KR102360881B1 - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. According to the present invention, Samchil skin stem cell proliferation promoting effect, NO production inhibitory effect, PPAR alpha activity excellent effect, filaggrin expression increase effect, skin brightness improvement effect, skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin It can be used in the manufacture of medicines, cosmetics or food for texture improvement. In addition, it promotes the proliferation of skin stem cells and exhibits a stem cell maintenance effect, so it can be used in the manufacture of cosmetics for promoting stem cell activity.

Description

Composition for improving skin

The present invention relates to a composition for skin improvement that exhibits skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing, skin texture improvement, and stem cell activation promoting effects.

Inflammation is the body's immune response to a wound or disease, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activate inflammatory factors, causing various diseases and aging of the skin. Vasoactive polypeptides such as kinin, plasmin, and complement act as vasodilators, contractiles, and chemotaxis, and in addition, lymphocytic 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, and mediates various inflammatory responses.

On the other hand, in order to eliminate inflammation, an anti-inflammatory agent that removes an inflammatory source, reduces biological reactions and symptoms. Substances used for anti-inflammatory purposes so far include flufenamic acid, ibuprofen, benzydamine, indomethacin, etc. as a non-steroidal type, and prednisolone as a steroid type. , and dexamethasone. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have limitations in usage due to safety issues for the skin, or have insignificant effects, so that no inflammatory relief effect can be expected. There is this.

In addition, the epidermis, which is located at the outermost part of the skin, performs a protective function of protecting against various external physical, chemical and mechanical stimuli and preventing excessive dissipation of body moisture through the skin. This protective function is made possible by the normal formation and maintenance of the stratum corneum composed of keratinocytes. Keratinocytes are cells that are formed through gradual changes in shape and function as basal cells that have continuously proliferated in the stratum basale move to the stratum corneum, and after a certain period of time, old keratinocytes The forming cells are removed from the skin, and the process of epidermis differentiation or keratinization is repeated in which new keratinocytes that rise from the lowermost layer of the epidermis take over their functions. In this keratinization process, keratinocytes generate natural moisturizing factor (NMF) and intercellular lipids such as ceramide, cholesterol, and fatty acids, and the stratum corneum acts as a blocking layer from the outside, thereby creating a skin barrier. retains its function as

In addition, dry skin, which is considered one of the major diseases in modern society, is one of the symptoms caused by abnormal skin barrier function. It is increasing more and more due to various factors such as excessive bathing culture, skin aging, etc., and the number of cases requiring treatment due to severe symptoms is also continuously increasing.

Recently, atopic dermatitis, which occurs in 10% of children, is also known to be the main cause of dry skin, more fundamentally, abnormal skin barrier function. In order to treat such atopic dermatitis, conventionally, many studies have been conducted to supply moisture from the outside or to minimize moisture loss from the body by focusing on maintaining adequate moisture in the skin, and in fact, ceramides ( Moisturizers such as ceramide) or derivatives thereof have been developed and are widely used in pharmaceutical or cosmetic fields.

However, most of the use of such a moisturizer is only a temporary relief of symptoms rather than a fundamental treatment, and thus does not exhibit sufficient effect on the treatment of dry skin including atopic dermatitis and skin barrier function abnormalities. Accordingly, there is an urgent need to develop a material that fundamentally regenerates the damaged skin barrier.

Recently, when PPAR alpha (Peroxisome Proliferator Activated Receptor-alpha) present in keratinocytes binds to its ligand and is activated, it is known that it has the effect of reconstructing the damaged skin barrier by promoting the differentiation of keratinocytes. In fact, when Clofibrate [Non-Patent Document 1] or WY14643 [Non-Patent Document 2], which are known agonists of PPAR alpha, 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 accelerated.

Also, to have fair and fair skin is the unchanging desire of everyone. It is genetically determined according to the concentration and distribution of melanin in the skin of a person, but it is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress. Melanin is produced through a non-enzymatic oxidation reaction after the enzyme tyrosinase acts on tyrosine, a kind of amino acid, to convert it into DOPA and dopaquinone. As such, the pathway by which melanin is made is known, but the mechanism of inducing melanin synthesis, which is the previous step of tyrosinase action, is still not fully elucidated.

