KR20130114493A - Cosmetic compositions comprising the extract of dendropanax morbiferum as an active ingredient - Google Patents

Abstract

PURPOSE: A cosmetics composition is provided to ensure an excellent antioxidant activity, improvement on skin wrinkles, and anti-aging effect. CONSTITUTION: A cosmetics composition having an antioxidant activity contains Dendropanax morbiferum leveille extract as an active ingredient. A composition for improving skin wrinkles and an anti-aging effect contains Dendropanax morbiferum leveille extract as an active ingredient. The Dendropanax morbiferum leveille extract is a Dendropanax morbiferum leveille stem extract. The Dendropanax morbiferum leveille extract is extracted by a supercritical fluid extraction or ethanol extraction. The Dendropanax morbiferum leveille extract is 80% of an ethanol extract. [Reference numerals] (AA) Dendropanax morbifera leveille (80% EtOH ext.); (BB,CC) Type 1

Description

Cosmetic composition containing Hwangchil wood as an active ingredient {COSMETIC COMPOSITIONS COMPRISING THE EXTRACT OF DENDROPANAX MORBIFERUM AS AN ACTIVE INGREDIENT}

The present invention relates to a cosmetic composition for skin wrinkle improvement to skin anti-aging.

Human skin structure is composed of the stratum corneum, epidermis, and dermis, and the epidermal cells such as keratinocytes, melanocytes, and Langerhans cells, and dermal cells such as fibroblasts, endothelial cells, mast cells, and collagen elastic fiber cells. consist of.

Antigens or biological response modifiers, such as collagenase, thrombospondin, and interleukin-1, which are overexpressed from senescent cells, may cause skin dermal thickness during aging of mitotic tissues such as the skin. It is known to play an important role in various functional changes such as decrease in elasticity, decreased elasticity, wrinkle formation, and weakened immunological function.

Reduction of cell mediation and humoral immunity with aging correlates closely with coral stress caused by reactive oxygen species and various biological activities including epidermal Langerhans cells and dermal antigen-transmitting cells and cytokines that regulate immune and inflammatory responses. Keratinocytes capable of producing and secreting response regulators play an important role in the immune response of the skin.

Keratinocytes are epidermal cells primarily responsible for protecting individuals from the outside by physical barriers, neuropeptides, and eicosae, including cytokines that regulate immunological responses as well as the construction of simple physical barriers. It is an element that regulates immune response and inflammatory response by generating and secreting various biological response modifiers such as eicosanoid.

Ultraviolet rays are the leading cause of skin aging. Photoaging factors such as reactive oxygen species, DNA damage, Maillard reaction, cytokines, enzymes, protein synthesis, metabolic activity, free radicals, hormones, Natural aging factors such as nutrition, sleep, stress and living environment.

Proliferation and differentiation of keratinocytes is directly related to aging, and when the turn-over of keratinocytes decreases and the thickening of the stratum corneum thickens, the water content decreases, resulting in hardening of the keratin, resulting in wrinkles. The function of the basal keratinocytes decreases, resulting in a slower cell division, resulting in dry skin.

Dermal-Epidermal Junction (DEJ) functions to support epidermal cell adhesion, transport of nutrients and control epidermal differentiation. Decreases epidermis and dermis due to the decrease of type IV collagen and type VII collagen with aging. Decreased supportive function and weakened selective permeation cause harmful ingredients to affect the dermis, causing final wrinkles, decreased collagen biosynthesis and increased denatured elastin due to deactivation of fibroblasts, MMP (Metrix Metalloproteinase) Decreased collagen due to the increase of solar scar in the dermis (solar scar) is also caused by the destruction of the epidermis and dermal boundary also causes skin wrinkles.

Aged skin is damaged by the generation of reactive oxygen species (ROS) caused by ultraviolet rays, and the degradation of enzyme function of antioxidant enzymes (SOD, Catalase, etc.) and of antioxidants (Vitamin C, E, etc.) The reduction may damage the biological defense system against external harmful environment, cause glycation of skin proteins, non-enzymatic cross-linking, collagen and elastin dysfunction, and cause skin wrinkles.

