KR20160001094A - Cosmetic composition comprising the extract of Lonicera coerulea var. edulis as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a Lonicera coerulea var. edulis extract as an active component. The cosmetic composition contains 0.0001-30.0 wt% of the Lonicera coerulea var. edulis extract as an active component for the total weight of the cosmetic composition. According to the present invention, the Lonicera coerulea var. edulis extract has an MMP-1 generation inhibiting effect, a bio-collagenesis effect, a melanin generation inhibiting effect, an inflammatory cytokein expression inhibiting effect, an anti-oxidation effect, a preservative effect, a skin moisturizing effect, a skin wrinkle reducing effect, and a skin anti-aging effect, thereby providing various functional cosmetics.

Description

The present invention relates to a cosmetic composition and a cosmetic process for preparing a cosmetic composition containing an extract of a Japanese persimmon tree as an active ingredient, and a cosmetic composition comprising the extract of Lonicera coerulea var. edulis as active ingredient}

The cosmetic compositions and cosmetic methods containing daengdaengyi tree extract as an active ingredient {Cosmetic composition comprising the extract of Lonicera coerulea there is . edulis as 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. Free radicals can be caused by activation of ultraviolet rays, stress, disease state, environmental factors, wounds, and aging. When such state is deepened, the antioxidant defense network existing in the living body is destroyed, And aging.

More 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 necessary to protect the cell membrane by destroying the free radicals mediated by free radicals, ultraviolet rays, and inflammation that occur during the metabolism of the body, and to regenerate already damaged cells by active metabolism, 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 No. 4-9320), hydroquinone (Japanese Patent Laid-Open No. 192062), kojic acid (Japanese Patent Laid-open No. 56-7710), arbutin Activity, and is used as a whitening cosmetic. However, since it is low in stability in cosmetic formulations, its use is restricted due to decomposition and coloring, generation of odor, efficacy at a living body level, unclear effect and safety problem. These substances have proved their tyrosinase inhibitory effect, but their effect is lower in experiments similar to actual living body 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.

The inventors of the present invention have studied the possibility of application to cosmetics in various natural products that have not been known until now. As a result, the extracts of the leaves and leaves of the buttocks were selected to produce MMP-1 production inhibitory effect, antioxidative effect, collagen biosynthesis The effect of inhibiting melanin formation, the inhibitory effect of inflammatory cytokine expression, the preservative effect, the moisturizing effect, the improvement of skin wrinkle, the skin elasticity improvement effect and the skin aging effect, I was able to find out.

It is an object of the present invention to provide a method for inhibiting MMP-1 production, a collagen biosynthesis effect, a melanin formation inhibitory effect, an antioxidative effect, an inflammatory cytokine expression inhibiting effect, a moisturizing effect, And to provide a cosmetic composition exhibiting an improvement effect.

It is also an object of the present invention to provide a cosmetic composition that produces supernatant, ultrasound, and fermented extracts to maximize the efficacy of Swallow's Tree extract, and exhibits efficacy through purification.

Another object of the present invention is to provide a makeup method using a cosmetic composition containing the above-described tree extract.

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

Preferably, the cosmetic composition is for improving skin wrinkles.

Further, preferably, the cosmetic composition is characterized by being for skin whitening.

Preferably, the cosmetic composition is characterized by being for antioxidation.

Preferably, the cosmetic composition is a preservative.

Preferably, the cosmetic composition is characterized in that it is for moisturizing.

Preferably, the cosmetic composition is for improving skin elasticity.

Further, preferably, the cosmetic composition is characterized by being anti-inflammatory.

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

In addition, preferably, the royal jellyfish 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 Butterfly tree extract is selected from the group consisting of (a) water, an anhydrous or hydric alcohol having 1-4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, (B) supercritical extraction, or (c) ultrasonic extraction, or (d) fermentation process, and the resulting mixture is subjected to a first extraction using an extraction solvent selected from the group consisting of dichloromethane, And (e) a purification step after the primary extraction.

