KR20100137113A - Composition for inhibition skin-aging comprising of 3', 4',7,8-tetrahydroxyflavanon-7-o-glucoside - Google Patents

Abstract

PURPOSE: A composition containing 3', 4',7,8-tetrahydroxyflavanone-7-O-glucoside for anti-aging is provided to suppress MMP-9(Matrix Metalloproteinase-9) and to prevent and treat aging. CONSTITUTION: A 3', 4',7,8-tetrahydroxyflavanone-7-O-glucoside is denoted by chemical formula 1. A pharmaceutical composition for preventing and treating aging or wrinkle contains 3', 4',7,8-tetrahydroxyflavanone-7-O-glucoside or pharmaceutically acceptable salt thereof as an active ingredient. A cosmetic composition for anti-aging or anti-wrinkling contains 4',7,8-tetrahydroxyflavanone-7-O-glucoside or pharmaceutically acceptable salt thereof as an active ingredient. The cosmetic composition is used in the form of lotion, essence, cream, pack, gel, ointment, or spray.

Description

 Composition for inhibition Skin-aging comprising of 3 ', 4', 7,8-tetrahydroxyflavanon-7 (3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside) -O-glucoside}

The present invention is a skin aging inhibitor composition comprising a 3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (hereinafter referred to as "flavanomare") compound represented by the formula (1) More specifically, by inhibiting the gene expression of Matrix Metalloproteinase-9 (MMP-9) induced by Tumor necrosis factor-α (TNF-α) inhibits the degradation of collagen, a component of the skin, skin aging The present invention relates to a composition for preventing skin aging and improving skin wrinkles, including the flavanomarene compound of Formula 1, which may improve and prevent the phenomenon.

Skin aging can be divided into physiological aging process over time and aging process due to environmental factors (Claude et al., Free radical Boil & Med , 26, 174, 1999).

As the skin ages, various metabolic activities are lowered, and cell activity is lowered, resulting in thinner, dry and wrinkled skin. The interest in anti-aging has been of major interest not only to researchers but also to many ordinary people, and research on this is expected to become more active over time.

The structure of the skin is divided into four categories from the surface to the stratum corneum, epidermis, dermis and subcutaneous tissue. At the top of the stratum corneum, dead cells are layered, and soft fibrous proteins called keratin fill the cells and protect the skin. Most of the epidermal cells are filled with keratinocytes that make up keratinocytes. The dermis is a connective tissue composed of glue fibers and elastic fibers, so it has toughness and elasticity. In addition, the skin tissue is a connective tissue layer under the dermis, and serves to connect the skin and fascia.

External factors affecting skin aging include humidity, tobacco smoke, pollution, wind, temperature and ultraviolet rays. In particular, aging due to ultraviolet rays is called photoaging. The shape seen in chronic sun-damaged skin is an abnormal deposition of elastic fibrous material in the upper collagen of the dermis, an increase in proteoglancan, and a significant decrease in collagen, the main protein of the dermis (Li et al., Cancer Research , 56, 284, 1996).

Collagen imparts strength and tension to the skin and thereby protects the skin from external irritation or force. Since collagen accounts for 90% of the dermis, collagen reduction is closely related to aging of the skin.

Matrix Metalloproteinase (MMPs) is a protein that is dependent on zinc ions and breaks down the extracellular matrix (ECM) present in the basement membrane and interstitial tissues (Steven D shapiro, Cell). Biol , 10, 602, 1998). After synthesis into inactive spheres, it is activated by proteolytic hydrolysis, and the activity of MMPs is known to be regulated by gene expression, inactivation process of enzymes, and inhibition of enzyme activity (Woodhouse et al., Cancer , 80, 1529, 1997; Wass et al., Br J Cancer , 86, 1876, 2002; Nagase et al., J Biol Chem , 274, 21491, 1999).

About 20 MMPs are known depending on substrate specificity. Unlike MMP-2, which is always expressed and functioning, MMP-9 is characterized by increased expression by stimulation of inflammatory mediators such as TNF-α and is regulated by specific transcription factors such as NFκB (Sanseau et al., J Biol Chem , 227, 35766, 2002). It can decompose most of the extracellular matrix, thus maintaining normal physiological functions during tissue reconstruction such as the development of wound healing fetus, neovascularization, menstruation, ovulation, and pregnancy, but if excessively expressed, skin aging, Involvement in several pathologies has been reported, including rheumatoid arthritis, chronic ulcers, periodontitis, arteriosclerosis, tumor growth, and tumor cell infiltration and metastasis (Linda et al., Chem Biol , 2, 466, 1998).

