KR20130140378A - Cosmetic composition comprising taraxacum platycarpum extract for preventing skin aging - Google Patents

Abstract

The present invention provides a cosmetic composition for preventing aging skin which contains a Ulmus davidiana var extract, a Cassia obtusifolia extract, and a dandelion extract as active ingredients. The Ulmus davidiana var extract, the Cassia obtusifolia extract, and the dandelion extract have remarkable a first type collagen expression improvement function and a type I collagenase (matrix metalloproteinase-1, MMP-1) expression reduction function, and have excellent anti-aging effect.

Description

Cosmetic composition for preventing skin aging comprising dandelion extract {Cosmetic Composition Comprising Taraxacum platycarpum Extract for Preventing Skin Aging}

The present invention relates to a skin anti-aging cosmetic composition comprising a plant extract, and more particularly, the skin comprising the extract of Ulmus davidiana var., Cassia obtusifolia and Dandelion (Taraxacum platycarpum) as an active ingredient It relates to a cosmetic composition for preventing aging.

The primary function of the skin is to protect the human body from external stimuli such as temperature, humidity and ultraviolet rays. However, when the skin is exposed to external stimuli such as strong ultraviolet rays, stress, and malnutrition, the skin may age, such as wrinkle formation, loss of elasticity, and keratinization.

The dermis of the skin occupies most of the volume of the skin and is mainly involved in the properties of the skin such as flexibility, elasticity and tension. The major component of connective tissue in the dermis is type 1 collagen and is known to be produced in fibroblasts (Bhogal RK, Stoica CM, McGaha TL, Bona CA: Molecular aspects of regulation of collagen gene expression in fibrosis. J. Clin. Immunol .25,592 (2005). TGF-β / Smad signaling system is involved in promoting type 1 collagen production (Verrecchia F., Mauviel A., Farge D .: Transforming growth factor-beta signaling through the Smad proteins: role in systemic sclerosis.Autoimmun.Rev .5,563 (2006)) When TGF-β1 binds to the TGF-β1 receptor on the surface of fibroblasts, an intracellular protein called the Smad family is activated. In particular, the activation of Smad2 and Smand3 promotes the production of type 1 collagen. When Smad7 is activated, collagen production is inhibited, and Smad proteins are involved in regulating collagen biosynthesis in a timely manner. Matrix metalloproteinase (MMP) is an important protein that breaks down the extracellular matrix and plays an important role in preventing excessive accumulation of collagen in the dermis. When MMP protein is excessively activated, collagen disintegration proceeds more than necessary, and Tissue inhibitor of MMPs (TIMP), which inhibits MMP activity, is timely expressed and is involved in homeostasis of type 1 collagen in the dermis. (Homebeck W .: Down-regulation of tissue inhibitors of matrix metalloprotease-1 (TIMP-1) in aged human skin contributes to matrix degradation and impaired cell growth and survival. Pathol. Biol. 51,569 (2003)). As such, the biosynthesis of collagen type 1 is precisely regulated by the interaction of TGF-β / Smad signaling system and MMP and TIMP-1.

Dandelion, the generic name Taraxacum , is not only used for food, tea, beverages, etc. (SchK., Carle R., Schieber A .: Taraxacum--a review on its phytochemical and pharmacological profile.J. Ethnopharmacol 107,313 (2006)), dandelion extracts are known to have anti-inflammatory and anti-allergic effects, making them a valuable ingredient that can be used in the development of therapeutic agents that inhibit inflammation (Cheong H., Choi EJ, Yoo GS, Kim. KM, Ryu SY: Desacetylmatricarin, an anti-allergic component from Taraxacum platycarpum.Planta.Med. 64,577 (1998)).

