KR101096206B1 - Compositions for Improving Skin Conditions Comprising Pterocladiella capillacea - Google Patents

Abstract

The present invention relates to a composition comprising a beet extract ( Pterocladiella capillacea ) as an active ingredient. An effective ingredient used in the composition of the present invention, beet extract has an excellent skin conditions (skin conditions) effect, the skin condition improvement includes wrinkle improvement, skin whitening improvement, hair growth promotion or hair loss prevention effect. Beetroot extract has an excellent skin whitening effect that inhibits melanin production by inhibiting the activity of intracellular tyrosinase, and improves wrinkles and enhances hair growth through molecular mechanisms such as inhibiting collagenase activity and promoting collagen synthesis. It has a promoting or preventing hair loss effect. In addition, beet extract has the effect of inhibiting the immune-related response associated with excellent inflammation and the effect of inhibiting obesity. And there is no cytotoxicity and skin side effects can be safely applied to cosmetics, pharmaceutical and food compositions.

Beetroot extract, skin whitening, wrinkle improvement, hair growth, anti-inflammatory, anti-obesity

Description

Composition for improving skin condition comprising cow radish extract as an active ingredient {Compositions for Improving Skin Conditions Comprising Pterocladiella capillacea}

The present invention relates to a composition for skin, and more particularly, excellent product stability and can be safely used without side effects on the skin, wrinkle improvement, skin whitening effect, anti-inflammatory effect, anti-obesity effect, hair loss prevention and hair growth effect It relates to a composition for skin comprising the extract of cow radish as an active ingredient.

Wrinkles are the result of aging of the skin, and aging skin is the aggregate of the natural changes associated with the process of aging. Skin aging is divided into two categories, physiological aging, which indicates a change in skin function, structure, or shape related to age, and photoaging by ultraviolet rays.

As the skin ages, the changes in the dermis appear remarkably, and dermis atrophy after 70 years is a representative aging phenomenon. Changes in the dermis are caused by changes in substances with large molecular weight in the extracellular matrix due to a decrease in the number of fibroblasts and their ability to synthesize. Specific changes include separation of collagen bundles, reduced point polysaccharide synthesis, decreased collagen and elastic fiber counts and diameters, grinding collagen and elastic fibers, and expansion of blood vessels. Generally, among the various factors such as skin moisture content, collagen content, and ability to immunize the external environment, the biggest influence on wrinkle formation is collagen, a collagen degrading enzyme that reduces collagen production and collagen content. It is the expression level and activity of genease. Although no effect on the collagen and collagenase enzyme of the radish extract was revealed, the results of this experiment showed that the radish extract increased collagen synthesis and inhibited the collagenase enzyme.

 Human skin color is determined by the concentration and distribution of melanin inside the skin. Melanin pigment produced by melanocytes of human skin is a phenolic polymer having a complex form of black pigment and protein, and plays an important role in preventing skin damage caused by ultraviolet rays. In melanin biosynthesis, the action of tyrosinase in melanocytes is reported to be the most important. Tyrosinase is a type of amino acid tyrosine (DOPA) and dopaquinone (dopaquinone) plays a key role in the skin blackening process.

Hormonal imbalances are aggravating due to environmental pollution and automobile exhaust. As a result, the incidence of hair loss increases, the age of incidence decreases, and the immunity of the skin immune system is caused, resulting in various skin diseases such as atopy and psoriasis. In addition, the population suffering from obesity is rapidly increasing despite the age of children due to the changes in the diet of meat and meat.

Therefore, the development of materials that can effectively solve various phenomena that are not only aesthetic but also serious social problems, such as endogenous aging and wrinkles caused by UV rays, blemishes or freckles, obesity, immune imbalance and hair loss, etc. Is being studied. Under these backgrounds, the present inventors searched for active substances in plant-derived compounds in order to find cosmetics having high safety and stability and excellent skin-related efficacy, and the extract of beetroot has various effects including wrinkles and anti-inflammatory, At the same time, it was confirmed that the safety and stability is excellent.

Throughout this specification many patent documents are referenced and their citations are indicated. The disclosures of the cited patent documents are incorporated by reference herein in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly described.

The present inventors have tried to develop a novel material having excellent wrinkle stability, skin whitening effect, hair loss prevention or hair growth effect, anti-obesity effect and anti-inflammatory effect and high stability without skin side effects.

The present invention was completed by confirming that the composition comprising the extract of cow radish as an active ingredient can provide a composition for improving skin condition, anti-inflammatory and anti-obesity having excellent efficacy and safety.

