KR102689774B1 - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. The mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekjaen extract according to the present invention exhibits a whitening effect by significantly reducing the total amount of melanin, promotes collagen synthesis in skin fibroblasts, and inhibits elastase activity. It improves wrinkles and improves elasticity, suppresses the production of nitric oxide (NO), has an anti-inflammatory effect, has an antioxidant effect by scavenging free radicals, and has an anti-glycation effect that suppresses glycation, thereby whitening skin, improving wrinkles, and improving elasticity. , can be used in the manufacture of medicines, cosmetics or foods for anti-inflammatory, antioxidant and/or anti-glycation purposes.

Description

Composition for improving skin {Composition for improving skin}

The present invention relates to a composition for skin improvement that exhibits skin whitening, elasticity enhancement, wrinkle improvement, anti-inflammatory, antioxidant, and anti-glycation effects.

It is a common wish to have white and fair skin. Skin color or brightness is genetically determined by the concentration and distribution of melanin in a person's skin, but is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, or stress. Melanin is made by sequentially converting tyrosine, a type of amino acid, into DOPA and dopaquinone through the enzyme tyrosinase as a catalyst, and then through a non-enzymatic oxidation reaction. Although the path by which melanin is produced is known, the cause of tyrosinase triggering in the mechanism that induces melanin synthesis has not yet been revealed in detail.

Meanwhile, commonly known whitening ingredients include substances that inhibit tyrosinase enzyme activity such as kojic acid or arbutin, hydroquinone, vitamin C (L-Ascorbic acid) or their derivatives, and various There are plant extracts. By inhibiting the synthesis of melanin pigment, they can not only brighten skin tone and achieve skin whitening, but also improve skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones, or genetics. However, when applied to the skin, there is a limit to the amount of use due to safety issues such as irritation and redness, or the effect is so small that no practical effect can be expected.

In addition, collagen is a major matrix protein produced by skin fibroblasts and exists in the extracellular matrix, and its important functions include mechanical rigidity of the skin, connective tissue resistance and tissue adhesion, support for cell adhesion, and cell division and differentiation ( It is known to induce growth of organisms or wound healing. This collagen decreases due to age and photoaging caused by ultraviolet irradiation, and this is known to be closely related to the formation of wrinkles in the skin. In addition, as extensive research on skin aging has developed in recent years, the important functions of collagen in the skin have been revealed.

There are known active ingredients that promote collagen synthesis and have a wrinkle-improving effect. For example, retinoic acid, TGF (transforming growth factor) [Non-patent Document 1], protein derived from animal placenta [Patent Document 1], betulinic acid [Patent Document 2], chlorella extract [ Patent Documents 3 and 4] are known as collagen synthesis promoting substances. However, the amount of the active ingredients is limited due to safety issues such as irritation and redness when applied to the skin, or the effect is so minimal that the effect of improving skin function by promoting collagen synthesis in the skin cannot be expected.

In addition, inflammation is the body's immune response to wounds or diseases, and oxidative stress such as ultraviolet rays, active oxygen, and free radicals activate inflammatory factors, causing various diseases and skin aging. Vasoactive polypeptides such as kinin, plasmin, or complement have vasodilation and constriction and chemotaxis effects, and lymphocytes such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized through two pathways, cyclooxygenase or lipooxygenase, into prostaglandins or leukotrienes, which are inflammatory mediators, and mediates various inflammatory reactions.

Anti-inflammatory drugs are called anti-inflammatory drugs that remove inflammation and reduce biological reactions and symptoms. Substances currently used for anti-inflammatory purposes include non-steroids such as flufenamic acid, ibuprofen, benzydamine, or indomethacin, and steroids such as prednisolone. or dexamethasone. In addition, allantoin, Azn or hydrocortisone are known to be effective in anti-inflammatory, but the amount of use of these substances is limited due to safety issues on the skin, or the effect is so minimal that a practical inflammation-relieving effect cannot be expected. There is.

Meanwhile, active oxygen flowing in from outside the body or generated within the body causes many problems, such as accelerating aging of the body or causing cancer. Therefore, much development and research is being conducted on antioxidant substances that inhibit oxidation by active oxygen. Antioxidants are widely distributed in the animal and plant kingdoms, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known to be present in fruits and vegetables. However, antioxidant substances that exist in nature cannot be expected to have a substantially sufficient effect when applied to the skin. Therefore, synthetic antioxidants that have excellent antioxidant power and are inexpensive are widely used, but their use is limited due to safety concerns such as side effects on the human body.

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 without the involvement of enzymes. Accumulation of advanced glycation end products changes proteins into a hard and more brittle state, and glycated collagen prevents collagen from forming an appropriate structure in the extracellular matrix of the dermal layer, causing skin to lose elasticity and promoting wrinkles [ Non-patent Document 2]. In addition, the advanced glycation end products produced by glycation are brown-colored substances and are thought to be the cause of the complexion gradually turning yellow as skin aging progresses. As certain advanced glycation end products accumulate, the reflected light reflected from the skin decreases. It is known that [Non-patent Document 3]. Aminoguanidine, Pyridoxamine, and Aspirin are known as substances that inhibit glycation, but these substances have limits on the amount of use due to safety issues on the skin, or their effects are so minimal that they are practically effective. There is a problem that cannot be expected.

