KR101176525B1 - Cosmetic Composition Comprising Acer tegmentosum Maxim as Active Ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising an extract of Acer tegmentosum Maxim as an active ingredient, characterized in that it contains 0.001 to 30.0% by weight as an active ingredient, according to the present invention, the bee extract is human In addition to the fibroblast activity effect, there is a skin wrinkle improvement effect, a whitening effect, moisturizing effect, skin damage relieving effect and irritation relieving effect can provide a variety of functional cosmetics.

Bee, whitening, wrinkle improvement, moisturizing, skin irritation, cosmetics

Description

Cosmetic composition containing bee extract as an active ingredient {Cosmetic Composition Comprising Acer tegmentosum Maxim as Active Ingredient}

The present invention relates to a cosmetic composition containing the bee extract as an active ingredient.

Human skin undergoes various physical and chemical changes during the aging process. The causes are largely divided into intrinsic aging and photo-aging, and research on this has been actively conducted. Free radicals can be caused by UV rays, stress, disease conditions, environmental factors, wounds, and aging, which intensify the body's ability to destroy antioxidant defenses and damage cells and tissues. And aging.

More specifically, lipids, proteins, polysaccharides, nucleic acids, and the like, which are major constituents of skin, are oxidized to destroy skin cells and tissues, resulting in skin aging.

In particular, the oxidation of the protein is a severe over-inflammatory reaction and skin elasticity by cutting the connective tissue collagen (collagen), hyaluronic acid (hyaluronic acid), elastin, proteoglycan, fibronectin (fibronectin) If this affects and worsens, mutations in the DNA can lead to mutations, cancer, and reduced immune function.

Therefore, to protect the cell membrane by eliminating the free radicals generated by the body's metabolic process, ultraviolet radiation, or inflammatory reactions, and to protect the cell membranes, the already damaged cells must be regenerated by active metabolism to proliferate the cells. The skin can recover quickly and maintain healthy skin.

Aging involves not only these free radicals but also an enzyme called matrix metalloproteinase (MMP), a collagen degrading enzyme. In other words, the synthesis and degradation of extracellular matrix such as collagen in vivo is properly controlled, but collagen synthesis decreases as aging progresses, and the expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, promotes the skin. Elasticity is reduced and wrinkles are formed. In addition, such degradation 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. Factors that determine skin color include basic or partial pigmentation, such as tanning due to blemishes, freckles, and sun exposure, other than acne, scars, and the distribution of acne and skin, except for differences in race, region, sex, and age. Blood circulation, stress, and health. Among the above factors, the most important factor is pigmentation.

Pigments that affect skin color include melanin, melanoids, carotene, and hemoglobin, the most important of which is melanin, the most important factor affecting the biosynthesis of melanin is the amount of ultraviolet rays and hormones in the body.

Melanin plays a major role in preventing or damaging the skin from ultraviolet rays by absorbing or scattering ultraviolet rays. There is no special maximum absorption wavelength and it 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 occurs through a series of oxidation processes, beginning with the conversion of tyrosine, an amino acid, from the melanocytes of melanin-forming cells into tyrocinase and into dihydroxy phenylalanine. It is formed from a polymer of eumelanin.

This biosynthesis process is carried out in a special type of brown cells in organelles called melanosomes. Melanosomes, including melanin granules, move from the periphery of the nucleus to the dendritic end of the dendritic projections and into the cytoplasm by the action of keratinocytes. Accumulate around the nucleus.

The synthesis of melanin, the number of melanosomes, and the migration to the surrounding keratinocytes are partially affected by hormones and ultraviolet rays, and primarily by genetic influences. In addition, it is known that metal ions such as cytokines, copper, zinc, and iron, interferon, prostaglandin, histamine, etc., which are involved in the expression of tyrosinase, synthesis and transfer of melanin, and cytokines, are known.

Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150), and some extracts of tyrosina Although it is used as a whitening cosmetic because of its anti-ageing activity, its use is limited due to its low stability in cosmetic formulations, which is degraded and colored, causing off-flavor, efficacy at the biological level, unclear effect, and safety problems. to be. The above substances have been demonstrated to be effective in inhibiting tyrosinase, but the effect is low in experiments similar to the actual biolevels. Therefore, the evaluation of the inhibitory effect by melanoma cell culture similar to the biological level is required. 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, cosmetics using natural products have recently been developed to reduce skin irritation caused by various chemicals. Natural ingredients have less side effects on the skin, and as the consumer's response to cosmetics using natural ingredients has recently increased, the development value of cosmetics as a cosmetic ingredient is increasing.