Yellow Chilli ( Dendropanax) morbifera ) is a genus of arboraceae and elusive tree, with about 75 species in East Asia, Malay Peninsula, Central and South America, and 1 species in Korea. The distribution map of Hwangchil trees in Korea is an evergreen broad-leaved arboreous tree that grows on Jeju Island, Jeonnam Wando, Bogildo, Daeheuksan Island, Geomundo, Jeonbuk Eocheong Island, and Gyeongsangnam-do Beach. It is called Hwangchil (黃 漆).

Since the Three Kingdoms, Hwangchil was used as a rare paint that emits golden color of emperor's armor, helmets, and other metal ornaments.

The components of the yellow lacquer consist of 66.7% of non-volatile components, 10.8% of aromatics, 8.1% of moisture, and 14.4% of solids, and are β-cubebene of sesquiterpenes as essential oil fractions. , β-selinene (selinene), δ-cardinene (cadinene) is composed of.

So far, research on yellow lacquer tree has been carried out to investigate the anti-cancer, antioxidant, hepatic cell regeneration, diabetes treatment, hard tissue cell regeneration, and aromatic components in the nervous system of paints used in natural gold crafts. It has a sedative and tonic effect, and it is known to help the whitening effect by inhibiting the activity of tyrocyanase, which produces melanin in the extract from Hwangchil.

The present inventors conducted a thorough study for the purpose of improving the quality of human life through the development of extraction methods, efficacy experiments and standardization of raw materials for commercialization of functional cosmetics that are effective for wrinkle improvement using only the stem part of Hwangchil-tree.

Republic of Korea Patent Publication No. 10-2005-0036093, published April 20, 2005 Republic of Korea Patent Publication No. 10-2011-0078527, published on July 7, 2011 Republic of Korea Patent Publication No. 10-2012-0012918, published February 13, 2012 Republic of Korea Patent Publication No. 10-2011-0082292, published 19 July 2011

Je-Chiuan Ye et al., "Extraction and analysis of β-sitosterol in herbal medicines", Journal of Medicinal Plants Research Vol. 4 (7), pp. 522-527 (2010) Ill-Min Chung et al., "Chemical composition and larvicidal effects of essential oil of Dendropanax morbifera against Aedes aegypti L.", Biochemical Systematics and Ecology 37, pp. 470-473 (2009) Do-Ik kim et al., "Screening of some crude plant extracts for their acaricidal and insecticidal efficacies", Journal of Asia-Pacific Entomol 8 (1), pp. 93-100 (2005) Chambers. A. F et al., "Changing views of the role of matrix metalloproteinases in metastasis", J Natl Cancer Inst 89, pp. 1260-1270 (1997) Fisher. G. J et al., "Molecular basis of sun-induced premature skin ageing and retinoid antagonism", Nature 379, pp. 335-339 (1996) Fisher. G. J et al., "Pathophysiology of premature skin aging induced by ultraviolet light", Nat Engl J Med 337, pp. 1419-1428 (1997) Jun-Chul Ahn, et al., "Selection of Hwangchil-Sap Salivary Sustained Individuals and Investigation of Aroma Essential Oils," Korea Journal of Medicinal Crop Sci 10 (2), pp. 126-131 (2002) Shin, Min-Min et al., "Antioxidant Effect of Soybean Extract by DPPH Method", Journal of Cosmetics and Public Health 6 (6), pp. 19-23 (2010) Moon-Kon Moon et al., "Study on the Antioxidant Functionality of Hwangchil Tree Extract", Ph.D. Dissertation, Inje University Graduate School Park, Sook-Kyung et al., "Materials of Cosmetic Material of Angelica gigas Nakai Root Extract", Korea Chem. Eng. Res 47 (5), pp. 553-557 (2009) Byung Suk Jung et al., "A Study on the Distribution of Hwangchil Tree and the Component Analysis of Hwangchil", Korea Journal of Biotechnol. Bioeng 10 (4), pp. 393-400 (1995)

The present invention is to provide a cosmetic composition for improving skin wrinkles to skin anti-aging, skin elasticity of natural products that are not toxic to the skin.