In addition, the skin texture, which is the shape of the skin surface, is characterized by a three-dimensional microstructure formed by fine lines and is significantly different by age and region. The skin texture is divided into 1st (about 20-100 μm), 2nd (about 5-40 μm), and 3rd (about 0.5 μm) lines according to the depth of the line. It has a characteristic that polygons and star formations formed by the intersection of lines forming a clear pattern [Non-Patent Document 3].

However, it is known that with age or as the skin is damaged, the network structure of dense fine lines collapses, so fine lines (secondary, tertiary, etc.) disappear and the primary lines become deeper and wrinkles are created [non-patented] Literature 4,5]. In other words, wrinkles, which are a typical symptom of aging skin, are the accumulation of minute changes that have already occurred over a long period of time.

It is known that all components of human skin are embryologically derived from ectoderm or mesoderm. The epidermis, hair follicles, sebaceous and sweat glands are derived from the ectoderm, while melanocytes, nerves and special sensory receptors are derived from the neuroectoderm. Depending on the embryonic period, embryonic stem cells repeatedly differentiate to 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. In the skin, stem cells exist mainly in two places. . The first is in the hair follicle. This is the cell before cell differentiation and is known to play an important role in the regeneration of the epidermis, and plays an important role in the regeneration and growth of hair. The second is the basal layer of the 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. The stem cells here are widely used in skin stem cell research because they have the advantage of being relatively abundant and easy to obtain. The skin is continuously regenerated, and stem cells existing in the epithelium of the skin, that is, epidermal stem cells of the skin, are involved in the repair of the epithelium after a wound [Non-Patent Document 6]. The epithelial stem cells of the skin are also referred to as skin stem cells, and when the skin stem cells are activated, it is possible to treat skin wounds such as trauma or promote wound healing, and wrinkle relief of the skin can also be expected. have. Integrin β1 and integrin α6 are used as indicators of skin stem cells, and it is reported that the maintenance of skin stem cells necessary for epithelial morphogenesis and differentiation is regulated by p63 protein.

Skin stem cells play an important role in maintaining skin health and physiological and biochemical homeostasis. Stem cells present in the skin also have malfunctions due to the effects of aging, which causes various problems as the homeostasis of the skin is broken. Therefore, it will be possible to improve various phenomena of skin aging through the activity of these stem cells [Non-Patent Document 7].

In addition, it is safe to the living body, the active ingredient is stable, and above all, it has anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement activities that are superior to the existing substances that have anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement effects. There is an urgent need for the development of ingredients with

On the other hand, Samchil is a perennial herb, the root of Panax pseudoginseng and P.notoginseng, and its appearance is conical, 1-6cm long and 1-4cm in diameter, with small hump-shaped projections around it, and the outer surface is grayish-brown-gray. It is yellow, with vertical wrinkles and traces of fine roots attached, and the cut surface is gray-green-yellow-green and gray-white with radial patterns, and the quality is heavy and hard. According to Donguibogam, it has been used for the treatment of various types of hyperemia, vaginal pain, and heart pain. According to the existing research on the identification of ingredients, compounds such as β-carotene, cryptoxanthine, meloside A, L saponin, and trilinolein have been reported as major components.

However, the mechanism of samchil on the skin is not known yet, and there is no research on it.

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 aging process, Catherin Niemann, Stem Cell Aging and Regenerative Medicine, November 2008, Pages 986-997

Accordingly, the present inventors found that the ginseng extract promotes skin regeneration by promoting the proliferation of skin stem cells, has an anti-inflammatory effect by suppressing NO production, and has excellent PPAR alpha activity to improve atopy, and increases filaggrin expression. To complete the present invention by confirming the effect of promoting skin moisturizing and improving skin texture, promoting the proliferation of skin stem cells, and maintaining stem cell characteristics became

Accordingly, an object of the present invention is to provide a composition for anti-inflammatory, atopic improvement, skin moisturizing, and skin texture improvement, which includes a ginseng 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 extract of ginseng as an active ingredient.