Accordingly, it is an object of the present invention to provide a composition for improving skin conditions.

       Another object of the present invention to provide a composition for inhibiting inflammation.

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Another object of the present invention to provide an anti-obesity composition.

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

The present invention provides a composition comprising a beet extract (Pterocladiella capillacea) as an active ingredient. The effective ingredient used in the composition of the present invention, beet extract has an excellent skin improvement effect, the skin improvement includes wrinkle improvement, skin whitening improvement, hair growth promotion or hair loss prevention effect. Beetroot extract has excellent anti-wrinkle effect through molecular mechanisms such as inhibition of collagenase activity and promotion of collagen synthesis, and excellent skin whitening effect of inhibiting melanin production by inhibiting the activity of intracellular tyrosinase and It has excellent hair growth promotion or hair loss prevention effect. In addition, beet extract has the effect of inhibiting the immune-related response associated with excellent inflammation and the effect of inhibiting obesity. And there is no cytotoxicity and skin side effects can be safely applied to cosmetics, pharmaceutical and food compositions.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

According to one aspect of the invention, the present invention provides a composition for improving skin conditions (skin conditions) comprising the extract as an active ingredient (Pterocladiella capillacea).

According to another aspect of the present invention, the present invention provides a composition for inhibiting inflammation, which comprises an extract of P. chinensis extract (Pterocladiella capillacea) as an active ingredient.

According to another aspect of the present invention, the present invention provides a composition for anti-obesity comprising the extract as an active ingredient (Pterocladiella capillacea).

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The present inventors have tried to develop a novel single substance for a naturally derived compound having excellent anti-wrinkle effect, skin whitening effect, hair loss prevention or hair growth effect, anti-inflammatory effect and anti-obesity effect and high skin stability without any side effects. . As a result, it was confirmed that the preparation of the composition comprising the extract of cow radish as an active ingredient can provide a composition for improving skin condition, anti-inflammatory and anti-obesity having excellent efficacy and safety.

The ox radish ( Pterocladiella capillacea ) belongs to the genus Locust and is distributed in various parts of Jeju Island, South and West Coast of Korea and inhabits Ulam and Kori on the East Coast. It is similar in shape to a locust, but the body is usually thin, biased, and wide. Eggplants are formed as either fresh or regenerated. The bases of the branches are thin, but the ends are usually blunt or thin. Used as a raw material for agar.

Beetroot ( Pterocladiella capillacea ) extract of the present invention can be obtained by using the following extraction method. Specifically, the above extract of beet may be obtained using various extraction methods known in the art, and preferably, (a) water, (b) anhydrous or hydrous lower alcohol (methanol) having 1 to 4 carbon atoms. , Ethanol, propanol, butanol, etc.), (c) the mixed solvent of the lower alcohol and water, (d) acetone, (e) ethyl acetate, (f) chloroform or (g) 1,3-butylene glycol It is a useful seaweed that can be obtained as a solvent.

On the other hand, it will be apparent to those skilled in the art that the beet extract of the present invention can be obtained extracts that exhibit substantially the same effect using not only the above-described extraction solvent but also other extraction solvents.

In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone a conventional purification process. Obtained by various additional purification methods, such as, for example, separation using ultrafiltration membranes having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). The fraction is also included in the extract of the present invention.

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

According to a preferred embodiment of the present invention, the skin condition improvement refers to a composition, characterized in that wrinkle improvement, skin whitening improvement, hair growth promotion or hair loss prevention.

The composition for improving skin conditions of the present invention has a wrinkle improvement use. The composition of the present invention exhibits excellent anti-wrinkle effect through molecular mechanisms such as inhibition of collagenase activity and promotion of collagen synthesis while having low cytotoxicity, which is clearly described in the following examples. . Wrinkle improvement, which is the use of the composition of the present invention, is understood to include conventional skin protection applications (prevention of wrinkles, removal of wrinkles, inhibition of skin aging, etc.).

In addition, the composition for improving skin conditions of the present invention has a skin whitening use. As used herein, the term "whitening" refers to the action of improving skin trouble by preventing blackening of the skin caused by the deposition of melanin pigment.

In the skin whitening composition of the present invention, the cow radish extract exhibits a whitening effect of inhibiting melanin production by inhibiting the activity of intracellular tyrosinase, has high stability, hardly changes in the content of ingredients over time, and skin irritation. There are few side effects such as induction.