Repeated UV exposure is known to reduce collagen synthesis and increase the expression of MMP-9, an enzyme that degrades collagen and other matrix proteins in cells through multiple signaling pathways (Fisher et al., Arch Dermatol , 138, 1462, 2002; Lee et al., J Korean Soc Agric Chem Biotechnol , 46, 60, 2003). In fact, even one UV irradiation increases the activity of MMP-9 in the skin and significantly breaks down collagen in the skin, which has been reported to play an important role in photoaging and MMP-9 affects collagen breakdown of the dermis. Lee et al., J. Dermatol Sci , 17, 182, 1998.

When collagen is reduced by MMP-9, the skin's strength and tension decreases, and the function of protecting the skin is significantly reduced, thereby accelerating skin aging. As such, the increased activity of MMP-9 breaks down the skin components and accelerates skin aging, and thus, develops a new leader that improves or prevents skin aging using compounds that inhibit the expression of MMP-9 genes. It is expected to be used as an indicator.

Accordingly, the present inventors have diligently tried to find a compound capable of suppressing skin aging phenomenon, and as a result, the 3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomarein) compound is MMP- By inhibiting the gene expression of 9 to inhibit the degradation of collagen, a component of the skin, and completed the present invention.

In conclusion, the present invention refers to 3 ′, 4 ′, 7,8-tetrahydroxyflavanone-7-O-glucoside (hereinafter referred to as “flavanomarein”) having inhibitory activity of Matrix Metalloproteinase-9 (MMP-9). The main purpose is to provide a compound and a novel use of the compound as an anti-aging agent.

In order to achieve the above object, the present invention inhibits the expression of Matrix Metalloproteinase-9 (MMP-9) gene to inhibit the degradation of collagen which is a component of skin 3 ', 4', 7,8-tetrahydroxyflavanone Provided is a -7-O-glucoside (flavanomare) compound and a skin aging inhibitor composition comprising the compound.

In the present invention, the skin aging inhibitor composition includes a pharmaceutical composition, cosmetic composition, and health functional food that can prevent skin aging and / or improve skin wrinkles.

Since the 3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomare) compound according to the present invention has an effect of inhibiting the expression of MMP-9 gene and inhibiting collagen degradation, It may be usefully used as an active ingredient of a composition for preventing skin aging and / or improving skin wrinkles caused by skin aging.

Hereinafter, the present invention will be described in detail.

The present invention provides a compound 3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (hereinafter referred to as "flavanomarein") represented by the following formula (1).

[Formula 1]

In the present invention, the flavanomarine compound inhibits the gene expression of Matrix Metalloproteinase-9 (MMP-9) induced by Tumor necrosis factor-α (TNF-α) to inhibit the breakdown of collagen, a component of skin. It is characterized by inhibiting.

In addition, the flavanomarine compound may be extracted from plants or separated or prepared by synthetic methods well known in the art. In one embodiment of the present invention, the flavanomarene compound was synthesized based on Bruce's method (Bruce A. Bohm, Harwood academic publishers, Introduction to flavanoids, 328-329, 1999).

The present invention also provides a method of inhibiting the degradation of collagen through the inhibition of gene expression of MMP-9 induced by TNF-α.

In another aspect, the present invention provides a new use of the 3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomare) compound of Formula 1 as a skin aging inhibitor.

Specifically, the present invention provides a pharmaceutical composition capable of preventing skin aging and / or treating skin wrinkles, including the flavanomare compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The compounds of the present invention can also be synthesized through conventional synthesis and fractionation methods of substituents (Herbert O. House, Modern Synthetic Reactions, 2nd Ed., The Benjamin / Cummings Publishing Co., 1972).

In addition, the compounds of the present invention may be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.

As salts are acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts are prepared by conventional methods, for example, by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water may be heated and then the mixture is evaporated to dryness or the precipitated salts may be suction filtered.

In this case, an organic acid and an inorganic acid may be used as the free acid, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, glycolic acid gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salt, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of Formula 1 include salts of acidic or basic groups which may be present in compounds of Formula 1 unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfates, hydrosulfates, phosphates, hydrogen phosphates, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.

The flavanomarine compound according to the present invention prepared as described above is excellent in the MMP-9 gene expression inhibitory effect induced by TNF-α, it can be used as an skin aging inhibitor.