Therefore, the present inventors have studied the possibility of applying to cosmetics in a variety of natural products that have not been known so far, as a result of selecting the maple of the maple and deciduous broad-leaved arborescent and extracts from it, the melanin production inhibitory effect, MMP-1 production inhibition Effects of enhancing collagen synthesis, improving wrinkles, inhibiting MMP-1 production by induction of UV rays, reducing cytotoxicity by UV irradiation, and moisturizing effects showed that the results were very good. I found it.

It is an object of the present invention to contain the extract of the bee, skin whitening and MMP-1 production inhibitory effect, MMP-1 production inhibitory effect by UV induction, collagen synthesis enhancement effect, such as skin wrinkle improvement effect and UV damage inhibition effect and skin It is to provide a cosmetic composition exhibiting a stimulating effect.

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

The object of the present invention as described above is achieved by a cosmetic composition for skin whitening comprising bee extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for improving skin wrinkles comprising bee extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for inhibiting skin damage by ultraviolet rays comprising a bee extract as an active ingredient.

Still another object of the present invention is achieved by a cosmetic composition for alleviating skin irritation, including a bee extract as an active ingredient.

Another object of the present invention is achieved by a skin moisturizing cosmetic composition comprising a bee extract as an active ingredient.

In addition, preferably, the bee extract is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, dichloromethane, hexane And an extract extracted using a solvent selected from the group consisting of the mixture, or extracted using a supercritical extraction method or an ultrasonic extraction method.

In addition, preferably, the bee extract is characterized in that it contains 0.001 to 30.0% by weight relative to the total weight of the cosmetic composition.

In addition, the cosmetic composition preferably inhibits melanin biosynthesis, inhibits MMP-1 production, inhibits MMP-1 production by induction of ultraviolet rays, enhances collagen synthesis, and improves whitening, wrinkle improvement, and cytotoxicity by UV irradiation. It is characterized by showing a skin damage inhibiting and irritating effect and a moisturizing effect.

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

Still another object of the present invention is to apply a cosmetic composition containing the bee extract to human skin, whitening effect, wrinkle improvement effect, cytotoxicity mitigating effect by UV irradiation, skin damage inhibitory effect, skin irritation easing effect or moisturizing It is achieved by the makeup method characterized by obtaining the effect.

The bee extract according to the present invention has a whitening effect such as melanin production inhibitory effect (Experimental Example 1), which is a substance causing blemishes, freckles and skin pigmentation.

In addition, the bee extract extracts MMP-1 by inhibiting MMP-1 production (Experimental Example 2), enhancing collagen synthesis (Experimental Example 3), and improving wrinkles (Experimental Example 6) and moisturizing effect (Experimental Example 7). 1 production inhibition (Experimental Example 4), to reduce the cytotoxicity by ultraviolet irradiation (Experimental Example 5), and has an excellent skin damage suppression effect or irritation relief effect.

Therefore, the cosmetic composition containing such bee extract has a whitening effect (Experimental Example 1), MMP-1 production inhibitory effect (Experimental Example 2), collagen synthesis enhancement effect (Experimental Example 3), UV-induced MMP-1 production inhibition It can be seen that the effect (Experimental Example 4), the cytotoxicity mitigating effect (Experimental Example 5) by the ultraviolet irradiation, the wrinkle improvement effect (Experimental Example 6) and the skin moisturizing effect (Experimental Example 7).

The present invention relates to a cosmetic composition for skin whitening, skin wrinkle improvement, skin damage suppression by ultraviolet rays, skin irritation relief, skin moisturizing comprising a bee extract as an active ingredient.