In addition, the composition for improving skin of the present invention works very effectively in promoting hair growth or preventing hair loss. As used herein, the term “anti-hair loss” or “promoting hair growth” is used in the same sense, which has the same meaning as another term wool or hair growth promotion used in the art.

The composition of the present invention has the effect of inhibiting the immune-related response associated with inflammation, including the extract of cow radish as an active ingredient. Specifically, the extract of cow radish, an active ingredient, inhibits interleukin-2 expression by reducing the activity of the interleukin-2 (IL-2) promoter, which has been reported to play an important role in the production of pro-inflammatory cytokines. As a result, it has an immunomodulatory function that effectively inhibits T cell-mediated inflammatory responses.

Therefore, the anti-inflammatory composition of the present invention can be applied to various diseases, diseases or abnormal conditions that can be prevented or treated by inhibiting an immune response. Inflammation-related diseases that can be prevented or treated by the compositions of the present invention include, for example, bullous scutaneous swelling, scar stool swelling, disc lupus, lupus erythematosus, psoriasis, systemic lupus erythematosus, lupus erythematosus, allergy And atopy, but is not limited thereto. The anti-inflammatory composition of the present invention is preferably used for the prevention or treatment of atopy.

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In addition, the beet extract as an active ingredient of the composition of the present invention has an excellent anti-obesity effect.

In a preferred embodiment of the present invention, the effective ingredient of the cow radish extract is 0.00001 to 15.0% by weight, more preferably 0.0001 to 10% by weight, most preferably 0.0001 to 5% by weight based on the total composition. When the weight of the active ingredient of the extract of the cow radish is less than 0.00001% by weight, the skin-related efficacy is weak, and when it exceeds 15.0% by weight, the increase in the effect of the increase of the content is very weak, and the stability of the formulation is not secured. There is a problem.

According to a preferred embodiment of the present invention, the composition of the present invention is a cosmetic composition.

The components included in the cosmetic composition of the present invention include components commonly used in cosmetic compositions in addition to the extract of beet as an active ingredient, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings, And a carrier.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a nourishing cream, astringent lotion, a flexible lotion, a lotion, an essence, a nourishing gel or a massage cream.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

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

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The composition of the present invention may be prepared as a pharmaceutical composition.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical compositions of the present invention may be administered orally or parenterally, preferably by parenteral administration, more preferably by topical application by application.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg / kg on an adult basis. In addition, in the case of external preparations, it is recommended to continue the application for 1 month to 5 times a day in an amount of 1.0 to 3.0 ml on an adult basis. However, the dosage does not limit the scope of the present invention.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

In addition, the composition of the present invention may be prepared as a food composition.

When the composition of the present invention is prepared as a food composition, as an active ingredient, as well as the extract of cow radish, it contains components that are commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors Includes the first. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

 For example, when the food composition of the present invention is prepared with a drink, the citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. Can be.

On the other hand, in a specific example of the present invention, the cumulative skin irritation test results for the extract of the cow radish was found to be a natural substance harmless to the human body. Therefore, the beet extract of the present invention has little toxicity and side effects, so it can be used safely even in long-term use, in particular can be safely applied to the cosmetics, pharmaceutical and food compositions as described above.

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

(Iii) The composition of the present invention contains the extract of the extract as an active ingredient.

(Ii) Oxygen extract, which is an active ingredient, has an excellent skin improvement effect, and the skin improvement includes wrinkle improvement, skin whitening improvement, hair growth promotion or hair loss prevention effect. Beetroot extract has an excellent skin whitening effect that inhibits melanin production by inhibiting the activity of intracellular tyrosinase, and improves wrinkles through molecular mechanisms such as inhibiting collagenase activity and promoting collagen synthesis. It has the effect of promoting hair growth or preventing hair loss.

(Iii) Oxygen extract, an active ingredient, has the effect of inhibiting the immune-related response associated with excellent inflammation and the effect of inhibiting obesity.

(Iii) In addition, there is no cytotoxicity and skin side effects, so that it can be safely applied to cosmetics, pharmaceutical and food compositions.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention to those skilled in the art. Will be self explanatory.

Example  1: Preparation of Extract

The entire dry bark was refluxed three times for 5 hours with a 95% ethanol aqueous solution and then cooled, and then filtered through Whatman filter paper. The filtered concentrated solution was concentrated under reduced pressure and lyophilized at 50 ° C or lower. It provides a cosmetic and pharmaceutical composition, characterized in that the reduced pressure concentrated and lyophilized is 0.0001 ~ 10% by weight. In addition, according to the method of Japanese Patent Laid-Open No. 2005-60232, wasot extract was prepared.

