KR20200054151A - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. The composition for improving skin containing a Panax notoginseng extract according to the present invention has effects of promoting the proliferation of skin stem cells, inhibiting NO production, having an excellent activity of PPAR alpha, increasing the expression of filaggrin, and improving skin brightness, thereby being able to be used for manufacturing medicine, cosmetics, or food for skin regeneration, ani-inflammation, atopy alleviation, skin moisturizing, and skin texture improvement. In addition, the composition has effects of promoting the proliferation of skin stem cells and maintaining stem cell properties, thereby being able to be used for manufacturing a cosmetic for promoting stem cell activity.

Description

Composition for improving skin

The present invention relates to a composition for skin improvement showing skin regeneration, anti-inflammatory, atopy improvement, skin moisturization, skin texture improvement, and stem cell activity promoting effect.

Inflammation is an immune response of the human body that responds to wounds and diseases, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activates inflammatory factors, causing various diseases and skin aging. Vascularly active polypeptides, such as kinin, plasmin, and complement, act to expand and contract blood vessels and chemotaxis, and other lymphokas such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to prostaglandin and leukotriene, which are inflammatory mediators, through two pathways: cyclooxygenase or lipooxygenase.

On the other hand, in order to eliminate inflammation, the action of reducing the inflammatory source, reducing biological reactions and symptoms is called anti-inflammatory. Materials that have been used for anti-inflammatory purposes to date include non-steroidal drugs such as flufenamic acid, ibuprofen, benzydamine, and indomethacin, and prednisolone as a steroid system. , Dexamethasone, and the like. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have limitations in the amount of use due to safety problems for the skin, or the effect is insignificant, so that the effect of alleviating inflammation cannot be expected. There is this.

In addition, the epidermis (epidermis) located on the outermost part of the skin serves to protect against various physical, chemical and mechanical stimuli from the outside and to prevent excessive dissipation of moisture in the body through the skin. This protective function is possible by forming and maintaining a stratum corneum composed of keratinocytes normally. Keratinocytes are cells formed by undergoing morphological and functional changes step by step as the basal cells, which were continuously proliferating in the stratum basale, migrate to the stratum corneum. The forming cells are removed from the skin, and new keratinocytes from the lowermost layer of the epidermis repeat the process of epidermis differentiation or keratinization in place of its function. During this keratinization process, keratinocytes produce natural moisturizing factor (NMF) and intercellular lipids such as ceramide, cholesterol, and fatty acids, so that the stratum corneum acts as a blocking layer from the outside, and thus a skin barrier. It retains its functions as.

In addition, skin dryness, which is considered as one of the major diseases in modern society, is one of the symptoms caused by skin barrier dysfunction. Recently, environmental pollution, increased dry environments such as apartments and high-rise buildings, increased social stress, is unique to our country. It is increasing more and more due to various factors such as excessive bath culture, skin aging, etc.

Recently, atopic dermatitis, which occurs in 10% of children, is also known to be a major cause of skin dryness and, more fundamentally, an abnormality of the skin barrier function. In order to treat such atopic dermatitis, a lot of research has been conducted to supply moisture from the outside or minimize moisture loss from the body by focusing on maintaining proper moisture in the skin, and ceramide having the ability to actually retain moisture ( ceramide) or a derivative thereof has been developed and is widely used in the pharmaceutical or cosmetic field.

However, the use of such moisturizers is only a temporary symptom relief rather than a fundamental treatment, and thus does not exhibit sufficient effects for the treatment of skin dryness and skin barrier function abnormalities including atopic dermatitis. Accordingly, there is an urgent need to develop a substance that fundamentally regenerates the damaged skin barrier.

Recently, it has been known that when PPAR alpha (Peroxisome Proliferator Activated Receptor-alpha) present in keratinocytes is activated in combination with its ligand, it promotes the differentiation of keratinocytes and reconstructs the damaged skin barrier. When a known agonist of PPAR alpha, clofibrate (non-patent document 1) or WY14643 (non-patent document 2), etc. is applied to the skin with damaged skin barrier, differentiation of keratinocytes is promoted It has been confirmed that the recovery of the skin barrier is promoted.

In addition, it is a constant desire of everyone to have a white skin. It is genetically determined according to the concentration and distribution of melanin in a person's skin, but is also affected by environmental or physiological conditions such as sun UV rays, fatigue, and stress. Melanin is produced by undergoing a non-enzymatic oxidation reaction after tyrosinase, an amino acid tyrosine, is converted to dopa (DOPA) or dopaquinone. The pathway by which melanin is produced is known, but the mechanism by which tyrosinase acts as a precursor for melanin synthesis is still unknown.

