KR101445643B1 - Cosmetic composition comprising the extract of Halenia corniculata (L.) Cornaz as active ingredient - Google Patents

Abstract

The present invention relates to an anthracnose corniculata (L.) Cornaz) as an active ingredient, wherein the anthocyanin extract is an effective ingredient in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. According to the present invention, an antharophyll extract can provide a variety of functional cosmetic compositions having an inhibitory effect on MMP-1 production, collagen biosynthesis effect, melanin formation inhibitory effect, preservative effect, skin wrinkle improvement effect, moisturizing effect and antioxidative effect.

Description

[0001] The present invention relates to a cosmetic composition containing an extract of anchovy flower as an active ingredient (Cosmetic composition comprising Halene corniculata (L.) Cornaz as active ingredient)

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

Skin protects the body from moisture, electrolytes, and proteins from being lost from the outermost part of the body. It is an important organ of the body that performs various roles such as body temperature control, skin sensory function, and immune function. . Skin aging is caused by endogenous aging, genetic factors, hormones, etc., and extrinsic aging caused by pollution, smoking, stress, drinking, and UV exposure.

The main cause of extrinsic aging is ultraviolet rays, so it is also called photoaging. Ultraviolet light is a light of a wavelength range of 200-400 nm. It reaches the double ground and affects the skin. UVA (320-400 nm) and UVB (290-320 nm) cause the generation of active oxygen causing skin damage, collagen fiber damage And deterioration of elasticity due to increase in decomposition and aging phenomenon such as wrinkle formation, pigmentation such as spots, black spots and freckles.

Another important factor in skin aging is reactive oxygen species (ROS). Active oxygen also occurs in normal metabolism, but it is overproduced in the body due to environmental pollution, chemicals, ultraviolet rays, blood circulation disorder, and stress. In general, active oxygen protects the body from bacteria or foreign substances. However, when it is overproduced, it attacks not only DNA, lipids, proteins, polysaccharides but also cells and tissues to lose or alter its function and cause various diseases and aging. Therefore, it is important to maintain the healthy skin by properly removing the active oxygen to prevent cell and tissue damage.

Elasticity and wrinkle formation are mainly caused by damages and changes in the dermis. Ultraviolet light and active oxygen increase the production of matrix metalloprotease (MMP), which inhibits the synthesis of collagen fibers, causes denaturation and degrades collagen fibers and elastic fibers. As a result, the normal structure and shape of the skin are maintained, and the skin tissue is tensioned to weaken the collagen fiber, which plays a role of strengthening the tissue, and the skin is dented and wrinkled. As the age increases, the body produces more dihydrotestosterone (DHT) hormones. Increasing levels of DHT reduce the production of elastic fibers in the dermal fibroblasts. In addition, ultraviolet light penetrates into the dermis and damages fibroblasts, thereby reducing the elasticity of the skin by reducing the production of elastic fibers that impart elasticity to the skin.

Glycosaminoglycan (GAG's), which is present in the dermis, is a structure in which disaccharide-type sugars are repeatedly linked, and includes hyaluronic acid, chondroitin sulfate and dermatan sulfate. GAG functions to fill void spaces between collagen fibers and elastic fibers, and recognizes changes in surrounding proteins or changes in surrounding cells to transmit signals to cells, thereby affecting growth, differentiation and function of cells, It has a strong binding force with water molecules and plays an important role in water retention. As you get older, the synthesis of GAG is reduced and affects the amount of water in the dermis, weakening collagen and reducing skin tension. The decrease of the dermis thickness and the change of the constitutional component of the dermis cause the skin elasticity to be reduced, so that the skin appears as a sagging or bold wrinkled skin.

BACKGROUND ART Retinoic acid, adenosine, and alpha hydroxy acids are used as anti-wrinkle agents mainly used in cosmetics. However, it is difficult to expect a practical wrinkle-reducing effect because the use is limited or its effect is insufficient because of poor stability or skin irritation. Therefore, it is necessary to develop materials capable of promoting the synthesis of collagen fibers and elastic fibers and controlling the expression and activity of MMPs, while ensuring safety and stability.

Melanin, carotene, and hemoglobin are the two most important determinants of human skin color. Melanin plays a major role in preventing skin damage from ultraviolet rays by absorbing or scattering ultraviolet light.