On the other hand, KR Patent Publication No. 10-2010-0057036 (name of the invention: anti-inflammatory composition using the dandelion extract) discloses an anti-inflammatory composition comprising the extract of the dandelion as an active ingredient, KR KR 10-2010-0057036 0033838 (name of the invention: anti-obesity and antioxidant composition containing dandelion extract) discloses an anti-obesity and antioxidant composition comprising ultrasonic dandelion extract as an active ingredient. However, studies on the effect of the dandelion extract on the components of the connective tissue of the skin is insufficient.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have made diligent research efforts to develop a cosmetic composition excellent in anti-aging activity by exhibiting a marked increase in collagen expression or type I collagenase expression. As a result, it was found that the anti-aging activity was remarkable when the extracts of the roots of the roots, the extracts of the leaves, and the extracts of dandelion were included as active ingredients. The invention was completed.

Therefore, an object of the present invention is to provide a cosmetic composition for preventing skin aging, including the extract of the roots of the root, the extract of the deficiency, and the dandelion extract.

In addition, another object of the present invention to provide a food composition or pharmaceutical composition for preventing skin aging comprising the extract.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

The present invention provides a cosmetic composition for preventing skin aging.

The present inventors have made diligent research efforts to develop a cosmetic composition excellent in anti-aging activity by exhibiting a marked increase in collagen expression or type I collagenase expression. As a result, it was confirmed that the anti-aging activity was remarkable when the extracts of the roots of the roots, the extracts of the deficiency, and the dandelion extract were included in the cosmetic composition as the active ingredient, showing a marked increase in the expression of collagen type 1 or type I collagenase expression.

According to an aspect of the present invention, the present invention provides a cosmetic composition for preventing skin aging, which comprises an extract of Ulmus davidiana var., Cassia obtusifolia, and Dandelion (Taraxacum platycarpum) as active ingredients.

The cosmetic composition of the present invention has the use of skin anti-aging. As used herein, the term "anti-aging of skin" has the meaning encompassing comprehensively the prevention and treatment of skin aging. On the other hand, the anti-aging of the skin, which is the use of the present invention, may include a condition of the skin derived from ultraviolet light exposure or the like, for example, inhibiting skin aging and the formation of wrinkles.

According to a preferred embodiment of the present invention, the extracts of the roots of the root, extract extract and dandelion extract may exhibit the ability to increase the type 1 collagen expression or type I collagenase (Matrix Metalloproteinase-1, MMP-1) expression.

The main cause of skin aging is collagen reduction, collagen reduction can be promoted by an increase in the type I collagenase, a collagen degrading enzyme, and also by a decrease in the type 1 collagen expression, the root skin extract in the present invention It is very important for the extract, Dandelion extract and dandelion extract to exhibit type 1 collagen expression increasing ability or type I collagenase expression reducing ability, which is specified in the following examples.

According to a preferred embodiment of the present invention, the roots of the root may be elm root bark.

According to a preferred embodiment of the present invention, the dandelion may be preferably selected from the group comprising dandelion's flowers, leaves, stems, fruits, seeds, roots and outposts, more preferably the leaves, stems, It may be selected from the group comprising fruits, roots and outposts, most preferably the outposts of dandelion plants. The reason why the most preferable extraction site of the plant in the present specification is the outpost is because it proved that the outpost of the dandelion was excellent in the anti-aging effect through the following examples.

As used herein, the term "extract" is used to refer to the roots of the root extract, the deflector extract and dandelion extract, as well as the extraction result obtained by treating the extract solvent on the roots of the root, the deficiency and dandelion, as well as to apply the roots of the root, the deficiency and dandelion itself to the skin. It is also meant to include workpieces that are formulated (eg, powdered) to make it possible.

If the extract used in the composition of the present invention is obtained by treating the extract solvent on the roots of the roots, the defect and dandelion, various extraction solvents may be used. Preferably, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (e.g. methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol and normal-butanol, etc.), (c) Mixed solvent of the lower alcohol and water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol, (h) hexane, (i) diethyl ether, or (j) Butyl acetate can be obtained by treating the roots of the roots of the roots, the leaves and dandelions.