Preferably, the cosmetic composition of the present invention is used in combination with at least one selected from the group consisting of MMP-1 inhibiting effect, collagen biosynthesis inhibiting effect, melanin inhibiting effect, inflammatory cytokine inhibiting effect, preservative effect, antioxidative effect, And an anti-aging effect on the skin.

The cosmetic composition may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing oil, powder foundation, emulsion liquid emulsion foundation, ≪ RTI ID = 0.0 > spray. ≪ / RTI >

Another object of the present invention is achieved by a cosmetic method using the cosmetic composition.

The extracts of Staphylococcus aureus 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), moisturizing effect (Experimental Example 6), preservative effect (Experimental Example 7), skin wrinkle improving effect (Experimental Example 8) and skin elasticity improving effect (Experimental Example 9).

According to the present invention, in order to maximize the efficacy of the Sweet Potato Tree extract, the extract is extracted through a process selected from among ultrasonic, supercritical extraction and fermentation processes, purified and then contained in a cosmetic composition to inhibit the production of MMP-1 Effect of collagen synthesis, inhibition of melanin production, antioxidative effect, inhibitory effect of inflammatory cytokine expression, moisturizing effect, preservative effect, and skin elasticity improvement effect.

In the cosmetic composition of the present invention, the swallowtail tree (danggang tree, honeyberry, honeyberry) used as an active ingredient is written in English, and its scientific name is Lonicera coerulea there is . edulis . It grows at an elevation of 700 to 2,300 m above sea level. It is 1.5 meters high, full of fawns, and small branches with many hairs. Leaves are opposite to each other, and are basically straight or inverted. They are 1 ~ 4cm long and 1cm wide. There are hairs on the front side of the leaf, no villus on the back side, no sawtooth, and petiole length is 1 ~ 6mm. In May ~ June, a yellowish white flower hangs on the axil, and the peduncle is 2 ~ 10mm long with hairs. The bract is 5 ~ 8mm long and has hairs. Calyx is divided into 5 pieces like sawtooth, corolla is yellowish white, cylindrical bell shape, length is 1.2 ~ 1.5cm and there are some hairs. Surgery is shorter than the style and two ovaries are combined. Fruits are elliptical berries which are black in autumn and covered with white powder. The fruit is eaten raw, and the juice is eaten by making a soft drink. It is distributed in Korea (Jeonnam, Gangwon, Pyungnam, Pyongbuk, Hamnam, Hambuk), Japan, Kuril Islands, Sakhalin, East Siberia and Kamchatka Peninsula. Surgery is shorter than the style and two ovaries are combined.

In the present invention, the Swallow's tree extract is obtained by extracting from various organs or parts (for example, leaves, branches, flowers, roots, stems, fruits and the like) of the buttock tree and is preferably an extract obtained from a seedling or a fruit , And most preferably from an outpost.

In addition, the farina tree extract of the present invention can be produced by using conventional solvents known in the art, that is, under the conditions of ordinary temperature and pressure, but preferably using (a) water, (Such as methanol, ethanol, propanol and butanol) having 1 to 4 carbon atoms, propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloro (B) a supercritical extraction method using reduced pressure and high temperature, or (c) an ultrasonic extraction method, or (d) a fermentation extraction method using a solvent selected from the group consisting of methanol, .

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

In the present invention, the Thyme extract is obtained from a variety of extraction solvents, for example, water, anhydrous or lower alcohols having 1-4 carbon atoms (such as methanol, ethanol, propanol and butanol), propylene glycol, It may be obtained by using at least one extraction solvent selected from the group consisting of glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, % (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 Staphylinothi japonica of the present invention can be obtained not only by the above-mentioned extraction solvent but also by using other extraction solvent.

The farina tree extract of the present invention includes extracts obtained by the conventional extraction and fermentation as well as the extracts obtained by the above-mentioned extraction solvents. 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 an extract obtained by fermentation products using natural or various microorganisms, are also 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.