In addition, it is safe for the body, the active ingredients are stable, and above all, it has better skin whitening, wrinkle improvement, elasticity promotion, and anti-inflammatory effects than existing substances that have skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, and/or antioxidant effects. There is an urgent need for the development of ingredients with and/or antioxidant activity.

Meanwhile, Cervi Cornu is the ossified horn of Cervus nippon Temminck, Cervus elaphus Linne, or Cervus canadensis Erxleben (Deer family Cervidae). It promotes blood circulation, eliminates stagnant blood, and helps kidney and liver functions. It also contains a lot of calcium, which helps strengthen bones.

In addition, Cuscuta japonica Chois is the seed of Saesam, an annual vine belonging to the Convolvulaceae family, and is also called Saesam seed. It is mainly known as a medicinal herb that protects the liver and kidneys, brightens the eyes, helps positive energy, and strengthens kidney function. It is effective for impotence in men caused by weak kidneys, spontaneous semen flow, and wet dreams. It strengthens the bones, strengthens the back, and treats pain in the back and knees due to weak kidney function. It is also known to cure urinary incontinence, difficulty urinating, and diarrhea, and is also effective in treating diabetes.

In addition, REHMANNIAE RADIX PREPARAT is made by processing the roots of Rehmannia glutinosa Liboschitz ex Steudel (Scrophulariaceae). Rehmannia glutinosa is the main medicinal ingredient in Samultang and is used to treat symptoms such as internal heat, dry throat, and thirst that appear due to weakness in the body among various chronic diseases.

In addition, Baekjain is a species of the encyclopedia plant Platycladus orientalis, Thuja orientalis (=Biota orientalis), and is a medicinal herb that stabilizes the mind and promotes bowel movement.

Japanese Patent Laid-Open No. 8-231370 Japanese Patent Laid-Open No. 8-208424 Japanese Patent Laid-Open No. 9-40523 Japanese Patent Laid-Open No. 10-36283

Cardinale G. et al, Adv. Enzymol., 41, p. 425, 1974 Dyer, D. G. et al, The Journal of Clinical Investigation., 9, p. 2463, 1993 Hiroshi, O. et al, Skin Research and Technology, 15, p. 496, 2009

Accordingly, the present inventors found that a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract showed a whitening effect by significantly reducing the total amount of melanin, promoted collagen synthesis in skin fibroblasts, and inhibited elastase activity. The present invention was completed by confirming that it improves wrinkles, improves elasticity, exhibits an anti-inflammatory effect by suppressing nitric oxide (NO) production, and has an antioxidant effect that scavenges free radicals and an anti-glycation effect that inhibits glycation.

Therefore, the object of the present invention is to provide a composition for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation, which contains as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract and Baekjain extract. It is provided.

As a means to solve the above problems, the present invention provides a skin whitening, wrinkle improvement, and elasticity product containing as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract for manufacturing medicine, cosmetics, or food. Provided are compositions for enhancing, anti-inflammatory, antioxidant and/or anti-glycation properties.

The mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekjaen extract according to the present invention exhibits a whitening effect by significantly reducing the total amount of melanin, promotes collagen synthesis in skin fibroblasts, and inhibits elastase activity. It improves wrinkles, improves elasticity, inhibits the production of nitric oxide (NO), has an anti-inflammatory effect, and has an antioxidant effect by scavenging free radicals and an anti-glycation effect by suppressing glycation, making it suitable for manufacturing medicine, cosmetics or food. You can use it.

Hereinafter, the configuration of the present invention will be described in detail.

In order for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation ingredients to have excellent effects when applied to actual skin, they must be highly active at low concentrations for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation properties. /or exhibit anti-glycation activity, have an excellent ability to penetrate and be absorbed into the skin, and have low volatility so that they remain for a sufficient period of time to exhibit skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant and/or anti-glycation effects. , it is desirable that the active ingredient is maintained stably in the composition or on the skin, that it is easy to manufacture into medicine, cosmetics, or food, and that it is safe on the skin. However, among known components, components that satisfy all of the above characteristics are rare. For example, some skin-whitening, wrinkle-improving, elasticity-promoting, anti-inflammatory, antioxidant and/or anti-glycation ingredients have skin-whitening, wrinkle-improving, elasticity-promoting, anti-inflammatory, antioxidant and/or anti-glycation properties even at low concentrations when tested in vitro. It is excellent, but its ability to penetrate and absorb through the skin is low, making it difficult to apply to actual skin. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, or foods. In addition, some skin whitening, anti-wrinkle, elasticity-enhancing, anti-inflammatory, antioxidant and/or anti-glycation ingredients may decompose or transform into other compounds when exposed to heat, light, or oxygen, prior to application to the skin. In some cases, the effect disappears.

As can be seen in the examples below, deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekjain extract have a significantly excellent total melanin reduction effect, anti-glycation effect, collagen synthesis promotion effect, anti-inflammatory, antioxidant, and/or anti-glycation effect at low concentrations. It can be used as an active ingredient in pharmaceutical, cosmetic, or food compositions.