As used herein, the term " extract " has the meaning conventionally used in the art as a crude extract, but broadly includes fractions obtained by further fractionation of the extract. That is, the root extract of the root, the extract of the deficiency, and the dandelion extract are not only obtained by using the above-mentioned extraction solvent, but also include those obtained by additionally applying a purification process. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through the purification method are also included in the extract of the root of the present invention, the extract of the deficiency and the dandelion extract.

The roots of the roots of the root of the present invention, the extract and the dandelion extract may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying.

According to a preferred embodiment of the present invention, the amount of the roots of the roots, the clarifier and the dandelion extract is preferably 0.00002-30% by weight, more preferably 0.0001-0.1% by weight, most preferably 0.0025-0.01 based on the total weight of the cosmetic composition. Weight percent.

In the composition of the present invention, the reason that the most preferable amount of the roots of the roots of the root, the deficiency and the dandelion extract is 0.0025-0.01% by weight is the effect of the invention as an anti-aging cosmetics when the content of the roots of the roots of the roots of the root, the deficiency and the dandelion is less than 0.0025% by weight. This is because there is a problem that can not be obtained sufficiently, if the amount exceeds 0.01% by weight is a problem of safety due to cytotoxicity.

According to a preferred embodiment of the present invention, the mixing ratio of the extract of the root of the root, the extract of the deficiency and the dandelion extract may be preferably 1: 0.1-10: 0.1-10: 0.1-10 based on the weight of the extract, more preferably 1: 0.3-5: 0.3-5: 0.3-5, and most preferably 1: 0.5-2: 0.5-2: 0.5-2.

According to a preferred embodiment of the present invention, the cosmetic composition is in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray ≪ / RTI >

According to a preferred embodiment of the present invention, the extracts of the roots of the root, the extract of the deficiency and the dandelion extract exhibiting anti-aging activity may be preferably included in a composition selected from the group comprising cosmetic compositions, pharmaceutical compositions and food compositions, most preferably May be used in the cosmetic composition. As the result of the KFDA food raw material search ( http://fse.foodnara.go.kr ), there is no problem in using it as a pharmaceutical composition or a food composition, and also has excellent effects such as atopy or diabetes. In view of what is known to be a plant that is present, it can be used in food or pharmaceutical compositions that exhibit anti-aging effects.

The cosmetic composition of the present invention may include other components in addition to the above-mentioned roots of the root extract, deficiency extract and dandelion extract. According to an embodiment of the present invention, the cosmetic composition of the present invention is obtained from the group comprising glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate and allantoin Further comprising at least one auxiliary component selected from the group consisting of glycerin, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, squalane and sodium citrate, and at least one auxiliary ingredient selected from the group consisting of butylene glycol, citric acid, panthenol and allantoin And most preferably at least one component selected from the group consisting of glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate and allantoin .

Since the cosmetic composition of the present invention is basically applied to the skin, it can be provided with reference to the cosmetic composition of the related art, for example, as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, But are not limited to, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. 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 .

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a 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, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

According to a preferred embodiment of the present invention, the composition of the present invention may be a pharmaceutical composition excellent in skin anti-aging effect.

When the composition of the present invention is manufactured from a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the formulation and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans in various routes such as oral or parenteral routes such as oral, rectal or intravenous, muscular, subcutaneous, intra-uterine, Can be administered by injection. Preferably, it is applied by transdermal administration during parenteral administration, more preferably by topical application by application.

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . The dose of the pharmaceutical composition of the present invention can be administered in an amount of 0.1-100 mg / kg on an adult basis once or several times a day in the case of an oral formulation, and in the case of an external preparation, It is preferable to apply it once to 5 times a day in an amount of 3.0 ml and continue for 1 month or longer. However, the dosage is not intended to limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulations may be in any form suitable for pharmaceutical preparations including oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories and sterile injectable solutions, , Dispersants, or stabilizers.