The bee (Acer tegmentosum Maxim), which is used as an active ingredient in the cosmetic composition of the present invention, grows in a height of 10 to 15 m as a plant that grows in a damp valley of highlands. The branches are named honeycomb because they are honeycomb-shaped. Another name is Sancheongmok, and it is known that folk cancer is cured for liver cancer, cirrhosis, hepatitis and leukemia. Ingredients in wood include 3,7,3 '', 4 ''-tetramethyl-quercetin (1), 5,3 ''-dihydroxy-3,7,4 ''-trimethoxy flavone (2), 2,6 -dimethoxy-p-hydroquinone (3), (-)-catechin (4), morin-3-O--L-lyxoside (5), p-hydroxy phenylethyl-O--D-glucopyranoside (6), 3, 5-dimethoxy-4-hydroxy phenyl-1-O--D-glucoside (7), fraxin (8), 3,5-dimethoxy-benzyl alcohol 4-O--D-glucopyranoside (9), 4- (2 , 3-dihydroxy propyl) -2,6-dimethoxy phenyl -D-glucopyranoside (10).

In the present invention, the bee extract means that obtained by extracting from various organs or parts of the bee (e.g., leaves, flowers, roots, stems, branches, bark and seeds, etc.), preferably leaves, stems, branches, bark Or an extract obtained from a root, more preferably an extract obtained from a leaf, a branch, a bark or a root, and most preferably an extract obtained from a branch or a bark.

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

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

In the present invention, the bee extract is a variety of extraction solvents, for example, water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol , At least one extraction solvent selected from the group consisting of glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, preferably 70% (v / v) obtained using ethanol or water, most preferably obtained using 70% (v / v) ethanol.

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

In addition, the bee extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone the conventional purification and fermentation process.

For example, decompression by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by ultrasonic extraction, separation using ultrafiltration membranes with constant molecular weight cut-off values, various chromatography (size, charge, hydrophobicity or affinity) The active fraction obtained through various purification and extraction methods additionally carried out, such as by separation by the sex) or by fermentation products using natural or various microorganisms, is also included in the extract of the present invention.

In addition, the present invention provides a cosmetic composition, characterized in that the extract is obtained by cold condensation at room temperature, heating and filtration, further obtained by concentrating the solvent under reduced pressure or lyophilization.

The decompression by carbon dioxide and the supercritical extraction by high temperature mean supercritical fluid extraction. Generally, the supercritical fluid is a liquid having a gas when the gas reaches a critical point under high temperature and high pressure. It is gaseous, chemically similar in polarity to nonpolar solvents, and because of its properties, supercritical fluids are used for extraction of fat-soluble substances (J. Chromatogr. A. 1998; 479: 200-205).

Carbon dioxide becomes a supercritical fluid with both liquid and gaseous properties as the pressure and temperature reach a critical point through the operation of a supercritical fluid device, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from the 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 the active ingredient by using a supercritical carbon dioxide or a mixed fluid in which a co-solvent is added to carbon dioxide.

The cosolvent may be used one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether.

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

In addition, the ultrasonic extraction method is an extraction method using the energy generated by the ultrasonic vibration, the ultrasonic wave can destroy the insoluble solvent contained in the sample in the water-soluble solvent, the reactants located around due to the high local temperature generated Since the kinetic energy of the particles is increased, sufficient energy required for the reaction is obtained, and the effect of ultrasonic energy is induced to increase the pressure by increasing the mixing effect of the substance and the solvent contained in the sample, thereby increasing the extraction efficiency.

As the extraction solvent that can be used in 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 vacuum filtered to recover the filtrate, and then removed by a vacuum evaporator, the extract can be obtained through a conventional extract preparation method of freeze drying.

The bee extract according to the present invention also includes the extract after the fermentation process, the fermentation extract of bee can be prepared as follows. Finely crush the extract area (leaf, flower, root, stem, branch, bark and seeds) of the bee tree to 100 ~ 500 mesh, and then add 1 ~ 50g / L to the conventional microbial culture solution and add yeast strain or lactic acid bacteria. Add microorganisms in amounts of 10,000-100,000 cfu / L. The culture temperature is incubated in the normal microbial culture conditions of 30 ~ 37 ℃. The pH is 5 to 7 and incubated for about 5 to 10 days under aerobic or anaerobic conditions. It can then be obtained through aging and filtration.

The cosmetic composition of the present invention is an effective amount of bee extract extracts showing melanin production inhibition, MMP-1 production inhibition, collagen biosynthesis enhancement, inhibition of MMP-1 production excessively generated by ultraviolet light, cytotoxicity caused by ultraviolet light and moisturizing effect Characterized in that it comprises a.