Therefore, the present invention provides a composition for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation, comprising as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract and Baekja extract. .

In the present invention, the deer antler is Plum Cervus nippon Temminck, Marok Cervus elaphus Linne or Cervus The ossified horn of canadensis Erxleben (Deer family Cervidae), its scientific name is Cervi Cornu. In addition, Tosaja is a seed of Cuscuta chinensis Lamark (Convolvulaceae), and its scientific name is Cuscuta japonica Chois. In addition, Rehmannia glutinosa is an herbal medicine made by steaming and drying the roots of Rehmannia glutinosa, and its scientific name is Rehmannia glutinosa (Gaertner) Libosch. Baekjain is a medicinal product made from the seeds of Thuja orientalis Linne (Cupressaceae), with the seed coat removed.

The deer antler extract, Tosa extract, Rehmannia glutinosa extract, or Baekja extract can be extracted by a method known in the art, and the method is not particularly limited. Alternatively, commercially available extracts can be used.

Preferably, the deer antler extract, Tosa extract, Rehmannia glutinosa extract, or Baekja extract can be used as an extract obtained by extracting antler, Tosa chinensis, Rehmannia glutinosa, or Baekjae extract with water and/or an organic solvent, and the organic solvent is a polar organic solvent, a non-polar organic solvent, or a non-polar organic solvent. It may be an organic solvent or a mixed solvent thereof. The polar organic solvent may be a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone, and the nonpolar organic solvent may be ether, chloroform, benzene, hexane, or dichloromethane. For example, the lower alcohol having 1 to 5 carbon atoms may be methanol, ethanol, propanol, butanol, or isopropanol.

In one embodiment, the deer antler extract, Tosa extract, Rehmannia glutinosa extract, or Baekjain extract may include a fraction obtained by fractionating the primary extract extracted using the above-described extraction solvent using an extraction solvent of different polarity. For example, deer antler extract, Tosa extract, Rehmannia glutinosa extract, or Baekjain extract may be fractions extracted with alcohol having 1 to 5 carbon atoms and then fractionated again with a solvent of different polarity such as ether, benzene, or hexane. Two or more types of solvents can be used during the fractionation, and extracts for each solvent can be prepared by using them sequentially or by mixing them depending on the polarity of the solvents.

In the present invention, the prepared extract or the fraction obtained by performing the fractionation process can be filtered, concentrated, or dried to remove the solvent, and the filtration, concentration, and drying can all be performed. Specifically, the filtration can be performed using filter paper or a vacuum filter, the concentration can be performed under reduced pressure using a vacuum concentrator, etc., and the drying can be performed using a freeze-drying method.

In addition, the prepared extract or the fraction obtained by performing the fractionation process may be subjected to silica gel column chromatography, thin layer chromatography, or high performance liquid chromatography. Additional purified fractions can be obtained by purification using various chromatographies.

The mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract of the present invention is preferably a mixture of the antler extract, Tosaja extract, Rehmannia glutinosa extract, and Baekja extract in a weight ratio of 1: 1-5: 1-5: 1-5. . In particular, it is more preferable that the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract are mixed at a weight ratio of 1: 1: 1: 1.

The compositions of the present invention can be used for preparing pharmaceutical formulations.

The pharmaceutical formulation may be in the form of a solution, suspension or emulsion in oil or aqueous medium, or in the form of extracts, powders, granules, tablets or capsules.

In addition, the composition may further contain one or more active ingredients that exhibit the same or similar functions. For example, it may include known skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation ingredients. When additional skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation ingredients are included, the skin regeneration, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation improvement effects of the composition of the present invention are reduced. It could be further improved. When adding the above ingredients, skin safety due to combined use, ease of formulation, and stability of the active ingredients can be taken into consideration. In one embodiment of the present invention, the composition includes skin regeneration ingredients known in the art, such as retinoic acid, TGF, protein derived from animal placenta, betulinic acid and chlorella extract, and antioxidant ingredients known in the art, such as tocopherol and selenium. , Vitamin C and phenolic compounds, whitening ingredients known in the art, substances that inhibit tyrosinase enzyme activity such as Kojic acid and Arbutin, Hydroquinone, Vitamin-C (L- Ascorbic acid) and its derivatives and various plant extracts may additionally contain one or two or more ingredients selected from the group consisting of. Additional ingredients may be included in an amount of 0.0001% to 10% by weight 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 active ingredient.

Additionally, the composition of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain several ingredients such as buffers, sterile water for injection, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various oral and parenteral formulations during clinical administration. When formulated, commonly used fillers and extenders are used. It can be prepared using diluents or excipients such as binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, such as starch, calcium carbonate, and water, in the pharmaceutical composition of the present invention. It can be prepared by mixing sucrose, lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogenatin, etc. can be used.

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

In the present invention, the 'wrinkle improvement effect' refers to suppressing or inhibiting the formation of wrinkles on the skin or alleviating wrinkles that have already been formed.

In the present invention, the 'elasticity enhancing effect' refers to increasing the elasticity of the skin, suppressing or inhibiting the loss of skin elasticity, or alleviating already reduced elasticity.