In addition, the skin texture, which is a form of the surface of the skin, is characterized by a three-dimensional microstructure formed by fine lines and is significantly different for each age and part. Skin texture is divided into 1st (about 20-100 μm), 2nd (about 5-40 μm), and 3rd (about 0.5 μm) lines depending on the depth of the line. It has the characteristic that the polygons and star formation that occur when the lines meet are distinct [Non-Patent Document 3].

However, it is known that fine lines (secondary, tertiary, etc.) disappear and wrinkles are generated as the primary line becomes deeper as the network structure of dense fine lines collapses with age or skin damage. Documents 4,5]. In other words, wrinkles, which are a typical sign of skin aging, are accumulations of minute changes that occur over a long period of time.

Genetically, it is known that all components of human skin are derived from ectoderm or mesoderm. The epidermis, hair follicles, sebaceous glands, and sweat glands originate from ectoderm, and melanocytes, nerves, and special sensory receptors originate from neuroectoderm. Depending on the developmental period, embryonic stem cells repeat differentiation and become cells with characteristics suitable for each tissue function, and even after becoming an adult, a certain number of stem cells remain in the tissue. Stem cells exist mainly in two places on the skin. . The first is in the hair follicle. This is a cell before cell differentiation and is known to play an important role in the regeneration of the epidermis and plays an important role in hair regeneration and growth. The second is the basal layer of epidermis. It has been found that the stem cells located here play an important role in protecting skin health by controlling not only the epidermis but also the fibroblasts of the dermis layer. Stem cells here are relatively large and have the advantage of being readily available, making them widely used in skin stem cell research. The skin is constantly renewed, and stem cells present in the epithelium of the skin, that is, epidermal stem cells of the skin, are involved in the restoration of the epithelium after wounding [Non-Patent Document 6]. The epithelial stem cells of the skin are also referred to as skin stem cells, and when activating the skin stem cells, it can treat skin wounds such as trauma or promote wound healing, and can also reduce wrinkles of the skin. have. Integrin β1 (integrin β1) and integrin α6 (integrin α6) are used as indicators of skin stem cells, and maintenance of skin stem cells required for epithelial morphogenesis and differentiation is reported to be regulated by the p63 protein.

Skin stem cells play an important role in maintaining the health and physiological and biochemical homeostasis of the skin. Stem cells present in the skin also cause abnormalities in function due to the effects of aging, and accordingly, various problems occur as the homeostasis of the skin is broken. Therefore, various phenomena of skin aging may be improved through the activity of these stem cells [Non-Patent Document 7].

In addition, it is safe for the living body, has an active ingredient that is stable, and above all, has anti-inflammatory, atopy-improving, skin-moisturizing and skin-improving activity that is superior to existing anti-inflammatory, atopy-improving, skin-moisturizing and skin-improving substances. Development of the ingredients possessed is urgently desired.

On the other hand, Samchil is a perennial herb, the root of the agaraceae plant Panax pseudoginseng, P.notoginseng. Its conical shape is 1-6cm long, 1-4cm in diameter, and has small lump-shaped protrusions around it. Yellow, with vertical wrinkles and fine roots, and the cut surface is grayish-yellowish green and grayish white with a radial pattern, and the quality is heavy and hard. According to Donguibogam, it has been used for the treatment of various types of hememia, vaginal swelling, cardiovascular disease, and is known to have cardiovascular diseases, immune-enhancing effects, and cell protection effects. According to the existing studies on the ingredient identification, compounds such as β-carotene, cry ptoxanthine, meloside A, L saponin, and trilinolein have been reported as major components.

However, the mechanism of Samchil's skin is not yet known, and no research has been done on this.

Feingold et al., J. Invest. Dermatol., 110, pp 368-375, 1998. Feingold et al., J. Invest. Dermatol., 115, pp 353-360, 2000. Journal of the European Academy of Dermatology and Venereology. 12: 103-114, 1999 The British journal of dermatology. 110: 129-138, 1984 Skin research and technology.5: 189-194, 1999 Epidermal Stem Cells of the Skin, Cedric Blanpain, Elaine Fuchs, Annual Review of Cell and Developmental Biology, November 2006, Vol. 22, Pages 339-373 Human skin stem cells and the ageing process, Catherin Niemann, Stem Cell Aging and Regenerative Medicine, November 2008, Pages 986-997

Thus, the inventors of the present invention promote the proliferation of skin stem cells by promoting the regeneration of skin, inhibiting NO production, and have an anti-inflammatory effect, and have excellent PPAR alpha activity, thereby improving atopy and increasing the expression of pilar green. It shows the effect of improving skin moisturizing, and improves skin texture due to improvement of skin brightness, promotes the proliferation of skin stem cells, and shows the effect of maintaining stem cells, thereby completing the present invention by confirming the effect of promoting stem cell activity. Became.