Melanin is synthesized by a reaction in which an amino acid, tyrosine, is oxidized by an enzyme called tyrosinase and then converted to dopaquinone. Two types of tyrosinase (TRP-1 and TRP-2) are known to be involved in the synthesis of melanin, and a series of processes are performed to synthesize eumelanin and peromelanin. Melanin is synthesized in melanocytes, the organelles in the melanocytes, and migrates to the surrounding keratinocytes through the dendrites. Melanocyte stimulation hormone (alpha-MSH), stress, and ultraviolet rays are involved in the proliferation of melanocytes and the activation of melanin synthesis.

There are several types of pigmented skin on the skin that are caused by localized melanin deposits. As you get older, melanocytes are getting smaller and your skin becomes more sensitive to ultraviolet light. In addition, the melanocyte site becomes lumpy and the coloring matter called spot is gathered. A hormonal imbalance that occurs as a result of age impairs the melanocyte and allows the remaining melanocyte mass to produce more melanin. In addition, ultraviolet light stimulates melanocytes, causing more skin pigment, damaging melanocyte DNA, and lengthening the melanocyte site to cause pigmentation such as spots, black spots, and freckles.

Melanin is used as a whitening agent for treating melanin as a treatment for dysentery caused by excessive increase of melanin in the epidermis. In order to alleviate the effect on melanocytes, whitening cosmetics include arbutin (Japanese Patent Laid-open No. 4-93150), kojic acid (Japanese Patent Laid-open No. 56-7710), vitamin C and derivatives thereof (Japanese Patent Laid- Japanese Patent Application Laid-Open No. 192962). The use thereof is limited due to poor stability in the formulation of cosmetics, coloration due to decomposition, generation of odor, unclear efficacy at the living body level, and safety.

Various materials have been developed to prevent aging of skin caused by aging or by ultraviolet rays and active oxygen. Among these, there are few side effects on the skin, development of natural materials having various effects such as antioxidation, wrinkle improvement, whitening, moisturizing and the like are active, and as consumers' interest in natural products increases, the value of development as cosmetic raw materials is further increasing.

Cosmetics can contain water, oils, surfactants, proteins, plant extracts, and other components, which can cause proliferation when microorganisms enter the 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 applicability of various natural products to cosmetics. As a result, the present inventors have selected an antharaceae which is a dragonfly and a plant, and produced an extract from the extract to inhibit MMP-1 production, collagen biosynthesis, The effect of preservation, the effect of improving wrinkles of skin, the effect of moisturizing, and the antioxidant activity of skin have been measured. As a result, it has been found that the efficacy as a cosmetic product can be expected.

It is an object of the present invention to provide a cosmetic composition which contains an anthony flower extract and exhibits an effect of inhibiting MMP-1 production, collagen biosynthesis effect, melanin formation inhibitory effect, preservative effect, skin wrinkle improving effect, moisturizing effect and skin antioxidative effect.

It is still another object of the present invention to provide a cosmetic method using the cosmetic composition containing the anthrax extract.

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

Preferably, the cosmetic composition is for improving skin wrinkles.

Preferably, the cosmetic composition is for skin whitening.

Preferably, the cosmetic composition is for nail.

Preferably, the cosmetic composition is for moisturizing.

Preferably, the cosmetic composition is for antioxidation.

Preferably, the cosmetic composition is for preventing skin aging.

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

Preferably, the cosmetic composition exhibits an effect of suppressing MMP-1 production, an effect of collagen biosynthesis, an effect of inhibiting melanin formation, a preservative effect, a skin wrinkle improving effect, a moisturizing effect and a skin antioxidative effect.

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 ≪ / RTI >

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

The anthrax extract according to the present invention is effective for inhibiting MMP-1 production (Experimental Example 1), collagen synthesis enhancing effect (Experimental Example 2), melanin formation inhibitory effect (Experimental Example 3), moisturizing effect (Experimental Example 4) (Experimental Example 5), wrinkle improving effect (Experimental Example 6), and skin antioxidative effect (Experimental Example 7).

Further, according to the present invention, in order to maximize the efficacy of the anthrax extract, the effect of inhibiting MMP-1 production, the effect of promoting collagen synthesis, the inhibitory effect on melanin production, and the skin antioxidative effect are further enhanced through ultrasound, supercritical extraction and fermentation processes give.