In the present invention, 'anti-inflammatory effect' refers to suppressing inflammation. The inflammation is one of the defense responses of biological tissues to certain stimuli, and refers to a complex lesion that occurs together with three factors: tissue degeneration, circulatory failure, exudation, and tissue proliferation. More specifically, inflammation is part of innate immunity, and as in other animals, human innate immunity recognizes patterns on the cell surface that are specific to pathogens. Phagocytes recognize cells with such a surface as non-self and attack the pathogen. If pathogens break through the body's physical barriers, an inflammatory response occurs. The inflammatory response is a non-specific defense function that creates a hostile environment for microorganisms invading the wound area. In an inflammatory response, when a wound occurs or an external infectious agent enters the body, white blood cells responsible for the initial immune response flock in and express cytokines. Therefore, the expression level of intracellular cytokines becomes an indicator of inflammatory response activation. Examples of skin diseases related to inflammation include atopic dermatitis, psoriasis, erythematous diseases caused by radiation, chemicals, or burns; Itching caused by acid burns, blistering skin disease, lichenoid disease, or allergies; Inflammatory hair loss, such as seborrheic eczema, acne rosae, pemphigus vulgaris, erythema multiforme, erythema nodosum, balanitis, vulvovaginitis, or alopecia areata; Cutaneous T-cell lymphoma, etc., but is not limited thereto.

In the present invention, 'antioxidant effect' refers to the damage to cells caused by highly reactive free radicals or reactive oxygen species (ROS) due to oxidative stress due to intracellular metabolism or the influence of ultraviolet rays. It refers to inhibiting oxidation and includes removing free radicals or reactive oxygen species to reduce damage to cells.

In the present invention, the 'anti-glycation effect' refers to preventing or suppressing the glycation phenomenon in which protein function in the body is reduced by the reaction between glucose and protein present in cells.

In the present invention, 'effective amount' refers to the ability to exhibit a damaged whitening effect, improve wrinkles, enhance elasticity, suppress inflammation, inhibit or alleviate cell oxidation, or inhibit the production of glycation. It refers to the amount of extract present. When the composition of the present invention contains an effective amount of a mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract, the desired skin whitening effect, wrinkle improvement effect, elasticity enhancement effect, anti-inflammatory effect, antioxidant effect, and anti-glycation effect are desirable. It can provide an effect. The effective amount of the mixed extract containing the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract 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 remains on the skin. . For example, when the composition is commercialized as a pharmaceutical formulation, it may contain the active ingredient at a higher concentration compared to when it is commercialized 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, and more preferably, based on the amount of the mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekjain extract. is 50 to 60 mg/kg and can be administered 1 to 6 times a day.

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

The pharmaceutical formulation of the present invention may include a skin topical formulation.

When the mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract is used as an external skin preparation, fatty substances, organic solvents, solubilizers, thickening agents, gelling agents, softeners, antioxidants, suspending agents, and stabilizers are added. topicals, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or it may contain adjuvants commonly used in the field of dermatology, such as lipophilic active agents, lipid vesicles or any other ingredients commonly used in topical preparations for the skin. Additionally, the ingredients may be introduced in amounts commonly used in the field of dermatology.

When the mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract is provided in the form of an external skin preparation, it is not limited thereto, but may have a formulation such as an ointment, patch, gel, cream, or spray.

In addition, the composition of the present invention for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation can be used for manufacturing cosmetic formulations.

The cosmetic formulation may be in the form of a general emulsified formulation or solubilized formulation. For example, lotion such as flexible lotion or nourishing lotion, emulsion such as facial lotion, body lotion, etc., cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, liquid such as nutritional cream, moisture cream, eye cream, etc. It can have formulations such as foundation type, solid type or spray type, powder, makeup remover such as cleansing cream, cleansing lotion, cleansing oil, cleansing foam, soap, body wash, etc.

In addition, the cosmetics contain fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, and foaming agents in addition to a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract. (foaming agent), fragrance, surfactant, water, ionic or non-ionic emulsifier, filler, sequestering agent and chelating agent, preservative, vitamin, blocking agent, wetting agent, essential oil, dye, pigment, hydrophilic or lipophilic It may contain auxiliaries commonly used in the field of cosmetology, such as active agents, lipid vesicles or any other ingredients commonly used in cosmetics.

The cosmetic formulation may contain a relatively high concentration of the active ingredient in the case of a wash-off type cosmetic such as a makeup remover or detergent in which the active ingredient stays on the skin for a short period of time. On the other hand, in the case of leave-on type cosmetics such as lotions, emulsions, creams, and essences, in which the active ingredients stay on the skin for a long period of time, the active ingredients may be contained at a lower concentration than wash-off type cosmetics. It will be fine. Although not limited thereto, in one embodiment of the present invention, the composition may include the active ingredient in an amount of 0.0001% by weight to 10% by weight (preferably 0.0001% by weight to 1% by weight) based on the total weight of the composition. . When the composition of the present invention contains less than 0.0001% by weight of a mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract, it provides sufficient skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-oxidant properties. The saccharification effect cannot be expected, and if it is included in excess of 10% by weight, unwanted reactions such as allergies may occur or skin safety problems may occur. This is to prevent this.

