KR102265411B1 - Cosmetic or pharmaceutical composition for skin whitening, elasticity, anti-wrinkle, or skin moisturizing comprising Acetylaconitine or a pharmaceutically acceptable salt thereof - Google Patents

Abstract

The present invention provides a composition for skin whitening, elasticity, wrinkle improvement, or moisturizing, in particular, a cosmetic composition, a pharmaceutical composition, and a health food comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The cosmetic or pharmaceutical composition of the present invention has few side effects and is safe for the human body and has excellent skin whitening, elasticity, wrinkle improvement, skin moisturizing effect, and the like.
[Formula 1]

Description

A cosmetic or pharmaceutical composition for skin whitening, elasticity, anti-wrinkle, or skin moisturizing comprising Acetylaconitine comprising acetyl aconitine or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable salt thereof}

The present invention relates to a cosmetic composition, a pharmaceutical composition, and a health food having excellent effects such as skin whitening, elasticity, wrinkle improvement, or moisturizing, in particular, having excellent skin-related effects. The present invention also relates to a cosmetic composition, a pharmaceutical composition, and a health food that can exhibit the above effects even in a small amount.

It is a common desire to have fair and fair skin. The color or brightness of the skin is genetically determined according to the concentration and distribution of melanin in the skin of a person, but it is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, or stress. Melanin is produced through a non-enzymatic oxidation reaction after sequentially changing into dopa and dopaquinone by an enzyme called tyrosinase acting as a catalyst in tyrosine, a kind of amino acid. As such, the pathway by which melanin is produced is known, but the cause of tyrosinase triggering in the mechanism of inducing melanin synthesis has not been elucidated in detail.

On the other hand, as a commonly known whitening ingredient, substances that inhibit tyrosinase enzyme activity, such as kojic acid or arbutin, hydroquinone, vitamin C (L-Ascorbic acid) or derivatives thereof and various There are plant extracts. By inhibiting the synthesis of melanin pigment, it is possible not only to realize skin whitening by brightening the skin tone, but also to improve skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity. However, when applied to the skin, there is a problem in that the amount of use is limited due to safety issues such as irritation and redness, or the actual effect cannot be expected because the effect is insignificant.

Collagen is a major matrix protein produced by fibroblasts of the skin and is present in the extracellular matrix, and its important functions include mechanical firmness of the skin, resistance of connective tissue and tissue bonding, support of cell adhesion, cell division and differentiation. Induction (at the time of growth of an organism or wound healing) and the like are known. Such collagen is reduced by age and photoaging caused by UV irradiation, and collagen reduction is promoted by collagenase enzyme activity that decomposes collagen. It is known that it is closely related to the formation of wrinkles on the skin.

In addition, elastin fibers form cross-links with collagen and are an important skin component for wrinkle generation, which is involved in skin elasticity. Deficiency and aggregation of elastin fibers and a marked increase in the activity of the elastin-degrading enzyme, Elastase, have been found to be one of the factors that cause skin wrinkles. Elastase is the only enzyme capable of decomposing elastin, and inhibition thereof is known to fundamentally reduce skin wrinkle improvement.

On the other hand, collagen and elastin fibers are matrix proteins that play an important role in retaining moisture in the dermis layer. These matrix proteins adsorb moisture, and increase moisture retention within their structures so that the skin can maintain a state of adequate moisture. It is known that it is involved in maintaining the elasticity of the skin through this.

Currently, retinoid, adenosine, animal placenta-derived protein, chlorella extract, and the like are known as wrinkle-improving cosmetics. Retinol, the most well-known, is a substance that promotes collagen synthesis and inhibits the elastase enzyme, but it is unstable, and its usage is limited due to safety issues such as irritation and redness when applied to the skin. It is known that it is difficult to expect a wrinkle improvement effect.