Accordingly, an object of the present invention is to provide a composition for improving anti-inflammatory, atopy, skin moisturizing and skin texture containing the Samchil extract as an active ingredient.

In addition, another object of the present invention is to provide a composition for promoting stem cell activity comprising the following samchi extract as an active ingredient.

As a means for solving the above problems, the present invention provides a composition for improving anti-inflammatory, atopic, skin moisturizing and skin texture, including as an active ingredient, a pharmaceutical extract for pharmaceutical, cosmetic or food production.

As another means for solving the above problems, the present invention provides a composition for promoting stem cell activity, which includes a samchi extract as an active ingredient for preparing a cosmetic formulation.

The Samchil extract according to the present invention promotes the proliferation of skin stem cells, promotes skin regeneration, has an anti-inflammatory effect by inhibiting NO production, and has an excellent activity of PPAR alpha, thereby improving atopy, and due to increased expression of pilar green In addition to the skin moisturizing improvement effect, by showing the skin texture improvement effect due to skin brightness improvement, it can be used in medicine, cosmetics, or food manufacturing.

In addition, the Samchil extract promotes the proliferation of skin stem cells and shows a stem cell maintenance effect, and thus can be used for preparing a cosmetic formulation for promoting stem cell activity.

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

Skin regeneration, anti-inflammatory, atopy improvement, skin moisturization and skin texture improvement In order to exert excellent effects when applied to actual skin, high activity at low concentrations of skin regeneration, anti-inflammatory, atopy improvement, skin moisturization and skin texture improvement activities It is excellent in the ability to absorb and penetrate through the skin, and has low volatility so that it can remain for a sufficient time to exhibit skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement effects, and the active ingredient is stable on the composition or skin It is preferably maintained, and is easy to manufacture in medicine, cosmetics or food, and is also safe for the skin. However, few known components satisfy all of the above properties. For example, some skin regeneration, anti-inflammatory, atopy-improving, skin-moisturizing and skin-improving ingredients have excellent skin regeneration, anti-inflammatory, atopy-improving, skin-moisturizing and skin-improving activity even at low concentrations in vitro experiments. It is difficult to apply to real skin due to its poor ability to penetrate and absorb. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicine, cosmetics, and food. In addition, some skin regeneration, anti-inflammatory, atopy improvement, skin moisturizing and skin texture improving ingredients are decomposed or transformed into other compounds when exposed to heat, light, or oxygen, so that the effect disappears before being applied to the skin. There are cases.

As can be seen in the examples below, the Samchil extract exhibits excellent skin proliferation effect, anti-inflammatory effect, PPAR alpha activity effect, pilarin expression effect, and skin brightness improvement effect at a low concentration, which is superior to skin regeneration. It can be used as an active ingredient in medicine, cosmetic or food composition for anti-inflammatory, atopy improvement, skin moisturization and skin texture improvement.

Therefore, the present invention regenerates the skin containing the extract of Samchil as an active ingredient. Provides a composition for improving anti-inflammatory, atopy, moisturizing skin and improving skin texture.

In the present invention, Samchil is the root of the Panax pseudoginseng, P.notoginseng, and is also called Jeonchil (田七).

The samchil extract may be extracted by a method known in the art, the method is not particularly limited. Alternatively, a commercially available extract can be used.

Preferably, the samchil extract may use an extract obtained by extracting samchil with water and / or an organic solvent, and the organic solvent may be a polar organic solvent, a non-polar 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 non-polar 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 samchil extract may include a fraction obtained by fractionating a primary extract extracted using the above-described extraction solvent using an extraction solvent having a different polarity. For example, the extract of samchil may be a fraction that is extracted with an alcohol having 1 to 5 carbon atoms, and then fractionated again with a solvent having different polarities such as ether, benzene, and hexane. At the time of fractionation, two or more solvents may be used, and each solvent extract may be prepared by sequentially using or mixing depending on the polarity of the solvent.