Additionally, the composition for skin whitening, wrinkle improvement, elasticity promotion, anti-inflammatory, antioxidant and/or anti-glycation effect of the present invention can be used to prepare food formulations.

The food formulation is made by adding a mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending the product. means.

Since the above food formulation can be consumed on a daily basis, it is very useful as it can be expected to have high skin whitening, wrinkle improvement, elasticity enhancement, anti-inflammatory, antioxidant and/or anti-glycation effects.

When using the active ingredient as a food additive, the mixed extract of the deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract may be added as is or used together with other foods or food ingredients, and may be appropriately prepared according to a conventional method. can be used The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when producing a 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 materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below 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 are no special restrictions on the types of foods above. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, etc. These include alcoholic beverages and vitamin complexes, and include all health foods in the conventional sense.

When the food formulation is a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, like regular beverages. The above-mentioned natural carbohydrates include 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 a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame 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, food formulations include various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonated drinks. It may contain carbonating agents used in. Additionally, the food formulation may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These ingredients can be used independently or in combination. The ratio 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.

Hereinafter, the present invention will be described in detail by examples. However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

Preparation Example 1: Preparation of deer antler extract

After the deer antler was well dried and cut into pieces, 100 g of the dry weight was placed in a flask and extracted by cold soaking with 1000 g of extraction solvent (distilled water) for 3 days. An antler extract was prepared by filtering the cold-soaked extract through a filter with a pore size of 0.2㎛.

Preparation Example 2: Preparation of Tosaja extract

After drying well and cutting into small pieces, 100g of the dry weight was placed in a flask and extracted by cold soaking with 1000g of extraction solvent (distilled water) for 3 days. Tosaja extract was prepared by filtering the cold-soaked extract through a filter with a pore size of 0.2㎛.

Manufacturing example 3: Preparation of Rehmannia glutinosa extract

After thoroughly drying and cutting Rehmannia glutinosa, 100 g of dry weight was placed in a flask and extracted by cold immersion with 1000 g of extraction solvent (distilled water) for 3 days. Rehmannia glutinosa extract was prepared by filtering the cold-soaked extract through a filter with a pore size of 0.2㎛.

Manufacturing example 4: Preparation of Baekjain Extract

After thoroughly drying and cutting Baekjain, 100 g of the dry weight was placed in a flask and extracted by cold soaking with 1000 g of extraction solvent (distilled water) for 3 days. Baekjain extract was prepared by filtering the cold-soaked extract through a filter with a pore size of 0.2㎛.

Manufacturing example 5: Deer antler, Tosaja, Rehmannia glutinosa and white person's Mixed extract preparation

10 g of the deer antler extract of Preparation Example 1, 10 g of the Tosa extract of Preparation Example 2, 10 g of the Rehmannia glutinosa extract of Preparation Example 3, and 10 g of the Baekja extract of Preparation Example 4 were mixed to prepare a mixed extract of the deer antler, Tosa persimmon, Rehmannia glutinosa, and Baekja extract.

Example 1: Melanin production inhibition effect

In order to confirm the whitening effect through inhibition of melanin production, in the culture medium of B-16 mouse melanoma cells according to the method described by Lotan R. et al. (Cancer Res. 40:3345-3350, 1980), The total amount of melanin was measured by adding the extract. During the experiment, toxicity was first evaluated on rat melanoma cells and whitening was evaluated at a non-toxic concentration. DMSO was used as a negative control, and arbutin was used as a positive control.

Specifically, the sample was added to the medium to a final concentration of 100 ppm, arbutin was added to the medium to a final concentration of 100 ppm, and the melanoma cells were cultured for 3 days. Afterwards, the cells were treated with trypsin, removed from the culture vessel, centrifuged, and melanin was extracted. To the removed cells, 1 ml of sodium hydroxide solution (1N concentration) was added and boiled for 10 minutes to dissolve the melanin. The amount of melanin produced was measured by measuring the absorbance at 400 nm using a spectrophotometer.

The amount of melanin was measured by expressing the absorbance per unit cell ( ¹ It was performed and expressed as an average value.

Effect of reducing the total amount of melanin at the cellular level according to concentration (number of repetitions = 3) sample Melanin production amount (abs) Inhibition rate (%) Control (DMSO, 10ppm) 0.33 - Positive control group (arbutin, 100ppm) 0.228 30.91 Preparation Example 1 (100 ppm) 0.318 3.64 Preparation Example 2 (100 ppm) 0.290 12.12 Preparation Example 3 (100 ppm) 0.302 8.48 Preparation Example 4 (100 ppm) 0.298 9.70 Preparation Example 5 (100 ppm) 0..230 30.30

As can be seen from the results in Table 1, the mixed extract of Preparation Example 5 showed an excellent effect of reducing the total amount of melanin even at low concentrations and could be used for whitening purposes.

Example 2: Effect of promoting collagen synthesis

The sample was added to the culture medium of human-derived fibroblasts to confirm the effect of promoting type 1 collagen synthesis at the cellular level. The synthesized collagen was quantified using the PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT).