As a means for solving the above problems, the present invention provides a composition for anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement comprising an extract of ginseng as an active ingredient for the manufacture of pharmaceuticals, cosmetics or food.

As another means for solving the above problems, the present invention provides a composition for promoting stem cell activity, comprising a ginseng extract as an active ingredient, for the preparation of a cosmetic formulation.

The ginseng extract according to the present invention promotes skin regeneration by promoting the proliferation of skin stem cells, has an anti-inflammatory effect by suppressing NO production, and has excellent PPAR alpha activity to improve atopy. In addition to the skin moisturizing improvement effect, by showing the skin texture improvement effect due to the improvement of skin brightness, it can be used in pharmaceuticals, cosmetics, or food production.

In addition, the ginseng extract promotes the proliferation of skin stem cells and exhibits a stem cell maintenance effect, so it can be used in the preparation of a cosmetic formulation for promoting stem cell activity.

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

In order for the skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement ingredients to exhibit excellent effects when applied to the actual skin, high-active skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement activities at low concentrations are required. It has excellent ability to penetrate and absorb the skin, and has low volatility so that it can stay for a sufficient time to show 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 desirable that it is maintained to be clean, easy to manufacture as a pharmaceutical, cosmetic or food, and is also safe for the skin. However, it is rare among known components that satisfy all of the above properties. For example, some skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement ingredients have excellent skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement activities even at low concentrations in in vitro experiments, but It has poor ability to penetrate and absorb, making it difficult to apply to actual skin. Another active ingredient has low hydrophilicity, making it difficult to formulate into medicines, cosmetics, and food. In addition, some skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement ingredients are decomposed or transformed into other compounds when exposed to heat, light, or oxygen, so that their effects disappear before application to the skin. In some cases.

As can be seen in the examples below, the ginseng extract exhibits excellent skin stem cell proliferation promoting effect, anti-inflammatory effect, PPAR alpha activation effect, filaggrin expression effect, and skin brightness improvement effect at a low concentration, thereby regenerating skin. It can be used as an active ingredient in a pharmaceutical, cosmetic or food composition for anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement.

Therefore, the present invention regenerates the skin comprising Samchil extract as an active ingredient. To provide a composition for anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement.

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

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

Preferably, the extract obtained by extracting ginseng extract with water and/or an organic solvent may be used, and the organic solvent may be a polar organic solvent, a non-polar organic solvent, or a mixture 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 the primary extract extracted using the above-described extraction solvent using an extraction solvent having a different polarity. For example, the Samchil extract may be a fraction obtained by extraction with an alcohol having 1 to 5 carbon atoms and then re-fractionation with a solvent having a different polarity, such as ether, benzene, or hexane. In the fractionation, two or more solvents may be used, and each solvent extract may be prepared by sequentially using or mixing them according to the polarity of the solvent.

In the present invention, the prepared extract or the fraction obtained by performing the fractionation process may be filtered or the solvent may be removed by performing a concentration or drying process, and all of the filtration, concentration and drying may be performed. Specifically, the filtration may be performed using filter paper or a reduced pressure filter, and the concentration may be concentrated under reduced pressure using a reduced pressure concentrator or the like, and drying may be performed by a freeze-drying method.

In addition, silica gel column chromatography, thin layer chromatography, or high performance liquid chromatography with respect to the prepared extract or the fraction obtained by performing the fractionation process, such as Further purified fractions can be obtained by purification using various chromatography methods.

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 extract, powder, granule, tablet or capsule.