According to a preferred embodiment of the present invention, the composition of the present invention may be a food composition excellent in skin anti-aging effect.

When the composition of the present invention is prepared with a food composition, it includes not only the active ingredient, but also ingredients normally added during the manufacture of the food, including, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings.

For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.

As a result of analyzing cell viability with respect to the active ingredient of the composition of the present invention in the concrete examples of the present invention, it was found that the active ingredient of the composition of the present invention is a harmless substance as a natural substance. Therefore, since the active ingredient of the present invention has little toxicity and side effects, it can be safely used even for long-term use, and can be safely applied to cosmetic, pharmaceutical and food compositions as described above.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a cosmetic composition for preventing skin aging comprising Ulmus davidiana var. extract, Cassia obtusifolia extract, and Taraxac platycarpum extract as active ingredients.

(b) The extracts of the roots of the roots of the present invention, the extracts of the locusts and the extracts of the dandelion showed a marked increase in the expression of collagen type 1 or type I collagenase (Matrix Metalloproteinase-1, MMP-1), an anti-aging effect. Is excellent.

Figure 1 shows the results of the mixed extract (AF-343) to the fibroblasts according to the treatment time.
Figure 2 shows the results according to the treatment concentration of the mixed extract (AF-343) to the fibroblasts.
Figure 3 shows the type 1 collagen mRNA increase in the fibroblasts when mixed extract (AF-343) treatment.
Figure 4 shows the expression changes of Smad proteins upon treatment of the mixed extract (AF-343) to fibroblasts.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

Example

Throughout this specification, "%" used to denote the concentration of a particular substance is intended to include solids / solids (wt / wt), solid / liquid (wt / The liquid / liquid is (vol / vol)%.

Example  1: Preparation of plant extract

Specifically, in the preferred embodiment of the present invention using an extractor equipped with a reflux device extracts while stirring for 72 hours using a 70% ethanol aqueous solution of 10 times with respect to the roots of the roots of the roots, Cultivator and dandelion or its dried, filtered and the filtrate Concentrated to obtain an ethanol extract, and, if necessary, an excipient such as maltodextrin was added and lyophilized or spray dried to obtain an extract powder.

Example  1-1: Yugunpi  Extract recipe

Ulmus davidiana var., Ie, the dry elm root bark, is shredded with a grinder and 2 to 200 times its volume, preferably 10 to 30 times the organic solvent, is added at 10 to 50 ° C. for 1 to 10 days, Preferably, the mixture may be extracted for 2 to 5 days, filtered, concentrated under reduced pressure, and prepared in a drying step. In this case, ethanol, methanol, dichloromethane, nucleic acid and butylene glycol may be used as the extraction solvent, and 40 to 90% aqueous ethanol solution is preferable.

Example 1-2: Clarifier extract preparation

After crushing Cassia obtusifolia, 2 to 200 times, preferably 10 to 30 times, organic solvent was added to the volume and extracted and filtered at 10 to 30 ° C. for 1 to 10 days, preferably 2 to 5 days. After concentration under reduced pressure and may be prepared by a drying step. In this case, ethanol, methanol, dichloromethane, nucleic acid and butylene glycol may be used as the extraction solvent, and 40% to 90% aqueous ethanol solution is preferable.

Example  1-3: Dandelion Extract Preparation

After grinding the Dandelion (Taraxacum platycarpum) starch, 2 to 200 times, preferably 10 to 30 times the amount of the organic solvent is added to the volume and extracted for 1 to 10 days, preferably 2 to 5 days at 10 to 30 ℃ After filtration and concentration under reduced pressure, it may be prepared in a drying step. In this case, ethanol, methanol, dichloromethane, nucleic acid and butylene glycol may be used as the extraction solvent, and 40 to 90% aqueous ethanol solution is preferable.