In another aspect, the present invention provides a pharmaceutical composition for the prevention and treatment of skin aging or / and wrinkles, including the flavanomarene compound of Formula 1 as an active ingredient, and a pharmaceutically acceptable carrier.

The pharmaceutical composition comprising the flavanomarine compound of the present invention may further comprise a suitable carrier, excipient or diluent according to conventional methods.

Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose And methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

Pharmaceutical compositions comprising a compound of the present invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, or the like, external preparations, suppositories, or sterile injectable solutions according to conventional methods. Can be used.

Specifically, when formulated, it may be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrating agents, surfactants, etc. which are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the compound, for example, starch, calcium carbonate, sucrose. ), Lactose, gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc may also be used. Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid parazin, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. May be included. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention will be appropriately selected by those skilled in the art depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration. However, for the desired effect, the compounds of the present invention may be administered in divided doses of 10 to 500 mg once to several times daily. The compound of the present invention in the composition should be present in an amount of 0.01 to 50% by weight relative to the total weight of the total composition.

In addition, the pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes, and all modes of administration can be expected, for example, oral, rectal or intravenous, muscular, subcutaneous. , Intrauterine dural or intracerebroventricular injection.

In another aspect, the present invention provides a cosmetic composition that can prevent the skin aging and / or improve the skin wrinkles comprising the flavano marin compound of Formula 1 as an active ingredient.

When using the flavanomarene compound according to the present invention as a cosmetic, including conventional cosmetic base components in addition to the compound, for example, conventional containing agents such as stabilizers, solubilizers, pigments and fragrances, and And a carrier. In addition, the cosmetics may be prepared in any formulation conventionally prepared in the art, and are not particularly limited, for example, cosmetic lotions, essences, lotions, pastes, surfactant-containing cleansing, creams, packs, gels, ointments, It may have a formulation of powder, patch or spray (spray).

In case the formulation of the cosmetic composition is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc and zinc oxide. One or more selected from the group can be used, and when the formulation is a lotion, an essence, a solvent, a solubilizing agent or an emulsifying agent is used as the carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, One or more selected from the group consisting of benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils, glycerol aliphatic esters, polyethylene glycols, fatty acid esters of sorbitan, and the like can be used.

In addition, when the formulation of the cosmetic composition is a powder or spray, one or more selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in particular, spray May further comprise propellants such as chlorofluorohydrocarbons, propane / butane or dimethyl ether. In addition, in the case where the formulation of the cosmetic composition is a surfactant-containing cleansing agent, as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, methyl taurate, sarcosinate, One or more selected from the group consisting of fatty acid amide ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like can be used.

At this time, the compound in the cosmetic is preferably added at 0.01 to 50% by weight, preferably 0.01 to 30% by weight of the total weight.

In addition, the present invention provides a health functional food composition that can prevent skin aging and / or improve the skin wrinkles comprising the flavanomare compound of Formula 1 as an active ingredient.

Examples of the food to which the compound of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods. It may also be added to food or beverages for the purpose of preventing cerebral disease symptoms.

At this time, the amount of the extract in the food or beverage is generally added to the functional food composition of the present invention 0.01 to 50% by weight, preferably 0.01 to 30% by weight of the total food weight, the health beverage composition is 100ml It can be added in a ratio of 0.01 to 5g, preferably 0.1 to 3g on the basis of.

The health functional food of the present invention includes the form of tablets, capsules, pills, liquids, and the like, and the health beverage composition of the present invention contains the compound as an essential ingredient in the indicated ratio, and there is no particular limitation on the liquid component. Like beverages may further contain various flavors or natural carbohydrates.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and the like. Sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the above, natural flavors, taumartins, stevia extracts such as rebaudioside A, glycyrrhizin and the like and synthetic flavors, saccharin, aspartame and the like can be advantageously used as flavoring agents.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. These components may be used independently or in combination, and the proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight relative to 100 parts by weight of the composition of the present invention.

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 examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example 1 Confirmation of the MMP-9 Gene Expression Inhibitory Effect of 3 ′, 4 ′, 7,8-tetrahydroxyflavanone-7-O-glucoside

Of 3 ′, 4 ′, 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomare) compounds In order to confirm the effect of inhibiting MMP-9 gene expression induced by TNF-α, a luciferase reporter essay measuring MMP-9 gene promoter activity was used.

