KR102040157B1 - Composition for skin cell regeneration, anti-wrinkle, antioxidant and skin whitening comprising linderalactone - Google Patents

Abstract

The present invention provides a composition for skin regeneration, anti-wrinkle, antioxidant and whitening, particularly a cosmetic, pharmaceutical or health food composition comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof. The cosmetic, pharmaceutical or health food composition of the present invention has fewer side effects and has excellent skin regeneration, wrinkle improvement, antioxidant and whitening effects while being safe for the human body.
[Formula 1]

Description

Composition for skin cell regeneration, anti-wrinkle, antioxidant and skin whitening comprising linderalactone

The present invention relates to a composition related to skin regeneration, wrinkle improvement, antioxidant and skin whitening, in particular, a composition for cosmetics, pharmacy, and health foods. It relates to a composition for health food.

Collagen is a major matrix protein produced by the fibroblasts of the skin and is present in the extracellular matrix.The main functions are the skin's mechanical firmness, the resistance of connective tissue and the adhesion of tissues, the support of cell adhesion, cell division and differentiation. Induction of growth or wound healing) is known. This collagen is reduced by age and light aging by ultraviolet irradiation, which is known to be closely related to the wrinkle formation of the skin. In addition, in recent years, extensive research on aging of the skin has been developed an important function of collagen in the skin.

Active ingredients are known that promote collagen synthesis and exhibit wrinkle improvement. For example, retinoic acid, transforming growth factor (TGF), protein from animal placenta, betulinic acid, chlorella extract and the like are known as collagen synthesis promoting substances. However, the active ingredients are limited in the amount of use due to safety problems such as irritation and redness when applying the skin, or there is a problem in that the effect of improving the skin function is not expected by substantially promoting collagen synthesis of the skin.

On the other hand, free radicals introduced from outside the living body or generated in the living body cause many problems such as promoting aging of the living body or generating cancer. Therefore, many researches and developments on antioxidants that inhibit oxidation by active oxygen have been made. Antioxidants are widely distributed in copper and plant systems, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known in fruits and vegetables. However, the antioxidants present in nature are not expected to have a substantial enough effect on skin application. Therefore, many synthetic antioxidants having excellent antioxidant power and low cost are used, but their use is limited due to safety concerns such as human side effects.

Also, to have white and fair skin is everyone's constant desire. It is genetically determined by the concentration and distribution of melanin in human skin, but is also influenced by environmental or physiological conditions such as sun ultraviolet rays, fatigue and stress. Melanin is a type of amino acid tyrosine (tyrosine) acts as an enzyme called tyrosinase (tyrosinase) is converted into dopa (DOPA), dopaquinone (dopaquinone) is produced through a non-enzymatic oxidation reaction. The pathway by which melanin is produced is known, but the mechanisms that induce melanin synthesis, the previous stages of tyrosinase action, are still unknown.

On the other hand, as a known whitening component, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, hydroquinone, vitamin-C (L-Ascorbic acid) and derivatives thereof and various There is a plant extract. By inhibiting the synthesis of melanin pigments, they not only brighten the skin tone to realize skin whitening, but also improve skin hyperpigmentation such as blemishes and freckles due to ultraviolet rays, hormones or heredity. However, when the skin is applied, there are limitations on the amount of use due to safety problems such as irritation and redness, or there is a problem in that a substantial effect cannot be expected due to a slight effect.

Therefore, there is an urgent need for the development of ingredients that are safe for the living body, stable in active ingredients and, above all, have superior skin regeneration, anti-wrinkle, anti-oxidation and whitening activities, compared to conventional skin regeneration, anti-wrinkle, antioxidant and whitening agents. It is requested.

Therefore, the present invention solves the above problems, and provides a new active ingredient, that is, the active use of the active ingredient, which has little side effects and is safe for the human body, and has excellent skin regeneration, wrinkle improvement, antioxidant and whitening effects.

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

In order to solve the above problems, the present invention is a composition for skin regeneration and anti-wrinkle, antioxidant or skin whitening, preferably a cosmetic composition comprising a Linder Lactone (linderalactone) or a pharmaceutically acceptable salt thereof as an active ingredient It provides a pharmaceutical composition, or a composition for health food.