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

In addition, silica gel column chromatography, thin layer chromatography or high performance liquid chromatography, etc. on the prepared extract or the fraction obtained by performing the fractionation process Further purified fractions can be obtained by purification using various chromatography.

The composition of the present invention can be used for the manufacture of pharmaceutical formulations.

The pharmaceutical formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an ex-agent, powder, granule, tablet or capsule.

In addition, the composition may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improving ingredients. If additional skin regeneration, anti-inflammatory, atopy improvement, skin moisturizing and skin texture improving ingredients are included, the skin regeneration, anti-inflammatory, atopy improvement, skin moisturizing and skin texture improving effects of the composition of the present invention may be further enhanced. When adding the above components, skin safety, ease of formulation, and stability of active ingredients according to the combined use may be considered. In one embodiment of the invention, the composition is a skin regeneration component known in the art, retinoic acid, TGF, animal placenta-derived protein, betulinic acid and chlorella extract, as an antioxidant component known in the art, tocopherol, selenium , Vitamin C and phenolic compounds, as whitening ingredients known in the art, substances that inhibit tyrosinase enzyme activity, such as kojic acid, arbutin, hydroquinone, vitamin-C (L- Ascorbic acid) and their derivatives and one or more components selected from the group consisting of various plant extracts. The additional components may be included in an amount of 0.0001% to 10% by weight based on the total composition weight, and the content range may be adjusted according to requirements such as skin safety and ease in formulating the samchi extract.

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

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

The composition of the present invention may be administered orally or parenterally, and may be administered in various forms of oral pharmaceutical and parenteral forms in the form of general pharmaceutical preparations, for example, clinical administration. , Can be prepared using diluents or excipients, such as binders, wetting agents, disintegrating agents, surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example, starch, calcium carbonate, water It can be prepared by mixing Sucrose, Lactose, or gelatin.

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

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents, suspension solvents include propylene glycol (Propylene glycol), polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

In the present invention, the term "skin regeneration effect" refers to the recovery of skin tissue against damage caused by external and internal causes of skin as the activity of skin stem cells is promoted. At this time, the damage caused by the external cause may include ultraviolet rays, external pollutants, wounds or trauma, and the damage caused by the internal cause may include stress.

In the present invention, the term 'anti-inflammatory effect' refers to suppressing inflammation, and the inflammation is one of biological tissues' defense responses to certain stimuli, and is associated with three types of tissue degeneration, circulatory disorder and exudation, and tissue proliferation. Complex lesions. More specifically, inflammation is part of innate immunity, and as in other animals, innate immunity in humans recognizes patterns on cell surfaces that are specifically present in pathogens. Phagocytes recognize cells with such a surface as non-magnetic and attack pathogens. If pathogens break through the body's physical barrier, an inflammatory reaction occurs. Inflammatory reactions are non-specific protective actions that create a hostile environment against microorganisms that have entered the wound. In an inflammatory reaction, when a wound or an external infectious agent enters the body, white blood cells that are in charge of the initial stage of immune reactions flock to express cytokines. Therefore, the amount of cytokine expression in the cells is an indicator of activation of the inflammatory response. Examples of skin diseases associated with inflammation include atopic dermatitis, psoriasis, radiation, chemicals, erythematous diseases triggered by burns, acid burns, blistering skin diseases, thyroid-like diseases, itching due to allergies, seborrheic eczema, rose acne, Inflammatory hair loss such as vulgar erythema, polymorphic exudative erythema, nodular erythema, laryngitis, vulvitis, alopecia areata, skin T-cell lymphoma, but is not limited thereto.

In the present invention, the term 'atopy improving effect' refers to the prevention and treatment effect of skin diseases such as atopy and dry skin, and refers to suppressing, inhibiting, or alleviating atopic symptoms.

In the present invention, the term "moisturizing effect" refers to inhibiting or suppressing the reduction of moisture in the skin or increasing the moisture content of the skin to smooth the skin surface and impart shine.

In the present invention, the term "skin improvement effect" refers to improving the skin surface smoothly, giving gloss, and improving the skin tone by inhibiting or inhibiting the roughness of the skin surface due to aging, stress, etc. .