The sample was added to the fibroblast culture medium (DMEM medium) at a final concentration of 10 ppm, cultured for 48 hours, the culture medium was taken, and the degree of type 1 collagen synthesis was measured at each concentration using a PICP EIA kit using a spectrophotometer at 450 nm. It was measured in .

To compare effects, the degree of collagen synthesis was measured in the same manner for samples to which untreated fibroblast culture medium (negative control) and vitamin C (positive control) were added to a final concentration of 52.85 ㎍/ml.

The rate of increase in collagen production was calculated as the ratio of collagen production relative to the negative control group, and the results are shown in Table 2 below. The experiment was performed four times and expressed as the average value.

Effect of promoting collagen synthesis (number of repetitions = 4) sample Type 1 collagen production (ng/ml) Collagen production increase rate (%) Negative control group 150.2 - Positive control group (vitamin C) 246.8 64.3 Preparation Example 1 (10 ppm) 200.7 33.6 Preparation Example 2 (10 ppm) 168.3 12.1 Preparation Example 3 (10 ppm) 218.5 19.4 Preparation Example 4 (10 ppm) 176.9 17.8 Preparation Example 5 (10 ppm) 246.3 64.0

As shown in Table 2, treatment with the mixed extract of Preparation Example 5 increased collagen synthesis, and showed a similar effect on collagen synthesis as when applying vitamin C, which is generally known to induce collagen synthesis.

Example 3: Elastase active inhibitory effect

The effect of inhibiting the activity of Elastase, an enzyme that decomposes elastin, was confirmed as follows.

Elastase derived from human white blood cells was used, and the synthetic substrate MeOSuc-Ala-Ala-Pro-Val-pNA was used as the elastase substrate. A 100 mM Tris (pH 7.5) solution was used as the buffer solution. Elastase was used in a final amount of 0.2 mU using a buffer solution. In addition, the synthetic substrate of elastase was made into a 100mM solution using DMSO and then diluted with a buffer solution to reach a final concentration of 0.5mM. At this time, the positive control was set to include Quercetin, known as an elastase inhibitor, at a concentration of 10 ppm. The elastase inhibition candidate was added to a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and reacted at room temperature for 20 minutes. Absorbance was measured at 405 nm at 1-minute intervals using a spectrophotometer, and the slope of absorbance versus time was determined to determine enzyme activity. The elastase inhibition rate was calculated according to Equation 1 below.

[Equation 1]

Elastase activity inhibition effect (number of repetitions = 3) sample enzyme activity Inhibition rate (%) Preparation Example 1 (10 ppm) 4.7 54.80 Preparation Example 2 (10 ppm) 4.9 52.88 Preparation Example 3 (10 ppm) 4.3 58.65 Preparation Example 4 (10 ppm) 5.1 50.96 Preparation Example 5 (10 ppm) 3.6 65.38 Positive control group (Quercetin, 10ppm) 3.5 66.35 Control (DMSO, 20ppm) 10.4 -

As can be seen in Table 3, when the mixed extract of Preparation Example 5 was treated, the effect was less than that of the positive control, but the inhibition of elastase activity was excellent. In other words, the mixed extract showed excellent elastase activity inhibition effect. Therefore, it was found that the mixed extract can be used to improve elasticity and improve wrinkles.

Example 4: Anti-inflammatory effect

To confirm the anti-inflammatory effect and skin trouble improvement effect, a nitric oxide (NO) production inhibition test was conducted using the GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are mouse macrophages, were subcultured several times, placed in a 24-well plate at 3×10 ⁵ cells per well, and cultured for 24 hours. Then, it was replaced with cell medium containing the sample at a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), an NO-production inhibitor, was treated as a positive control and cultured for 30 minutes, and 1 μg of LPS (Lipopolysaccharide) was treated as a stimulant and cultured for 24 hours. Take 100 ㎕ of the supernatant and transfer it to a 96-well plate, add 100 ㎕ of GRIESS solution each and react at room temperature for 10 minutes. The NO inhibition effect is determined by measuring the absorbance at 540 nm, and the NO production inhibition rate (%) is It was calculated using Equation 2 below and shown in Table 4 below, and the experiment was performed three times each and expressed as the average value.

[Equation 2]

NO production inhibition rate (%) = {(absorbance of negative control group - absorbance of each extract)/absorbance of negative control group} x 100

NO production inhibition effect (number of repetitions = 3) sample NO production inhibition rate (%) Control (DMSO, 10 ppm) - L-NMMA (positive control, 10 ppm) 41.02 Preparation Example 1 (10 ppm) 26.96 Preparation Example 2 (10 ppm) 25.39 Preparation Example 3 (10 ppm) 27.99 Preparation Example 4 (10 ppm) 30.15 Preparation Example 5 (10 ppm) 39.98

As can be seen in Table 4, the mixed extract of Preparation Example 5 has low activity compared to L-NMMA, a representative anti-inflammatory medicinal substance, but shows excellent activity as a natural substance, and has an anti-inflammatory effect in improving skin troubles. By acting as an antagonist, a synergistic effect can be expected.

Example 5: Antioxidant effect - free radical scavenging rate

To confirm the antioxidant activity of the sample, free radical scavenging activity was measured. Free radical scavenging activity was measured using DPPH (Blois, Nature 181, 1190, 1958).