Datkkot used as an active ingredient of the cosmetic composition of the present invention (datkkot, hwamyo, datkkot pool, dot kkotpul) is Halenia is in English representation is the scientific name corniculata (L.) It is labeled as Cornaz. Yongdam This is a 1-year-old or 2-year-old herb with Yongdam. It grows in sunny meadow. It is 10 ~ 60cm in height. The whole abandon has no hair, and the stem is long and straight. There are four ridges with lots of branches. Leaves are opposite, long oval or narrow ovate, 2 ~ 6cm long, 1-2 ~ 2.5cm wide. It has 3 ~ 5 veins, the tip is pointed and the lower part becomes like petiole. There is a curvature at the back vein and edge. The flowers bloom in June to August in light yellowish green, hanging on the tip of the stem and axillary axilla. Small peduncle is 1 ~ 4cm long and bracts like leaves. The calyx is divided into 4 pieces, and the cut pieces are in the shape of a line. The corolla is bell-shaped, deeply divided into 4 branches, and a base of 3 ~ 7mm in length. There are 4 stamens and the stigma divides into 2 bifurcations. The fruit is barcosiform, divided into two pieces, ripened in September-October. Contains smooth seeds. Korea (Gyeonggi Province, Gyeonggi Province), Japan, China, Heilong River, Sakhalin Island, Kamchatka Peninsula.

 In the present invention, an anthrax flower extract means an extract obtained from various organs (for example, leaves, flowers, roots, stems, etc.) of anchovy flowers, preferably an extract obtained from branches, shells or roots, ≪ / RTI >

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

In the present invention, the anthrax extract can be obtained from various extraction solvents, for example, water, anhydrous or lower alcohols having 1-4 carbon atoms (e.g., methanol, ethanol, propanol and butanol), propylene glycol, And 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. (v / v) ethanol or water, and most preferably 70% (v / v) ethanol.

It is apparent to those skilled in the art that an extract of Anchovy flower of the present invention can be obtained not only by the above-mentioned extraction solvent but also by using other extraction solvent, which exhibits substantially the same effect.

In addition, the anthrax extract of the present invention includes extracts obtained by the conventional extraction and fermentation processes as well as extracts obtained by the above-mentioned extraction solvents. For example, decompression by carbon dioxide, extraction by supercritical extraction at high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cut-off 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 anthrax extract according to the present invention includes an extract obtained through fermentation, and the fermented extract of anthrax can be prepared as follows. The anthracnose is finely crushed to a size of about 100 to 500 mesh, then 1 to 50 g / L of a conventional microorganism culture 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 anthrax flower extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, and more preferably, the anthrax extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition do. When the content of the extract is less than 0.0001% by weight, the effect of improving the skin is not exhibited. When the content of the extract is more than 30.0% by weight, the degree of improvement of the skin against the increase of the content is insignificant, It is not even.

Therefore, the cosmetic composition of the present invention comprising anchovy extract having various functions of the present invention has excellent collagen synthesis promoting effect, MMP-1 production inhibiting effect, skin wrinkle improving effect, skin whitening effect, melanin formation inhibiting effect, preservative effect, , A skin antioxidant effect, and a skin aging prevention effect.

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.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / 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, methyl taurate, 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. In addition, the cosmetic composition according to the present invention can be used in accordance with a conventional method of use, and the number of times of use may be varied depending on the skin condition or taste of the user.

By carrying out the cosmetic method using the cosmetic composition comprising an anthrax extract of the present invention, it is possible to obtain an effect of inhibiting MMP-1 production, an effect of promoting collagen synthesis, an effect of inhibiting melanin formation, a preservative effect, a moisturizing effect, .

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: Aquatic flowers  Extract preparation

The shrubs of the shrubby anchovy flowers were refluxed for 3 hours with 70% (v / v) aqueous ethanol solution for 5 hours, cooled, and filtered through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or lower.

Manufacturing example  2 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. Aquatic flowers  Extract MMP -1 production inhibitory effect

In Experimental Example 1, the effect of anthocyanin extract obtained in Production Examples 1, 2, and 3 on the production of collagenase, MMP-1, was measured.