The farina tree extract according to the present invention also includes a fermented extract, and the fermented extract of the buttock tree can be prepared as follows. After finely crushing the buttock tree to a size of about 100 to 500 mesh, 1 to 50 g / L of a 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 to 37 ° C. The pH is 5 to 7 and aerobic or anaerobic conditions are maintained for about 5 to 10 days. Which can be obtained through aging and filtration.

According to the present invention, the Butterfly tree extract is contained in an amount of 0.0001 to 30.0 wt% with respect to the total weight of the cosmetic composition, more preferably, the Butterfly tree extract is contained in an amount of 0.01 to 10 wt% with respect to the total weight of the cosmetic composition . 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 comprising the various functions of the soya tree extract of the present invention is excellent in collagen synthesis enhancing effect, MMP-1 formation inhibitory effect, inflammatory cytokine expression inhibiting effect, antioxidant effect, skin wrinkle improving effect, A skin whitening effect, a melanin formation inhibiting effect, a preservative 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. The surfactant-containing cleansing formulation may be a cleansing foam, a cleansing water, a cleansing towel and a cleansing pack. The surfactant-free cleansing formulation may be a cleansing cream, , 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. (Change order in context)

The inventive cosmetic composition comprising the extract of Stalagrand's tree of the present invention can be used for cosmetic compositions which are effective for inhibiting MMP-1 production, for enhancing collagen synthesis, for inhibiting melanin formation, for inhibiting the expression of inflammatory cytokines, Effect of improving skin wrinkles and improving skin elasticity 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: Butt  Tree extract manufacturing

The shrubs of the shrubs were washed with a 70% (v / v) aqueous ethanol solution three times 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 to 3. Supercritical  Manufacture of fluid extracts and ultrasonic extracts

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). Extracts were obtained by extraction using ultrasound (Super Sonic Ultrasonic Device at 25 ° C for 2 hours at 30 ° C). In the case of Production Example 3, ethanol was used as a co-solvent.

Experimental Example  One. Butt  Tree extract MMP -1 production inhibitory effect

In Experimental Example 1, the effect of inhibiting the production of MMP-1, a collagenase, was determined by the extracts of the sole obtained from 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. Then, the cells were further cultured in serum-free DMEM medium supplemented with the above-described Preparative Examples 1, 2, and 3, and serum-free DMEM medium containing no extracts of foreskin for 48 hours. 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 Hemai tree
extract 10ppm 22.79 24.17 23.82 50 ppm 34.93 37.55 36.95 100ppm 49.74 52.81 54.14 500ppm 74.57 77.23 78.55 1000ppm 85.12 87.19 89.51 TGF-beta (200 nM) 76.8

As shown in the results of Table 1, it was confirmed that the herpes simplex extract of the present invention suppresses the production of MMP-1 (inhibits MMP-1 activity) in a concentration-dependent manner. Even considering the difference between the concentration (200 nM) of TGF-beta, a substance known to have an inhibitory effect on MMP-1 production, and the concentration (500 ppm) of the farina tree extract of the present invention, The inhibition of MMP-1 production (antioxidant effect) is shown by the Swallow's tree extract of the present invention.

Experimental Example  2. Butt  Enhancement of Collagen Synthesis in Tree 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 in serum-free DMEM medium supplemented with the above-described Preparation Examples 1, 2, and 3 and in serum-free DMEM medium containing no extracts of foreskin for 48 hours. 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 Butterfly Tree Extract 2.5 ppm 12.2 13.7 12.7 5 ppm 22.0 25.9 21.8 10ppm 42.4 47.5 44.5 25 ppm 94.1 103.9 97.8 50 ppm 150.4 166.5 162.1 TGF-beta (200 nM) 165.5

According to Table 2, it was confirmed that the collagen biosynthesis rate of the purified Butterfly tree extract prepared according to 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 Examples 1 and 3.