Example  2: Ox radish  Wrinkle improvement effect of extract

The anti-wrinkle effect can usually be measured as collagen biosynthesis and collagenase degradation inhibitory effect and clinical trial in humans. Human normal fibroblasts (Pacific) were inoculated into 6-well microplates containing DMEM medium (2 × 10 ⁵ cells / well) and stored at 37 ° C. in a 5% CO ₂ incubator. Incubated for 24 hours. After 24 hours, the medium was removed from each well, samples were treated by concentration and then incubated again for 24 hours. After 24 hours, cell media were collected to prepare samples.

The amount of collagen synthesized was measured using the collagen measurement kit (Procollagen Type I C-peptide EIA kit (MK101), Takara, Kyoto, Japan). Was measured. Detailed test method was performed according to the kit instructions of Takara Co., Ltd.

An antibody against collagenase was used as a method for measuring the activity of collagenase, an enzyme that degrades collagen. Porbolol myristate acetate (PMA) (Sigma) was treated with a substance that induces collagenase activity. The entire test was performed according to the vendor's protocol. Type I collagenase assay kit (Amersham Biosciences, RPN2629) was used and absorbance was measured with an ELISA reader (Bio-Tek ELx808 ™ Series Ultra Microplate Reader, UK). The measured standard values were expressed in the form of mean ± standard deviation, and the significance was tested by t-test using the SPSS / PC + program. The results are shown in Table 1 below.

Test Items Wild Radish Extract
(1 ppm) Lobster Extract
(1 ppm) Wild Radish Extract
(10 ppm) Lobster Extract
(10 ppm) Wild Radish Extract
(50 ppm) Lobster Extract
(50 ppm) Collagen Synthesis Growth Rate 17 4 19 7 25 9 Collagenase inhibition rate 10 2 18 5 24 6

As can be seen in Table 1, it can be seen that the beet extract inhibits collagenase activity while increasing collagen synthesis. In addition, it showed significantly higher potency than the extract of wormwood prepared according to the method of Japanese Patent Laid-Open No. 2005-60232. Eventually, it can be seen that the beet extract has the effect of improving wrinkles and the efficacy is very excellent compared to the extract of the same genus. In addition, this increase in collagen synthesis and inhibition of collagenase activity appeared to be dependent on the concentration of the radish extract.

Example  3: Wrinkle improvement effect measurement

The anti-wrinkle effect of the cosmetics containing the extract of wild radish was measured through clinical trials. The nourishing cream prepared in Preparation Example A (nutrition cream containing 0.01%, 0.05% and 1% of beet extract or Loch fern extract, respectively) and Comparative Preparation Example (nutrition cream containing purified water) was used.

Wrinkle improvement effect was evaluated by measuring skin elasticity change. Skin elasticity was measured in 30 healthy women (25 to 35 years old) under the condition that the temperature is maintained at a temperature of 24 to 26 ° C. and a humidity of 38 to 40%. Each subject's face was used twice a day for 3 months, and then measured using Cutometer SEM 474 (Courage + Khazaka, Cologne, Germany). The relative value was compared by setting 5 as. The test results are shown in Table 2 below.

Test Items Cream containing 0.01% of radish extract Cream of Buttercup Extract 0.01% Cream containing 0.05% of radish extract Cream containing 0.05% of Buttercup Extract Cream containing 1% of radish extract Cream containing 1% of Buttercup Extract Comparative Preparation Example
Cream of Buttercup Extract 0% Skin elasticity 2.68 2.57 2.79 2.61 3.05 2.72 2.55

As can be seen in Table 2, the cream extract containing cow beet of the present invention was much better wrinkle improvement effect compared to the comparative preparation and cream containing the wood starch extract, skin elasticity was also improved as the concentration of the extract of beet extract increased.

Preparation Example A Preparation of Cream Containing Extract of Beet Radish or Beetroot Extract

The composition of the nourishing cream containing the radish or radish extract is shown in Table 3 below. The aqueous phase purified water, triethanol amine and propylene glycol were heated and dissolved at 70 占 폚, and the oily fatty acid, oil component, emulsifier, and preservative were heated to 70 占 폚 to dissolve the solution. After the emulsification was completed the solution was cooled to 45 ° C., the beet extract and flavor were added and dispersed and then cooled to 30 ° C.