Human normal skin cells were inoculated into a 48-well microplate (Nunc, Denmark) at a concentration of 1 × 10 ⁶ cells per well and cultured in DMEM medium (Sigma, USA) and 37 ° C For 24 hours, and further cultured in serum-free DMEM medium supplemented with anthocyanin extracts of Preparation Examples 1, 2 and 3 and in serum-free DMEM medium without anthocyanin extract for 48 hours as a control.

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

[ Equation  One]

Name of sample Treatment concentration (Production Example 1)
MMP-1
Production inhibition rate (%) (Production Example 2)
MMP-1
Production inhibition rate (%) (Production Example 3)
MMP-1
Production inhibition rate (%) Anthurium extract
10ppm 21.59 23.77 22.52 50 ppm 34.83 39.59 37.59 100ppm 48.22 53.21 50.24 500ppm 75.67 78.73 76.15 1000ppm 86.18 89.59 87.01 TGF-beta   (200 nM) 74.6

As shown in the results of Table 1, it was confirmed that the anthocyanin extract of the present invention inhibits the production of MMP-1 (inhibits MMP-1 activity) in a concentration-dependent manner. Even considering the difference between the concentration of TGF-β (200 nM), which is a substance having an inhibitory effect on MMP-1 production, and the concentration of anthocyanin extract (500 ppm) of the present invention, Showed that the anthrax extract of the present invention had an inhibitory effect on MMP-1 production.

Experimental Example  2. Aquatic flowers  Enhancement of collagen synthesis of extract

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. Serum-free DMEM medium supplemented with anthocyanin extract prepared in Preparation Examples 1, 2 and 3, and control group were further cultured in serum-free DMEM medium without anthocyanin extract 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.

[ Equation  2]

Name of sample Treatment concentration (Production Example 1)
Collagen biosynthesis rate (%) (Production Example 2)
Collagen biosynthesis rate (%) (Production Example 3)
Collagen biosynthesis rate (%) Anthurium extract 2.5 ppm 11.9 13.5 12.1 5 ppm 18.9 22.8 20.5 10ppm 41.8 49.2 44.7 25 ppm 95.7 103.5 97.9 50 ppm 158.5 169.8 161.3 TGF-beta (200 nM) 165.3

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

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

In Experimental Example 3, to examine the whitening effect of the anthocyanin extract obtained in Production Examples 1, 2, and 3, the degree of inhibition of melanin formation on B16F1 melanocyte-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 melanocyte-forming cells were divided into 6 well plates at a concentration of 2 X 10 ⁵ per well. After attaching the cells, the anthracycline extract according to Preparation Examples 1 to 3 was cultured for 72 hours at a concentration not causing toxicity. After incubation for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Lotan (Cancer Res., 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin, and the absorbance of melanin was measured at 405 nm with a microplate reader. The inhibition rate (%) of melanin formation was measured. Melanin formation inhibition rate (%) of B16F1 melanin-forming cells was calculated by the following equation (3), and the results are shown in Table 3 below.

[ Equation  3]

Melanin formation inhibition rate (%) = [(A-B) / A] 100

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

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

Name of sample Treatment concentration (%) Melanin formation inhibitory effect (%) Production Example 1 Production Example 2 Production Example 3 Anthurium extract
(Production Example 2) 0.001 21.57 23.47 22.51 0.005 39.17 40.69 41.36 0.01 51.23 56.12 55.52 0.05 70.46 76.51 72.92 Arbutin 200ppm 67.65

As can be seen from the results in Table 3, it was found that the purified anthrax extract prepared according to Production Examples 1, 2, and 3 of the present invention significantly inhibited melanin production in B16F1 melanin-forming cells.