Glycation generally refers to a reaction in which a simple sugar such as glucose or fructose forms a covalent bond with a protein or fat that occurs without the involvement of an enzyme. Glycosylation occurs through a series of complex chemical reactions known as Amadori reactions and Maillard reactions, resulting in the formation of advanced glycation endproducts (AGEs).

On the other hand, the glycosylation process of proteins occurs slowly because enzyme action is not involved, and as a result, proteins with long half-lives such as collagen have a greater effect. As a result of analyzing the final glycated products, glycated collagen was accumulated as the age increased, and the accumulation of final glycated products changed the protein to a harder and more brittle state, and glycated collagen was released from the extracellular matrix of the dermal layer. By preventing collagen from forming an appropriate structure, the skin loses its elasticity and promotes wrinkle formation (Dyer, DG et al, The Journal of Clinical Investigation., 9, p. 2463, 1993).

In addition, the final saccharification product produced by saccharification is a brownish substance and is known to accumulate on the upper dermis layer as skin aging progresses. As skin aging progresses, it is thought to be the causative agent of gradually turning yellow in the face, and it is known that the more specific final glycation products accumulate, the less reflected light reflected from the skin. While the amount of melanin in the skin has a weak relationship with skin aging, the final glycation products accumulated in the dermal layer of the skin gradually increase as the skin ages and can make the face dull (Hiroshi, O. et al, Skin). Research and Technology, 15, p. 496, 2009)

Aminoguanidine, Pyridoxamine, Aspirin, etc. are known as substances that inhibit glycation. However, these substances have a limited amount of use due to safety concerns on the skin or are practically effective due to insignificant effects. There is a problem that cannot be expected.

Therefore, the development of ingredients with whitening, elasticity, wrinkle improvement, or moisturizing effects that are safe for the body, stable in active ingredients, and more effective than existing skin whitening, elasticity, wrinkle improvement, or moisturizing effects is desperately needed.

Therefore, the present invention solves the above problems, and provides a new active ingredient, that is, a useful use of the active ingredient, which has few side effects and is safe for the human body and has excellent whitening, elasticity, wrinkle improvement, or moisturizing effect.

In other words, the problem to be solved by the present invention is to provide a composition comprising an active ingredient having excellent efficacy as an active ingredient.

In order to solve the above problems, the present invention provides a composition for skin whitening, elasticity enhancement, wrinkle improvement, or moisturizing comprising acetyl aconitine or a pharmaceutically acceptable salt thereof as an active ingredient, preferably provides a cosmetic composition, pharmaceutical composition or health food.

That is, the present invention provides a new use of acetylaconitine or a pharmaceutically acceptable salt thereof for skin whitening, elasticity enhancement, wrinkle improvement, and moisturizing.

The inventors of the present invention confirmed the fact that acetylaconitine inhibits collagenase and elastase activity of the skin, promotes skin regeneration by exhibiting an anti-glycosylation effect, and has moisturizing, skin elasticity and anti-wrinkle effects. and completed the present invention. In addition, it was confirmed that the acetyl aconitine inhibits melanin synthesis to exhibit a whitening effect, thereby completing the present invention.

In the present invention, the 'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity.

In the present invention, 'improving elasticity or skin elasticity' means alleviating the degree of sagging or sagging skin. In addition, the elasticity means maintaining the elasticity of the skin in a state in which elastin and collagen are sufficiently present.

In the present invention, the term 'wrinkle improvement effect' refers to suppressing or inhibiting the formation of wrinkles on the skin, or alleviating the already generated wrinkles.

In the present invention, "moisturizing or moisturizing the skin" means to increase the feeling of moisture in the skin and to maintain a moist state. The skin moisturizing effect can help to improve wrinkles and increase elasticity of the skin.

The present invention provides a composition comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof in order to solve the above problems and achieve the object of the present invention.

[Formula 1]

The compound represented by Formula 1 has a molecular formula C ₃₆ H ₄₉ NO _{12 , a} molecular weight of 687.778, and is named acetyl aconitine (Acetylaconitine).