That is, the present invention provides a new use for skin regeneration and wrinkle improvement, antioxidant or skin whitening of linderalactone (linderalactone) or a pharmaceutically acceptable salt thereof.

The inventors of the present invention promote the regeneration of the skin and improve wrinkles by linderalactone promotes collagen synthesis of fibroblasts of the skin and inhibits collagenase activity, and it has an antioxidant effect by eliminating free radicals and melanin The present invention was completed by confirming that the synthesis was suppressed to exhibit a whitening effect.

In the present invention, the 'skin regeneration effect' refers to the recovery of skin tissue against damage caused by external and internal causes. The damage caused by the external causes may include ultraviolet rays, external pollutants, wounds, traumas, and the like, and the damage caused by the internal causes may include stress.

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

In the present invention, the 'antioxidative effect' refers to a cell caused by free radicals or reactive oxygen species (ROS) that are highly reactive due to oxidative stress caused by intracellular metabolism or ultraviolet light. Refers to the inhibition of oxidation, including the removal of free radicals or reactive oxygen species, thereby reducing damage to cells.

In the present invention, the term "whitening effect" means not only brightening skin tone by inhibiting the synthesis of melanin pigments, but also improving skin hyperpigmentation such as blemishes and freckles due to ultraviolet rays, hormones or heredity.

Skin regeneration, wrinkle improvement, anti-oxidation and whitening ingredients have excellent skin regeneration, wrinkle improvement, anti-oxidation and whitening activity at low concentration, and have excellent ability to penetrate and absorb skin. Low volatility, long active to maintain skin regeneration, wrinkle improvement, antioxidant and whitening effect, keep active ingredient stable on composition or skin, easy to formulate into medicine or cosmetic, and also safe for skin It is preferable. However, among the known components, components that satisfy all of the above properties are rare. For example, some skin regeneration, anti-wrinkle, antioxidant and whitening ingredients have excellent skin regeneration, anti-wrinkle, antioxidant and whitening activity even at low concentrations in vitro. It is difficult. Other active ingredients are less hydrophilic and difficult to formulate in medicine or cosmetics. In addition, some skin regeneration, anti-wrinkle, antioxidant and whitening ingredients may decompose or transform into other compounds when exposed to heat, light, or oxygen, and lose their effects before application to the skin.

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

In addition, the present invention can provide a composition comprising a glycoside or non-glycoside of the compound represented by the following formula (1).

[Formula 1]

The compound represented by Chemical Formula 1 is called 6H-4,7-Methenofuro (3,2-c) oxacycloundecin-6-one, also known as linderaclactone.

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

"Pharmaceutically acceptable salts" of the present invention may be acid addition salts formed using organic or inorganic acids, which may be, 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, succinate 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, aminooxy Acetic acid, benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid salts and inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid salts. It may preferably be in the form of hydrochloride or acetate, more preferably in the form of hydrochloride.

The above-mentioned acid addition salts may be a) directly mixing the compound of formula 1 and an acid, b) dissolving and mixing one of them in a solvent or a hydrous solvent, or c) solvent or submerging the compound of formula 1 It is prepared by a general salt preparation method which is placed in an acid in a solvent and mixed with them.

In addition to the above, salts that can be additionally salted are Gabar salt, Gabapentin salt, Pregabalin salt, Nicotinate, Adipate salt, Hemimalonate, Cysteine salt, Acetylcysteine salt, Methionine salt, Arginine salt, Lysine salt, Ornithine salt, Aspartate.

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

In addition, when using the compound of Formula 1 of the present invention as a medicine, it may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin regeneration, wrinkle improvement, antioxidant and skin lightening ingredients. The inclusion of additional skin rejuvenation, wrinkle improvement, antioxidant and skin lightening ingredients will further enhance the skin regeneration, wrinkle improvement, antioxidant and skin lightening effects of the compositions of the present invention. When the ingredient is added, skin safety, ease of formulation, and stability of the active ingredients may be considered. In one embodiment of the invention, the composition is a skin regeneration or wrinkle improvement component known in the art, Retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract; Antioxidant ingredients known in the art include tocopherol, selenium, vitamin C and phenolic compounds; As a whitening component known in the art, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, hydroquinone, vitamin-C (L-Ascorbic acid); And one or two or more components selected from the group consisting of derivatives and various plant extracts thereof may be further included. Additional ingredients may be included from 0.0001% to 10% by weight relative to the total composition weight, and the content range may be adjusted according to requirements such as skin safety, ease of formulating the compound of Formula 1, and the like. .

