Adenosine) 20 6 4 86.7

As can be seen from the results of Table 9, the cosmetic composition of Formulation Example 2 containing the fruit extract of the present invention showed a wrinkle-improving effect of about 2.7 times higher than Comparative Formulation Example 1 containing moisturizing agent.

In addition, similar results were obtained with the cosmetic preparation of Comparative Formulation Example 2 containing adenosine, which is known to have a wrinkle-reducing effect instead of the wicker-extract, indicating that the wicker extract has excellent wrinkle-reducing effect.

Experimental Example  8: Measurement of skin elasticity improvement effect

In Experimental Example 8, in order to examine the skin elasticity improving effect of the cosmetic composition according to the formulation examples and the comparative formulations including the walnut extract obtained in Production Examples 1, 2, and 3, 40 Were measured. 10 groups were divided into four groups, and the first group had Formulation Example 1, the second group had Formulation Example 2, the third group had Formulation Example 3, and the fourth group had Comparative Formulation Example 1 in the morning, morning and evening The skin elasticity was measured after 12 weeks. The elasticity of the facial region was measured using a cutometer SEM575 (MPA 580, Courage + Khazaka GmbH, Germany), and the R2 (biological elasticity) indicating skin elasticity was measured before and after application . The results are shown in Table 10 below, and the results are an average of the degree of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 24.8 Formulation Example 2 26.5 Formulation Example 3 23.8 Comparative Formulation Example 1 5.5

As shown in Table 10, Formulation Examples 1, 2 and 3 containing a winkle extract showed significantly improved skin elasticity as compared with Comparative Formulation Example.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (9)

A cosmetic composition comprising a fennel extract as an active ingredient. The cosmetic composition according to claim 1, wherein the cosmetic composition has an effect of improving skin wrinkles. The cosmetic composition according to claim 1, wherein the cosmetic composition has a skin whitening effect. The cosmetic composition according to claim 1, wherein the cosmetic composition has an antioxidative effect. The cosmetic composition according to claim 1, wherein the cosmetic composition has skin moisturizing effect. The cosmetic composition according to claim 1, wherein the cosmetic composition has an anti-aging effect. The cosmetic composition according to claim 1, which contains a walnut extract as a preservative. The cosmetic composition according to any one of claims 1 to 7, wherein the walnut extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. [Claim 7] The method according to any one of claims 1 to 7, wherein the fruit extract comprises (a) water, an anhydrous or a lower alcohol having 1-4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, (B) a supercritical extraction method, or (c) an ultrasonic extraction method, wherein the solvent extraction method is an extraction method using an extraction solvent selected from the group consisting of butyl acetate, diethyl ether, dichloromethane, hexane and mixtures thereof.