The present invention is not particularly limited to the method for obtaining acetyl aconitine, and may be chemically synthesized by a method known in the art, or a commercially available material may be used.

In the cosmetic composition, pharmaceutical composition and health food according to the present invention, the content of acetyl aconitine is preferably 0.00001 to 10% by weight based on the total weight of the cosmetic composition, pharmaceutical composition and health food.

The "pharmaceutically acceptable salt" of the present invention refers to salts prepared using organic or inorganic acids with little or no toxicity.

The organic acid is, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid , glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxyacetic acid, benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid salts, and inorganic acids include, for example, hydrochloric acid. , hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid salts. Preferably, it may be in the form of hydrochloride or acetate, and more preferably in the form of hydrochloride.

The above-mentioned acid addition salt can be prepared by a) directly mixing the compound of formula 1 and an acid, b) dissolving one of them in a solvent or a hydrous solvent and mixing, or c) dissolving the compound of formula 1 in a solvent or water. It is prepared by a general method of preparing salts by placing them in an acid in a solvent and mixing them.

In addition to the above, additional salts available include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt, Aspartate, etc.

Acetylaconitine of the present invention may exist in unsolvated as well as solvated forms including hydrates, ethanolates, and the like. Acetylaconitine of the present invention may exist in crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.

The cosmetic composition for skin whitening, elasticity enhancement, wrinkle improvement, and skin moisturizing according to the present invention is a solution, external ointment, cream, foam, nourishing lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleaning Group consisting of cosmetics, bath products, sunscreen cream, sun oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray It may be prepared in a formulation selected from, but is not limited thereto.

In addition, the cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and conventional ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, A thickener, a chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately mixed, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation. When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures thereof may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier component. In particular, in the case of a spray, additional chlorophyll propellants such as fluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3 -butylglycol oil, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

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

When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars, etc. may be used as carrier components. can

The present invention also provides a method for skin whitening, elasticity enhancement, wrinkle improvement, and skin moisturizing, comprising the step of applying a compound represented by Formula 1 to the skin of an individual. The subject includes, without limitation, mammals including rats, livestock, humans, and the like.

According to another embodiment of the present invention, the present invention provides a pharmaceutical composition for skin whitening, elasticity enhancement, wrinkle improvement, or skin moisturizing containing the compound of Formula 1 as an active ingredient.

The composition comprising the compound of the present invention may be in various oral or parenteral formulations, but preferably parenteral formulations. In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include at least one excipient in one or more compounds, for example, starch, calcium carbonate, sucrose or lactose ( lactose), gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, and emulsions. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The present invention may also provide a formulation of an external preparation for skin for skin whitening, elasticity enhancement, wrinkle improvement, and skin moisturizing effect comprising the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as an external preparation for skin, in addition, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant , water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or common for external applications for the skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as a dermatome. In addition, the ingredients may be introduced in an amount generally used in the field of dermatology.

When the compound of Formula 1 is provided as an external preparation for skin, it is not limited thereto, but may have a formulation such as an ointment, a patch, a gel, a cream, or a spray.

The pharmaceutical composition of the present invention can be particularly preferably used as a parenteral preparation, for example, the external preparation for skin includes a suitable pharmaceutically acceptable base such as vaseline, stearyl alcohol; suitable pharmaceutically acceptable surfactants such as polysorbate and sorbitan sesquioleate; pharmaceutically acceptable suitable moisturizers such as glycerin; a suitable pharmaceutically acceptable solvent; And it can be prepared by a conventional method for preparing an external preparation for skin in which a flavoring agent, a colorant, a stabilizer, a viscous agent, and the like are homogeneously mixed.