Experimental Example  3: Butt  Inhibitory Effect of Tree Extracts on B16F1 Melanogenesis Cells (Whitening Effect)

In Experimental Example 3, in order to examine the whitening effect of the soya tree extract obtained in the above Production Examples 1, 2 and 3, the inhibitory effect of melanin formation on the B16F1 melanin-forming cells was examined to determine the whitening effect. 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 melanocytes were plated at a concentration of 2 × 10 ⁵ per well on a 6-well plate, and the cells were adhered and treated with the extracts of the foreskins of Preparative Examples 1 to 3 at a concentration not causing toxicity and cultured for 72 hours. 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 well without addition of sample

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

Name of sample Treatment concentration (%) Melanin formation inhibitory effect (%) Production Example 1 Production Example 2 Production Example 3 Butterfly Tree Extract
(Production Example 2) 0.001 20.17 20.15 21.13 0.005 37.45 40.12 38.11 0.01 53.43 54.52 53.42 0.05 72.86 74.91 75.82 Arbutin 200ppm 68.25

As can be seen from the results shown in Table 3, the purified butt jar tree extract prepared according to Production Examples 1, 2, and 3 of the present invention was found to have a whitening effect that significantly inhibits melanin production in B16F1 melanin-forming cells.

Experimental Example  4: Butt  Tree extract DPPH Radical Scatters  Analysis (antioxidant effect)

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 extracts of the foreskins 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 0.025 Production Example 2 0.010 Production Example 3 0.020

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.

Experimental Example  5: Butt  Inhibitory Effect of Tree Extracts on Inflammatory Cytokine Expression by UV Irradiation

Experimental Example 5 was conducted to evaluate the inhibitory effect of inflammatory cytokine expression expressed by ultraviolet irradiation on the extracts of Stingidae obtained in Production Examples 1, 2 and 3, and fibroblasts isolated from human epidermal tissue Each 5 X 10 ⁴ -well plate was attached to a 24-well test plate and allowed to attach for 24 hours.

Each well was washed once with PBS and 500 占 퐇 of PBS was added to each well. After irradiating the fibroblasts with 10 mJ / cm 2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan), PBS was removed and the cell culture medium ≪ / RTI > medium).

After the treatment, the plants were treated for 5 hours. 150 [mu] l of the culture supernatant was measured to quantitate IL-1 [alpha], thereby determining the effect of inhibiting the expression of inflammatory cytokines in the tree extract. The amount of IL-1α was quantitated using an enzyme-linked immunosorbent assay, and the inhibition rate (%) of inflammatory cytokine (IL-1α) was calculated by the following equation (5).

Bo: IL-1? Production in wells without UV irradiation

Bt: IL-1? Production in wells irradiated with ultraviolet light and not treated with the sample

St: IL-1α production in wells irradiated with ultraviolet light and treated with the sample

Name of sample Treatment concentration Inflammatory cytokine
Expression inhibition rate (%) Manufacturing Example) Production Example 2 Production Example 3 Butterfly Tree Extract 10ppm 21.11 24.79 23.53 50 ppm 35.52 37.48 34.12 100ppm 48.15 61.25 58.75 500ppm 54.12 65.92 64.77 1000ppm 75.28 85.06 77.10 Indomethacin 100 ppm 61.23

According to Table 5, it can be seen that the extract prepared according to Production Example 2 of Swallow's Tree extract inhibits the production of IL-1α, which is an inflammatory cytokine caused by ultraviolet rays, by 87.26% at a concentration of 1000 ppm. The extracts of Stag beetle according to the present invention showed an excellent inflammatory cytokine expression ratio at the same concentration as that of indomethacin, which is an inhibitor of inflammatory cytokine expression. Therefore, it can be seen that the herringbone extract of the present invention effectively inhibits the inflammation caused by ultraviolet irradiation even at a low concentration.