The present inventors have confirmed that the Hwangchil-tree extract exhibits antioxidant activity, from which it can show skin wrinkle improvement and skin anti-aging effect and completed the present invention.

The present invention provides a cosmetic composition having an antioxidant effect containing an active ingredient, Hwangchil-tree extract (Dendropanax morbiferum).

The present invention also provides a cosmetic composition for improving skin wrinkles to anti-aging skin containing the extract of Hilchi chinensis.

The hwangchil wood is meant to include all or part of the hwangchil wood, preferably the stem portion of the hwangchil wood.

The hwangchil wood extract according to a conventional method known in the art, for example, using a solvent extraction method using a conventional solvent, under reduced pressure by carbon dioxide and supercritical extraction by high temperature, etc. under the conditions of the usual temperature and pressure You can get it.

The extraction process may be performed by extracting with the primary extraction solvent about 2 to 20 times, preferably about 5 to 10 times the weight of the yellow lacquer tree. Extraction may be performed by an extraction method known in the art, such as cold soaking, hot water extraction, ultrasonic extraction, reflux cooling extraction, but is not limited thereto. The extraction temperature may be adopted by those skilled in the art in various temperature ranges suitable for the extraction method, for example, may be carried out at 20 ℃ to 100 ℃, etc., but is not limited thereto. In addition, the extraction time is different depending on the extraction method, a person skilled in the art can adopt an appropriate extraction time, but is not limited to this, can be performed in a single or multiple times in the range of about 1 hour to several days. The extract obtained by performing the extraction with the primary extraction solvent may be obtained in the form of a liquid in which impurities are removed by filtration according to a conventional method, or the obtained liquid form extract may be concentrated under reduced pressure and / Can be obtained.

Preferred extraction solvents in the present invention can be used ethanol, more preferably 80% (v / v) aqueous ethanol.

In addition, the extraction process may further comprise the step of obtaining a fraction having a high content of the active ingredient, if necessary. That is, by dispersing the extract obtained by extraction with the primary extraction solvent in water, and then extracting with an appropriate secondary extraction solvent, for example, the alcohol of saturated C4, to further increase the active ingredient content in the resulting extract Can be.

In addition, the hwangchil wood extract of the present invention may include not only the extract by the above-described extraction solvent, but also the extract through 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 by various purification and extraction methods additionally carried out, such as separated by the sex) or by fermentation products using natural or various microorganisms, is also included in the extract of the present invention.

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 impact effect of the ultrasonic energy, the mixing efficiency of the substance and solvent contained in the sample is increased and the extraction efficiency is increased.

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 cosmetic composition of the present invention comprises about 0.0001 to 60% by weight, preferably about 0.01 to 30% by weight, of the Hilchi chinensis extract, based on the total weight of the composition.

The cosmetic composition of the present invention is an oil, powder, surfactant, humectant, thickener, lower alcohol, coating agent, ultraviolet absorber, metal ion sequestrant, organic amines, pH adjuster, active ingredient, sugars, preservatives, vitamins, antioxidants, perfumes , Water and the like.