In addition, when the compound of Formula 1 of the present invention is used as a pharmaceutical, it may further contain one or more active ingredients exhibiting the same or similar function. For example, it may include known skin whitening, elasticity enhancement, wrinkle improvement, and moisturizing ingredients. When additional skin whitening, elasticity enhancement, wrinkle improvement, and moisturizing ingredients are included, the skin whitening, elasticity enhancement, wrinkle improvement, and moisturizing effects of the composition of the present invention may be further enhanced. When adding the above ingredients, skin safety according to complex use, ease of formulation, and stability of active ingredients can be considered. In one embodiment of the present invention, the composition is a skin elasticity or wrinkle improving component known in the art, retinoic acid, TGF, animal placenta-derived protein, betulinic acid and chlorella extract; As a whitening ingredient known in the art, kojic acid (Kojic acid), a substance inhibiting tyrosinase enzyme activity such as arbutin (Arbutin), hydroquinone (Hydroquinone), vitamin-C (L-Ascorbic acid); and one or two or more components selected from the group consisting of derivatives thereof and various plant extracts. Additional ingredients may be included in an amount of 0.0001 wt% to 10 wt% based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety and ease of formulation of the compound of Formula 1 .

When the pharmaceutical composition of the present invention contains an effective amount of the compound of Formula 1, it can provide desirable skin whitening effect, elasticity enhancing effect, wrinkle improvement effect, and moisturizing effect.

In the present invention, the term “effective amount” refers to an amount of a compound capable of exhibiting a skin whitening effect, enhancing elasticity and wrinkle improvement, or exhibiting a moisturizing effect.

The effective amount of the compound of Formula 1 included in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the compound is applied to the skin, and the time it stays on the skin. For example, when the composition is commercialized as a pharmaceutical, the compound of Formula 1 may be included at a higher concentration than when commercialized as a cosmetic that is routinely applied to the skin. Accordingly, the daily dose is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, and more preferably 50 to 60 mg/kg, based on the amount of the compound of Formula 1, and 1 to It can be administered 6 times.

According to another embodiment of the present invention, there is provided a health food for skin whitening effect, elasticity enhancement effect, wrinkle improvement effect, and moisturizing effect comprising the compound of Formula 1 above.

As used herein, the term 'health food' refers to food prepared by adding the compound of Formula 1 to food materials such as beverages, teas, spices, gum, and confectionery, or encapsulating, powdering, suspension, etc. It means to bring a specific effect, but unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long period of time by using food as a raw material. The health food of the present invention obtained in this way is very useful because it can be ingested on a daily basis, and thus high skin whitening effect, elasticity enhancing effect, wrinkle improving effect, and moisturizing effect can be expected.

When the compound of Formula 1 is used as a food additive, the compound of Formula 1 may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. . There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense. The health beverage composition of the present invention may contain various flavoring agents or natural carbohydrates as an additional component like a conventional beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention. In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Although not limited thereto, in one embodiment of the present invention, the composition may include the compound of Formula 1 in an amount of 0.00001 wt% to 10 wt% based on the total weight of the composition.

The composition containing acetylaconitine or a pharmaceutically acceptable salt thereof of the present invention exhibits a skin whitening effect.

The composition containing acetylaconitine or a pharmaceutically acceptable salt thereof of the present invention exhibits an effect of enhancing skin elasticity or improving wrinkles.

The composition containing acetylaconitine or a pharmaceutically acceptable salt thereof of the present invention exhibits a skin moisturizing effect.

The composition containing acetyl aconitine or a pharmaceutically acceptable salt thereof of the present invention can be used as a safe and excellent cosmetic raw material and pharmaceutical ingredient.

Hereinafter, in order to explain the present invention in more detail, the following examples and the like will be described. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the embodiments described below. The embodiments of the present invention are provided by way of example to help the specific understanding of the present invention.

Reference example One: Acetylaconithine ( Acetylaconitine ) substance information

[Formula 1]

Substance name: Acetylaconitine

CAS No. : 77181-26-1

Molecular Formula: C ₃₆ H ₄₉ NO ₁₂

Molecular Weight: 687.778

Where to buy: Tauto Biotech (China)

Example One: collagenase activity inhibitory effect

The inhibitory effect on the activity of collagenase, an enzyme that decomposes collagen, was confirmed as follows.