Example 1-4: Preparation of Mixed Extract

In addition to increasing the protein expression of type 1 collagen and significantly reducing the expression of MMP-1, three plant extracts that can help skin aging, i.e. The mixed extract (AF-343) was completed. As a result of measuring the protein expression increase amount of collagen type 1 using the mixed extract, the effect was superior to using each plant extract alone.

Test Example: Confirmation of Anti-Aging Effects of Mixed Plant Extracts

Experimental Method

Cell line and cell culture

Fibroblasts were treated with 5% CO ₂ , 37 ° C cells using Dulbecco's modified Eagle's medium (DMEM, GibcoBRL, Braunschweig, Germany) medium with 10% fetal bovine serum (FBS) and penicillin / streptomycin (10 ul / ml). While culturing in an incubator, mainly 3-7 generation fibroblast line was used for the experiment.

Mixed extract treatment

The fibroblasts were treated with concentrations of 0, 0.1, 0.5, 1 and 2 mg / ml and after 48 hours of cell culture, the proteins were used for extraction experiments.

Western blot  analysis

0.2 M PMSF (phenyl-methyl-sulfonyl-fluoride) in lysis buffer [10 mM Tris-Cl (pH7.4), 5 mM EDTA (pH 8.0), 130 mM NaCl, 1% TritonX-100] in cultured cells, Proteolytic enzyme inhibitors (0.02 mM aprotinin, 2 mM leupeptin, 5 mM phenanthroline, 28 mM benzamidine-HCl) were extracted and quantified at 30 μg concentration. Protein expression patterns were analyzed using the respective primary agents for type 1 collagen, Smad2 / 3, MMP-1 and TIMP-1 (Santa Cruz Co., CA, USA).

CAT Assay

Glycerol for 1 minute after standing for 4 hours on calcium chloride / DNA coprecipitation method in fibroblasts inoculated with CAT-promoter plasmid (COL1A2) of collagen type 1 promoter at a density of 1 × 10 ⁵ cells / mL (15%) shock and transfection. Calcium chloride precipitate was prepared at a plasmid DNA concentration of 25 μg / mL. Cells were incubated for 24 hours in curcumin treated experimental media and then digested by three cycles of freeze-thaw in 0.25 mol Tris-HCl at pH 7.8. The protein extracted from each was used for the measurement of CAT activity using [14C] -chloramphenicol as a substrate. Acetylated and nonacetylated forms of chloramphenicol were separated by thin layer chromatography and subjected to autoradiography.

Experiment result

Increased Expression of Type 1 Collagen with Mixed Extract Treatment Time and MMP -1 decrease

To investigate the effect of the mixed extract (AF-343) treatment on the expression of collagen type 1 in fibroblasts, the mixed extract (2 μg / ml) was treated with fibroblasts and subjected to Western blot analysis 48 hours later. 24 hours after the mixed extract treatment, the protein expression of type 1 collagen increased, and the expression of MMP-1 decreased significantly (see FIG. 1 below). No significant difference in expression was observed for MMP-2 and TIMP-1.

Expression of collagen type 1 was increased according to the concentration of mixed extract and MMP -1 reduced expression

To determine whether the expression of collagen type 1, MMP-1, MMP-2 and TIMP-1 changes in the fibroblasts in different concentrations, the mixed extracts were prepared by concentration (0, 0.1, 0.5, 1 and 2 μg / ml). Fibroblasts were treated and Western blot 48 hours later. The expression of collagen type 1 was increased with increasing the concentration of the mixed extract, whereas the expression decreased gradually with increasing the concentration of the mixed extract. TIMP-1, which has a function of inhibiting MMP-1, was slightly increased as the concentration of the mixed extract increased. In the case of MMP-2 and TIMP-1, no significant expression change was observed in comparison with type 1 collagen and MMP-1 (see FIG. 2 below).