Examples of the oil include jojoba oil, olive oil, avocado oil, castor oil, palm oil, tallow, hydrogenated oil, natural oils and derivatives thereof such as liquid lanolin, lead such as carnauba wax, beeswax and lanolin, liquid paraffin, micro crystalline wax , Hydrocarbons such as squalene and petrolatum, higher fatty acids such as stearic acid, higher alcohols such as cetyl alcohol and stearyl alcohol, glycerin monostearic acid ester, glyceryl monoglyceride, glycerin monooleate and isopropyl acid Esters such as propyl, diisostearyl, di2-heptyl undecanoic acid glycerin, tri2-ethylhexyl acid trimetholpropane, trioctanoate trimetholol propane, sebacic acid di2-ethylhexyl, peppermint oil, rose oil And essential oils such as citroneral, silicone oils such as dimethylpolysiloxane and decamethylcyclopentasiloxane. The content of the oil component in the cosmetic composition is appropriately selected depending on the form, the formulation, and the like of the cosmetic, but can usually be 0.1 to 95% by weight in the total weight of the cosmetic composition.

Examples of the powder include talc, mica, kaolin, silica, galvanized, mica titanium, titanium oxide, iron oxide, nylon powder, and the like.

Examples of the surfactant include polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene sorbitan fatty acid ester, glycerin fatty acid ester, polyoxyethylene hardened castor oil, polyoxyethylene sorbitol fatty acid ester, polyoxyalkylene-modified polysiloxane, and the like. Nonionic surfactants, anionic surfactants such as sodium palmitate, cationic surfactants such as stearyl trimethylammonium chloride, and amphoteric surfactants such as betaine, amidebetaine, sulfobetaine, and imidazolinium. Can be.

Examples of the moisturizing agent include glycerin, 1,3-butylene glycol, polyethylene glycol, dipropylene glycol, sorbitol, and the like.

Examples of the thickener include water-soluble polymers such as carboxyvinyl polymer, carboxymethyl cellulose and polyvinyl alcohol, and clay minerals such as bentonite.

Examples of ultraviolet absorbers include paraamino benzoic acid (abbreviated as PABA), glyceryl PABA, ethyldihydroxypropyl PABA, octylmethoxycinnamate, 2-ethoxyethyl-p-methoxycinnamate, 2,4- Dihydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4-methylbenzophenone, 2-hydroxy-4-methoxy-4-methylbenzophenonesul Fonates, urokanoic acid ethyl esters, 2-phenyl-5-methylbenzokizazole, 4-methoxy-4-t-butyldibenzoylmethane, paramethoxycinnamic acid ethylhexyl, and the like.

As an example of a metal ion blocking agent, tetrasodium edetic acid, citric acid, etc. are mentioned. Examples of the lower alcohols include ethanol and the like. Examples of the organic amines include monoethanolamine, triethanolamine, and the like. As an example of a pH adjuster, buffers, such as a lactic-sodium lactate and a citric acid-sodium citrate, are mentioned.

Examples of the active ingredient include pantothenyl ethyl ether, glycidyl acid salt, and the like. Examples of the vitamins include vitamin E or derivatives thereof. Examples of the antioxidant include tocopherols, dibutylhydroxytoluene, propyl gallate, and the like.

Examples of the saccharides include erythritol, sucrose, hyaluronic acid and the like. As an example of a preservative, ethyl paraben, butyl paraben, sodium benzoate, etc. are mentioned. In addition, it can also select and mix | blend among the components which can be mix | blended with the medicine and food mentioned later.

The present invention may be in the form of an external preparation for skin, and such external preparation for skin may include various additives such as excipients, stabilizers, moisturizers, emulsifiers, absorption accelerators, pH adjusters, surfactants, diluents, and carriers. Specific examples of these additives include starch, sugars such as lactose, cellulose derivatives such as magnesium sulfate, talc, gelatin, hydroxypropyl cellulose, vegetable oils such as soybean oil, sesame oil, animal oils or synthetic oils, rubber, physiological Water such as saline, alcohols such as ethanol, 1,3-butylene glycol, polyalkylene glycol, and the like.

The cosmetic composition to the skin external preparation of the present invention can be prepared in any form and formulation by mixing the active ingredient of the present invention with one or two or more kinds of the above-mentioned arbitrary compounding ingredients.