DPPH is a relatively stable free radical. When it exists in a radical state, it shows maximum light absorption at 517 nm and loses light absorption when the radical is eliminated. DPPH was used from Sigma.

First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg/ml) was prepared. Then, make a 1000 μg/ml (0.1%) vitamin C solution using tertiary distilled water and then serially dilute it by 1/2 to obtain 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, 0.98. , a vitamin C sample with a concentration of 0.49 μg/ml was prepared. In addition, dilutions of Preparation Examples 1 to 5 were prepared at the same concentration as vitamin C. Next, the sample and the standard DPPH solution were added in equal proportions, stirred well, reacted at 37°C for 30 minutes, and the absorbance at 517 nm was measured using a microplate reader (BioTek EL-340) to determine IC _50. The results were obtained and shown in Table 5 below.

IC ₅₀ (half maximal inhibitory concentration) is the concentration of the substance required to remove 50% of free radicals in the no-added control group (in this case, DPPH), and is a general way to express free radical scavenging ability.

Free radical scavenging effect (number of repetitions = 3) sample IC ₅₀ (%w/v) Positive control group (vitamin C) 0.0005 Manufacturing Example 1 0.0007 Production example 2 0.0018 Production example 3 0.0022 Production example 4 0.0017 Production example 5 0.0006

As shown in Table 5, the mixed extract of Preparation Example 5 had excellent free radical scavenging ability compared to Preparation Examples 1 to 4, and was confirmed to have an antioxidant effect similar to that of vitamin C, a positive control group.

Example 6: antiglycation effect

To confirm the anti-glycation effect, the glycation inhibition activity was measured using L-arginine and glucose.

First, 1M L-arginine and 1M glucose were prepared by dissolving them in 1M phosphate buffer solution (pH 7.4), and the sample was diluted to 50 ppm using 1M phosphate buffer solution. 1M L-arginine and 1M phosphate buffer solution were mixed at a ratio of 1 to 4, and then 80 μl each was dispensed into a 96-well plate. Here, 100 μl each of the sample diluted to 50ppm and 0.01M aminoguanidin to be used as a positive control were added. These samples were mixed well, and finally, glucose diluted with 1M phosphate buffer solution was added so that the final concentration of glucose was 0.1M, and then reacted at 70°C for 4 hours. The degree of glycosylation was measured by measuring the absorbance of the 96-well plate at 420 nm using a spectrophotometer.

The Glycation experimental group in Equation 1 below is an experimental group in which 1M L-arginine and 1M glucose were added to induce glycation. To measure the absorbance of the sample itself, only 1M L-arginine and the sample were added without adding glucose and the absorbance was measured at 420 nm. did.

Glycosylation inhibition activity can be obtained using Equation 3 below. Each experiment was performed three times and expressed as the average value.

[Equation 3]

Sample (number of repetitions = 3) Glycosylation inhibition rate (%) Control (DMSO, 50 ppm) - Positive control (Aminoguanidine, 55 ppm) 54.89 Production Example 1 (50 ppm) 47.28 Production example 2 (50 ppm) 48.33 Production example 3 (50 ppm) 50.68 Production example 4 (50 ppm) 37.84 Production example 5 (50 ppm) 67.97

As can be seen from the results in Table 6, the mixed extract of Preparation Example 5 according to the present invention has a more excellent anti-glycation effect compared to Aminoguanidine, a known anti-glycation substance. In other words, the mixed extract exhibits an excellent anti-glycation effect, so when it comes to whitening or wrinkle improvement, the anti-glycation effect acts as an auxiliary role, so a synergistic effect can be expected.

Example 1: Preparation of pharmaceutical preparations

1. Preparation of tablets

0.2 mg of mixed extract of Preparation Example 5

Corn starch 100mg

Lactose 100mg

Magnesium stearate 2mg

After mixing the above ingredients, tablets were manufactured by tableting according to a conventional tablet manufacturing method.

Example 2: Manufacturing of cosmetics

1. Manufacturing of nutritious cream

As shown in the following composition, a nutritional cream containing the mixed extract of Preparation Example 5 as an active ingredient was prepared according to a conventional method.

0.2% by weight of mixed extract of Preparation Example 5

Beta-1,3-glucan 5.0% by weight

Beeswax 10.0% by weight

Polysorbate 60 1.5% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitan sesquioleate 0.5% by weight

Liquid paraffin 10.0% by weight

Squalane 5.0% by weight

Caprylic/capric triglyceride 5.0% by weight

Glycerin 5.0% by weight

Butylene glycol 3.0% by weight

Propylene glycol 3.0% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Fragrance 0.05% by weight

Purified water to 100% by weight

Example 3: Preparation of external skin preparations

1. Preparation of ointment

As shown in the composition below, an ointment containing the mixed extract of Preparation Example 5 as an active ingredient was prepared according to a conventional method.