Ingredients and Contents of Nutritious Cream Containing Radish Extract ingredient Content (% by weight) Beetroot Extract or Wood Starch Extract 0.01, 0.05 or 1.00 Jojoba oil 5.0 Floating paraffin 7.0 Cetylaryl alcohol 2.0 Polyglyceryl-3 Methyl Glucose Disteare 2.0 Glyceryl stearate 0.5 Squalane 3.0 Propylene glycol 4.0 glycerin 5.0 Triethanolamine 0.3 Carboxy Vinyl Polymer 0.3 Tocopheryl acetate 0.2 Preservative, incense a very small amount Purified water Balance Sum 100

Example  4: Determination of melanin production inhibitory effect

Rat B16 melanoma cells (Korea Cell Line Bank) were used to measure the inhibitory effect of melanin production by cow radish extract and compared with the inhibitory effect of melanin production by arbutin known to inhibit melanin production.

This experiment was performed in 6-well plates using murine melanoma (B-16 F10) cells (Korea Cell Line Bank) in DMEM medium containing 10% fetal bovine serum (FBS). Inoculated at 1 × 10 ⁵ per cell and incubated under 5% CO ₂ and 37 ° C. until cells were at least about 80% attached to the bottom of the well. The medium was then removed and the sample was replaced with medium diluted to the appropriate concentration and incubated for 3 days under 5% CO ₂ and 37 ° C. Concentration ranges of bark radish or radish extract were determined to be 1 ppm, 10 ppm and 100 ppm without cytotoxicity. The cells from which the medium was removed were washed with PBS (phosphate buffered saline) and treated with trypsin to recover the cells. The recovered cells were measured by using a hematocytometer (Tiefe Depth Profondeur 0.100 mm, Paul Marienfeld GmbH & Co. KG, Germany), the cell number was measured, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and then the upper layer. The liquid was removed to obtain pellets. The cell pellet was dried at 60 ° C., and 100 ml of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60 ° C. thermostat. This was measured by absorbance at 490 nm with a microplate reader (Bio-Tek ELx8081U, United States of America) to determine the amount of melanin per cell. The results are shown in Table 4 below.

sample Treatment Concentration (ppm) Melanin production inhibition rate (%) Arbutin 100 29 Wild Radish Extract 10 11 50 21 100 33 Lobster Extract 10 4 50 8 100 13

As shown in Table 4, it can be seen that the beet extract at the treatment concentration has a higher melanin production inhibition rate than the arbutin and wormwood extract. In addition, it can be seen that the melanin production inhibitory activity of the extract of beetroot is concentration-dependent.

Example  5: Ox radish  By extract Intracellular Tyrosinase  Activity Inhibition Effect Measurement

Intracellular tyrosine by arbutin, known to inhibit melanin production, by measuring the inhibitory effect of tyrosinase on the activity of wild boar extract using rat B16 melanoma cells (Korea Cell Line Bank). It was compared with naze activity inhibitory effect.

This experiment was performed in 6-well plates using murine melanoma (B-16 F1) cells (Korea Cell Line Bank) in DMEM medium containing 10% fetal bovine serum (FBS). Inoculated at 1 × 10 ⁵ per cell and incubated at 5% CO ₂ and 37 ° C. until cells were at least about 80% attached to the bottom of the well. The medium is then removed and the sample replaced with medium diluted to the appropriate concentration, followed by 5% CO ₂ and Incubated at 37 ° C. for 3 days. Concentration ranges of the extract of cow radish were determined to be 1 ppm, 10 ppm and 100 ppm without cytotoxicity. The cells from which the medium was removed were washed with PBS (phosphate buffered saline) and treated with trypsin to recover the cells. The recovered cells were measured using a hematocytometer (Tiefe Depth Profondeur 0.100 mm, Paul Marienfeld GmbH & Co. KG, Germany) and the cell number was measured and centrifuged at 5,000 to 10,000 rpm for 10 minutes, and then the supernatant. Was removed to obtain pellets. The cell pellet was crushed using lysis buffer and centrifuged at 12,000 rpm for 10 minutes to collect supernatant. Tyrosinase activity per cell constant was determined by measuring absorbance at 492 nm with a microplate reader (Bio-Tek ELx8081U, United States of America). The results are shown in Table 5 below.

sample Treatment Concentration (ppm) Inhibition rate of intracellular tyrosinase activity (%) Arbutin 100 25 Wild Radish Extract 10 15 50 24 100 37 Lobster Extract 10 3 50 7 100 11

As shown in Table 5, it was found that the extract of bark was higher in intracellular tyrosinase inhibition rate than Arbutin and Locust extract. In addition, it was found that the degree of inhibition of tyrosinase activity of the radish extract was concentration-dependent.