In addition, the pharmaceutical composition for skin regeneration, anti-wrinkle, antioxidant and skin whitening of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain various components such as buffers, sterile water for injection, general saline or phosphate buffered saline, sucrose, histidine, salts, polysorbates and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and may be administered in the form of a general pharmaceutical preparation, for example, in various dosage forms, orally and parenterally, during clinical administration. , Diluents or excipients such as extenders, binders, wetting agents, disintegrants, surfactants and the like.

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 pharmaceutical composition of the present invention, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.

Liquid preparations for oral use include suspensions, solutions, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are commonly used simple diluents.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester 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.

The pharmaceutical composition of the present invention may provide a desirable skin regeneration effect, wrinkle improvement effect, antioxidant effect and whitening effect when an effective amount of the compound of Formula 1 is included. In the present invention, the term 'effective amount' refers to an amount of a compound that may promote regeneration of damaged skin, improve wrinkles, inhibit or alleviate oxidation of cells, or exhibit a whitening 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, how the compound is applied to the skin, the time it stays on the skin, and the like. For example, when the composition is manufactured as a medicine, it may include the compound of Formula 1 in a higher concentration than when manufactured as a cosmetic that is routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, 1 to 1 day, based on the amount of the compound of Formula 1 It may be administered six times.

The pharmaceutical composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

The present invention can also be provided in the formulation of the skin external preparation for skin regeneration, wrinkle improvement, antioxidant and whitening comprising the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as an external preparation for skin, it is further added to fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants. Common in water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for the skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as. The ingredients may also be introduced in amounts 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, and may have a formulation such as an ointment, a patch, a gel, a cream, or a spray.

The present invention can also be provided in the formulation of a cosmetic for skin regeneration, wrinkle improvement, antioxidant and whitening comprising the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as a cosmetic, the cosmetic prepared by containing the compound of Formula 1 as an active ingredient may be prepared in the form of a general emulsion formulation and solubilized formulation. For example, lotions such as flexible lotions or nourishing lotions, emulsions such as facial lotions, body lotions, creams such as nourishing creams, moisture creams, eye creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, liquids Formulations such as foundations, powders, cleansing creams, cleansing lotions, makeup removers such as cleansing oils, cleansing foams, soaps, body washes and the like.

In addition, the cosmetics are in addition to the compound of Formula 1, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water Commonly used in ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics It may contain adjuvants conventionally used in the cosmetic field, such as any other ingredient.

The compound of Formula 1 may be a relatively high concentration of the compound of Formula 1 in the case of a wash-off type cosmetic such as a make-up remover, a detergent, etc., in which the active ingredient stays on the skin within a short time when it is commercialized. It may include. On the other hand, in the case of a leave-on type cosmetic such as a lotion, an emulsion, a cream, an essence, etc., in which the active ingredient stays on the skin for a long time, the compound of Chemical Formula 1 has a lower concentration than the wash-off type cosmetic. It may be included. Although not limited thereto, in one embodiment of the present invention, the composition may include the compound of Formula 1 in an amount of 0.0001% to 10% by weight (preferably 0.0001% to 1% by weight) based on the total weight of the composition. Can be. When the composition of the present invention contains less than 0.0001% by weight of the compound of Formula 1, sufficient skin regeneration, anti-wrinkle, antioxidant and whitening effects cannot be expected. This is to prevent the reaction may occur or there may be a problem with the skin safety.

The present invention also relates to a health food for skin regeneration, wrinkle improvement, antioxidant and whitening comprising the compound of Formula 1.