In the present invention, the term 'effective amount' refers to the amount of extract that can promote regeneration of damaged skin, suppress inflammation, improve skin dryness such as atopy, moisturize effect, or improve skin texture. . When the composition of the present invention includes an effective amount of the above-mentioned samchil extract, it is possible to provide a desirable skin regeneration effect, anti-inflammatory effect, atopy improvement effect, moisturizing effect and skin texture improvement effect. The effective amount of the Samchil extract contained 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 length of time it remains on the skin. For example, when the composition is commercialized as a pharmaceutical formulation, it may contain the extract of the swallowtail at a higher concentration than when commercialized as a cosmetic that is applied to the skin on a daily basis. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, and 1 to 6 times a day, based on the amount of the Samchil extract. Can be administered.

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

The pharmaceutical formulation of the present invention may include an external preparation for skin.

When the above-mentioned samchil extract is used as an external preparation for skin, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water , Ionic or nonionic emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or external preparations for skin Can contain adjuvants commonly used in the field of dermatology such as any other ingredients. In addition, the ingredients may be introduced in an amount commonly used in the field of skin science.

When the extract is provided as an external preparation for skin, it is not limited thereto, and may have a formulation such as ointment, patch, gel, cream or spray.

In addition, the composition for skin regeneration, anti-inflammatory, atopy improvement, skin moisturization and skin texture improvement of the present invention can be used for the preparation of cosmetic formulations.

The cosmetic formulation may be in the form of a general emulsifying formulation and a solubilizing formulation. For example, lotion such as soft lotion or nutrition lotion, emulsion such as facial lotion, body lotion, nutrition cream, moisture cream, cream such as eye cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, liquid It may have a formulation such as foundation type, solid type or spray type, powder, cleansing cream, cleansing lotion, makeup remover such as cleansing oil, cleansing agent such as cleansing foam, soap, body wash, and the like.

In addition, in addition to the Samchil extract, the cosmetics include fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, and ionic forms Or nonionic emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other commonly used in cosmetics. It may contain adjuvants commonly used in the field of cosmetics such as ingredients.

The cosmetic formulation may include the relatively high concentration of the swallowtail extract in the case of a wash-off type cosmetic such as a makeup remover, detergent, etc., in which the active ingredient stays on the skin within a short period of time. On the other hand, in the case of a leave-on type cosmetic such as lotion, emulsion, cream, essence, etc., in which the active ingredient stays on the skin for a long period of time, even if it contains a lower concentration of the swallowtail extract than the wash-off type cosmetic, It will be okay. Without being limited thereto, in one embodiment of the present invention, the composition may include the trillium extract in an amount of 0.0001% to 10% by weight (preferably 0.0001% to 1% by weight) based on the total composition weight. . When the composition of the present invention contains less than 0.0001% by weight of the above-mentioned samchil extract, sufficient skin regeneration, anti-inflammatory, atopy improvement, skin moisturizing and skin texture improvement effects cannot be expected, and when it contains more than 10% by weight This is to prevent unwanted reactions such as allergies or skin safety problems.

In addition, the composition for skin regeneration, anti-inflammatory, atopic improvement, skin moisturization, and skin texture improvement of the present invention can be used for preparing food formulations.

The food formulation refers to a food prepared by adding the extract of the samchil to food materials such as beverages, teas, spices, chewing gum, confectionery, or by encapsulation, powdering, suspension, or the like.

The food formulation is very useful because it can be ingested on a daily basis, and thus it is possible to expect high skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing, and skin texture improvement effects.

When the extract is used as a food additive, it can be added as it is or used with other foods or food ingredients, it can be suitably used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to its purpose of use (prevention, health or therapeutic treatment). Generally, 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, with respect to the raw materials in the production of a food or beverage. However, in the case of long-term intake for health and hygiene purposes or for 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 particular restrictions on the type of food. Examples of foods to which the above substances can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and all of the health foods in the ordinary sense.

When the food formulation is a beverage, it may contain various flavoring agents, natural carbohydrates, and the like as additional ingredients, like a normal beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and 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, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonated drinks It may contain a carbonizing agent used in. Other food formulations may contain flesh for the manufacture of natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients may be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a composition for promoting stem cell activity, which includes the above-mentioned samchil extract as an active ingredient.

In the present invention, the term “stem cell” is a cell capable of cell division by itself and capable of differentiating into a wide variety of specific cell types. The type of stem cells is not particularly limited, and in one embodiment, the stem cells may be skin stem cells. The 'skin stem cells' refers to stem cells capable of differentiating into cells forming the skin (epidermis, dermis and subcutaneous fat layer). The cells constituting the skin include keratinocytes, melanocytes, and fibroblasts present in the dermis (mainly responsible for the biosynthesis of collagen and elastin).