Example  6: Evaluation of the whitening effect at the animal level

Similar to humans, the whitening effect of beet extract was measured using brown guinea pigs (Tortoiseshell guinea pigs), which are known to cause pigmentation by ultraviolet rays.

In order to cause pigmentation due to ultraviolet rays (UV) in the brown guinea pigs, a light shielding aluminum foil having a square window of 3 × 3 cm ² is adhered to the skin from which the hairs of the brown guinea pigs are removed, and then SE lamp ( Ultraviolet rays were irradiated with a wavelength of 290-320 nm (Toshiba) (total amount of irradiation energy = 1350 mJ / cm ² ). After UV irradiation, the aluminum foil was peeled off and a sample (beet extract and arbutin) was applied in the following manner. Pigmentation appeared 2-3 days after UV irradiation and reached a peak after about 2 weeks, from which each sample was applied.

The number of coatings was continued once or twice a day for 50 days. Samples were dissolved and diluted using a specific solvent (Propylene Glycol: Ethanol: Water = 5: 3: 2 mixed solvent), and a control site was prepared using a cotton swab and necessarily applying the solvent to other sites. In this case, the cumulative stimulus was observed along with the effect determination.

The effect was determined by measuring the degree of black and white of the skin using a color difference meter (CR2002 chromameter, Minolta, Japan), the results are shown in Table 6 below. L ^* A ^* B ^* color system is used to display color, and L ^* value was used as an index in this invention. L ^* value was calibrated with a standard whiteboard, and L ^* value was measured 5 times or more at one site, and pigmentation was equalized. The difference (ΔL ^* ) of the skin color at the start of application and at the end of application was determined and the effect was determined using this value. The results are shown in Table 6 below.

△ L ^* = L ^* value after 00 days application - L ^* value of the coating start date

Comparing the ΔL ^* value with respect to the sample application site and the control site, the effect of the whitening material can be seen.

sample Treatment Concentration (%) Whitening effect (△ L) Wild Radish Extract 0.01 0.30 0.1 0.39 Lobster Extract 0.01 0.25 0.1 0.29 Arbutin 1.0 0.33 Control group - 0.24

As shown in Table 6, the beet extract showed a superior whitening effect than arbutin and wormwood extract, the cumulative stimulation was not observed during the test application test results showed excellent safety.

Example 7 Safety Confirmation Experiment of Human Radish Extract on Human Skin

In order to confirm that the radish extract is safe for human skin, skin safety verification experiments were performed. To this end, a cumulative skin irritation test was performed.

Squalene-based formulations were added with 1%, 5%, and 10% of beet extract, respectively, and a total of nine 24-hour cumulative patches were applied to the upper forearm every other day for 30 healthy adults. It was determined whether the beet extract irritates the skin. The patching method used a fin chamber (Finn chamber, Epitest Ltd, Finland). 15 µl of each of the external skin preparations was added dropwise to the chamber, followed by patching. The degree of response to the skin each time was scored using Equation 2 below, and the results are shown in Table 7 below.

Average responsiveness = [[{(response x responsiveness) / (total number of subjects x highest score (4 points))}] x 100] ÷ number of tests (9)

In the reactivity, ± gives 1 point, + gives 2 points, and ++ gives 4 points, which are judged as safe compositions when the average reactivity is less than 3.

Test substance Number of subjects with response Average reactivity 1 week 2 weeks 3 weeks Primary Secondary 3rd 4th 5th 6th 7th 8th 9th ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + ± + + + Control group (squalene) One - - 0 - - --- --- --- --- --- --- --- 0.09 Wild Radish Extract (1%)
[Test Group 1] 0 - - 0 - - --- --- --- --- --- --- --- 0.00 Beetroot Extract (5%) [Test Group 2] 0 - - 0 - - --- --- --- --- --- --- --- 0.00 Wild Radish Extract (10%)
[Test Group 3] 2 - - 0 - - --- --- --- --- --- --- --- 0.18

In Table 7, the number of persons corresponding to ±, +, and ++ in test groups 1, 2, and 3 was 0, 0, and 2, respectively, and the average reactivity was 0.00, 0.00, and 0.18, respectively. Therefore, the average reactivity was less than 3, so the extract of beetroot was determined to be a safe substance for human skin that does not show a distinct cumulative stimulus pattern.