Collagen Type 1 by Mixed Extract Treatment mRNA  increase

To investigate the effect of the mixed extract on the mRNA expression of collagen type I in fibroblasts, the mixed extract (2 mg / ml) was treated with fibroblasts and subjected to CAT analysis by time zone. The CAT activity of type 1 collagen was slightly increased (see FIG. 3) when the mixed extract was treated, indicating that the type 1 collagen expression was increased at the mRNA level when the mixed extract was treated.

By mixed extract treatment Smad  Expression changes of proteins

To determine if Smad2 / 3 activation is involved in the increase of collagen expression by type 1 extracts, the mixed extracts were treated with fibroblasts at different concentrations (0, 0.1, 0.5, 1, 2 μg / ml) 48 hours after Western treatment. As a result of the blot, as the concentration of the mixed extract increased, the expression of phosphor-Smad2 / 3 indicating the activation of Smad2 / 3 slightly increased (see FIG. 4 below).

Review

In the present invention, the effect of the mixed extract (AF-343) on the signaling system involved in the type 1 collagen biosynthesis of fibroblasts, the mixed extract is expressed in type 1 collagen through smad2 / 3 activation in fibroblasts In addition to the increase, it was confirmed that the expression of MMP-1 that degrades collagen is decreased.

80% of the connective tissue collagen in the dermis of human skin is type 1 collagen and is mainly produced in fibroblasts. Collagen is not only involved in elasticity and flexibility of skin tissue, but also biosynthesis and degradation of collagen are important in skin aging and wound regeneration (Wulf HC, Sandby-Møller J., Kobayasi T., Gniadecki R .: Skin aging and natural photoprotection.Micron 35,185 (2004)) (Metcalfe AD, Ferguson MW: Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration.JR Soc.Interface. 4,413 (2007) )). Smad, AP-1, and SP-1 are the major transcriptional regulators in the production of type 1 collagen, and the TGF-β / Smad pathway is recognized as the most important in the production of type 1 collagen. Cutroneo KR: TGF-beta-induced fibrosis and SMAD signaling: oligo decoys as natural therapeutics for inhibition of tissue fibrosis and scarring.Wound.Repair.Regen. 15, S54 (2007). MMP-1 and MMP-2 proteins are involved in the degradation of collagen, and TIMP-1 protein maintains homeostasis by appropriately inhibiting MMP function to prevent excessive MMP activation (Davis GE, Saunders WB: Molecular balance of capillary tube formation versus regression in wound repair: role of matrix metalloproteinases and their inhibitors.J. Investig. Dermatol. Symp. Proc. 11, 44 (2006)).

Dandelion (Taraxacum platycarpum) is a perennial plant of the Asteraceae family named Pogongyoung, which is used for oriental medicine, such as fever, diarrhea, diuresis, dryness, expectoration, detoxification, and in the West, it is used for antioxidant, antibacterial, bile secretion, antirheumatic and diuretic. It is one of the medicinal plants that have long been used by folks for various diseases in the East and West (Chang JK: Seasonly wild flowers of Korea.Doseochulpan Nexus, Seoul, 1997). Dandelion contains Vitamin H (Biotin), Choline, Gluten, Gum, Inositol, Inulin, Iron, Lactpicrine, Linolenic Acid, Magnesium, Niacin, Para-aminobenzoic acid (PABA), Phosphorus, Potassium, Protein, Resin, Sulfur, Zinc, Vitamin It is known to contain components such as A, B1, B2, B5, B6, B9, B12, C, E, and P (Leu YL, Shi LS, Damu AG: Chemical constituents of Taraxacum formosanum.Chem.Pharm.Bull (Tokyo). 51,599 (2003). In previous studies, the chemical composition, antimicrobial and anti-tumor properties of dandelion extract (Jeong JY, Chung YB, Lee CC, Park SW, Lee CK: Studies on immunopotentiating activities of antitumor polysaccharide from aerial parts of Taraxacum platycarpum.Archi.Pharm.Res. 14,68 (1991)), anti-inflammatory (Kim HM, Shin HY, Lim KH, Ryu ST, Shin TY, Chae HJ, et al. Taraxacum officinale inhibits tumor necrosis factor-alpha production from rat astrocytes.Immunopharmacol. Immunotoxicol. 22,519 ( (Seo SW, Koo HN, An HJ, Kwon KB, Lim BC, Seo EA, et al. Taraxacum officinale protects against cholecystokinin-induced acute pancreatitis in rats.World.J.Gastroenterol.11,597 (2005)), antioxidant (Hu C., Kitts DD: Dandelion (Taraxacum officinale) flower extract suppresses both reactive oxygen species and nitric oxide and prevents lipid oxidation in vitro.Phytomedicine. 12,588 (2005)). Food or anticancer drugs There is a drug being developed.