The type of skin stem cells is not particularly limited. The skin stem cell used in the present invention can be used regardless of where it originated. For example, the skin stem cells may be obtained from a known skin stem cell source, for example, the base layer of the hair follicle or epidermis, and the animal to be harvested may be a mammal. In one embodiment, the mammal may include, but is not limited to, a human, mouse, rat, guinea pig, rabbit, monkey, pig, horse, cow, sheep, antelope, dog or cat. Preferably, the mammal can be a human. Methods for obtaining skin stem cells from such a stem cell source are well known in the art.

In the present invention, the term "stem cell activity promoting effect" refers to a stem cell proliferation promoting effect, and / or a specific indicator molecule of stem cells and an effect of maintaining stem cellularity, which is an auto-dividing property.

In addition, the composition of the present invention can be used for preparing cosmetic formulations. These cosmetic formulations are the same as those described in the description of the cosmetic formulations containing the Samchil extract.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Preparation Example 1: Preparation of Samchil extract

After washing, powdering, and cooling for 5 days in a 10-fold amount of ethanol solvent, the extracted stock solution was filtered to obtain a concentrated extract. Here, a mixed solution of purified water and ethanol was added to dissolve, and then filtered to prepare a final extract.

Preparation Example 2: Culture of skin stem cells under serum-free medium conditions

Human epidermal stem cells purchased from Cellntec were added to a 48-well plate (6 × 10 ³ cells / well), and bovine pituitary extract (BPE) similar to fetal bovine serum was added. CNT-57 medium (Cellntec) was incubated at 5% CO ₂ and 37 ° C. for 24 hours. Thereafter, the culture solution was removed through a suction tube, and then a medium solution was removed using a PBS solution (GibcoBRL), and 0.0005% by weight of Samchil extract was treated with CNT-57 medium without BPE to treat 5% CO ₂ and 37. Incubated for 72 hours at ℃ condition.

Experimental Example 1: Skin stem cell proliferation promoting effect through CCK-8 evaluation method

CCK-8 (Cell counting kit-8) was evaluated on the skin stem cells cultured in Reference Example 2. CCK-8 evaluation is an analytical method that indirectly indicates the density of living cells by measuring the absorbance of Formazan produced by dehydrogenase decomposition of Tetrazolium Salt in the intracellular electron transport system.

The cells were treated with CCK-8 solution (treated with 1/10 of the medium volume) at 37 ° C. for 2 hours, and then measured spectroscopically by measuring absorbance at a wavelength of 450 nm. The effect of promoting the proliferation of the skin stem cells of the Samchil extract was calculated from the measured absorbance value (%) compared to the control group (CNT-57 medium added with BPE) according to the following equation (1), the values are shown in Table 1 below.

[Equation 1]

Proliferation rate (%) = (absorbance of the sample treatment group / absorbance of the control group) × 100

Effect of promoting the proliferation of skin stem cells Addition sample density Cell proliferation rate compared to control (%) Samchil extract 0.0005% (w / w) 129% Control - 100%

As shown in Table 1, the skin stem cells in the medium condition treated with the Samchil extract showed excellent proliferation promoting effect.

Experimental Example 2: Confirmation of the effect of maintaining stem cells on stem cells

The degree of expression of p63 was evaluated in order to confirm the effect of maintaining the stem cellularity of the skin stem cells cultured in Reference Example 2. p63 is an intracellular transcription factor, and is well known as a maintenance marker for skin stem cells. Cell RNA was isolated to synthesize cDNA, and real-time PCR was performed with Taqman staining solution to measure the expression level of p63. In this experiment, the concentration of RNA was normalized to S16 ribosomal RNA.

The experimental results are shown in Table 2 below. In the following Table 2, the value of the increase in p63 fold refers to the fold relative to the control (CNT-57 medium added with BPE).

Stem cell maintenance effect (repeated number = 3) Addition sample density p63 increase multiple (times) Samchil extract 5 μg / mL 2.09 Control - 1.0

As shown in Table 2, the medium condition treated with the Samchil extract promotes p63 expression of skin stem cells, and thus has an excellent stem cell maintenance effect.