The skin cosmetic composition of the present invention and the skin external preparation are preferably applied directly to the skin individually.

The area to be treated may mean the entire skin surface of the subject or only the area required. Application of the composition should be repeated periodically to maintain wrinkle improvement or anti-aging of the skin.

The cosmetic composition and the external preparation for skin of the present invention preferably further include a cosmetically acceptable amount of a local carrier capable of delivering the active ingredient to the keratinocyte cell skin layer under in vivo conditions. The carrier may comprise a solution, suspension, emulsion or other form capable of delivering the reagent to the keratinocyte cell skin layer under in vivo conditions. Preferably, the carrier should not substantially interact with the reagents so that the active ingredient performs the same function as described herein.

The component and content of the carrier depend on the amount of the active ingredient used in the composition of the present invention and the like, and the carrier may be included in an amount of about 0.1% to about 99% by weight based on the total weight of the composition. Preferably the carrier comprises an alcohol. Alternatively, the carrier may be liposomes or hydrated lipidic lamellar phases as known to those skilled in the art.

Preferred combinations of carriers include alcohols (eg methanol, ethanol or isopropanol) and thickeners such as propylene glycol, polyethylene glycol (PEG) or carbopol and penetration enhancers such as transcutol.

The cosmetic composition of the present invention may be topically applied in various ways to infiltrate the activating material through the skin to improve wrinkles of the skin and to prevent skin aging. Preference is given to gel, lotion or solution forms which can rub the composition by hand on the skin. Another method that is applicable is the use of aerosol sprays or applicators. Typically applied on the skin at a dosage of 0.3-1 ml / 5-50 cm 2. This formulation can be frequently applied in 1 hour-2 hours-6 hours-8 hours-12 hours cycles to the areas requiring application. Dosage described in the present invention means the amount and frequency of application of the composition to the skin. The composition may comprise a film former such as polyvinyl pyrrolidone or polymethylpyrrolidone.

The cosmetic composition to the skin external preparation of the present invention is acceptable for cosmetics and treatments containing other solvents, humectants, wetting agents, oils, emulsifiers, thickeners, diluents, surface activators, flavoring agents, preservatives, antioxidants, vitamins and minerals. Other carriers or developers may further be included.

The cosmetic compositions to the skin external preparations of the present invention may also additionally include substances commonly used in skin care products, such as liquid or solid emollients, silicone oils, solvents, thickeners, sunscreens, skin whitening agents, propellants, and other special skin-benefit agents. And active agents.

Formulation diagram of the cosmetic composition of the present invention, solution, solubilizing, emulsifying, powder, powder dispersion, emulsion, gel, ointment, aerosol, water-oil 2-layer, water-oil-powder 3-layer, etc., It may vary and includes, for example, ointments, solutions, creams, emulsions, lotions, lotions, gels, essences (cosmetics), foundations, pack masks, rouges, sticks, baths, bath or shower gels or soaps (including liquid soaps) And various other products.

In addition, the cosmetic composition to the skin external preparation of the present invention may be formulated in a general formulation in the art, and the ingredients to be combined with the active ingredient may be appropriately selected and combined by those skilled in the art according to the formulation.

In addition to the above description, those skilled in the art can appropriately modify or apply the embodiments of the invention from the description herein, and are also included within the scope of the present invention.

Hwangchil-tree extract of the present invention exhibits excellent antioxidant activity, and improves skin wrinkles and prevents skin aging.