Dandelion extract has been found to have anticoagulant efficacy by binding to thrombin. In particular, desacetylmatricarin has been reported to have antiallergic function. Therefore, anti-inflammatory and anti-allergic agents have been developed using dandelion extract (Seo SW, Koo HN, An). HJ, Kwon KB, Lim BC, Seo EA, et al. Taraxacum officinale protects against cholecystokinin-induced acute pancreatitis in rats.World.J. Gastroenterol. 11,597 (2005)).

As a result of analyzing the expression pattern of collagen type 1 after treating the mixed extracts on the skin fibroblasts cultured in the present invention, the type 1 collagen mRNA expression was slightly increased in CAT assay, which is the type 1 collagen in Western blot. The results were consistent with the results of increased protein expression. These results suggest that the mixed extract may play an important role in the development of drugs that promote the aging process or wound regeneration associated with collagen reduction in the future.

The TGF-β / Smad signaling system is an important upstream signaling system for the production of type 1 collagen. In this study, it was confirmed that the increased expression of collagen type 1 by the mixed extract was due to the inhibition of the TGF-β / Smad signaling system, and the phosphorylation of Smad2 and Smad3 proteins was slightly increased when the mixed extract was treated. It means the activation of smad3, and activated smad2, smad3 move into the nucleus to promote the transcription process of type 1 collagen.

There are few reports on the effect of the mixed extract on the expression of MMP-1, MMP-2, which degrades collagen, and TIMP-1, which is involved in inhibiting collagen degradation, in fibroblasts. In this study, the mixed extract reduced the expression of MMP-1 protein in a concentration-dependent manner in fibroblasts and did not significantly affect the expression of MMP-2 and TIMP-1 proteins. MMP-1 is known to be stimulated by mitogen activated protein kinases (MAPK), such as p38 or extracellular signal-regulated kinase (ERK), which is further studied by investigating whether p38 and ERK MAPK are inhibited when mixed extracts are applied to fibroblasts. Detailed regulatory mechanisms are expected to be elucidated.

In conclusion, the mixed extract of the present invention increased the production of type I collagen by activating Smad2 and Smad3 in cultured fibroblasts, and at the same time, the expression of MMP-1 was decreased, which was used in the future. It is suggested that the mixed extract of the present invention can be used as a major component in the development of anti-aging and wound regeneration formulations.

conclusion

Treatment of the mixed extract on the skin fibroblasts cultured in the present invention resulted in an increase in the expression of collagen type 1 protein by the activation of Smad 2 and Smad 3 proteins, and the expression of MMP-1 protein was decreased. It is considered that the mixed extract is an active ingredient that can be used for anti-aging and skin regeneration in the future.