In addition, the composition may further contain one or more active ingredients exhibiting the same or similar function. For example, it may include known skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement ingredients. When additional skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement components are included, the skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement effects of the composition of the present invention may be further enhanced. When adding the above ingredients, skin safety according to combined use, ease of formulation, and stability of active ingredients can be considered. In one embodiment of the present 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 a whitening ingredient known in the art, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, etc. Hydroquinone, vitamin-C (L- Ascorbic acid) and derivatives thereof and one or two or more components selected from the group consisting of various plant extracts may be further included. Additional components 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 ginseng extract.

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

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

The composition of the present invention may be administered orally or parenterally, and may be administered in the form of general pharmaceutical preparations, for example, oral and parenteral various formulations during clinical administration. , a binder, a wetting agent, a disintegrant, a diluent such as a surfactant, or an excipient.

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

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral use include suspensions, 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 solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

In the present invention, the 'skin regeneration effect' refers to the recovery of skin tissue against damage caused by external and internal causes of the skin as the activity of skin stem cells is promoted. In this case, 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 the defense responses of a living tissue to a certain stimulus, which causes tissue deterioration, circulatory disturbance and exudation, and tissue proliferation. complex lesions. More specifically, inflammation is part of innate immunity, and as in other animals, human innate immunity recognizes patterns on the surface of cells that are specific to pathogens. Phagocytes recognize cells with such a surface as non-self and attack pathogens. If pathogens break through the body's physical barriers, an inflammatory response occurs. The inflammatory response is a non-specific defense action that creates a hostile environment for microorganisms that invade the wound site. In the inflammatory response, when there is a wound or an external infectious agent enters the body, the white blood cells responsible for the initial stage immune response flock and express cytokines. Therefore, the expression level of intracellular cytokines is an indicator of inflammatory response activation. Examples of skin diseases related to inflammation include atopic dermatitis, psoriasis, erythematous disease triggered by radiation, chemicals, burns, etc., acid burn, bullous dermatosis, lichenoid type disease, allergic itch, seborrheic eczema, rose acne, pemphigus vulgaris, erythema multiforme, erythema nodosum, balanitis, vulvitis, inflammatory hair loss such as alopecia areata, cutaneous T-cell lymphoma, and the like.

In the present invention, the term 'atopic improvement effect' refers to preventing or treating skin diseases such as atopic dermatitis and dry skin, and inhibiting, inhibiting, or alleviating atopic symptoms.

In the present invention, the 'moisturizing effect' refers to inhibiting or suppressing a decrease in moisture in the skin or increasing the moisture content of the skin to smooth the skin surface and to impart gloss.

In the present invention, the 'skin texture improvement effect' refers to smoothing the skin surface, imparting gloss, and brightening the skin tone by suppressing or inhibiting the roughening of the skin surface due to the effects of aging, stress, etc. .

In the present invention, the term 'effective amount' refers to an amount of the extract capable of promoting regeneration of damaged skin, inhibiting inflammation, improving skin dryness such as atopic dermatitis, moisturizing effect, or improving skin texture . When the composition of the present invention includes an effective amount of the ginseng extract, it is possible to provide a desirable skin regeneration effect, anti-inflammatory effect, atopic improvement effect, moisturizing effect and skin texture improvement effect. The effective amount of the ginseng 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 time it stays on the skin. For example, when the composition is commercialized as a pharmaceutical formulation, it may contain the Samchil extract at a higher concentration than when commercialized as a cosmetic to be applied to the skin on a daily basis. Therefore, the daily dose is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, and more preferably 50 to 60 mg/kg, based on the amount of the ginseng extract, 1 to 6 times a day may be administered.

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

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

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

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

In addition, the composition for skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement of the present invention may be used for preparing a cosmetic formulation.

The cosmetic formulation may be in the form of a general emulsified formulation and a solubilized formulation. For example, lotions such as flexible lotions or nourishing lotions, emulsions such as facial lotions and body lotions, creams such as nourishing creams, moisture creams, and eye creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, liquids It may have formulations such as foundation, powder, cleansing cream, cleansing lotion, and makeup remover such as cleansing oil, cleansing foam, soap, body wash, and the like.