In the present specification, 'health food' is a food prepared by adding the compound of Formula 1 to food materials such as beverages, teas, spices, gums, confectionery, or the like, encapsulated, powdered, suspensions, and the like when ingested It means to bring a certain effect, but unlike the general medicine has the advantage that there are no side effects that can occur when taking a long-term use of the drug as a raw material.

Since the health food of the present invention thus obtained can be consumed on a daily basis, high skin regeneration, wrinkle improvement, antioxidant and whitening effects can be expected and are very useful.

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 with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .

There is no particular limitation on the kind of food. Examples of the food to which the substance may be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of said 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, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. In addition, the health food of the present invention may contain a flesh for preparing natural fruit juice, fruit juice drink and vegetable drink. These components can be used independently or in combination. The proportion of such additives is not critical 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.

The composition containing linderalactone or a pharmaceutically acceptable salt thereof of the present invention exhibits skin regeneration and wrinkle improvement.

The composition containing linderolactone of the present invention or a pharmaceutically acceptable salt thereof has an antioxidant effect or a skin lightening effect.

The composition containing linderlalactone of the present invention or a pharmaceutically acceptable salt thereof may be used as a safe and excellent cosmetic raw material, pharmaceutical ingredient, health functional food raw material.

Hereinafter, the following examples and the like will be described in order to describe the present invention in more detail. However, embodiments according to the present invention can be modified in many different forms and the scope of the present invention should not be construed as being limited to the embodiments described below. Embodiments of the present invention are provided by way of example in order to facilitate a specific understanding of the present invention.

Reference Example  One: Linderalactone substance  Information

[Formula 1]

linderalactone

CAS No .: 728-61-0

Where to buy: Tauto Biotech Co., Ltd.

Example  1: collagen synthesis effect

The substance of Formula 1 was added to the culture solution of human-derived fibroblasts to test the collagen synthesis promoting effect at the cellular level.

The synthesis of collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT). Evaluation of cytotoxicity at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm in human-derived fibroblasts before the experiment (method of culturing fibroblasts and performing MTT test [Reference: Mossman T. (1983) Rapid Colorimetric Assay for Cellular Growth & Survival: application to proliferation & cytotoxicity assays.Journal of Immunological Methods 65, 55-63].

In the experiment, the final concentration of the substance of Formula 1 was set to 1 ppm and 10 ppm, and each sample was added to the culture medium of human fibroblasts and incubated for 2 days, and then the culture medium was taken and collagen synthesis was performed at each concentration with the PICP EIA kit. The extent was measured at 450 nm using a spectrophotometer. For comparison of the effects, the degree of collagen synthesis was measured by the same method for the culture medium (control) of the fibroblasts to which nothing was added and the sample to which the vitamin C was added at a final concentration of 52.8 μg / ml. Collagen production was measured by UV absorbance, the collagen production increase rate was calculated as the ratio of collagen production relative to the control group, the results are summarized in Table 1 below.

Collagen Biosynthesis Effect of Substance of Formula 1 (Repeat Number = 3) sample Absorbance average Collagen Growth Rate (%) 1 ppm of substance of Formula 1 521 129.3 Substance of Formula 1
10 ppm 671 166.5 Control
(No addition) 403 - Vitamin c
52.8 μg / ml 519 128.8

As can be seen from the results of Table 1, the material of Formula 1 exhibited better collagen synthesis effect at a lower concentration than when vitamin C, which is generally known to have collagen synthesis ability, was applied.

Example  2: Collagenase  Active inhibitory effect

The inhibitory effect of collagenase activity on the substance of Chemical Formula 1 was confirmed as follows.

Evaluation of cytotoxicity at concentrations of 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, and 0.001 ppm in human-derived fibroblasts before the experiment (method of culturing fibroblasts for MTT test [Reference: Mossman T. (1983) Rapid Colorimetric Assay for Cellular Growth & Survival: application to proliferation & cytotoxicity assays.Journal of Immunological Methods 65, 55-63].

Fibroblasts, which are human normal skin cells, were seeded in a 24-well microplate at 2.5 × 10 ⁴ cells per well, incubated for 24 hours at 10% serum DMEM medium and 37 ° C., and then 10% serum DMEM medium was removed. After washing once with phosphate buffer solution, the mixture was further cultured in serum-free DMEM medium containing the substance of Formula 1 and serum-free DMEM medium without the substance of Formula 1 as a control for 30 minutes.