Formulation Example  One

Formulation examples of the flexible lotion in the cosmetic containing the mixed extract of Examples 1-4 of the present invention are as follows.

ingredient Content (% by weight) Mixed extract 0.5 1,3-butylene glycol 5.2 Oleyl alcohol 1.5 ethanol 3.2 Polysorbate 20 3.2 Benzophenone-9 2.0 Carboxyl vinyl polymer 1.0 glycerin 3.5 incense a very small amount antiseptic a very small amount Purified water Balance system 100

Formulation Example  2

Formulation example of the milk lotion in the cosmetic containing the mixed extract of Examples 1-4 of the present invention is as follows.

ingredient Content (% by weight) Mixed extract 0.6 glycerin 5.1 Propylene glycol 4.2 Tocopheryl acetate 3.0 Liquid paraffin 4.6 Triethanolamine 1.0 Squalane 3.1 Macadamia nut oil 2.5 Polysorbate 60 1.6 Sorbitan sesquiterate 1.6 Propylparaben 0.6 Carboxyl vinyl polymer 1.5 incense a very small amount antiseptic a very small amount Purified water Balance system 100

Formulation Example  3

Formulation examples of the nourishing cream in the cosmetic composition containing the mixed extract of Examples 1-4 of the present invention are as follows.

ingredient Content (% by weight) Mixed extract 1.0 glycerin 4.0 vaseline 3.5 Triethanolamine 2.1 Liquid paraffin 5.3 Squalane 3.0 Wax 2.6 Tocopheryl acetate 5.4 Polysorbate 60 3.2 Carboxyl vinyl polymer 1.0 Sorbitan sesquioleate 3.1 incense a very small amount antiseptic a very small amount Purified water Balance system 100

Formulation Example  4

Formulation example of the massage cream in the cosmetic containing the mixed extract of Examples 1-4 of the present invention is as follows.

ingredient Content (% by weight) Mixed extract 0.5 glycerin 4.0 vaseline 3.5 Triethanolamine 0.5 Liquid paraffin 24.0 Squalane 3.0 Wax 2.1 Tocopheryl acetate 0.1 Polysorbate 60 2.4 Carboxyl vinyl polymer 1.0 Sorbitan sesquioleate 2.3 incense a very small amount antiseptic a very small amount Purified water Balance system 100

Formulation Example  5

Formulation examples of the pack in the cosmetic containing the mixed extract of Examples 1-4 of the present invention are as follows.

ingredient Content (% by weight) Mixed extract 1.0 Ethyl alcohol 3.0 EDTA-2Na 0.02 Propylene glycol 5.1 Glycerin 4.5 Carbopol 1.0 Polyoxide 0.1 antiseptic a very small amount incense a very small amount Purified water Balance system 100

Claims (10)

A cosmetic composition for preventing skin aging, which comprises an extract of Ulmus davidiana var., Cassia obtusifolia, and Dandelion (Taraxacum platycarpum) as active ingredients.
The method of claim 1,
The extracts of the roots of the root, extract extracts and dandelion extracts, characterized in that the type 1 collagen expression increasing ability or type I collagenase (Matrix Metalloproteinase-1, MMP-1) expression reducing ability.
The method of claim 1,
The root of the skin is a cosmetic composition, characterized in that the elm root bark.
The method of claim 1,
The dandelion is a cosmetic composition, characterized in that selected from the group consisting of dandelion flowers, leaves, stems, fruits, seeds, roots and outposts.
The method of claim 1,
Extraction solvent of the extract is a cosmetic composition, characterized in that selected from the group comprising water, ethanol, methanol, methylene chloride, ethyl acetate, butanol, hexane, butylene glycol, chloroform and mixtures thereof.
The method of claim 5, wherein
Extraction solvent of the extract is a cosmetic composition, characterized in that ethanol.
The method of claim 1,
The amount of the extract is a cosmetic composition, characterized in that 0.00002-30% by weight relative to the total weight of the cosmetic composition.
The method of claim 1,
The cosmetic composition may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray Or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition for preventing skin aging, which comprises an extract of Ulmus davidiana var., Cassia obtusifolia and dandelion (Taraxacum platycarpum) as an active ingredient.
A skin composition for preventing skin aging comprising Ulmus davidiana var. Extract, Cassia obtusifolia extract, and Dandelion (Taraxacum platycarpum) extract as active ingredients.

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