Figure 1 is a graph showing the results of high-speed liquid chromatograph confirming the β-sitosterol content in the Hwangchil wood extract.
Figure 2 is a graph showing the results of high-speed liquid chromatography of β-sitosterol as an indicator.
Figure 3 is a graph showing the MMP-1 inhibitory activity of Hwangchil-tree extract (20 ppm).
Figure 4 is a graph showing the MMP-1 inhibitory activity of Hwangchil-tree extract (200 ppm).
Figure 5 is a graph showing the MMP-1 inhibitory activity of Hwangchil-tree extract (2000 ppm).
Figure 6 is a graph showing the fibroblast proliferation activity by the Hwangchil-tree extract.
Figure 7 is a photograph showing the shape of fibroblasts treated with Hwangchil-tree extract.
8 is a graph showing DPPH inhibitory activity of α-tocopherol in comparison with Hwangchil-tree extract.
Figure 9 is a graph showing the similar antioxidant activity of the hwangchil-tree extract and the comparison group α-tocopherol.

Hereinafter, the present invention will be described in more detail through examples and test examples. However, these Examples and Test Examples are for illustrating the present invention, and the scope of the present invention is not limited to these Examples and Test Examples.

[ Example ]

< Example  1> Preparation of Hwangchil Tree Extract

Hwangchil tree, native to Heuksan-myeon, Sinan-gun, Jeollanam-do, Jeollanam-do, purchased dried material, crushed with a grinder (HMF-502, Hanil Electric), and then used 20g of powder through 14 mesh powder as an experimental material. It was.

As shown in the figure below, the crushed yellow lacquer trees were extracted by hot water extraction method, solvent (80%, 50%, 20% ethanol) extraction method and supercritical extraction method for natural products, and the extraction conditions were established. 80% ethanol extraction was performed.

The extracted sample was transferred to a 20 ml round flask, concentrated using a rotary vaccum evaporator, and dried using a lyophilizer and used for testing.

[Table 1]

[ Experimental Example ]

< Experimental Example  1> Indicator Components of Hwangchil Tree (β- Sitosterol A) content analysis

Hwangchil-tree is composed of 66.7% of non-volatile components, 10.8% of aromatic components, 8.1% of moisture, and 14.4% of solids. Especially, it is an essential oil fraction of β-Cubebene and β of Sesquiterpene. -Selinene, δ-cardinene (Cadinene). Β-sitosterol, widely known as a sterol component of plants, is an anti-inflammatory active substance used to treat prostatic hyperplasia, prostatitis, neurostatic prostate disease, bladder dysfunction before and after prostate surgery (urination disorder, nocturnal enuresis, urinary incontinence, etc.) It is known as a major component of prostatic hyperplasia ("cytonyl": modern drug). In this experiment, β-sitosterol, which is effective for anti-inflammatory, was set as an index component and quantified by the following quantitative method.

In the above example, 0.1 mg of the concentrated extract extract concentrated under reduced pressure was accurately used, and 10 ml of 80% ethanol was accurately used as a sample solution.

10 μl each of the sample solution and the standard solution were tested according to High Performance Liquid Chromatography under the following conditions, and the peak areas A _T and A _s of the sample solution and the standard were measured.

Amount of β-sitosterol (C ₂₉ H ₅₀ O) (mg) = Amount of β-sitosterol (mg) X A _T / A _s

<HPLC (Waters, LC Module I plus) measurement conditions>

Detector: UV absorbance photometer (wavelength 196 nm)

Column: SunFire ^TM C ₁₈ (5 μm, 4.6 × 250 mm, Waters)

Flow rate: 1.0 ml / min

Injection volume: 5 μl

Column oven temperature: 25.0 ℃

Mobile phase: A-Ethanol: Acetonitrile (15: 85 v / v)

[Table 2]

The results are shown in Table 3 below and FIG. 1. HPLC measurement peaks for the indicator β-sitosterol are shown in FIG.

[Table 3]

< Experimental Example  2> of yellow lacquer extract MMP -1 inhibition effect

The MMP-1 fluorimetic drug discovery kit (MMP-1 fluorimetic drug discovery kit) -BML-AK405 kit (QuantiZyme ^™ Assay System) is a useful kit for screening MMP-1 inhibitors. OmniMMP fluorescent quantitative substrate (Quinimetric substrate) is the same principle as shown in the following figure, if the MMP-1 inhibitor does not show catalytic activity for the substrate, the quenching phenomenon (vice versa) shows the fluorescence MMP-1 Inhibition rate can be efficiently classified.