Thirty minutes after stimulation with TNF-α (tumor necrosis factor-α) 50ng / ml, a substance known to produce MMP-1 after 30 minutes of sample treatment. At this time, the TNF-α untreated group and the treated group were treated as the untreated group not treated with TNF-α and the treated group treated with TNF-α among the controls not containing the substance of Formula 1.

The supernatant of each well was collected to measure the amount (ng / ml) of newly synthesized MMP-1 using an MMP-1 assay kit (Amersham, USA), and the collagenase activity inhibition rate was determined according to Equation 1 below. -1 generation inhibition rate (%) was calculated, the results are shown in Table 2 below.

The amount of MMP-1 in the control group refers to the amount of MMP-1 in the TNF-α treatment group, and the amount of MMP-1 in the experimental group refers to the group to which the substance was added at each concentration.

[Equation 1]

Inhibitory Effect of the Chemical Formula 1 on Collagenase Activity (Repeat Number = 4) sample Amount of MMP-1 (ng / mL) % Inhibition 10 ppm of a substance of Formula 1 8.15 49.72% 1 ppm of substance of Formula 1 13.43 17.15% Negative Control (TNF-α No Treatment) 7.97 Positive control group (TNF-α treatment) 16.21

As can be seen from the results of Table 2, the substance of Formula 1 showed an effect of inhibiting the collagenase activity increased by TNF-α even at a low concentration of 10ppm.

Example 3: whitening effect - confirm melanin production inhibitory effect

The compound of formula 1 was added to the culture solution of mouse melanoma cells (B-16 mouse melanoma cells) to test the whitening effect at the cellular level. (Lotan R., Lotan D. Cancer Res. 40: 3345-3350, 1980).

Before the experiment, rat melanoma cells were assessed for toxicity, and non-toxic concentrations were selected for whitening evaluation.

The compound of formula 1 was tested in the culture medium so that the final concentration was 1 μg / ml, 2.5 μg / ml, 5 μg / ml, 10 μg / ml, 20 μg / ml, and the control arbutin was 200 μg / ml. Each medium was treated with B-16 melanoma cells and cultured for 3 days.

Then, cells were trypsin-treated (trypsin) was removed from the culture vessel and centrifuged to extract melanin. The detached cells were added with 1 ml of sodium hydroxide solution (1N), boiled for 10 minutes to dissolve the melanin, and the absorbance was measured at 400 nm using a spectrophotometer to measure the amount of melanin produced.

The melanin amount was measured by the absorbance of the unit cell count (10 ⁶ cells). The relative melanin production relative to the control group was calculated as the inhibition rate (%) and the results are summarized in Table 3. In addition, the experiment was repeated three times.

Inhibitory effect of melanin production at the cellular level sample Melanin production amount (Abs.) % Inhibition Control group (no addition) 0.681 - Control 1: Arbutin (200 μg / ml) 0.539 20.85 Compound of Formula 1 (20 µg / ml) 0.432 36.56 Compound of Formula 1 (10 μg / ml) 0.329 51.69 Compound of Formula 1 (5 μg / ml) 0.557 18.21

As can be seen from the results in Table 3, the compound of Formula 1 has a superior melanin production inhibitory ability against the cultured mouse melanoma cells compared with the known whitening material arbutin.

Example  4: antioxidant effect Free radical  Erasure rate

Free radical scavenging activity was measured to confirm the antioxidant activity of the substance of formula (1). Free radical scavenging activity was measured using DPPH. DPPH was purchased from Sigma Co., Ltd, USA. First, a 1.5 mM (0.06 mg / ml) standard DPPH ethanol solution was made. Ethanol was added to the ascorbic acid, an antioxidant, as a compound of Formula 1 and a reference material, respectively, to prepare samples at concentrations of 50 µg / ml, 25 µg / ml, 12.5 µg / ml, 6.25 µg / ml, and 3.125 µg / ml. Then, the sample and the standard DPPH solution were added in the same ratio and stirred well, followed by reaction at 37 ° C. for 30 minutes, and absorbance at 520 nm. At this time, ethanol was added instead of the sample as a control. The free radical scavenging ability was obtained from IC ₅₀ , which is a half maximal inhibitory concentration, and the results are shown in Table 4 below. IC ₅₀ is a general method of expressing free radical scavenging ability as a concentration of ascorbic acid and a compound of formula (1) required to remove 50% of free radicals from no additive control.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (µg / ml) Ascorbic acid 4.98 Compound of Formula 1 20.97

As can be seen from the results in Table 4, the compound of Formula 1 showed an excellent antioxidant effect of scavenging free radicals at a concentration of μg / ml.