0.5% by weight of mixed extract of Preparation Example 5

Beta-1,3-glucan 10.0% by weight

Beeswax 10.0% by weight

Polysorbate 60 5.0% by weight

PEG 60 hydrogenated castor oil 2.0% by weight

Sorbitan sesquioleate 0.5% by weight

Vaseline 5.0% by weight

Liquid paraffin 10.0% by weight

Squalane 5.0% by weight

Shea butter 3.0% by weight

Caprylic/capric triglyceride 5.0% by weight

Glycerin 10.0% by weight

Propylene glycol 10.2% by weight

Triethanolamine 0.2% by weight

Preservative 0.05% by weight

Pigment 0.05% by weight

Fragrance 0.05% by weight

Purified water to 100% by weight

Claims (20)

A composition for skin whitening containing as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract.
According to claim 1,
Antler extract is an extract obtained by extracting deer antler with one or more selected from the group consisting of water and organic solvents,
The soil extract is an extract obtained by extracting the soil soil with one or more selected from the group consisting of water and organic solvents,
Rehmannia glutinosa extract is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents,
Baekjain extract is a composition for skin whitening obtained by extracting Baekjain with one or more selected from the group consisting of water and organic solvents.
According to claim 2,
The organic solvent may be a polar solvent containing lower alcohols having 1 to 5 carbon atoms, ethyl acetate, or acetone; Non-polar solvents including ether, chloroform, benzene, hexane, or dichloromethane; Or a composition for skin whitening that is a mixed solvent thereof.
According to claim 1,
The composition is a composition for skin whitening, which is intended for manufacturing medicine, cosmetics or food for skin whitening.
A composition for improving skin elasticity or improving wrinkles, comprising as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract.
According to claim 5,
Antler extract is an extract obtained by extracting deer antler with one or more selected from the group consisting of water and organic solvents.
The soil extract is an extract obtained by extracting the soil soil with one or more selected from the group consisting of water and organic solvents,
Rehmannia glutinosa extract is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents,
Baekjain extract is a composition for promoting skin elasticity or improving wrinkles, which is an extract obtained by extracting Baekjain with one or more selected from the group consisting of water and organic solvents.
According to claim 6,
The organic solvent may be a polar solvent containing lower alcohols having 1 to 5 carbon atoms, ethyl acetate, or acetone; Non-polar solvents including ether, chloroform, benzene, hexane, or dichloromethane; Or a composition for improving skin elasticity or improving wrinkles, which is a mixed solvent thereof.
According to claim 5,
The composition is a composition for improving skin elasticity or improving wrinkles, which is intended for the manufacture of medicines, cosmetics, or foods for improving skin elasticity or improving wrinkles.
An anti-inflammatory composition containing as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract.
According to clause 9,
Antler extract is an extract obtained by extracting deer antler with one or more selected from the group consisting of water and organic solvents,
The soil extract is an extract obtained by extracting the soil soil with one or more selected from the group consisting of water and organic solvents,
Rehmannia glutinosa extract is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents,
Baekjae extract is an anti-inflammatory composition obtained by extracting Baekjae with one or more selected from the group consisting of water and organic solvents.
According to claim 10,
Organic solvents include polar solvents including lower alcohols having 1 to 5 carbon atoms, ethyl acetate, or acetone; Non-polar solvents including ether, chloroform, benzene, hexane, or dichloromethane; Or an anti-inflammatory composition that is a mixed solvent thereof.
According to clause 9,
The composition is an anti-inflammatory composition for the manufacture of anti-inflammatory medicine, cosmetics or food.
An antioxidant composition containing as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract.
According to claim 13,
Antler extract is an extract obtained by extracting deer antler with one or more selected from the group consisting of water and organic solvents,
The soil extract is an extract obtained by extracting the soil soil with one or more selected from the group consisting of water and organic solvents,
Rehmannia glutinosa extract is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents,
Baekjae extract is an antioxidant composition obtained by extracting Baekjae with one or more selected from the group consisting of water and organic solvents.
According to claim 14,
The organic solvent may be a polar solvent containing lower alcohols having 1 to 5 carbon atoms, ethyl acetate, or acetone; Non-polar solvents including ether, chloroform, benzene, hexane, or dichloromethane; Or an antioxidant composition that is a mixed solvent thereof.
According to claim 13,
The composition is an antioxidant composition for the manufacture of antioxidant medicine, cosmetics or food.
An anti-glycation composition containing as an active ingredient a mixed extract of deer antler extract, Tosa extract, Rehmannia glutinosa extract, and Baekja extract.
According to claim 17,
Antler extract is an extract obtained by extracting deer antler with one or more selected from the group consisting of water and organic solvents,
The soil extract is an extract obtained by extracting the soil soil with one or more selected from the group consisting of water and organic solvents,
Rehmannia glutinosa extract is an extract obtained by extracting Rehmannia glutinosa with one or more selected from the group consisting of water and organic solvents,
Baekjae extract is an anti-glycation composition obtained by extracting Baekjae with one or more selected from the group consisting of water and organic solvents.
According to claim 18,
Organic solvents include polar solvents including lower alcohols having 1 to 5 carbon atoms, ethyl acetate, or acetone; Non-polar solvents including ether, chloroform, benzene, hexane, or dichloromethane; Or a composition for anti-glycation that is a mixed solvent thereof.
According to claim 17,
The composition is an anti-glycation composition for the manufacture of anti-glycation drugs, cosmetics or foods.