Specifically, genometics software (www.genomatix.de) was used for gene sequencing, and the transcriptional regulatory region (base sequence -925 to +13 base pair) of the human MMP-9 gene was synthesized by PCR and lucifer PGL4 / MMP-9-Luc (-925 / + 13) vector was prepared by inserting into multiple cloining site of pGL4-basic vector (Promega, Medison, Wisconsin, USA) containing luciferase gene It was. At this time, the primers used for sequencing are as follows.

forward primer (-925F): 5'-ccatacctggccctgaatct-3 '(SEQ ID NO: 1)

reverse primer (+ 13R): 5'-ggcagaggtgtctgactgcag-3 '(SEQ ID NO: 2)

Using the analyzed results, 0.5 μg of pGL4 / MMP-9-Luc (-925 / + containing MMP-9 binding sequence in MDA-MB231, a human breast cancer cell line purchased from American Type Culture Collection, USA) was used. 13) Plasmid was injected into the cell using fuGENE6, and then 48 hours after transfection, the cell culture solution had a concentration of 10 ng / ml. TNF-α alone (control) or at a concentration of 10 ng / ml Treatment of TNF-α with flavanomarene at a concentration of 20 μg / ml (experimental group) confirmed the inhibitory effect of flavanomarene on TNF-α activity.

Cells treated with TNF-α and flavanomarein were incubated for another 12 hours, harvested, lysed, and then luminescent luciferin fluorescence was measured by adding 50 μL of luciferin as a substrate to the cell solution protein. Cell lysates and substrate luciferin were obtained from the Dual-Glo Luciferase Assay System Kit purchased from Promega (Madision, WI, USA), and the luciferin fluorescence specificity was determined using Centro LB 960 lumino. Meter (Centro LB 960 Luminometer, Berthold Technology, German).

As shown in FIG. 1, when treated with TNF-α alone (control), luciferase enzyme activity was increased two-fold, which means that expression of MMP-9 gene was increased by TNF-α. However, in the experimental group treated with TNF-α and the flavanomarene of the present invention, the activity of luciferase by TNF-α was completely inhibited.

Thus, the 3 ′, 4 ′, 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomare) of the present invention inhibits the expression of MMP-9 gene activated by TNF-α at the transcriptional level. I could confirm that.

As described above, specific portions of the contents of the present invention have been described in detail, and for those skilled in the art, these specific techniques are merely preferred embodiments, and the scope of the present invention is not limited thereto. Will be obvious. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

1 shows the inhibitory effect of TNF-α induced MMP-9 gene expression of 3 ′, 4 ′, 7,8-tetrahydroxyflavanone-7-O-glucoside (flavanomare) compounds of the present invention. will be.

<110> Konkuk University Industrial Cooperation Corp. <120> Composition for inhibition skin-aging comprising of          3 ', 4', 7,8-tetrahydroxyflavanon-7-O-glucoside <130> P09-E175 <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for identifying human MMP-9 promoter <400> 1 ccatacctgg ccctgaatct 20 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer for identifying human MMP-9 promoter <400> 2 ggcagaggtg tctgactgca g 21  

Claims (11)

3 ', 4', 7,8-tetrahydroxyflavanone-7-O-glucoside (hereinafter, referred to as "flavanomare") compound represented by the following formula (1). [Formula 1]

The method of claim 1, The compound inhibits the expression of Matrix Metalloproteinase-9 (MMP-9) induced by Tumor necrosis factor-α (TNF-α), thereby inhibiting the breakdown of collagen, a component of skin. Lane compound. The method of claim 1, Flavanomarein compound, characterized in that the compound is used as a skin aging inhibitor. Claim 1 flavanomarein compound or a pharmaceutically acceptable salt thereof as an active ingredient skin aging prevention and treatment pharmaceutical composition.  A pharmaceutical composition for preventing and treating skin wrinkles, comprising the flavanomarene compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The method according to claim 4 or 5, The composition is a pharmaceutical composition for the prevention and treatment of skin aging or wrinkles, characterized in that it further comprises a pharmaceutically acceptable carrier. A cosmetic composition for preventing and improving skin aging comprising the flavanomarine compound of claim 1 as an active ingredient.  A cosmetic composition for preventing and improving skin wrinkles comprising the flavanomarene compound of claim 1 as an active ingredient. The method according to claim 7 or 8, The cosmetic composition is a cosmetic composition, characterized in that the lotion, essence, lotion, cream, pack, gel, ointment, powder, patch or spray.  Claim 1 flavanomarein compound for preventing skin aging and improving health functional composition comprising as an active ingredient.  Claims 1 Flavanomarein compound for preventing skin wrinkles and improving health functional food composition comprising as an active ingredient.

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