Formulation example  1: Preparation of Pharmaceutical Formulations

1. Preparation of powder

Compound of Formula 1 0.001g Lactose 1 g

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

Compound of Formula 1 0.2mg Corn starch 100mg Lactose 100mg Stearic acid  magnesium 2mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

Compound of Formula 1 0.2mg Corn starch 100mg Lactose 100mg Stearic acid  magnesium 2mg

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

4. Manufacture of rings

Compound of Formula 1 0.003 g Lactose 1.5 g glycerin 1 g Xylitol 0.5 g

After mixing the above components, it was prepared to be 4g per ring in a conventional manner.

5. Preparation of Granules

Compound of Formula 1 2 mg Soybean Extract 50 mg glucose 200 mg Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60 ° C. to form granules, and then filled in fabric.

Formulation example  2: manufacture of cosmetics

1. Preparation of Soft Cosmetics (Skin Lotion)

Like the following composition, a flexible longevity containing the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 1.0 wt% Butylene glycol 2.0 wt% Propylene glycol 2.0 wt% Carboxy Vinyl Polymer 0.1 wt% Fiji-12 nonylphenyl ether 0.2 wt% Polysorbate 80 0.4 wt% ethanol 10.0 wt% Triethanolamine 0.1 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

2. Preparation of Nutrients (Milk Lotion)

As shown in the following composition, nutrient longevity containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 1.0 wt% Beeswax 4.0 wt% Polysorbate 60 1.5 wt% Sorbitan sesquioleate 1.5 wt% Liquid paraffin 0.5 wt% Caprylic / Capric Triglycerides 5.0 wt% glycerin 3.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Carboxy Vinyl Polymer 0.1 wt% Triethanolamine 0.2 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

3. Preparation of nutrition cream

As in the following composition, a nourishing cream containing the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.2 wt% Beta-1,3-glucan 5.0 wt% Beeswax 10.0 wt% Polysorbate 60 1.5 wt% Sebum 60 Cured Castor Oil 2.0 wt% Sorbitan sesquioleate 0.5 wt% Liquid paraffin 10.0 wt% Squalane 5.0 wt% Caprylic / Capric Triglycerides 5.0 wt% glycerin 5.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Triethanolamine 0.2 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

4. Preparation of Massage Cream

As in the following composition, a massage cream containing a compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 3.0 wt% Beeswax 10.0 wt% Polysorbate 60 1.5 wt% Sebum 60 Cured Castor Oil 2.0 wt% Sorbitan sesquioleate 0.8 wt% Liquid paraffin 40.0 wt% Squalane 5.0 wt% Caprylic / Capric Triglycerides 4.0 wt% glycerin 5.0 wt% Butylene glycol 3.0 wt% Propylene glycol 3.0 wt% Triethanolamine 0.2 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

5. Manufacture of pack

As in the following composition, a pack containing the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.2 wt% Beta-1,3-glucan 1.0 wt% Polyvinyl alcohol 13.0 wt% Sodium Carboxymethyl Cellulose 0.2 wt% glycerin 5.0 wt% Allantoin 0.1 wt% ethanol 6.0 wt% Fiji-12 nonylphenyl ether 0.3 wt% Polysorbate 60 0.3 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

Formulation example  3: External skin preparations  Produce

1. Preparation of Gel

As shown in the following composition, a gel comprising the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.1 wt% Beta-1,3-glucan 0.1 wt% Ethylenediamine Sodium Acetate 0.05 wt% glycerin 5.0 wt% Sebum 60 Cured Castor Oil 0.5 wt% Carboxy Vinyl Polymer 0.3 wt% ethanol 5.0 wt% Triethanolamine 0.3 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