Formulation Example  One : Aquatic flowers  Extract-containing Cosmetics  Preparation of composition

Cosmetic preparations containing anchovy extract were prepared in the form of cosmetic compositions (Formulation Examples 1, 2 and 3) containing anchovy extracts of Preparation Examples 1 to 3, respectively. For comparison of efficacy, glycerin and 1,3butyl (Comparative formulation example 1) containing glycerin and sorbitol (comparative formulation example 2), cosmetic composition (comparative formulation example 3) containing neither extract nor moisturizer were shown in the following table Respectively.

division Component (% by weight) Formulation Example 1 Formulation Example 2 Formulation Example 3 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 A Anthurium extract
(Production Example 1) 5.0 - - - - Anthurium extract
(Production Example 2) - 5.0 - - - Anthurium extract
(Production Example 3) 5.0 glycerin 5.0 5.0 1,3-butylene glycol - - 5.0 - - Sorbitol - - - 5.0 - 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  4 : Aquatic flowers  Moisturizing effect of extract

The clinical moisturizing effects of the cosmetic compositions of Formulation Examples 1, 2 and 3 were measured as follows. A, B, and C groups were divided into 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 anthocyanin extracts, and a cosmetic composition comprising glycerin and sorbitol as a moisturizing agent (Comparative Example 2) cosmetic compositions were applied to the face for 2 weeks each for 8 weeks. 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 5 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 22.3 38.3 46.8 Formulation Example 2 21.9 39.5 48.5 Formulation Example 3 23.1 41.8 50.8 Comparative Formulation Example 1 23.1 34.1 43.2 Comparative Formulation Example 2 23.3 33.9 42.5 Comparative Formulation Example 3 23.1 24.8 25.3

As shown in the above Table 5, the moisturizing effect of the cosmetic compositions (Formulation Examples 1, 2 and 3) containing the anthrax extract of the present invention is superior to the cosmetic compositions containing other moisturizers (Comparative Formulations 1 and 2) I could confirm.

Experimental Example  5: Aquatic flowers  Preservative effect of extract

In order to examine the preservative effect of the anthuria extract, the cosmetic compositions of Formulation Example 4-11 containing the anthrax extracts of Preparation Examples 1, 2 and 3 at various ratios in the composition of the following Table 6, the comparative formulation containing purified water instead of the anthrax flower extract The cosmetic of Comparative Formulation Example 5 containing methylparaben and imidazolidinyl urethane as a preservative instead of the cosmetic and anchovy extract of Comparative Example 4 was prepared.

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

The cosmetic effects of the formulation examples and comparative formulations of Table 6 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 (ATCC 9027) and Staphylococcus aureus (ATCC 6538) 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 7 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, And the results are shown in Table 7 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 850000 40000 5000 500 <100 - Formulation Example 5 1 x 10 ⁷ 720000 17000 1500 250 <100 - Formulation Example 6 1 x 10 ⁷ 320000 5000 450 <100 <100 - Formulation Example 7 1 x 10 ⁷ 780000 34000 3000 400 <100 - Formulation Example 8 1 x 10 ⁷ 670000 11000 850 150 <100 Formulation Example 9 1 x 10 ⁷ 285000 4100 300 <100 <100 Formulation Example 10 1 x 10 ⁷ 750000 24000 2500 500 <100 - Formulation Example 11 1 x 10 ⁷ 650000 8000 700 300 <100 - Comparative Formulation Example 4 1 x 10 ⁷ 8700000 8200000 7100000 6400000 5700000 +++ Comparative Formulation Example 5 1 x 10 ⁷ 360000 500 450 <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 7, the cosmetic composition of Comparative Example 4, which did not use the preservative, showed molds on the surface and the entire surface within 4 weeks, and the bacterial counts were remarkably increased even after 1 day. However, the cosmetic compositions of Formulation Examples 4 to 11 containing the anthracyca extract of the present invention exhibited a concentration-dependent antiseptic effect on bacteria and fungi, and after about 28 days, 2.0% (w / v) The preservation force similar to or better than Comparative Formulation Example 5 in which the preservative methylparaben and imidazolidinyl urea were mixed was secured. When the content of the preservative was judged to be 2.0% or more, It was found that a far superior buoyant force was obtained as compared to Comparative Formulation Example 5 in which methylparaben, which is methylparaben, and imidazolidinyl urea were mixed. In particular, it was confirmed that the anthocyanin extract prepared in Preparation Examples 2 and 3 was more excellent in flotation power than Preparation Example 1.

Experimental Example  6. Aquatic flowers  Effect of extracts on skin wrinkles

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

That is, in the clinical experiment, the cosmetic composition of Comparative Formulation Example 1 containing Formulation Example 2 containing an anthrax extract of Preparation Example 2 in Table 4 and glycerin and 1,3-butylene glycol as a moisturizing agent in place of an anthrax extract was applied to human The effect of improving the skin wrinkles due to the use of cosmetics was confirmed and compared.