Example 1: anti-inflammatory effect-NO production inhibitory effect

In order to confirm the anti-inflammatory effect and the effect of alleviating skin troubles of the Samchil extract, a nitric oxide (NO) formation inhibitory force experiment was conducted by the GRIESS method using a RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are macrophages of mice, were passaged several times, placed in a 24-well plate to enter 3 × 10 ⁵ cells in one well, and then cultured for 24 hours. Subsequently, the extract of Samchil was diluted to the concentration shown in Table 6 and replaced with a cell medium containing the same. At this time, 20 µg / mL of L-NMMA (L-NMMA), a NO-producing inhibitor, was treated together as a positive control and cultured for 30 minutes, and treated with 1 µg of LPS (Lipopolysaccharide) as a stimulator for 24 hours. Cultured for a while. 100 µl of the supernatant was transferred to a 96-well plate, 100 µl of the GRIESS solution was reacted for 10 minutes at room temperature, and the absorbance at 540 nm was measured to determine the NO inhibitory effect of Compound 1, and the results were as follows. It is shown in Table 3.

NO production suppression effect sample NO production inhibition rate (%) L-NMMA (positive control) 38.68 Samchil extract (0.1%) 49.42 Samchil extract (1%) 29.94

As shown in Table 3, it can be seen that the Samchil extract has the same level of NO production inhibitory activity as the previously known anti-inflammatory substance L-NMMA.

Example 2: Validation effect of promoting PPAR alpha activation

In order to confirm the effect of promoting the PPAR alpha activation of the Samchil extract, the following experiment was performed using the Samchil extract. C2C12 cells (ATCC CRL-1772), a mouse-derived myoblast cell line, were passaged and cultured in Dulbeco's Modified Eagle's Medium (DMEM) medium containing 10% fetal calf serum. As the vector used here,

1) DNA fragments encoding Ser ₁₆₇ to Tyr ₄₆₈ in the human transcription factor GAL4-DBD (DNA Binding Domain) and human PPAR alpha LBD (Ligand Binding Domain) in the SV40 promoter of pZEO vector (Invitrogen) (genetic bank information, NM 005036);

2) a GAL4 response sequence inserted into the multiple restriction enzyme recognition site of the pGL3 vector (Promega) expressing luciferase, and 3) β-galactosidase as a transfection internal control ( A vector expressing β-galactodisase) was used.

The cultured cells were smeared on a 24-well plate at a concentration of 4 × 10 ⁴ , and then cultured for 12 hours, and the three types of plasmid genes were transiently transfected using Lipopectamine. . After 8 hours, after treating the Samchil extract and incubating for 12 hours, washed with 1 × PBS and lysed the cells with 1 × reporter lysis buffer, Luciferase Assay Kit (Promega) and β- The activity of luciferase was measured using a galactosidase assay kit (Promega). That is, the PPAR alpha activity was measured as 'luciferase activity / β-galactose activity'.

In this experiment, Wy-14,643 (Calbiochem), which is known to be the most potent of the ligands of PPAR alpha, was used as a positive control, and 0.05% of DMSO used to melt the sample was used as a negative control.

According to the above experimental method, the activity of PPAR alpha according to the concentration change was measured by treating the Samchil extract with C2C12 cells, and the results are shown in Table 7 below. In Table 4, the numerical value of PPAR alpha activity refers to a percentage of the negative control (DMSO 0.05%).

PPAR alpha active sample PPAR alpha activity (%) DMSO (0.05%) 100 Wy-14,643 (0.5 μM) 301 Wy-14,643 (1 μM) 937 Samchil extract (0.05%) 630 Samchil extract (0.5%) 953

As shown in Table 4, compared to the positive control group, it was confirmed that the swallowtail extract exhibits very strong PPAR alpha activity.

Example 3: Effect of promoting expression of Filaggrin

After 5 days / day, the extract of Samchil was added to the culture medium of the human keratinocyte cell line HaCaT cells at 5 µg / mL and 10 µg / mL, and then the cells were separated from RNA, synthesized cDNA, and then treated with Taqman stain. time PCR was performed. RNA concentration was normalized to S16 ribosomal RNA.

Effect of pilar green expression according to concentration (repeated number = 3) Addition sample Increase in pilar green compared to the control group Control (no additives) 1.00 Samchil extract (5 ㎍ / ml) 28.4 Samchil extract (10 ㎍ / ml) 53.5

As shown in Table 5, the Samchil extract has the ability to promote the expression of pilargreen in the human keratinocyte line HaCaT cells.

Example 4: Efficacy for improving turn-over

For the nutritional cream of Preparation Example 2, the effect of improving exfoliation was tested on healthy women in their 20s and 50s as follows.