According to the present invention, the bee extract is contained 0.001 to 30.0% by weight based on the total weight of the cosmetic composition, more preferably, the bee extract is contained 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 0.001% by weight, no skin improvement effect is observed, and when the content of the extract is more than 30.0% by weight, the degree of improvement of the skin improvement effect against the increase of the content is insignificant, and there is a problem in the safety and stability of the formulation and economics. Neither ever

The cosmetic composition of the present invention containing the bee extract has melanin production inhibitory effect (Experimental Example 1), MMP-1 production inhibitory effect (Experimental Example 2) and collagen synthesis enhancement effect (Experimental Example 3), MMP- by UV induction 1 It is excellent in the production inhibitory effect (Experimental example 4), the cytotoxicity relieving effect by the ultraviolet irradiation (Experimental example 5), the skin wrinkle improvement effect (Experimental example 6), and the skin moisturizing effect (Experimental example 7).

Therefore, the cosmetic composition of the present invention comprising a bee extract with such various functions is to have an excellent skin damage suppression and stimulation relieving effect by relieving cytotoxicity by ultraviolet irradiation with excellent whitening effect, wrinkle improvement effect, moisturizing effect It features.

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

Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, 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.

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

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

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

In addition, the present invention by applying a cosmetic composition comprising the extract of the bee extract of the present invention as an active ingredient to the skin of the human being, excellent whitening effect, anti-wrinkle effect, moisturizing effect and relieve cytotoxicity by ultraviolet irradiation to inhibit excellent skin damage And it provides a makeup method characterized in that it exhibits a stimulating effect.

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

The cosmetic composition of the present invention may be used alone or in duplicate, or may be used in combination with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention may be used according to a conventional method of use, and the number of times of use may vary depending on the skin condition or taste of the user.

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

Performing a cosmetic method using a cosmetic composition comprising the extract of the bee tree of the present invention, whitening effect, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, UV-induced MMP-1 production inhibitory effect, cytotoxicity by ultraviolet irradiation It can be used to alleviate the effects of excellent skin damage, improve wrinkles, or moisturize.

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: Bee  Extract manufacturer

The dried and dried burl branches were refluxed three times for 5 hours with an aqueous 70% (V / V) ethanol solution, cooled, and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 캜 or lower. The beech extract was prepared by dispersing / dissolving using a 50% butylene glycol solution such that the vacuum concentrate and the freeze-dried were 1% by weight.

Experimental Example  One: Bee  Inhibitory Effects of Extracts on Melanin Production Using B16F1 Melanocytes

Experimental Example 1 is to determine the whitening effect by looking at the degree of inhibition of melanin production on B16F1 melanin forming cells in order to confirm the whitening effect of the bee extract obtained in Preparation Example 1. B16F1 melanin forming cells used in Experimental Example 1 are cell strains derived from mice, and cells that secrete a black 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).

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

B16F1 melanin forming cells were divided into 6 well plates at a concentration of 2 X 10 ⁶ per well, and the cells were attached and incubated for 72 hours by treating the bee extract according to Preparation Example 1 at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, and the cells were counted and centrifuged to recover the cells. Quantification of intracellular melanin was performed with a slight modification of the method of Rotan: Cancer Res., 40: 3345-3350, 1980. The cell pellet was washed once with PBS, and then 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added to vortex 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. The melanin production inhibition rate (%) of B16F1 melanin forming cells was calculated by the following Equation 1, the results are shown in Table 1. At this time, hydroquinone and arbutin, which are known to have melanin production inhibitory effects, were used as a comparison group.

A: Melanin amount in wells without sample

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

Name of sample Treatment concentration (%) Melanin production inhibition rate (%) Beetle Extract (Manufacturing Example 1) 0.1 0.57 Hydroquinone 0.1 0.03 Arbutin 0.1 0.20

As can be seen from the results of Table 1, it can be seen that the bee extract of the present invention significantly inhibits melanin production in B16F1 melanin forming cells, compared to hydroquinone and arbutin, which are known to have melanin production inhibitory effects. The bee extract showed a melanin production inhibition rate of 0.57%, 17 times that of hydroquinone and 2.85 times that of arbutin. Therefore, it can be seen that the cosmetic composition comprising the bee extract of the present invention has an excellent effect on inhibiting melanin production.