Example 8: Evaluation of anti-inflammatory efficacy

In order to examine the anti-inflammatory effect, COX (cyclooxygenase) activity inhibitory activity was performed according to a conventional method on THP-1 cells (Korea Cell Line Bank). COX activity was measured according to Method in Enzymology 43: 9 (1994) published by FJ Van de Ouderaaa and Muytenhek. The COX assay cultured THP-1 cell line and then dispensed the cells into 24-well plates. Adjust the reaction volume to 500 μl per well, add 2 μl of sample compound dissolved in lipopolysaccharide (LPS) (1 μg / ml) (Sigma) and the solvents shown in Table 5 below, and then 24-48 hours. Incubated under the same conditions. After 24-48 hours, add 1 µl of calcium ionophore (Sigma) and [1-14C] arachidonic acid (in EtOH, 0.1 µCi / ml, Sigma) to each well and incubate for 10 minutes under the same conditions. It was. After incubation, citric acid was added to each well and shaken to adjust the pH to 3.5. 500 μl of each culture solution was aliquoted into a microcentrifuge tube, and 700 μl of ethyl acetate was added thereto, followed by shaking extraction for 10 minutes. 500 µl of ethyl acetate layer was taken and concentrated for 20 minutes using a speed vacuum dryer, 20 µl of the concentrated residue was dissolved in ethyl acetate, and then applied to an TLC plate with an authentic standard. Radioactivity bands were identified by authentic eicosanoid standards, and the radioactivity of the identified bands was measured using a BAS 2000 bio-imaging analyzer (Fuji, Japan).

sample COX activity (%) LPS (1 μg / ml) 100 LPS (1 μg / ml) + Beet Extract (1 ppm) 88 LPS (1 μg / ml) + Beet Extract (10 ppm) 74 LPS (1 μg / ml) + Beet Extract (50 ppm) 60

As shown in Table 8, it can be seen that the beet extract effectively inhibits the activity of COX concentration-dependently. It can be seen that it can exhibit the inhibitory efficacy.

Example 9: Antioxidant Effect

DPPH (1,1-diphenyl-2-picrylhydrazyl) (Sigma) is a colored radical that can directly identify the radical removal ability of a sample by color change. First, 20 μl of the sample dissolved in the solvent was put into a 96 well plate, and 180 μl of 200 μM DPPH solution was added thereto, followed by incubation for 30 minutes at room temperature. The amount of DPPH remaining was determined by absorbance measurement at 540 nm. Antioxidant results of the extract of cow radish are shown in Table 9 below. Briefly, the effect was judged the presence or absence of antioxidant function by observing the free radical removal rate by the sample.

sample Treatment Concentration (ppm) Free radical scavenging rate (%) Wild Radish Extract 10 24 50 77 100 85

As shown in Table 9, it can be seen that the beet extract has a concentration-dependent antioxidant effect.

Example 10 Anti-Obesity Effect

Obesity inhibition test was generally carried out by the following method using a well-known animal. The results are shown in Table 10 below.

The method for measuring obesity inhibitory activity is as follows:

Crj: 7 week old ICR male mice (Higgs River, Japan) were pre-cultured for 1 week and used in experiments with 7 groups of 1 animals. Animals are kept in a constant temperature room set at a temperature of 23 ± 1 ° C, a humidity of 55 ± 5% and an illumination time of 12 hours / day, using Rabo MR (manufactured by Nippon Agricultural Co., Ltd.) for feeding, and freely ingesting water. It was made. Beet extract was prepared in liposome formulation (dissolved in 5% lecithin) to 0.1% and 1%. Each sample solution was adjusted to a concentration of 0.1 ml per 10 g of the mouse to doses of 1.5 g / kg and 1 g / kg. In addition, the control group was a 5% lecithin emulsion. Mice were fasted from the day before administration and were forcibly administered once the next day. The test period was 2 weeks and body weight and general symptoms were measured and observed.

sample Days Weight (g) Untreated group Wild Radish Extract (0.1%) Wild Radish Extract (1%) Wild Radish Extract
Liposomal formulation 5 22 21 21 10 30 27 26 15 39 34 32

As described in Table 10, the extract of bark showed the effect of inhibiting obesity, the higher the concentration was observed that the efficacy increases.