20% EtOH extract, 50% EtOH extract, 80% EtOH extract, H ₂ O (hot water) extract, CO ₂ (supercritical) extract of each of the Hwangchil trees was concentrated by the method described in Examples, 20 ppm, 200 ppm, MMP-1 catalyst activity was confirmed at a concentration of 2000 ppm.

After 10 minutes of sample treatment, the fluorescence was measured and expressed in Relative fluorescence (unit), and converted into enzymatic activity.

The results are shown in FIGS. 3 to 5 and Table 4.

[Table 4]

When all samples were at a concentration of 20 ppm, when the control group, which was not treated as shown in FIG. 3, was 100%, the treatments of Hwangchil-tree 20% EtOH extract and Hwangchil-tree CO ₂ extract showed 48.7% and 16.8%, respectively.

< Experimental Example  3> Confirmation of Fibroblast Proliferation Activity of Hwangchil Tree

(1) Superculture of human skin fibroblasts and culture of cells

Primary cultures of human dermal fibroblasts were obtained from adult foreskin containing 10% fetal calf serum, 2 mM glutamine, penicillin (100 U / mL) and streptomycin (100 μg / mL). 37 ° C., 5% CO ₂ using Dulbecco's modified eagle medium (DMEM) Lt; / RTI &gt; Fibroblasts were passaged when grown to about 90%, cells were used in the experiment since the fifth generation.

Fibroblasts were passaged every 3 to 6 days under conditions of 37% 5% CO ₂ using FBS-containing complete medium, and then seeded with a certain amount of each well plate for cell viability experiments.

Cell lines were injected into banking vials (banking vials), stored in a liquid nitrogen tank and used for the experiment.

(2) measuring cell viability

20 ml of the sample extracted in Example was transferred to a circular flask and concentrated by a rotary vacuum evaporator to obtain the following amount.

[Table 5]

Samples of the Hwangchil-tree extract were treated with fibroblasts at a concentration of 200 ppm with respect to five extracts as shown in the above table, and cytotoxicity experiments were performed. Supercritical extracts and 80% EtOH extracts were observed in detail at concentrations of 200, 100 and 50 ppm.

The number of fibroblasts was taken in a certain amount, stained with trypan-blue, counted using a hematocytometer, and seeded in a well plate. Incubated for a certain period of time with or without treatment.

20μl of MTT ([3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) solution (5 mg / ml) was added to the cells in culture to measure the viability of the cells after a certain period of time. After reaction at 37 ° C. for 4 hours, the supernatant was removed, and 200 μl of dimethyl sulfoxide was added to dissolve the formazan product.

This was measured for absorbance at 570 nm using an ELISA reader. The experimental results were expressed as a percentage of the absorbance compared to the control, and repeated 6 times was expressed as mean ± SEM. As a control, a medium to which no sample was added was used, or a solvent in which the sample was dissolved was treated under the same conditions.

Cell survival test (MTT) for 9 sample treatments showed that at the same concentration of 200 ppm, each treatment showed 20%, 40%, 67%, 79%, 90% viability and supercritical extraction> 80% Toxicity was confirmed in the order of EtOH extraction> 50% EtOH extraction> 20% EtOH extract. Supercritical extracts showed about 30% toxicity even at low concentrations of 50 ppm, but 80% EtOH extract was found to reduce toxicity at low concentrations of 50 ppm (FIG. 6).

(3) cell morphology observation

Fibroblasts in culture were treated with a specific concentration of the sample after measuring the cell viability and the shape of the cell was observed using a microscope (Olympus CKX31, Japan). Observations were made at 40 × and 100 × magnifications and organized using image analysis (IMT I-solution capture). The trends for the cell types according to each extraction method are shown in FIG. 7.