Experimental Example  2: Bee  Extract MMP -1 suppression effect

Experimental Example 2 is a substance known to have an effect of inhibiting the production of MMP-1 in order to determine whether the bee extract obtained in Preparation Example 1 has an effect of inhibiting the production of collagen degrading enzyme MMP-1 Phosphorus TGF-β was used as a comparative group to determine the effect of inhibiting the production of MMP-1.

Fibroblasts (Korean Cell Line Bank, South Korea), which are human normal skin cells, were seeded in 48-well microplates (Nunc, Denmark) at 1 × 10 ⁶ cells per well, and treated with DMEM medium (Sigma, United States) and 37 ° C. After culturing for 24 hours at 24 hours was further incubated in serum-free DMEM medium containing the beech extract of Preparation Example 1 to a final concentration of 200 ㎍ / ㎖ and serum-free DMEM medium containing no bee extract as a control.

After incubation, the supernatants of each well were collected and the amount of ng / ml newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, United States of America), and MMP-1 production according to Equation 2 below. The percent inhibition was calculated and the results are shown in Table 2.

At this time, TGF-β ( 100 ng / ml , Roche, United States), a substance known to have an effect of inhibiting MMP-1 production, was used as a comparison group.

Test substance Inhibition rate of MMP-1 production (%) Beetle Extract (Manufacturing Example 1) 63.7 TGF-β 23.5

As shown in the results of Table 2, the bee extract of the present invention exhibited an inhibition rate of 63.7% when the MMP-1 production inhibition rate was measured at a concentration of 200 µg / ml, and TGF-β showed an inhibition rate of 23.5%, MMP-1 inhibition was about 2.7 times better than TGF-β. Even if the Nadogae persimmon extract of the present invention takes into account the difference between the concentration of TGF-β (100 ng / ml), which is known to have an inhibitory effect on MMP-1 production, and the concentration of the bee extract of the present invention (200 ㎍ / ml), The above-described degree of inhibition of MMP-1 at the level of the extracts of the mixed substances shows that the bee extract of the present invention has an inhibitory effect on the production of MMP-1.

Experimental Example  3. Bee  Enhancement of Collagen Synthesis by Extracts

Fibroblasts, human normal epithelial cells, were seeded in 48-well microplates at 1 × 10 ⁶ cells per well and incubated in DMEM medium for 24 hours. The bee extract was further incubated for 48 hours in serum-free DMEM medium added to a final concentration of 100 ㎍ / ㎖ and serum-free DMEM medium containing no bee extract as a control. After incubation, the supernatant of each well was collected and measured in ng / ml by measuring the amount of procollagen (procollagen) type I C-peptide (PICP) using a collagen kit (Takara, Japan). Measured. In the comparison group, TGF-β (100ng / ml, Roche, United States), a substance known to have an effect of enhancing collagen synthesis, was used as a comparison group, and the extract of bee was extracted at 0.01% (w / v) concentration. Was carried out. Collagen biosynthesis increase rate was calculated according to the following equation (3), the results are shown in Table 3.

Test substance Increase in collagen biosynthesis (%) Beetle Extract (Manufacturing Example 1) 77.4 TGF-β 69.6

Table 3 shows the results of measuring the collagen synthesis enhancement effect of the cultures added with bee extract and TGF-β using fibroblasts. According to Table 3, the bee extract of the present invention showed a biosynthesis rate of 77.4% when the collagen biosynthesis increase rate was measured at 0.01% concentration, and TGF-β, a substance known to enhance collagen synthesis, was 69.6% biosynthesis. It can be seen that the rate. Even if the concentration of TGF-β (100 ng / ml), which is known to enhance collagen synthesis, and the concentration (0.01% (w / v)) of the bee extract of the present invention, the extracts are mixed The collagen biosynthesis rate as described above at the level shows that the bee extract of the present invention has a collagen synthesis enhancing effect.