Example 11 Hair Loss Prevention and Hair Growth Effect

In order to find out about the hair loss prevention and hair growth effect of the extract according to the present invention, the following experiment was performed.

1. Preparation of Hair Regrowth Composition

The hair repellent composition containing the extract of the cow radish as a component content of Table 11 to produce a hydrogel base. Test groups 1 and 2 are hair repellent compositions each comprising a beet extract.

ingredient content(%) Test group 1 Test group 2 Wild Radish Extract 0.1 1.0 Preservative (Gramben) 0.8 0.1 Thickener (Xanthan-Gum) 0.3 0.1 Total weight 100 100

2. Hair loss prevention and hair growth effect measurement

From 40's to 60's, 40 patients were divided into 4 groups, including alopecia areata alopecia, typical male alopecia patients, and acute alopecia areata, and 10 patients per group. The hair restorer prepared in 1 was applied to the bald area of the alopecia patient twice a day for 3 months each for 3 months. Hair loss and hair growth were observed on a monthly basis. A commercial moxidyl (Hanmi Pharm) was used as a comparison group, and only 50% ethanol was applied as a control group.

The criteria are as follows:

4 : High effect = newborn hair is visible

3 : moderate effect = visible newborn hair (down)

2 : slightly effective = the number of hair loss is reduced

1 : no effect = no change at all

The results are shown in Table 12.

- Control group Comparison Experiment 1 Experiment 2 Efficacy / effect One 3.5 2.4 3.5

As shown in Table 12, in the alopecia patients using the hair growth agent containing the cow radish extract of the present invention, new hair of bristles including fluffy began to appear from one month or two months of treatment, and at least sixty percent of treatments were found at six months of treatment. The hair growth effect appeared in the patient. In addition, it was observed that new hair grew continuously and no hair loss was found.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation Example 1  Cosmetic preparations

1. Flexible cosmetics (content: wt%)

Wild Beet Extract 0.01

Glycerin 3.0

Butylene Glycol 2.0

Propylene Glycol 2.0

Carboxy Vinyl Polymer 0.1

Ethanol 10.0

Triethanolamine 0.1

Preservatives, trace pigments, trace fragrances and traces

Total 100.0

2. Nutritional Cosmetics (Content: Weight%)

Wild Beet Extract 0.01

Beeswax 4.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.5

Liquid Paraffin 5.0

Squalane 5.0

Caprylic / Capric Triglycerides 5.0

Glycerin 3.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Carboxy Vinyl Polymer 0.1

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances and traces

Total 100.0

3. Nutrition Cream (Content: Weight%)

Foxtail Extract 0.01

Beeswax 10.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.5

Liquid Paraffin 10.0

Squalane 5.0

Caprylic / Capric Triglycerides 5.0

Glycerin 5.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances and traces

Total 100.0

4. Massage cream (content: wt%)

Wild Beet Extract 0.01

Beeswax 10.0

Polysorbate 60 1.5

Sorbitan Sesquioleate 0.8

Liquid Paraffin 40.0

Squalane 5.0

Caprylic / Capric Triglycerides 4.0

Glycerin 5.0

Butylene Glycol 3.0

Propylene Glycol 3.0

Triethanolamine 0.2

Preservatives, trace pigments, trace fragrances and traces

Total 100.0

5. Pack (Content: Weight%)

Wild Beet Extract 0.01

Polyvinyl Alcohol 13.0

Sodium Carboxymethylcellulose 0.2

Allantoin 0.1

Ethanol 5.0

Nonylphenyl Ether 0.3

Preservatives, trace pigments, trace fragrances and traces

Total 100.0

Formulation Example 2  : Pharmaceutical Formulations

1. Preparation of powder

2g of radish extract

1g lactose

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of Tablets

Beet Extract 100 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

3. Preparation of Capsule

Beet Extract 100 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Claims (10)

A composition for improving skin condition selected from wrinkle improvement and skin whitening, comprising: a beet extract ( Pterocladiella capillacea ) as an active ingredient cosmetic composition for improving skin conditions (skin conditions). delete delete delete delete delete A composition for improving skin condition selected from wrinkle improvement and skin whitening, comprising: a fermented radish extract ( Pterocladiella capillacea ) as an active ingredient, a pharmaceutical composition for improving skin conditions. The composition of claim 1 wherein the cosmetic composition consists of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation and a spray A composition characterized by having a formulation selected from the group. delete delete