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

The cosmetic formulation may include a relatively high concentration of the ginseng extract in the case of wash-off type cosmetics such as makeup removers and detergents in which the active ingredient stays on the skin within 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, even if it contains a lower concentration of the ginseng extract compared to the wash-off type cosmetics. will be free Although not limited thereto, in one embodiment of the present invention, the composition may include 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) of the ginseng extract based on the total weight of the composition . When the composition of the present invention contains the Samchil extract in an amount of less than 0.0001% by weight, sufficient skin regeneration, anti-inflammatory, atopic 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 problems with skin safety.

In addition, the composition for skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing, and skin texture improvement of the present invention may be used for preparing a food formulation.

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

Since the food formulation can be consumed on a daily basis, it is very useful because high skin regeneration, anti-inflammatory, atopic improvement, skin moisturizing and skin texture improvement effects can be expected.

When the ginseng extract is used as a food additive, the ginseng extract may be added as it is or used with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. .

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense.

When the food formulation is a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. 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 the sweetener, natural sweeteners such as taumartin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition to the above, food formulations include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonated beverages It may contain a carbonation agent, etc. used for Other food formulations may contain the pulp for the production of natural fruit juices, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. Although the proportion of these additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a composition for promoting stem cell activity comprising the ginseng extract as an active ingredient.

In the present invention, the term 'stem cell' refers to a cell capable of self-dividing and having the ability to differentiate into a wide variety of specific cell types. The type of these stem cells is not particularly limited, and in one embodiment, the stem cells may be skin stem cells. The 'skin stem cells' refer to stem cells that can be differentiated into cells constituting the skin (epidermal, dermis, and subcutaneous fat layer). Cells constituting the skin include keratinocytes, melanocytes, and fibroblasts (mainly responsible for the biosynthesis of collagen and elastin) present in the epidermis and dermis.

The type of the skin stem cells is not particularly limited. The skin stem cells used in the present invention can be used regardless of where they are derived. For example, skin stem cells may be obtained from a known skin stem cell source, for example, hair follicles or the basal layer of the epidermis, and the animal to be harvested may be a mammal. In one embodiment, a mammal can 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 may be a human. Such, a method for obtaining skin stem cells from a skin stem cell source is well known in the art.

In the present invention, the term 'stem cell activation promoting effect' refers to including the effect of maintaining the stem cell proliferation promoting effect, and/or stem cell specific indicator molecule and the self-dividing property of stem cell property.

In addition, the composition of the present invention can be used for the preparation of cosmetic formulations. For this cosmetic formulation, it is the same as described in the description of the cosmetic formulation containing the extract of ginseng extract.

Hereinafter, the present invention will be described in detail by way of 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 Samchil extract

After washing and pulverizing the ginseng chil, the extract was filtered and concentrated after cooling in a 10-fold amount of ethanol solvent for 5 days to obtain a concentrated extract. Herein, a mixed solution of purified water and ethanol was added and dissolved, followed by filtration to prepare a final extract.

Preparation Example 2: Culturing of skin stem cells in serum-free medium conditions

Human epidermal stem cells purchased from Cellntec were placed in a 48-well plate (6×10 ³ cells/well), and BPE (Bovine pituitary extract) similar to fetal bovine serum was added. CNT-57 medium (Cellntec) was used for 24 hours and cultured at 5% CO ₂ and 37° C. conditions. Thereafter, the culture medium is removed through a suction tube, and then the medium components are removed using a PBS solution (GibcoBRL), and 0.0005 wt% of the ginseng extract is treated in CNT-57 medium without BPE to 5% CO ₂ and 37 Incubated for 72 hours at ℃ condition.

Experimental Example 1: Effect of promoting skin stem cell proliferation through CCK-8 evaluation

CCK-8 (Cell counting kit-8) was evaluated on the skin stem cells cultured in Reference Example 2. CCK-8 evaluation is an analysis method that indirectly indicates the density of living cells by measuring the absorbance of Formazan, which is produced by dehydrogenase in the electron transport system in the cell decomposing tetrazolium salt.