Experimental Example  4. Bee  By UV induction of extract MMP -1 suppression effect

In general, MMP-1 is known to increase its amount by ultraviolet light. Therefore, an experiment was conducted to determine whether the bee extract of the present invention inhibits the amount of MMP-1, which is increased by UV light. MMP-1 production inhibition effect measurement by UV induction was carried out in the same manner as in Experiment 2 except that a certain amount of UVB treatment prior to sample treatment. As a bee extract used in this experiment, the one obtained in Preparation Example 1 was used. MMP-1 production inhibition rate of the bee extract was calculated according to Equation 2, the results are shown in Table 4 below. At this time, TGF-β (100 ng / ml, Roche, United States), a substance known to have an inhibitory effect on the production of MMP-1, was used as a comparison group, and the bee extract was tested at a concentration of 0.01% (w / v).

Test substance Inhibition rate of MMP-1 production (%) Beetle Extract (Manufacturing Example 1) 70.3 TGF-β 32.5

Table 4 shows the results of measuring the amount of collagen degrading enzyme MMP-1 secreted in the culture solution to which the bee extract and TGF-β culture medium were added after UVB irradiation to the cultured fibroblasts. Inhibition of the production of MMP-1 (%) is about 2.16 times better than TGF-β, a substance known to inhibit MMP-1 production by UV rays, at the concentration of 0.01% (w / v). Indicated.

Experimental Example  5. Bee  Cytotoxicity-Reducing Effect of Extracts by UV Irradiation

Experimental Example 5 was carried out to evaluate the alleviation effect of cytotoxicity by ultraviolet irradiation of the bee extract obtained in Preparation Example 1.

Fibroblasts (fibroblasts) were placed in 24-well test plates 1 X 10 ⁵ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with UV 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B15W, Sankyo Dennki, Japan), after which PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. It was incubated for 24 hours after treatment with bee extract. After 24 hours, the medium was removed, and 500 μl of cell culture medium and 60 μl of MTT solution (2.5 mg / ml) were added to each well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04N) was added thereto. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, followed by measurement of 565 nm absorbance in a microplate reader. Cell viability (%) was measured by Equation 4 below, and cytotoxicity relaxation rate (%) by UV was calculated by Equation 5.

Bo: Absorbance at 565 nm of the well of the cell culture medium alone

Bt: Absorbance at 565 nm of a well that underwent chromogenic reaction in a sample not treated with the sample

St: 565 nm absorbance of the wells that reacted the sample treated wells

Bo: cell viability of wells without UV irradiation and no sample treatment

Bt: Cell viability of wells irradiated with ultraviolet light and not treated with the sample

St: Cell survival rate of wells treated with UV

Name of sample Treatment concentration Cytotoxic Relaxation Rate (%) Bee extract 0.1% 34.7 0.2% 45.0

According to Table 5, the cytotoxicity alleviation rate (%) of the bee extract extract increased by 34.7% at 0.1% concentration and 45.0% at 0.2% concentration. It was found that it effectively defends against cytotoxicity caused by ultraviolet rays. Through the experimental results, it can be seen that the bee extract of the present invention effectively protects cell damage by ultraviolet rays even at low concentrations.

Hereinafter, based on the results of the above experimental examples, to present a cosmetic composition containing a bee extract as an active ingredient, but the formulation of the present invention is not necessarily limited thereto.

Formulation Example  1. Flexible Cosmetics

Formulation examples of the flexible longevity was prepared as shown in Table 6 in accordance with a conventional method.

ingredient Content (unit: weight%) Bee extract
glycerin
1.3-butylene glycol
PEG 1500
Allantoin
DL-panthenol
E.T.A.-2NA
Benzophenone-9
Sodium hyaluronate
ethanol
Octicdodeceth-16
Polysorbate 20
Preservative, fragrance, coloring
Distilled water 3.0
5.0
3.0
1.0
0.1
0.3
0.02
0.04
5.0
10.0
0.2
0.1
a very small amount
Balance Sum 100

Formulation Example  2. Converging Cosmetics

Formulation examples of convergent longevity was prepared as shown in Table 7 in accordance with a conventional method.

ingredient Content (unit: weight%) Bee extract
glycerin
1.3-butylene glycol
Allantoin
DL-panthenol
E.T.A.-2NA
Benzophenone-9
Sodium hyaluronate
ethanol
Polysorbate 20
Witch hazel extract
Citric acid
Preservative, fragrance, coloring
Distilled water 1.5
2.0
2.0
0.2
0.2
0.02
0.04
3.0
15.0
0.3
2.0
a very small amount
a very small amount
Balance Sum 100