Collagenase derived from mouse pancreas was used, and a synthetic substrate, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala, was used as the collagenase substrate. A buffer solution in which 50 mM Tricine, 10 mM CaCl ₂ ·H ₂ O, and 400 mM NaCl (pH 7.5) were dissolved was used. The synthetic substrate was dissolved at a concentration of 10 mM using a buffer and finally diluted to 75 μM. Collagenase was dissolved in 4 degree cold tertiary distilled water to a concentration of 1 unit/ml, and finally diluted with a buffer solution to 0.05 unit/ml. Collagenase inhibitory candidates were tested at final concentrations of 25 and 50 ppm. In this case, the positive control group was set to have a final concentration of 50 ppm of epigallocatechin gallate (EGCG), which is known to have a collagenase inhibitory effect. The reaction was carried out in a 96-well plate, and reacted at room temperature for 10 minutes. The absorbance was measured at 345 nm at 1-minute intervals using a spectrophotometer, and the maximum slope of the absorbance versus time was determined, and the enzyme activity was determined. The collagenase inhibition rate was calculated as follows.

Collagenase activity inhibitory effect of the compound of Formula 1 (number of repetitions = 3)

sample Enzyme activity Inhibition rate (%) Compound of Formula 1 (50ppm) -0.3 57.14 Compound of Formula 1 (25ppm) -0.6 14.28 Positive control (EGCG, 50ppm) -0.6 14.28 Control (DMSO, 50 ppm) -0.7 -

As can be seen from the results of Table 1, it can be seen that the compound of Formula 1 has a superior effect on inhibiting collagenase activity than the positive control EGCG. That is, the compound of Formula 1 exhibited an excellent inhibitory effect on collagenase activity. Therefore, it was found that the compound of Formula 1 can be used for skin regeneration and wrinkle improvement.

Example 2: elastase activity inhibitory effect

The inhibitory effect on the activity of Elastase, an enzyme decomposing elastin, was confirmed as follows.

Elastase used elastase derived from human leukocyte cells, and a synthetic substrate MeOSuc-Ala-Ala-Pro-Val-pNA was used as a substrate for elastase. A buffer solution of 100 mM Tris (pH 7.5) was used. Elastase was finally used in 0.2 mU using a buffer solution. In addition, the synthetic substrate of elastase was diluted with a buffer solution to a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, the positive control was set to put quercetin (Quercetin) known as an elastase inhibitor at a concentration of 10ppm. Elastase inhibitory candidates were added so that the final concentration was 10 or 20 ppm. The reaction was carried out in a 96-well plate, and reacted at room temperature for 20 minutes. The absorbance was measured at 405 nm at intervals of 1 minute using a spectrophotometer, and the slope of the absorbance versus time was obtained and determined as the enzyme activity. Elastase inhibition was calculated as follows.

Elastase activity inhibitory effect of the compound of Formula 1 (number of repetitions = 3)

sample Enzyme activity Inhibition rate (%) Compound of Formula 1 (20ppm) 2.4 76.92 Compound of Formula 1 (10 ppm) 4.2 59.61 Positive control (Quercetin, 10ppm) 3.5 66.35 Control (DMSO, 20 ppm) 10.4 -

As can be seen from the results of Table 2, when the compound of Formula 1 was treated, the effect was less than that of the positive control, but it was found that the elastase activity inhibition was excellent. That is, the compound of Formula 1 exhibited an excellent elastase activity inhibitory effect. Therefore, it was found that the compound of Formula 1 can be used for skin regeneration and wrinkle improvement.