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

[Equation 1]

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

Skin stem cell proliferation promoting effect additive sample density Cell proliferation rate (%) compared to control ginseng extract 0.0005% (w/w) 129% control - 100%

As shown in Table 1, skin stem cells in the medium condition treated with ginseng extract exhibited an excellent proliferation-promoting effect.

Experimental Example 2: Confirmation of stemness maintenance effect of skin stem cells

In order to confirm the stem cell maintenance effect of the skin stem cells cultured in Reference Example 2, the expression level of p63 was evaluated. p63 is an intracellular transcription factor and is well known as a maintenance marker of skin stem cells. Cell RNA was isolated to synthesize cDNA, and then, 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 Table 2 below, the value of the p63 increase fold means the fold compared to the control (CNT-57 medium with BPE added).

Stemness maintenance effect (number of repetitions = 3) additive sample density p63 increase multiplier (fold) ginseng extract 5 μg/mL 2.09 control - 1.0

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

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

In order to confirm the anti-inflammatory effect and skin trouble alleviation effect of ginseng extract, nitric oxide (NO) formation inhibition experiment was conducted by GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are macrophages of mice, were subcultured several times, placed in a 24-well plate so that 3×10 ⁵ cells were put into each well, and then cultured for 24 hours. Then, it was replaced with the cell medium containing the dilution of the Samchil extract to the concentration shown in Table 6 below. At this time, 20 μg/mL of L-NMMA (L-NG-Monomethylarginine), which is a NO-production inhibitor, was treated together as a positive control and incubated for 30 minutes, and LPS (Lipopolysaccharide) was treated as a stimulant by 1 μg for 24 hours. incubated during Take 100 μl of the supernatant and transfer it to a 96-well plate, add 100 μl of GRIESS solution, react at room temperature for 10 minutes, and measure the absorbance at 540 nm to determine the NO inhibitory effect of compound 1, and the results are shown below. Table 3 shows.

NO production inhibitory 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 ability as L-NMMA, which is a known anti-inflammatory substance.

Example 2: Verification of PPAR alpha activation promoting effect

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

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

2) the GAL4 response sequence was 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 plated on a 24-well plate at a concentration of 4×10 ⁴ , cultured for 12 hours, and the three types of plasmid genes were transiently transfected using Lipopectamine. . After 8 hours, the ginseng extract was treated and incubated for 12 hours, washed with 1×PBS and lysed with 1× reporter lysis buffer, followed by a luciferase assay kit (Promega) and β- The activity of luciferase was measured using a galactosidase assay kit (Promega). That is, PPAR alpha activity was measured as 'luciferase activity/β-galactose activity'.

As a positive control in this experiment, Wy-14,643 (Calbiochem), which is known to be the strongest of the ligands of PPAR alpha, was used as a positive control, and as a negative control, 0.05% of DMSO used for dissolving the sample was used.

According to the experimental method, C2C12 cells were treated with ginseng extract to measure the activity of PPAR alpha according to the concentration change, and the results are shown in Table 7 below. In Table 4 below, the value of PPAR alpha activity means a percentage compared to the negative control (DMSO 0.05%).

PPAR alpha activity 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, it was confirmed that the ginseng extract exhibited very strong PPAR alpha activity.

Example 3: Effect of promoting expression of Filaggrin

After adding the extract to 5 μg/mL and 10 μg/mL to the culture medium of the human keratinocyte line HaCaT cells, respectively, and culturing for 1 day, RNA of the cells was isolated, cDNA was synthesized, and Real- with Taqman staining solution time PCR was performed. RNA concentrations were normalized to S16 ribosomal RNA.

Effect of filaggrin expression according to concentration (number of repetitions = 3) additive sample Factor of increase in filaggrin compared to control group Control (no additive) 1.00 Samchil extract (5 μg/ml) 28.4 Samchil extract (10 μg/ml) 53.5

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

Example 4: Efficacy for improving turn-over

For the nutritional cream of Formulation Example 2 below, the effect of improving keratin exfoliation was tested on healthy women in their 20s to 50s.