Formulation Example  3. Nutritional Cosmetics

Formulation example of nutrient cosmetics was prepared as shown in Table 8 in accordance with a conventional method.

ingredient Content (% by weight) Bee extract
Stearyl alcohol
lanolin
Polysorbate 60
Sorbitan stearate
Cured Vegetable Oil
Mineral oil
Squalane
Trioctanoine
Dimethicone
Tocopherol Acetate
Carboxy Vinyl Polymer
glycerin
1.3-butylene glycol
Sodium hyaluronate
Tri ethanolamine
Preservative, fragrance, coloring
Distilled water 1.5
1.5
1.5
1.3
0.5
1.0
5.0
3.0
2.0
0.8
0.5
0.12
5.0
3.0
5.0
0.12
a very small amount
Balance Sum 100

Formulation Example  4. Nutrition Cream

Formulation of nutrition cream was prepared according to the conventional method as shown in Table 9.

Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation Example 2 ingredient Content (% by weight) Content (% by weight) Bee extract
Retinol
Lipophilic monostearate
Cetearyl Alcohol
Stearic acid
Wax
Polysorbate 60
Sorbitan stearate
Cured Vegetable Oil
Squalane
Mineral oil
Trioctanoine
Dimethicone
Sodium magnesium silicate
glycerin
Betaine
Triethanolamine
Sodium hyaluronate
Preservative, fragrance, coloring
Purified water 3.0
-
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance -
-
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance -
3000 IU
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance Sum 100 100 100

Formulation Example  5. Massage Cream

Formulation example of the massage cream was prepared according to the conventional method as shown in Table 10 below.

ingredient Content (unit: weight%) Bee extract
Chin type glycerin monosuccinic acid
Stearyl alcohol
Stearic acid
Polysorbate 60
Sorbitan stearate
Isostearyl Isosterate
Squalane
Mineral oil
Dimethicone
Hydroxyethyl Cellulose
glycerin
Triethanolamine
Preservative, fragrance, coloring
Distilled water 3.0
1.5
1.5
1.0
1.5
0.6
5.0
5.0
35.0
0.5
0.12
6.0
0.7
a very small amount
Balance Sum 100

Formulation Example  6. Essence

Formulation examples of the essence was prepared according to the conventional method as shown in Table 11.

ingredient Content (unit: weight%) Bee extract
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
E.T.A.-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxy Vinyl Polymer
Triethanolamine
Octyldodecanol
Octyldodeceth -16
ethanol
Preservative, fragrance, coloring
Distilled water 5.0
10.0
5.0
2.0
0.1
0.3
0.02
0.04
0.1
8.0
0.2
0.18
0.3
0.4
6.0
a very small amount
Balance Sum 100

Formulation Example  7. Pack

Formulation examples of the pack was prepared according to the conventional method as shown in Table 12 below.

ingredient Content (unit: weight%) Bee extract
Polyvinyl alcohol
Cellulose gum
glycerin
Fiji 1500
Cyclodextrin
DL-Panthenol
Allantoin
Monomethylammonium Glycyric Acid
Nicotinamide
ethanol
Fiji 40 Cured Castor Oil
Incense, pigment, preservative
Distilled water 3.0
15.0
0.15
3.0
2.0
0.15
0.4
0.1
0.3
0.5
6.0
0.3
a very small amount
Balance Sum 100

Experimental Example  6. Bee  Containing extract Cosmetic  Skin wrinkle improvement

Clinical trials were carried out on the skin wrinkle improvement effect of the cosmetic containing the extract of the bee of the present invention. Clinical trials were evaluated through the actual use test of each cosmetic.

The experiment was carried out by comparing the nutrient cream of Formulation Example 4, which contains the bee extract, and the cream obtained by replacing the bee extract with purified water, as Comparative Formulation Example 1, and a retinol which is a substance known to have an anti-wrinkle effect. As 2, the human body was identified and compared with the skin wrinkle improvement effect due to the use of cosmetics.