Example 3: Whitening effect - Melanin total reduction effect

The whitening effect was confirmed by adding the compound of Formula 1 to the culture medium of B-16 mouse melanoma cells and measuring the total amount of melanin at the cellular level (Lotan R., Lotan D. Cancer). Res . 40:3345-3350, 1980). Before the experiment, toxicity was evaluated for melanoma cells in mice, and a non-toxic concentration was selected for whitening evaluation.

The compound of Formula 1 was added to the culture medium to have final concentrations of 1 ppm and 10 ppm, and arbutin as a control was added to the medium to 100 ppm, treated with B-16 melanoma cells, respectively, and cultured for 3 days.

Thereafter, the cells were treated with trypsin, removed from the culture vessel, centrifuged, and then melanin was extracted. To the detached cells, 1 ml of sodium hydroxide solution (1N concentration) was added and boiled for 10 minutes to dissolve melanin. Using a spectrophotometer, absorbance was measured at 400 nm to measure the amount of melanin produced.

The amount of melanin was measured by a method indicated by the absorbance per unit cell number (1×10 ⁶ cells), the total amount of melanin relative to the control was calculated as the inhibition rate (%), and the results are summarized in Table 3.

Effect of reducing the total amount of melanin at the cellular level according to the concentration

sample melanin production
(abs) Inhibition rate (%) Control (DMSO, 10 ppm) 0.33 - Positive control (arbutin, 100ppm) 0.228 31.06 Formula 1 (10ppm) 0.218 33.89 Formula 1 (1ppm) 0.269 18.57

As can be seen from the results in Table 3, the compound of Formula 1 has better melanin production inhibition rate even when a lower concentration is applied to the cultured mouse melanoma cells when compared to arbutin, which is known as a whitening material. , it could be confirmed that the total amount of melanin was smaller.

Therefore, it was found that the compound of Formula 1 can be used for whitening purposes.

Example 4: anti-glycosylation effect

In order to confirm the anti-glycation efficacy, the glycation inhibitory activity was measured using L-arginine and glucose.

First, prepare by dissolving 1M L-arginine and 1M glucose using 1M phosphate buffer (pH 7.4), and dilute the sample to 50 and 100ppm using 1M phosphate buffer. Mix 1M L-arginine and 1M phosphate buffer in a ratio of 1 to 4, then dispense 80 μl of each in a 96-well plate. To this, add 100 μl each of a sample diluted to 50 and 100 ppm and 0.01M aminoguanidin to be used as a positive control. After mixing these samples well, finally add glucose diluted with 1M phosphate buffer so that the final concentration of glucose is 0.1M. The reaction was carried out at 70°C for 4 hours. The degree of glycosylation was measured by measuring the absorbance at 420 nm of the 96-well plate using a spectrophotometer.

The glycation experimental group of the following formula is an experimental group in which glycation was induced by adding 1M L-arginine and 1M glucose. In order to measure the absorbance of the sample itself, only 1M L-arginine and the sample were added and the absorbance was measured at 420 nm. did. The glycation inhibitory activity can be calculated as follows.

Anti-glycosylation effect of compound of formula 1

sample Inhibition rate (%) Control (DMSO, 50 ppm) - Positive control (Aminoguanidine, 55ppm) 54.89 Compound of Formula 1 (50ppm) 52.88 Compound of Formula 1 (25ppm) 25.63

As can be seen from the results of Table 4, the compound of Formula 1 exhibits excellent anti-glycosylation effects, and thus is thought to have a whitening or anti-wrinkle effect.

Claims (9)

A cosmetic composition for skin whitening comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A cosmetic composition for improving skin elasticity or wrinkles comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A cosmetic composition for improving dull skin through inhibition of glycation comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

delete The cosmetic composition according to any one of claims 1 to 3, wherein the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is included in an amount of 0.00001 to 10% by weight based on the total weight of the cosmetic composition. A health food composition for skin whitening comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A health food composition for improving skin elasticity or wrinkles comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A health food composition for anti-glycosylation comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

A pharmaceutical composition for wrinkle improvement comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]