A 20% to 50 year old female 20 test sites were coated with DHA (Dihydroxyacetone, Sigma Aldrich, USA) 1.5% solution for 8 hours, followed by deposition, and then, a preparation example nutritional cream was applied twice daily to the test site. 4 days after application, 8 days after application, and 12 days after application, photographs were taken using a device measurement for skin brightness at the deposition site using a Chromameter CR-400 (Minolta, Japan) and DSLR. The measured value was evaluated by obtaining the average value of 3 times excluding the maximum value and the minimum value, and the higher the skin brightness improvement in the deposition site, the better the exfoliation effect. The results are shown in Table 6 below.

Skin brightness improvement effect division 4 days after application 8 days after application 12 days after application Improvement rate (%) 1.32 5.34 13.07

As shown in Table 6, when using the nutrient cream according to the present invention was found to have a keratin peeling improvement effect.

Formulation Example 1: Preparation of pharmaceutical preparation

1. Preparation of tablets

Samchil extract 0.2mg

Corn starch 100mg

Lactose 100mg

Magnesium stearate 2mg

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

Formulation Example 2: Preparation of cosmetics

1. Preparation of nutrition cream

As shown in the following composition, a nutrient cream containing the Samchil extract as an active ingredient was prepared according to a conventional method.

Samchil extract 0.2% by weight

Beta-1,3-glucan 5.0 wt%

Wax 10.0 wt%

Polysorbate 60 1.5 wt%

PAGE 60 Cured Castor Oil 2.0 wt%

Solbitan sesquioleate 0.5 wt%

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

Caprylic / Capric Triglyceride 5.0 wt%

Glycerin 5.0 wt%

Butylene glycol 3.0 wt%

Propylene glycol 3.0 wt%

Triethanolamine 0.2% by weight

Preservative 0.05 wt%

Pigment 0.05 wt%

Fragrance 0.05% by weight

Purified water to 100% by weight

Formulation Example 3: Preparation of external preparation for skin

1. Preparation of ointment

As shown in the following composition, an ointment containing the Samchil extract as an active ingredient was prepared according to a conventional method.

Samchil extract 0.5% by weight

Beta-1,3-glucan 10.0 wt%

Wax 10.0 wt%

Polysorbate 60 5.0 wt%

PAGE 60 Cured Castor Oil 2.0 wt%

Solbitan sesquioleate 0.5 wt%

Vaseline 5.0 wt%

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

Shea butter 3.0 wt%

Caprylic / Capric Triglyceride 5.0 wt%

Glycerin 10.0 wt%

Propylene glycol 10.2 wt%

Triethanolamine 0.2 wt%

Preservative 0.05 wt%

Pigment 0.05 wt%

Fragrance 0.05% by weight

Purified water to 100% by weight

Claims (10)

A cosmetic composition for skin regeneration according to the promotion of stem cell activity, which includes the Samchil extract as an active ingredient.
According to claim 1,
Samchil extract is a cosmetic composition for skin regeneration according to the promotion of stem cell activity, which is an extract obtained by extracting samchil with one or more selected from the group consisting of water and organic solvents.
According to claim 2,
The organic solvent includes a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or a polar solvent containing acetone; Non-polar solvents including ether, chloroform, benzene, hexane or dichlorohexane; Or a cosmetic composition for skin regeneration according to the promotion of stem cell activity as a mixed solvent of these.
According to claim 1,
The stem cell activity is a cosmetic composition for skin regeneration according to the stem cell activity promotion characterized by promoting stem cell proliferation or maintaining stem cellity.
An external preparation for skin regeneration according to the promotion of stem cell activity, which includes the Samchil extract as an active ingredient.
The method of claim 5,
The stem cell activity is an external preparation for skin regeneration according to the promotion of stem cell activity characterized by promoting stem cell proliferation or maintaining stem cellity.
Food composition for skin regeneration according to the promotion of stem cell activity, including Samchil extract as an active ingredient.
The method of claim 7,
The stem cell activity is stem cell proliferation or stem cell activity, characterized in that the stem cell activity promoting skin composition for regenerative skin.
Pharmaceutical composition for the treatment of skin wounds or wounds according to the promotion of stem cell activity, including Samchil extract as an active ingredient.
The method of claim 9,
The stem cell activity is a pharmaceutical composition for the treatment of skin wounds or wounds according to the promotion of stem cell activity characterized by promoting stem cell proliferation or maintaining stem cellity.