< Experimental Example  4> Antioxidant Activity of Sacred Tree ( DPPH Low performance ) Confirm

DPPH (1,1-diphenyl-2-picrylhydrazyl) is a dark purple compound which itself is a very stable free radical and exhibits specific light absorption at 517 nm. It is oxidized by quantitative decolorization by an antioxidant with radical scavenging activity. Activity can be easily measured. The scavenging activity of this radical has been widely used for antioxidant search because it has a correlation with antioxidant activity including lipid peroxidation inhibitory activity.

※ Comparison with antioxidant active extract and α-tocopherol

Radical scavenging activity was measured by modifying the Blois DPPH method. That is, each of the extract and α-tocopherol were diluted stepwise to the concentration of 0.1, 0.2, 0.4, 0.6, 0.8, 1 mg / ml using EtOH. After 0.2 ml DPPH 0.8 ml was added to each 0.2 ml sample diluted in steps, the reaction was carried out for 25 to 30 minutes, and then measured at 517 nm using a spectrophotometer (Spectrophotometer, optizen 2120UV). The control group was added to 0.8 ml of DPPH in 0.2 ml of DMSO and measured in the same manner as above. IC _{50 was used} to determine the amount of sample (mg / ml) required for extract and α-tocopherol to reduce the absorbance of the control (untreated group) by 1/2, and the final concentration of each sample was calculated.

DPPH radical scavenging activity = (A control-A sample) / A control × 100

A control group: absorbance of extract-free

A sample: absorbance of extract added

The results are shown in FIG. 8 and Table 6. Antioxidant effect of 80% -ethanol extract of Hwangchil-tree extract IC ₅₀ was 39.952%, indicating a relatively good evaluation of 0.2 mg / ml, which means that the concentration required for inhibition rate of 64.115% in the α-tocopherol control group was 0.025 mg / As ㎖, it was found that about 8 times difference of Hwangchil wood appeared. Therefore, it can be seen that the extract of Hwangchil-tree has a relatively high antioxidant activity.

TABLE 6

< Experimental Example  5> Oxidation SOD - like ) Check activity

SOD-like activity was added to 0.3 ml of the extract diluted to 0.1, 0.2, 0.4, 0.6, 0.8, 1 mg / ml by adding 0.3 ml Tris-HCl buffer (PH 8.5) and 0.3 ml of 7.2 mM pyrogallol to 25 to 45 After the minute reaction, the absorbance was measured at 405 nm using a spectrophotometer (Optizen 2120UV). The treated group was treated with 0.3 ml of DMSO to measure absorbance. The SOD-like activity of each sample was calculated by the following equation, and the amount of sample (mg / ml) required for extract and α-tocopherol to reduce the absorbance of the control (untreated group) by 1/2 was measured. SC ₅₀ , a concentration that inhibits galol by 50%, was obtained and the final concentration of the sample was calculated. The results are shown in Fig. The SOD-like activity IC ₅₀ values for the Hwangchil-tree extract samples were shown in Table 7 below. As a result, it was confirmed that the Hwangchil-tree extract showed excellent pseudosulfuric activity.

[Table 7]

Claims (6)

Cosmetic composition having an antioxidant activity containing an extract of Hwangchil (Dendropanax morbiferum) as an active ingredient. Cosmetic composition for skin wrinkle improvement or skin anti-aging containing hwangchil wood extract as an active ingredient. The cosmetic composition according to claim 1 or 2, wherein the extract of Hwangchil-tree is Hwangchil-tree stem extract. The cosmetic composition according to claim 1 or 2, wherein the hwangchil tree extract is extracted by a supercritical extraction method or an ethanol extraction method. The cosmetic composition according to claim 4, wherein the hwangchil tree extract is 80% ethanol extract. The cosmetic composition according to claim 1 or 2, which is in the form of an ointment, solution, cream, emulsion, lotion, lotion, gel, essence, foundation, pack mask, stick, bath, bath or shower gel, or soap.

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