Twenty patients (20 to 35 years of age) were applied to Formulation Example 2 on the right side of the face and Comparative Formulation Example 1 on the left side of the face for 2 consecutive months, respectively.

(Comparative Formulation Example 1) treatment group containing glycerin and 1,3-butylene glycol as moisturizing agents, anchovy extract was added to adenosine 3% (v / v) (Comparative Formulation Example 6, no content was shown in Table 4) and an experimental group of Formulation Example 2 containing anchovy flower extract. Using the two sides of the face, the wrinkle improvement effect after 6 weeks 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 8 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 8 below.

Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 2
(Anthocyanin extract) 21 4 3 90.0 Comparative Formulation Example 1
(Moisturizer) 5 7 18 40.0 Comparative Formulation Example 6
(Adenosine) (19) (5) (6) (80.0)

As can be seen from the results of Table 8, the cosmetic composition of Formulation Example 2 containing an anthrax extract of the present invention showed a wrinkle-improving effect about 2.2 times higher than Comparative Formulation Example 1 containing a moisturizer.

In addition, similar results were obtained with the cosmetic preparation of Comparative Example 6 containing adenosine, which is known to have a wrinkle-reducing effect instead of anthocyanin extract, showing that anchovy extract has excellent wrinkle-reducing effect.

Experimental Example  7: DPPH Antioxidant effect measurement

In Experimental Example 7, the antioxidative effect (free radical scavenging ratio) was measured using the DPPH method in order to measure the antioxidative effect of the anthocyanin extract obtained in Production Examples 1, 2, and 3.

The DPPH method measures the antioxidative effect by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The degree of free radical scavenging (%) at a wavelength of 560 nm is measured by comparing the degree of decrease of the absorbance by DPPH reduction with the test substance (anthocyanin extract) and the absorbance of the blank test solution.

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 rate (%) were calculated according to equation (4), and the results are shown in Table 9 below.

[ Equation  4]

Free radical scavenging ratio (%) = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after free radical scavenging of the sample

Bt: absorbance at 560 nm after free radical scavenging of the blank test solution

So: the absorbance at 560 nm before the reaction in the absence of the free radical of the sample

Bo: absorbance at 560 nm before the reaction in the absence of the free radical of the blank test solution

Name of sample Treatment concentration (%) (Production Example 1)
Antioxidative effect(%) (Production Example 2)
Antioxidative effect(%) (Production Example 3)
Antioxidative effect(%) Anthurium extract 0.0001 12.0 14.2 15.7 0.0005 28.1 32.2 31.5 0.001 51.1 56.8 59.2 0.005 75.4 79.8 80.1 0.010 91.3 94.1 93.9 Vit-C 0.05  87.5

As shown in Table 9, in the free radical scavenging experiment using DPPH, the anthocyanin extract according to the present invention showed an increase in antioxidative effect in a concentration-dependent manner, and the anthocyanin extract showed an antioxidative effect (free radical scavenging effect) at a concentration of 0.01% Which is known to have an antioxidative effect than the vitamin C antioxidant effect.

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 (8)

A cosmetic composition for improving skin wrinkles containing an anthracite extract as an active ingredient. A cosmetic composition for skin whitening comprising an anthracite extract as an active ingredient. A cosmetic composition for skin containing an anthracite extract as an active ingredient. A cosmetic composition for moisturizing containing an anthracite extract as an active ingredient. An antioxidative cosmetic composition comprising an anthraquinone extract as an active ingredient. A cosmetic composition for preventing skin aging comprising an anthracite extract as an active ingredient. The cosmetic composition according to any one of claims 1 to 6, wherein the anthocyanin extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. The anthrax extract according to any one of claims 1 to 7, wherein the anthocyanin extract is selected from the group consisting of (a) water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, (B) a supercritical extraction method, or (c) an ultrasonic extraction method. The cosmetic composition according to any one of claims 1 to 5, wherein the cosmetic is characterized in that it is extracted with an extraction solvent selected from the group consisting of butyl acetate, diethyl ether, dichloromethane, Composition.