Formulation Example  One : Butt  Tree extract Cosmetics  Preparation of composition

Cosmetic preparations containing each of the extracts of Swallowtail tree were prepared as Cosmetic preparations (Formulation examples 1, 2 and 3) each containing the Swallow's tree extracts of Preparation Examples 1 to 3. For comparison of efficacy, glycerin and 1 , Cosmetic composition (Comparative Formulation Example 1) containing glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 2), cosmetic composition (Comparative Formulation Example 3) containing neither extract nor moisturizer, Are shown in Table 6 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 Butterfly Tree Extract
(Production Example 1) 5.0 - - - - Butterfly Tree Extract
(Production Example 2) - 5.0 - - - Butterfly Tree 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  6: Butt  Moisturizing effect of tree 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 five groups (A, B, C, D, and E) by a randomly selected group of 25 healthy adult men and women who felt skin dry through the questionnaire. (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as moisturizing agents instead of Swallow's Tree extract, Cosmetic composition containing no Swallow's Tree extract and Moisturizing agent (Comparison Formulation Example 3) was 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 7 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 22.4 35.6 44.7 Formulation Example 2 23.1 37.7 48.9 Formulation Example 3 24.1 38.5 47.7 Comparative Formulation Example 1 23.7 36.7 44.5 Comparative Formulation Example 3 23.5 24.4 23.9

As shown in Table 7, it was confirmed that the moisturizing effect of the cosmetic compositions (Formulation Examples 1, 2 and 3) containing the extract of Swallowtailia japonica according to the present invention was superior to the cosmetic compositions containing other moisturizing agents (Comparative Formulation Example 1) I could.

Experimental Example  7: Butt  Preservative effect of tree extract

In order to examine the preservative effect of the Japanese persimmon tree extract, the cosmetics of Formulation Example 4-11 containing various amounts of Swanberry extracts of Preparation Examples 1, 2 and 3 in the composition shown in Table 8 below, The cosmetic composition of Comparative Formulation Example 5 containing the methylparaben and imidazolidinyl urethane as a preservative instead of the cosmetic composition of Comparative Formulation Example 4 and the tree herb extract was prepared. The experimental results are shown in Table 8 below.

ingredient Formulation Example Comparative Formulation Example 4 5 6 7 8 9 10 11 4 5 Content (unit:% by weight) Butterfly Tree Extract
(Production Example 1) 0.5 1.0 2.0 - - - - - - Butterfly Tree Extract
(Production Example 2) - - - 0.5 1.0 2.0 - - - - Butterfly Tree 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 formulation examples in Table 8 were used to measure the preservative effect against 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 9 below.

Next, in the case of mold, Candida albicans ( Candida albicans ; ATCC 10231), Penicillium citrinum ; KCTC2123) and Aspergillus niger (ATCC16404) were added at a concentration of 10 ⁷ cfu / g per sample and cultured at 25 ° C in a thermostatic bath at intervals of 7 days, The results are shown in Table 9 below.

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 shown in Table 9, the cosmetic composition of Comparative Example 4, which did not use any preservative, showed molds on the surface and on the entire surface within 4 weeks, and the bacterial bacterium was remarkably increased even after 1 day. However, the cosmetic compositions of Formulations 4 to 11 of the present invention containing the farina tree extract showed a concentration-dependent preservative effect on bacteria and fungi, and after about 28 days, 2.0% (w / v) ), A repellency similar to or better than Comparative Formulation Example 5 in which methylparaben, which is a preservative, and imidazolidinyl urea were mixed was obtained, and when it was judged based on the whole culture day, when it was used in a content of 2.0% or more It was found that a far superior buoyancy was obtained as compared to Comparative Formulation Example 5 in which the preservative methylparaben and imidazolidinyl urea were mixed. In particular, it was confirmed that the farinae extracts prepared in Preparative Examples 2 and 3 were superior to those of Preparative Example 1 in antibacterial activity.