The cosmetics of Formulation Example 4 and Comparative Formulation Example 1 used in this comparative evaluation experiment are in the form of a cream, the composition of which is shown in Table 9. For 60 females (20-35 years old), the experimental group was divided into three groups, each of 20 groups of Formulation Example 4, Comparative Formulation Example 1, and Comparative Formulation Example 2, and using both sides of the face. After two months, the wrinkle improvement effect was visually evaluated to confirm the degree of wrinkle improvement.

After completion of the experiment, the skin wrinkle improvement effect was evaluated by visual observation by an experienced doctor before and after using the product for three months. The experimental results are shown in Table 13 below and FIG. 1.

Wrinkle improvement Effective rate (%) Great slightly none Formulation Example 4 24 4 2 85.0 Comparative Formulation Example 1 3 4 23 25.0 Comparative Formulation Example 2 (Retinol) 22 4  4 86.6

According to the experimental results, the cosmetic containing the bee extract (Formulation Example 4) showed a significant wrinkle improvement effect 3.4 times higher than the cosmetic (Comparative Example 1) containing no bee extract. It was found that the wrinkle improvement effect was superior to that of the cream of Comparative Formulation Example 2 containing retinol, a substance known to have an anti wrinkle effect.

Moreover, the skin irritation could not be observed in the skin of the subjects who apply the cosmetic composition of the present invention to the skin.

Experimental Example  7. Bee  Containing extract Cosmetic  Moisturizing effect

In order to evaluate the moisturizing power of the cosmetic preparation prepared in Formulation Example 4 containing the bee extract, the moisture retention ability of the skin to which each cosmetic was applied was measured as follows.

In the constant temperature and humidity room of 22 ° C. and 45% RH, each of the cosmetic formulations of Formulation Example 4 and Comparative Example 1, which does not contain the extract, was divided into half of 24 test subjects' faces. The water content after 4 weeks was measured while apply | coating a product (2 times / day).

As a measuring device, a moisture content measuring device (Corneometer CM825, Courage + Khazaka, Germany) was used to measure the moisturizing power by measuring the electric capacity of the skin according to the moisture content of the skin. The results are shown in Table 14 below.

Moisture content Use Formulation Example 4 Comparative Formulation Example 1 0 Week 23.61 ± 9.34 23.44 ± 11.36 4 Week 34.61 ± 7.09 27.38 ± 5.58 4 Week-0Week 11.00 ± 2.25 3.94 ± 5.78

As shown in the results of Table 14, the cosmetic composition containing the bee extract of the present invention (Formulation Example 4) as a result of measuring the moisturizing effect after 4 weeks of use, the cosmetic content does not contain the bee extract as 11.00 increase in skin moisture content It can be seen that the skin moisturizing effect is superior to 3.94, which is an increase in skin moisture content of the composition (Comparative Formulation Example 1).

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

1 is a result of measuring the wrinkle improvement effect of the cosmetic composition (Formulation Example 4) containing a bee extract in the embodiment of the present invention.

Claims (9)

Cosmetic composition for skin whitening comprising bee extract as an active ingredient. Cosmetic composition for improving skin wrinkles containing bee extract as an active ingredient. Cosmetic composition for preventing skin aging due to ultraviolet rays containing an extract of bee trees as an active ingredient. Cosmetic composition for skin moisturizing containing bee extract as an active ingredient. The cosmetic composition according to any one of claims 1 to 4, wherein the bee extract is contained in an amount of 0.001 to 30.0 wt% based on the total weight of the cosmetic composition. The method according to any one of claims 1 to 4, wherein the bee extract is (a) water, anhydrous or hydrous lower alcohol of 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, Solvent extraction using an extraction solvent selected from the group consisting of butyl acetate, diethyl ether, dichloromethane, hexane and mixtures thereof, (b) supercritical extraction, (c) ultrasonic extraction or (d) fermentation or natural Cosmetic composition, characterized in that extracted by fermentation using fermentation metabolism. The cosmetic composition according to any one of claims 1 to 4, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion emulsion, wax foundation Cosmetic composition, characterized in that it has a formulation selected from the group consisting of a pack, a massage cream and a spray. A cosmetic method according to any one of claims 1 to 4, wherein the cosmetic composition according to any one of claims 1 to 4 is applied to human skin to obtain a skin whitening effect, a wrinkle improvement effect, an anti-aging effect by ultraviolet rays, or a skin moisturizing effect. delete

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