2. Manufacture of ointments

As shown in the following composition, an ointment containing the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5 wt% Beta-1,3-glucan 10.0 wt% Beeswax 10.0 wt% Polysorbate 60 5.0 wt% Sebum 60 Cured Castor Oil 2.0 wt% Sorbitan sesquioleate 0.5 wt% vaseline 5.0 wt% Liquid paraffin 10.0 wt% Squalane 5.0 wt% Shea Butter 3.0 wt% Caprylic / Capric Triglycerides 5.0 wt% glycerin 10.0 wt% Propylene glycol 10.2 wt% Triethanolamine 0.2 wt% antiseptic 0.05 wt% Pigment 0.05 wt% Spices 0.05 wt% Purified water to 100 wt%

3. Preparation of Topical Administration Agent (Gel Ointment)

As the following composition, a gel ointment comprising the compound of formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5 wt% Beta-1,3-glucan 10.0 wt% Polyacrylic Acid (Carbopol 940) 1.5 wt% Isopropanol 5.0 wt% Hexylene glycol 25.0 wt% Triethanolamine 1.7 wt% Deionized water to 100 wt%

4. Preparation of medicament (patch) for topical administration

As in the following composition, a patch containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

Compound of Formula 1 0.5 wt% Beta-1,3-glucan 3.0 wt% Hexylene glycol 20.0 wt% Diethylamine 0.7 wt% Polyacrylic Acid (Carbopol 934P) 1.0 wt% Sodium sulfite 0.1 wt% Polyoxyethylene lauryl ether (E.O = 9) 1.0 wt% Polyhydroxyethylene cetyl stearyl ether (Cetomacrogol 1000) 1.0 wt% Viscous Paraffin Oil 2.5 wt% Caprylic Acid Ester / Capric Acid Ester (Cetiol LC) 2.5 wt% Polyethylene Glycol 400 3.0 wt% Deionized water to 100 wt%

Formulation example  4: manufacture of food

Food containing the compound of formula 1 of the present invention was prepared as follows.

1. Preparation of flour food

0.05 to 1.0 parts by weight of the compound of Formula 1 was added to the flour, and using the mixture to prepare bread, cake, cookies, crackers and noodles to prepare a health food.

2. Manufacturing of Dairy Products

0.2 parts by weight of the compound of Formula 1 was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

3. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh. Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh. The compound of Chemical Formula 1 was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The dry powder of the grains, seeds and the compound of Formula 1 prepared in the above was prepared by blending in the following ratio with respect to 100 parts by weight of the mixed powder.

Cereals (30 parts by weight brown rice, 15 parts by weight brittle, 20 parts by weight of barley),
Seeds (7 parts by weight perilla, 8 parts by weight black beans, 7 parts by weight black sesame seeds),
Compound of formula 1 (0.1 parts by weight),
Ganoderma lucidum (0.5 parts by weight),
Foxglove (0.5 part by weight)

Formulation example  5: manufacture of beverages

1. Preparation of health drinks

0.1 mg of compound of Formula 1

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

After mixing the above components in accordance with the conventional healthy beverage production method and purified water was added to prepare a total 900ml, and after stirring and heating at 85 ℃ for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2L container sealed sealing sterilization After refrigerated storage it was used to prepare a healthy beverage composition of the present invention.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

2. Preparation of Vegetable Juice

1 g of the compound of Chemical Formula 1 of the present invention was added to 1,000 mL of tomato or carrot juice to prepare vegetable juice for health promotion.

3. Preparation of Fruit Juice

1 g of the compound of Formula 1 was added to 1,000 mL of apple or grape juice to prepare fruit juice for health promotion.

Claims (7)

delete To the cosmetic composition for antioxidant comprising a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

delete delete The composition of claim 2, wherein the compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof is included in an amount of 0.0001 to 10 wt% based on the total weight of the composition. The health functional food composition for antioxidant containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
[Formula 1]

The health functional food composition according to claim 6, wherein the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof is included in an amount of 0.0001 to 10% by weight based on the total weight of the composition.