Experimental Example  8. Butt  Effect of tree extracts on skin wrinkles

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

That is, the clinical trial was conducted in the same manner as in the formulation example 2 containing Preparation Example 2 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 farina tree extract, Comparative Example 2, which contains adenosine 3% (v / v) in Daesan, was investigated and compared with human skin wrinkle improving effect of cosmetic use.

Sixty women (30 to 50 years old) were divided into 20 experimental groups, each of which was treated with a cosmetic composition (Comparative Formulation Example 1) containing glycerin and 1,3 butylene glycol as a moisturizing agent in place of the soya extract, (Comparative Formulation Example 2) treated with adenosine 3% (v / v), and the experimental group of Formulation Example 2 containing the extract of Buttwilia japonica, and after 6 weeks of wrinkle improvement using both sides of the face The effect was visually evaluated to confirm the degree of wrinkle improvement. 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 10 below.

Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 2
(Swallow's tree extract) 21 6 3 90.0 Comparative Formulation Example 1
(Moisturizer) 5 5 20 33.3 Comparative Formulation Example 2
(Adenosine) (20) (6) ( 4 ) (86.7)

As can be seen from the results of Table 10 above, the cosmetic composition of Formulation Example 2 containing the extract of the royal jellyfish extract of the present invention contains a moisturizing agent, Which was about 2.7 times higher than Comparative Formulation Example 1,

In addition, similar results were obtained with the cosmetic preparation of Comparative Formulation Example 2 containing adenosine, which is known to have a wrinkle-improving effect instead of the Swallow's Tree extract, and it was found that the Swallow's Tree extract exhibits excellent wrinkle-improving effect.

Experimental Example  9: Measurement of skin elasticity improvement effect

In Experimental Example 9, in order to examine the skin elasticity improvement effect of the cosmetic composition according to the formulation examples and comparative formulations obtained from Preparation Examples 1, 2, and 3, 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 11 below, and the results are an average value of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 24.5 Formulation Example 2 27.1 Formulation Example 3 25.8 Comparative Formulation Example 1 5.5

As shown in Table 11, Formulations 1, 2, and 3, which contain the crude extract of Stinging Tree, showed significantly improved skin elasticity compared to the comparative formulation.

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 (12)

A cosmetic composition comprising asparagus extracts as an active ingredient.
The cosmetic composition for improving skin wrinkles according to claim 1, wherein the composition comprises a farina tree extract as an active ingredient.
The cosmetic composition for whitening skin according to claim 1, wherein the composition comprises a tree frog extract as an active ingredient.
The antioxidative cosmetic composition according to claim 1, wherein the composition is an antioxidant. The cosmetic composition according to claim 1, wherein the composition comprises a farina tree extract as an active ingredient. The moisturizing cosmetic composition according to claim 1, wherein the composition comprises a jellyfish extract as an active ingredient. The cosmetic composition for improving skin elasticity according to claim 1, wherein the composition contains a tree frog extract as an active ingredient. The anti-inflammatory cosmetic composition according to claim 1, wherein the composition comprises an extract of a Japanese thistle tree as an active ingredient. The cosmetic composition for preventing aging of skin according to claim 1, which contains the extract of Stem tree as an active ingredient. 10. The cosmetic composition according to any one of claims 1 to 9, wherein the sea mustard extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. . 11. The method according to any one of claims 1 to 10, wherein the Swallow's Tree 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) supercritical extraction or (c) ultrasonic extraction or (d) fermentation from a microorganism, or (c) extraction with a solvent selected from the group consisting of acetone, methyl ethyl ketone, butyl acetate, diethyl ether, dichloromethane, hexane and mixtures thereof. (E) Purification process (liquid-liquid extraction method using solubility difference, precipitation and filtration, using polarity and molecular weight, etc.) after the first extraction and fermentation extraction using natural fermentation metabolism Saponin, cardiac glycosides, polyterpenes, and dyes are removed from the cosmetic composition by means of chromatography. A cosmetic method using a cosmetic composition containing the above-mentioned tree extract obtained according to any one of claims 1 to 10.