After depositing a 1.5% solution of DHA (Dihydroxyacetone, Sigma Aldrich, USA) with a closed patch for 8 hours on the test site of 20 women aged 20 to 50 years, the preparation nutrient cream was applied to the test site twice daily. After 4 days of application, 8 days after application, and 12 days after application, using a Chromameter CR-400 (Minolta, Japan) to measure the skin brightness of the deposition site and take pictures using a DSLR. The measured values were evaluated by obtaining the average value of three times excluding the maximum and minimum values, and the higher the improvement in skin brightness at the deposition site, the greater the effect of keratin exfoliation improvement. The results are shown in Table 6 below.

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

As shown in Table 6, when the nutritional cream according to the present invention was used, it was found that there was an effect of improving keratin exfoliation.

Formulation Example 1: Preparation of a pharmaceutical formulation

1. Preparation of tablets

ginseng extract 0.2mg

Corn Starch 100mg

Lactose 100mg

Magnesium stearate 2mg

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

Formulation Example 2: Preparation of cosmetics

1. Manufacture of nourishing cream

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

0.2% by weight of ginseng extract

Beta-1,3-glucan 5.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 1.5 wt%

Sebum 60 hydrogenated castor oil 2.0 wt%

0.5 wt% of sorbitan sesquioleate

Liquid paraffin 10.0 wt%

5.0 wt% of squalane

5.0 wt% caprylic/capric triglyceride

glycerin 5.0 wt%

Butylene glycol 3.0 wt%

Propylene glycol 3.0 wt%

0.2 wt% of triethanolamine

Preservative 0.05 wt%

dye 0.05% by weight

Flavor 0.05 wt%

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 Samchil extract as an active ingredient was prepared according to a conventional method.

0.5% by weight of ginseng extract

Beta-1,3-glucan 10.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 5.0 wt%

Sebum 60 hydrogenated castor oil 2.0 wt%

0.5 wt% of sorbitan sesquioleate

Vaseline 5.0% by weight

Liquid paraffin 10.0 wt%

5.0 wt% squalane

Shea Butter 3.0 wt%

5.0 wt% caprylic/capric triglycerides

10.0 wt% glycerin

Propylene glycol 10.2 wt%

0.2% by weight of triethanolamine

Preservative 0.05 wt%

dye 0.05% by weight

Flavor 0.05 wt%

Purified water to 100% by weight

Claims (14)

A cosmetic composition for improving keratin exfoliation by promoting stem cell activity, comprising Samchil extract as an active ingredient.
The method of claim 1,
The ginseng extract is an extract obtained by extracting ginseng extract with one or more selected from the group consisting of water and an organic solvent. A cosmetic composition for improving keratin exfoliation by promoting stem cell activity.
3. The method of claim 2,
The organic solvent is a polar solvent including a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; non-polar solvents including ether, chloroform, benzene, hexane or dichlorohexane; Or a cosmetic composition for improving keratin exfoliation by promoting stem cell activity, which is a mixed solvent thereof.
The method of claim 1,
The stem cell activity is a cosmetic composition for improving keratin exfoliation according to stem cell activity promotion, characterized in that stem cell proliferation promotion or stem cell maintenance.
A skin external preparation for promoting keratin exfoliation by promoting stem cell activity, containing Samchil extract as an active ingredient.
6. The method of claim 5,
The stem cell activity is a skin external preparation for promoting keratin exfoliation according to stem cell activity promotion, characterized in that stem cell proliferation promotion or stem cell maintenance.
A food composition for improving keratin exfoliation by promoting stem cell activity, comprising Samchil extract as an active ingredient.
8. The method of claim 7,
The stem cell activity is a food composition for improving keratin exfoliation according to stem cell activity promotion, characterized in that stem cell proliferation promotion or stem cell maintenance.
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